CN100582774C - Detection method of treating prostatitis formulation - Google Patents

Detection method of treating prostatitis formulation Download PDF

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CN100582774C
CN100582774C CN200610200511A CN200610200511A CN100582774C CN 100582774 C CN100582774 C CN 100582774C CN 200610200511 A CN200610200511 A CN 200610200511A CN 200610200511 A CN200610200511 A CN 200610200511A CN 100582774 C CN100582774 C CN 100582774C
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solution
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methyl alcohol
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CN1876050A (en
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周霞
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GUIZHOU YIBAI WOMAN BIG PHARMACEUTICAL FACTORY
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Yunyanxichuang Medicinal Science And Technology Development Co Ltd Guiyang C
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Abstract

Disclosed is a medicinal preparation for treating prostatitis, process for preparation and quality control method thereof, wherein the preparation is made mainly from Chinese medicinal herbs including corktree bark, radix paeoniae rubrathe, root of red rooted saliva, peach kernels and lycopus, or their extracts of corresponding weight parts.

Description

The detection method of treatment prostatitis pharmaceutical preparation
Technical field:
The present invention is a kind of detection method for the treatment of prostatitic medicinal preparation of QIANLIE ANTONG, belongs to technical field of Chinese medicine.
Technical background:
In between twenty and fifty period, the incident disease of prostate is mainly acute and chronic prostatitis.Trace it to its cause, between twenty and fifty period is the male's sexual animated period just, and sexuality is frequent, easily causes prostatic hyperemia repeatedly under the stimulation of heat, brings out inflammation.Secondly, be the most vigorous period of prostate secretion between twenty and fifty period, for the growth of bacterium provides good condition.If do not note personal hygiene, infect at low or other position of Abwehrkraft des Koepers, and pathogen just can enter prostate, forms acute and chronic inflammation.According to domestic statistical data, prostatitis is the highest with between twenty and fifty people's morbidity rate, and the number of going to a doctor because of prostatitis in policlinic accounts for the prescription on individual diagnosis patient's male sex 25%～30%.Nervous and the busy city male sex more should guard against prostatitis, bring inconvenience with exempt from customs examination work, life, study.Long-term clinical practice proves that the prostatitic curative effect of Chinese medicine is definite.Wherein commercially available ANLIETONG PIAN has clearing heat and promoting diuresis, and is promoting blood circulation and removing blood stasis, is used for syndrome of stagnant dampness-heat, and disease is seen: frequent micturition, and urgent urination, it is not smooth to urinate, lower abdominal distention pain etc.But this tablet is through extrusion modling, and disintegration is slow, and drug effect speed is slower; So there is the researcher that it should be capsule, as number of patent application is that " 200410023397.2 ", name are called " prostatitis-treating capsule and preparation method thereof ", and number of patent application is that " 200410000135.4 ", name are called the application of " a kind of pure oral preparation of Chinese traditional medicinal for the treatment of prostatic disorders and preparation method thereof "; But we find in the process of studying: the former has ignored this product is the very strong characteristics of the water extracted immersing paste, hydroscopicity, does not have rational technology and strict detection method, causes the easy moisture absorption bonding of product, the quality instability; Though the latter extracts the medicine that contains volatile oil, directly sprays into, cause volatile oil to be easy to loss, and big production can only obtain aqua aromatica, and not only extraction ratio is low to that is to say our volatile oil, and loss is serious, increase cost, do not increased curative effect; In view of such circumstances, in order to improve stability of drug, need to seek a kind of result of treatment ideal, reliable in quality, formulation reasonably effectively the medicine preparation overcome problem that prior art exists, enrich the formulation kind, satisfy the needs in market.And, make said preparation reach satisfactory effect, and can standard production management, and it is done further research and more new development, at first said preparation will have the quality of stable and controllable; And the detection method of existing prostatitis-treating preparation is comparatively rough, is difficult to reach modern medicines requirement stable and controllable for quality; Therefore, the new method that needs research prostatitis-treating preparation to detect.
Summary of the invention:
The objective of the invention is to: a kind of detection method for the treatment of prostatitic medicinal preparation of QIANLIE ANTONG is provided; Comprise capsule, pill and their preparation method and method of quality control with characteristics such as covering bitter taste, easy to carry, good mouthfeel, absorption are fast, steady quality.
The present invention constitutes like this: treat prostatitic pharmaceutical preparation, it is made into effervescent tablet, parenteral solution, powder pin, freeze-dried powder, gel, dispersing tablet, capsule, soft capsule, microcapsules, granule, pill, micropill, powder, pill, sustained release preparation, controlled release preparation, gel, oral liquid, soft extract, extract and film by golden cypress 200g, radix paeoniae rubrathe 200g, red sage root 100g, peach kernel 140g, Herba Lycopi 120g, root of three-nerved spicebush 120g, seed of cowherb 80g and root of Dahurain angelica 80g or with their extract of corresponding weight portion.
Concrete preparation is capsule or pill.
The preparation method who treats prostatitic pharmaceutical preparation is: get golden cypress, the fine powder that was ground into 80 mesh sieves is standby; When the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb and the root of Dahurain angelica add 8 times of water and are heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction, filter, relative density was 1.05～1.10 thick paste when filtrate was condensed into 60 ℃, added the golden cypress fine powder, made different preparations then respectively.
Capsule in the preparation prepares like this: get golden cypress, the fine powder that was ground into 80 mesh sieves is standby; When the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb and the root of Dahurain angelica add 8 times of water and be heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction filtered, relative density was 1.05～1.10 thick paste when filtrate was condensed into 60 ℃, and drying under reduced pressure is ground into fine powder, add golden cypress fine powder and starch, talcum powder, amount of talc is 5%, mixing, incapsulate, should control relative humidity during packing below 65.0%, promptly.
Pill in the preparation prepares like this: get golden cypress, the fine powder that was ground into 80 mesh sieves is standby; The radix paeoniae rubrathe, the red sage root, the Herba Lycopi, the root of three-nerved spicebush, when the seed of cowherb and the root of Dahurain angelica add 8 times of water and are heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction, filter, relative density was 1.05～1.10 thick paste when filtrate was condensed into 60 ℃, added the golden cypress fine powder, was matrix with the Macrogol 4000, according to medicine: the part by weight of matrix=1: 2 adds Macrogol 4000, mixing, the employing internal diameter is 3.0mm, external diameter is the dropper of 4.0mm, drips 80 ℃ of system temperature, dripping speed is 20～30d/min, dripping distance is 5cm, splash in the long cooling stud of 130cm, be liquid coolant again with the methyl-silicone oil, pill, promptly.
The detection method of the prostatitic pharmaceutical preparation of described treatment: comprise following all or part of content:
(1) one or more differential test method in Berberine hydrochloride, golden cypress, the radix paeoniae rubrathe, Paeoniflorin, the red sage root, danshensu or its sodium salt, protocatechualdehyde, tanshin polyphenolic acid B or its magnesium salts, the root of Dahurain angelica, peach kernel, amygdalin, Herba Lycopi, the ursolic acid;
(2) one or more content test method in Berberine hydrochloride, Paeoniflorin, tanshin polyphenolic acid B or its magnesium salts, the protocatechualdehyde;
The discrimination method of preparation comprises following all or part of content:
A. one or both thin-layer chromatography discrimination method in Berberine hydrochloride, the golden cypress in the preparation:
It is an amount of to get this preparation, adds diethyl ether or sherwood oil or ethyl acetate or chloroform extraction, discards extract, and the dregs of a decoction volatilize, and add 20～100% methyl alcohol or alcohol extract again, the preparation need testing solution; Or it is an amount of to get this preparation, directly adds 20～100% methyl alcohol or alcohol extract, the preparation need testing solution; Get the golden cypress medicinal material, with reference to a kind of control medicinal material solution of making among the need testing solution preparation method; Other gets the Berberine hydrochloride reference substance, adds 20～100% methyl alcohol or ethanol and makes reference substance solution; Test according to thin-layered chromatography, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively on same silica G or silica gel H thin layer plate, with benzene or toluene or dimethylbenzene-ethyl acetate or ethyl formate or sour acid butyl ester-isopropyl alcohol or normal butyl alcohol-methyl alcohol or ethanol-dense ammoniacal liquor=1～12: be developping agent at 1～6: 0～3: 0～3: 0～1, take out, dry, put under the uviol lamp and inspect, in the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
B. one or both thin-layer chromatography discrimination method in the radix paeoniae rubrathe, the Paeoniflorin in the preparation:
It is an amount of to get this preparation, adds 20～100% ethanol or methanol extraction, the preparation need testing solution; Get radix paeoniae rubrathe medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Paeoniflorin reference substance, adds 20～100% ethanol or methyl alcohol and makes reference substance solution; According to the thin-layered chromatography test, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with chloroform or methylene chloride-ethyl acetate or ethyl formate or sour acid butyl ester-methyl alcohol or ethanol-formic acid or acetate=10～40: be developping agent at 1～5: 2～12: 0～2, launches, take out, dry, spray is with vanillic aldehyde sulfuric acid or ethanol solution of sulfuric acid, and it is clear to be heated to spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
C. one or more thin-layer chromatography discrimination method in the red sage root, danshensu or its sodium salt, the protocatechualdehyde in the preparation:
It is an amount of to get this preparation, extracting in water, and extract is regulated pH1～4 with acid solution, and with ethyl acetate or chloroform extraction, extract evaporate to dryness, residue add methyl alcohol or ethanol makes dissolving, as need testing solution; Or it is an amount of to get this preparation, adds 20～100% ethanol or methanol extraction, the preparation need testing solution; Get red sage root control medicinal material,, make control medicinal material solution with reference to one of need testing solution preparation method; Get danshensu or its sodium salt, protocatechualdehyde, add 20～100% methyl alcohol or ethanol and make reference substance solution; According to the thin-layered chromatography test, draw one or more each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with chloroform or methylene chloride or benzene or ethyl acetate-acetone or butanone or ethyl acetate-formic acid or acetate=5～25: be developping agent at 1～10: 0～4, launch, take out, dry, spray is inspected under the visible light with the mixed solution of liquor ferri trichloridi or the ferric trichloride and the potassium ferricyanide, or with the ammonia steam smoked after, under uviol lamp, inspect; In the test sample chromatogram, with control medicinal material chromatogram, the corresponding position of reference substance chromatogram on, show the spot of same color;
D. one or more thin-layer chromatography discrimination method in the red sage root, root of red-rooted salvia phenolic acid B or its magnesium salts in the preparation:
It is an amount of to get this preparation, extracts the preparation need testing solution with 10～100% methyl alcohol or ethanol jolting; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; Root of red-rooted salvia phenolic acid B or its magnesium salts are made reference substance solution; According to the thin-layered chromatography test, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with toluene or benzene or dimethylbenzene-chloroform or methylene chloride-ethyl acetate or ethyl formate or sour acid butyl ester-methyl alcohol or ethanol-formic acid or acetate=0.2～2: be developping agent at 0.5～3: 0.5～4: 0～1: 0～2, launch, take out, dry, inspect under the uviol lamp, or spray is with ethanol solution of sulfuric acid, heating is inspected under the visible light; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
E. the thin-layer chromatography discrimination method of the root of Dahurain angelica in the preparation:
It is an amount of to get this preparation, extracting in water, and extract concentrates with chloroform or ethyl acetate extraction, extract, as need testing solution; Or it is an amount of to get this preparation, extracts the preparation need testing solution with chloroform or ethyl acetate jolting; Other gets root of Dahurain angelica control medicinal material, with reference to one of need testing solution preparation method, makes control medicinal material solution; Test according to thin-layered chromatography, draw each 1～30 μ l of above-mentioned two kinds of solution, put respectively on same silica G or silica gel H thin layer plate, with chloroform or methylene chloride or ethyl acetate-methyl alcohol or ethanol or acetone=1～9: 0.2～3 is developping agent, put in the vapour-saturated expansion cylinder of ammonia, launch, take out, dry, put under the ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
F. one or both thin-layer chromatography discrimination method in peach kernel, the amarogentin in the preparation:
It is an amount of to get this preparation, adds 20～100% methyl alcohol or alcohol extract, again by macroporous adsorptive resins, with 0～80% ethanol elution, collects the part eluent after extract is handled, and evaporate to dryness, residue add 10～100% methyl alcohol or ethanol makes dissolving, as need testing solution; Get the peach kernel medicinal material, add diethyl ether or Petroleum ether extraction, discard extract, add 20～100% methyl alcohol or alcohol extract again, preparation control medicinal material solution; Other gets the amarogentin reference substance, adds 20～100% methyl alcohol or ethanol and makes reference substance solution; Draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform or methylene chloride-ethyl acetate or ethyl formate or sour acid butyl ester-methyl alcohol or alcohol-water=0～15: 0～40: 1～22: 1～10 lower floor's solution 0～10 ℃ of placement is developping agent, launch, take out, spray is with phosphomolybdic acid sulfuric acid solution or ethanol solution of sulfuric acid, and it is clear to be heated to colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
G. one or both thin-layer chromatography discrimination method in Herba Lycopi, the ursolic acid in the preparation:
It is an amount of to get this preparation, adds diethyl ether or methenyl choloride or ethyl acetate or ethanol or methyl alcohol or acetone extraction the preparation need testing solution; Other gets Herba Lycopi's control medicinal material, shines medicinal material solution in pairs with legal system; Get the ursolic acid reference substance again, add methyl alcohol or methenyl choloride or ethyl acetate or ethanol and make dissolving, in contrast product solution; According to the thin-layered chromatography test, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with cyclohexane or normal hexane-methenyl choloride or methylene chloride-ethyl acetate or butyl acetate or ethyl formate-formic acid or acetate=0～20: be developping agent at 1～5: 1～8: 0～1, launches, take out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
H. the high performance liquid chromatography discrimination method of protocatechualdehyde in the preparation:
It is an amount of to get this preparation, extracting in water, and water liquid prepares need testing solution with ether or ethyl acetate or chloroform jolting extraction; Or it is an amount of to get this preparation, adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the protocatechualdehyde reference substance, adds 20～100% methyl alcohol or ethanol and is made as reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-acetate or formic acid or phosphoric acid=10～50: 30～95: 0.01～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
I. the high performance liquid chromatography discrimination method of tanshin polyphenolic acid B in the preparation:
It is an amount of to get this preparation, adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 10～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or ethanol-acetonitrile-formic acid or acetate or phosphoric acid-water=10～30: 0～30: 0～5: 50～95 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
J. the high performance liquid chromatography discrimination method of Paeoniflorin in the preparation:
It is an amount of to get this preparation, adds 20～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the Paeoniflorin reference substance, adds 20～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate=5～50: 20～60: 0～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
K. the high performance liquid chromatography discrimination method of Berberine hydrochloride in the preparation:
This preparation is an amount of before getting, and adds hydrochloric acid-methyl alcohol or ethanol=0～5: 100 solution extracts, the preparation need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; With water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate-acetonitrile or methyl alcohol=0～60: be moving phase at 0～5: 5～50; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Specifically: the discrimination method of preparation comprises following all or part of content:
A. one or both thin-layer chromatography discrimination method in Berberine hydrochloride, the golden cypress in the preparation:
It is an amount of to get this preparation, the 20ml that adds diethyl ether, and sonicated 20 minutes discards ether solution, and the dregs of a decoction volatilize, and add methyl alcohol 20ml, and sonicated 20 minutes filters, and filtrate is concentrated into 2ml, as need testing solution; Get the golden cypress medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Berberine hydrochloride reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate-methyl alcohol-isopropyl alcohol-strong ammonia solution=4: 3: 2: be developping agent at 1.5: 0.5, launches, take out, dry, put under the uviol lamp 365nm and inspect, in the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
B. one or both thin-layer chromatography discrimination method in the radix paeoniae rubrathe, the Paeoniflorin in the preparation:
It is an amount of to get this preparation, gets content, adds ethanol 25ml, and sonicated 20 minutes filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to thin-layered chromatography, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, respectively with same silica gel g thin-layer plate on, with chloroform-ethyl acetate-methyl alcohol-formic acid=40: 5: 12: 0.2 was developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ dry by the fire to the spot colour developing clear; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
C. one or more thin-layer chromatography discrimination method in the red sage root, danshensu or its sodium salt, the protocatechualdehyde in the preparation:
It is an amount of to get this preparation, adds water 20ml, and ultrasonic 30 minutes, centrifugal, supernatant was regulated pH value to 2 with watery hydrochloric acid, extracted with ethyl acetate 20ml jolting, and extract evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets red sage root control medicinal material 0.5g, and it is an amount of to add water, decocts 30 minutes, adds water to 20ml, shines medicinal material solution in pairs with legal system; Get danshensu or its sodium salt, protocatechualdehyde, add methyl alcohol and make reference substance solution; Test according to thin-layered chromatography, draw one or more each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-acetone-formic acid=25: 10: 4 was developping agent, launch, take out, dry, spray is placed about 30 minutes to clear spot with 2% ferric trichloride ethanolic solution; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
D. one or more thin-layer chromatography discrimination method in the red sage root, root of red-rooted salvia phenolic acid B or its magnesium salts in the preparation:
It is an amount of to get this preparation, and 75% methyl alcohol jolting is extracted, and extract concentrates, as need testing solution; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; Root of red-rooted salvia phenolic acid B or its magnesium salts add 75% methyl alcohol and make reference substance solution; According to the thin-layered chromatography test, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put in same silica G F respectively 254On the thin layer plate, with toluene-chloroform-ethyl acetate-methyl alcohol-formic acid=2: 3: 4: be developping agent at 0.5: 2, launches, and takes out, and dries, and 254nm inspects under the uviol lamp; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
E. the thin-layer chromatography discrimination method of the root of Dahurain angelica in the preparation:
It is an amount of to get this preparation, and content adds water 20ml, and ultrasonic 30 minutes, centrifugal, supernatant was regulated pH value to 12 with sodium hydroxide test solution, extracted with chloroform 20ml jolting, and extract is concentrated into 1ml, as need testing solution; Other gets root of Dahurain angelica control medicinal material 0.5g, adds chloroform 10ml, and ultrasonic 30 minutes, filter, filtrate is concentrated into 1ml, in contrast medicinal material solution; According to thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with chloroform-methanol=9: 1, put in the vapour-saturated expansion cylinder of ammonia, launch, taking-up is dried, and puts that 254nm inspects under the ultraviolet lamp; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
F. one or both thin-layer chromatography discrimination method in peach kernel, the amarogentin in the preparation:
It is an amount of to get this preparation, adds methyl alcohol 20ml sonicated 20min, filters, filtrate evaporate to dryness, residue add water 5ml low-grade fever makes dissolving, puts cold, by the D101 type macroporous adsorptive resins of the internal diameter 1.5cm of water 20m prewashing, long 8cm,, discard water liquid, with ammonia solution 30ml wash-out, discard ammoniacal liquor, again water 10ml wash-out, discard water liquid, continue, collect eluent with 20% ethanol 30ml wash-out, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving as need testing solution; Get the peach kernel medicinal material, the 20ml that adds diethyl ether refluxes, and discards ether solution, adds methyl alcohol 20ml again and refluxes, and filters, and filtrate concentrates, in contrast medicinal material solution; Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution; Draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-methanol-water=15: 40: 22: 10 was developping agent at 5～10 ℃ of lower floor's solution of placing 12h, launch, take out, spray immediately with the phosphomolybdic acid sulfuric acid solution, at 105 ℃ of about 10min of baking; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
G. one or both thin-layer chromatography discrimination method in Herba Lycopi, the ursolic acid in the preparation:
It is an amount of to get this preparation, adds acetone 30ml, and sonicated 10 minutes is put coldly, filter, and the filtrate evaporate to dryness, residue soaks with sherwood oil 10ml, and the sherwood oil that inclines, residue add absolute ethyl alcohol 1ml makes dissolving, as need testing solution; Other gets Herba Lycopi's control medicinal material, shines medicinal material solution in pairs with legal system; Get the ursolic acid reference substance again, add absolute ethyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, with cyclohexane-methenyl choloride-ethyl acetate-formic acid=20: 5: 8: 0.1 was developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the spot of same color;
H. the high performance liquid chromatography discrimination method of protocatechualdehyde in the preparation:
It is an amount of to get this preparation, adds water 50ml, sonicated 30 minutes, take out, put, filter to room temperature, subsequent filtrate extracts 3 times with the ether jolting with 10% salt acid for adjusting pH value to 2, each 10ml, merge ether extracted liquid, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and constant volume, filters with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, puts in the brown measuring bottle, adds methyl alcohol and makes the solution that contains 12 μ g among every 1ml, in contrast product solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-1% glacial acetic acid aqueous solution=13: 87 is a moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
I. the high performance liquid chromatography discrimination method of tanshin polyphenolic acid B in the preparation:
It is an amount of to get this preparation, adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 75% methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-formic acid-water=30: 10: 1: 59 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
J. the high performance liquid chromatography discrimination method of Paeoniflorin in the preparation:
It is an amount of to get this preparation, adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance that is dried to constant weight, adds methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.05mol/L potassium dihydrogen phosphate=40: 65 is moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
K. the high performance liquid chromatography discrimination method of Berberine hydrochloride in the preparation:
It is an amount of to get this preparation, adds the mixed solution sonicated 30 minutes of hydrochloric acid-methyl alcohol=1: 100, takes out, and puts to room temperature, supplies the solvent of loss, shakes up, and filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With the 0.05mol/L sodium dihydrogen phosphate-acetonitrile of phosphorus acid for adjusting pH value to 3.0=75: 25 was moving phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
The content assaying method of this preparation comprises following all or part of content:
A. the high-efficient liquid phase chromatogram process measuring method of preparation Central Plains catechu aldehyde:
It is an amount of to get this preparation, and accurate the title decides, extracting in water, and water liquid prepares need testing solution with ether or ethyl acetate or chloroform jolting extraction; Or it is an amount of to get this preparation, adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the protocatechualdehyde reference substance, adds 10～100% methyl alcohol or ethanol and is made as reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-acetate or formic acid or phosphoric acid=10～50: 30～95: 0.01～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures, and calculates with one point external standard method or calibration curve method; Content limit should be: must not be lower than 108 μ g with containing the red sage root in this preparation in protocatechualdehyde on 1st;
B. the high-efficient liquid phase chromatogram process measuring method of content of danshinolic acid B in the preparation:
It is an amount of to get this preparation, and accurate the title decides, and adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 10～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or ethanol-acetonitrile-formic acid or acetate or phosphoric acid-water=10～30: 0～30: 0～5: 50～95 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures, and calculates with one point external standard method or calibration curve method; Content limit should be: must not be lower than 10.8mg with containing the red sage root in this preparation in tanshin polyphenolic acid B on 1st;
C. the high-efficient liquid phase chromatogram process measuring method of paeoniflorin content in the preparation:
It is an amount of to get this preparation, and accurate the title decides, and adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the Paeoniflorin reference substance, adds 10～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate=5～50: 20～60: 0～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method or calibration curve method; Content limit should be: must not be lower than 13.2mg with containing the radix paeoniae rubrathe in this preparation in Paeoniflorin on 1st;
D. the high-efficient liquid phase chromatogram process measuring method of content of berberine hydrochloride in the preparation:
It is an amount of to get this preparation, and accurate the title decides, and adds hydrochloric acid-methyl alcohol=0～5: 100 solution extracts, the preparation need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; With water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate-acetonitrile or methyl alcohol=20～60: be moving phase at 0～5: 5～50; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures, and calculates with one point external standard method or calibration curve method; Content limit should be: must not be lower than 36mg with containing golden cypress in this preparation in Berberine hydrochloride on 1st.
Specifically, the content assaying method of this preparation comprises following all or part of content:
A. the high-efficient liquid phase chromatogram process measuring method of preparation Central Plains catechu aldehyde:
It is an amount of to get this preparation, and accurate the title decides, and puts in the tool plug conical flask, precision adds water 50ml, and weight, sonicated 30 minutes decided in accurate title, take out, put to room temperature, weight decided in accurate title, add water and supply the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 10ml that draws, with 10% salt acid for adjusting pH value to 2, extract 3 times each 10ml with the ether jolting, merge ether extracted liquid, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and quantitatively is transferred in the 10ml measuring bottle, adds methyl alcohol to scale, shake up, filter with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, and accurate the title decides, and puts in the brown measuring bottle, adds methyl alcohol and makes the solution that contains 12 μ g among every 1ml, in contrast product solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-1% glacial acetic acid aqueous solution=13: 87 is a moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 216 μ g with containing the red sage root in this preparation in protocatechualdehyde on 1st;
B. the high-efficient liquid phase chromatogram process measuring method of content of danshinolic acid B in the preparation:
It is an amount of to get this preparation, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, and accurate the title decides, and adds 75% methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-formic acid-water=30: 10: 1: 59 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 21.6mg with containing the red sage root in this preparation in tanshin polyphenolic acid B on 1st;
C. the high-efficient liquid phase chromatogram process measuring method of paeoniflorin content in the preparation:
It is an amount of to get this preparation, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance that is dried to constant weight, and accurate the title decides, and adds methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.05mol/L potassium dihydrogen phosphate=40: 65 is moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 26.4mg with containing the radix paeoniae rubrathe in this preparation in Paeoniflorin on 1st;
D. the high-efficient liquid phase chromatogram process measuring method of content of berberine hydrochloride in the preparation:
It is an amount of to get this preparation, accurate claims surely, adds the mixed solution sonicated 30 minutes of hydrochloric acid-methyl alcohol=1: 100, takes out, and puts to room temperature, supplies the solvent of loss, shakes up, and filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With the 0.05mol/L sodium dihydrogen phosphate-acetonitrile of phosphorus acid for adjusting pH value to 3.0=75: 25 was moving phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 72mg with containing golden cypress in this preparation with Berberine hydrochloride on 1st.
Compared with prior art, product provided by the invention has clearing heat and promoting diuresis, and effect promoting blood circulation and removing blood stasis is used for syndrome of stagnant dampness-heat, and disease is seen: frequent micturition, and urgent urination, it is not smooth to urinate, and lower abdominal distention pains etc. have reasonable result of treatment for acute and chronic prostatitis; That the preparation of preparation has is easy to carry, cover bitter taste, characteristics such as absorbing soon can be good, good anti-moisture ability, and the preparation method is rationally feasible, and production cost is suitable, and economic benefit is more satisfactory; Detection method can guarantee the quality and the curative effect of product; Overcome the problem that prior art, product exist; Reached the purpose of invention.
The applicant under study for action with amount of water as factor, the varying level of high spot reviews factor is taken all factors into consideration in conjunction with aspects such as production cost, the energy decocting the influence of extraction effect.For the moulding process of capsule, the relative humidity that the applicant should control during to its auxiliary material, packing is investigated.For pill, dressing can increase moisture resistance, but also can influence disintegration rate, so the selection of coating material is just very crucial.The dressing auxiliary material of the most suitable this product and consumption, ratio, process conditions etc. have been found by experiment; Guarantee its science, reasonable, feasible; The preparation that obtains has superperformance and result of treatment.
Experimental example 1: Study on extraction
1.1 volatile oil extracts experiment
(1) testosterone propionate is caused the influence of rat prostate hyperplasia
50 of Wistar rats, male, be divided into five groups at random by body weight: normal control group, model control group, estradiol control group, drug extract group (not extracting volatile oil), drug extract group (extraction volatile oil).Rat excision bilateral testes begins the hypodermic injection testosterone propionate after 1 week of recuperating, and begins administration simultaneously, continuous 1 month.The last administration was put to death animal after 24 hours, separate to take out prostate, weighed and carried out between each group relatively.The result shows: the model group prostate is heavy apparently higher than the normal control group, and estradiol control group prostate is heavy significantly to be reduced with model control group.Drug extract group (not extracting volatile oil) does not have marked difference with drug extract group (extraction volatile oil) effect.
(2) P-xylene causes the influence of mice ear
Get body weight 18-22g mouse, irritate stomach and give pharmaceutical composition of the present invention, every day 1 time, continuous 7 days, be coated with dimethylbenzene 0.1ml in the mouse auris dextra back of the body in 30 minutes after the last administration, mouse is put to death in the cervical vertebra dislocation after 2 hours, lays round auricle with card punch in left and right sides ear same area, weighs, ratio with interaural difference and left ear is the swelling degree, compare each group difference, the result shows: drug extract group (not extracting volatile oil) does not have marked difference with drug extract group (extraction volatile oil) effect, sees the following form.
Group Number of animals Contrast Cause scorching ear weight (%)
Control group 10 100.0 189.6±39.7
Aspirin 10 100.0 132.9±28.5
Drug extract group (extraction volatile oil) 10 100.0 121.1±31.6
Drug extract group (not extracting volatile oil) 10 100.0 120.8±12.5
1.2 decocting process research
(1) factor is selected: the decocting for Chinese herbal medicine extraction effect is subjected to the influence of factors such as amount of water, extraction time, extraction time.Prior art is investigated extraction time, extraction time, so when the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb, the root of Dahurain angelica and peach kernel medicinal material decoct investigation, choose amount of water as factor, the varying level of high spot reviews factor is to decocting the influence of extraction effect.Take all factors into consideration the selection factor level in conjunction with aspects such as production cost, the energy.
(2) index is determined: select medicinal extract recovery rate and protocatechualdehyde content as evaluation index, its reason and assay method are as follows:
1. medicinal extract recovery rate: medicinal extract is the material base of solid pharmaceutical preparation performance curative effect, and its yield height directly influence preparation process, is reasonable, effective control device so be chosen as the extraction index.
2. assay: medicinal extract recovery rate height can not reflect fully that effective ingredient extracts situation, so measure the contained protocatechualdehyde content of the red sage root simultaneously as the decoction screening index.With reference to relevant document, adopt high-efficient liquid phase technique to measure the content of protocatechualdehyde.
Test method: take by weighing radix paeoniae rubrathe 200g, red sage root 100g, Herba Lycopi 120g, root of three-nerved spicebush 120g, when the seed of cowherb 80g and the root of Dahurain angelica 80 add water and are heated to 80 ℃, add peach kernel 140 again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction filtered, filtrate concentrating adjusted and is settled to 1000ml, precision is measured 50ml, puts in the evaporating dish that is dried to constant weight, behind evaporate to dryness in the water-bath, in 105 ℃ of dryings 3 hours, move in the exsiccator, cooled off 30 minutes, weight decided in accurate rapidly title, calculate paste-forming rate, measure protocatechualdehyde content simultaneously.
(3) test: test arrangement and the results are shown in following table.
Amount of water is investigated table as a result
As seen from the above table, it is that 10,10,10 times medicinal extract recovery rate difference is little with amount of water that amount of water is 8,8,8 times, but being 8,8,8 times from protocatechualdehyde content amount of water, to be better than amount of water be 10 times of amounts, therefore considered with becoming originally from energy savings, the optimised process of amount of water is boiling three times, and amount of water is 8,8,8 times.And carry out confirmatory experiment according to this condition.
Table is as a result investigated in the amount of water checking
As seen from the above table, medicinal extract recovery rate and protocatechualdehyde content difference of each group are little, and the condition that screening be described is stablized feasible, so the optimised process of definite amount of water is boiling three times, add 8 times of amounts at every turn.And carry out confirmatory experiment according to this condition.
In sum, the optimum extraction process of the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb, the root of Dahurain angelica and peach kernel is that the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb and the root of Dahurain angelica are when adding 8 times of water and being heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time.
Experimental example 2 capsule Study on Forming
2.1 hydroscopicity is investigated
Golden cypress is ground into fine powder in the prior art, and we carry out hydroscopicity investigation test to the mixture that extraction ointment, ointment add behind the golden cypress medicinal powder, the results are shown in following table.
The wettability test result
Investigate by wettability test, think that the medicinal extract that adds the crude drug powder has anti-preferably hygroscopic effect.
2.2 thinning agent is selected
The material properties of considering extraction is basic identical, so we select for use starch to adjust prescription weight as thinning agent.
2.3 mobile the investigation
For guaranteeing that divided dose is accurate, require the good flowability of fill material tool, so to measure the flowability of angle of repose method examination mixture.
Fixed funnel method: get 3 funnel series connection, lowermost end is apart from horizontal positioned coordinate paper 1.5cm place, carefully fill material is poured into along hopper walls in the funnel of going up most till the particle cone tip that bottom funnel forms touches the funnel end opening, measure the diameter of conical base by coordinate paper, (tg α=H/R/2), result of calculation sees the following form to calculate the angle of repose.
The angle of repose α of fill material
Working sample The angle of repose
1 53.5°
2 52.3°
3 54.8°
As seen from the above table, mixture angle of repose〉40 °, show that promptly the mixture property that flows is bad, can not satisfy the branch reload request.
2.4 glidant is investigated
From top angle of repose measurement result as can be seen, mixture mobile relatively poor, divided dose is accurate when guaranteeing that capsule is loaded, and needs to increase the flowability of fill material, and we screen glidant for this reason.Owing to used starch as thinning agent in the mixture, still no longer consider to add again its consumption; Talcum powder can increase lubricity and flowability, and possess hydrophilic property, can not influence the disintegration of capsule, thus we to adopt talcum powder be glidant, and its consumption is investigated, test findings sees the following form.
The amount of talc investigation table
Sequence number Amount of talc (%) The angle of repose (°)
1 2 43.6
2 5 32.7
3 8 31.8
From top result as can be seen, consumption was at 2% o'clock, angle of repose to mixture improves to some extent, but can't meet the demands, consumption is 5% and can reaches requirement at 8% o'clock, but amount of talc is 8% o'clock, and the angle of repose decline scope is not clearly, so we determine that amount of talc is 5%.
2.5 the fill material bulk density is measured and capsulae vacuus is selected
Generally according to the difference of crystal formation, fineness, density and the dosage of medicine, particularly bulk density is determined in the selection of capsulae vacuus specification.Adopt the graduated cylinder method to measure the mixture bulk density, the potpourri that is about to precision weighing is packed in the dry 10ml graduated cylinder, about jolting gently, 20 times back and forth, recording capacity calculates, and the results are shown in following table.
The bulk density measurement result
The result records the about 0.60 (g/cm of filler particles bulk density 3), according to the relation of capsulae vacuus number and approx. volume, select the packing of No. 0 hungry area softgel shell, promptly every capsules loading amount is about 0.45g (being equivalent to 1.04g crude drug/grain).
2.6 critical relative moisture (CRH) is measured
Material can be loaded smoothly in order to guarantee to produce, and the humidity that need control environment so that make material have good mobility, makes loading amount even, and we carry out equilibrium hygroscopicity test, i.e. moisture equilibrium at dry side curve determination to mixture for this reason.Get totally six parts of the about 1g of mixture, put in the measuring cup, the accurate title, decide, the weighing bottle cap is opened, put into relative humidity respectively and be 20%, 33%, 43%, 60%, 75%, 92% environment, in 25 ℃ of constant incubators, placed 84 hours, take out measuring cup, it is fixed to add a cover the accurate title in back, calculating the moisture absorption percent, is horizontal ordinate with relative humidity, and the moisture absorption percent is an ordinate, draw the moisture equilibrium at dry side curve and see accompanying drawing 1, test findings sees the following form.
The moisture absorption percent (%) of filler particles
The saturated salt solution kind Relative humidity (%) Moisture absorption percent (%)
CH 3COOK·1.5H 2O 20 3.11
MgCl 2·6H 2O 33 4.62
K 2CO 3·2H 2O 43 7.24
NaBr·2H 2O 60 13.20
NaCl 75 22.22
KNO 3 92 42.97
As can be known from the above table, capsule is under different relative humidity environment, and its water absorbing capacity does not wait, and is about 65.0% when following in relative humidity, and hydroscopicity is less, so should control relative humidity below 65.0% during packing.
Experimental example 3 dripping pill Study on Forming
3.1 matrix screening
According to bibliographical information, dripping pill matrix commonly used has Macrogol 4000 and Macrogol 6000, thus our decision also to adopt polyglycol be matrix, and the two is compared test.The polyglycol of different model is put in the small beaker, be heated to 80-90 ℃, after treating whole fusions, add extract powder, investigate matrix and extract powder the fusion situation, select fusion situation dripping system (the system condition: expect warm 80 ℃ of writing out a prescription preferably, cooling medium is a dimethyl silicon oil, drip apart from 6cm, drip 30～40 droplets/minute of speed), the results are shown in following table.
The fusion situation of matrix and main ingredient relatively
The prescription number Prescription 1 Prescription 2 Prescription 3 Prescription 5 Prescription 6 Prescription 7
Extract powder (g) 10 10 10 10 10 10
Macrogol 4000 (g) 10 15 20 ---- ---- ----
Macrogol 6000 (g) ---- ---- ---- 10 15 20
Main ingredient: matrix 1∶1 1∶1.5 1∶2 1∶1 1∶1.5 1∶2
The fusion situation of main ingredient and matrix Main ingredient can merge with matrix, but system does not have flowability Main ingredient can merge with matrix, and the system flowability is fine Main ingredient can merge with matrix, and the system flowability is fine Main ingredient and matrix merge relatively poor Main ingredient can merge with matrix, but system does not have flowability Main ingredient can merge with matrix, and system flows better
The dripping pill outward appearance ---- Smooth, roundness is good Smooth, roundness is good ---- Roundness is poor, serious hangover Roundness is poor, hangover
Dripping pill hardness ---- Hardness is better Hardness is better ---- Hardness is better Hardness is better
The ball method of double differences is different ---- 4.3％ 6.8％ ---- ---- 11％
Dissolve scattered time limit (min) ---- 6～8 7～9 ---- ---- 12～18
The above results shows, the good fluidity of the 2 fusion soups of writing out a prescription, and the dripping pill good moldability, smooth, mellow and full, the ball method of double differences is different little, and molten loosing comparatively fast is so select No. 2 prescriptions to be optimal condition.
3.2 cooling medium is selected
The get it filled extract powder 10g of material, Macrogol 4000 15g mixes, and is heated to 80-90 ℃, treat whole fusions after, get in right amount, splash into respectively in dimeticone and the whiteruss cooling medium, be index with the moulding situation of dripping pill, the results are shown in following table.
Cooling medium is selected
Figure C20061020051100271
Last table shows, is that cooling medium dripping pill roundness is good with the dimeticone, forming.
3.3 coolant temperature is selected
Get the extract powder 10g of three kinds of medicinal materials, Macrogol 4000 15g mixes, and is heated to 80-90 ℃, treat whole fusions after, get in right amount, splash into respectively in the dimeticone cooling medium of different temperatures, observe dripping pill moulding situation, the results are shown in following table.
Coolant temperature is selected
Coolant temperature Drip distance Drip speed The material temperature Dripping pill moulding situation
10℃ 6cm 30～40d/min 80℃ Roundness is good, forming
20℃ 6cm 30～40d/min 80℃ Roundness is good, forming
The gradient cooling 6cm 30～40d/min 80℃ Roundness is good, forming
Annotate: the gradient cooling means is: top is 10～20 ℃, and the bottom is 5～10 ℃.
Last table shows that under above-mentioned three kinds of chilling temperatures, the mouldability of this product is all good, is easy operation, is 10～20 ℃ so select coolant temperature.
3.4 disintegration time is investigated
Disintegration time (min)
Group 123
Dripping pill 29.3 29.8 29.7 of the present invention
ANLIETONG PIAN 52.3 50.3 49.8
The result as can be known, technology gained preparation of the present invention has disintegration effect preferably, is beneficial to the absorption of medicine.
Experimental example 4: to the research of prostatitis treating effect
50 of SD rats, anesthesia back routine disinfection cuts about skin 1.5cm.Isolate prostate, 40 rats are wherein caused non-bacterial chronic prostatitis animal model from 2% agar solution 0.1ml of left lobe of prostate, each injection sterilization of lobus dexter, all the other 10 rats are respectively injected sterile saline 0.1ml as the normal control group at left lobe of prostate, lobus dexter.Sew up muscle, skin then respectively, operation back 24h, with the rat random packet, 10 every group.Normal control group: distilled water 1ml/100g body weight; Model control group: distilled water 1ml/100g body weight; Commercially available ANLIETONG PIAN group, Capsules group of the present invention, Capsules group of the present invention: give the relative medicine suspension respectively, the administration volume is the 1ml/100g body weight, and drug dose is equivalent to be grown up clinical plan with 10 times of dosage.Each treated animal gastric infusion every day (or distilled water) 1 time is treated 45d continuously.24h after the last administration behind the title the weight of animals, puts to death the disconnected neck of rat, peels off prostate rapidly, after weighing, puts into the liquid-solid 48h of deciding of FAA, ethanol dehydration step by step, and dimethylbenzene is transparent, waxdip, paraffin embedding, conventional section 4 μ m, HE dyes.Quantity and morphological change situation with matter inflammatory cell and body of gland and lumen of gland between pattern analysis instrument connection Computer Analysis processing rat prostate.
1. to the influence of chronic prostatitis rat prostate weight coefficient due to the agar, concrete outcome sees the following form:
Influence to chronic prostatitis rat prostate weight coefficient due to the agar
Group n dosage (m/gkg -1) weight of prostate coefficient (m/mg100g -1)
Normal control group 10-138.5 ± 26.7
Model control group 10-228.9 ± 36.4
ANLIETONG PIAN group 10 3.2 184.6 ± 45.7
Capsules group 10 3.2 169.1 ± 42.6 of the present invention
Dripping pill group 10 3.2 158.7 ± 26.8 of the present invention
From last table as seen, use ANLIETONG PIAN, capsule of the present invention, dripping pill of the present invention treatment 45d, its weight of prostate coefficient ratio model control group all reduces, and the effect of capsule of the present invention, dripping pill is better than commercially available ANLIETONG PIAN.
2. to the influence of matter inflammatory cell quantity and form between prostate, concrete outcome sees the following form:
Influence to matter inflammatory cell quantity and form between prostate
Dosage number of inflammatory cells inflammatory cell area summation area average
Group (m/gkg -1) (individual) (A/ μ m 2) (A/ μ m 2)
The normal control group----
Model control group-142.5 ± 59.4 104632.5 ± 28214.1 957.2 ± 432.7
ANLIETONG PIAN group 3.2 148.4 ± 74.5 55624.8 ± 30483.4 486.4 ± 158.8
Capsules group 3.2 146.8 of the present invention ± 26.4 48276.1 ± 25631.5 456.2 ± 125.9
Dripping pill group 3.2 148.2 of the present invention ± 62.8 45684.4 ± 15324.6 395.7 ± 265.4
From last table as seen, behind application ANLIETONG PIAN, capsule of the present invention, the dripping pill treatment 45d, inflammatory cell area summation, average area reduce.Inflammatory cell quantity and model control group are relatively, do not see minimizing, even slightly increase, this is owing to number of inflammatory cells in the matter between the model control group prostate is more, area is bigger, normal adhesion slabbing, and computing machine will connect to the reason that a plurality of inflammatory cell identifications of sheet are calculated as 1 inflammatory cell.
3. to the influence of prostate body of gland quantity and form
Influence to chronic prostatitis rat prostate body of gland quantity due to the agar and form
Area average girth average maximum gauge average minimum diameter average
Group (A/ μ m 2) (A/ μ m 2) (A/ μ m 2) (A/ μ m 2)
Normal control group 11265 ± 4,837 513.8 ± 59.4 359.5 ± 162.4 86.2 ± 26.7
Model control group 12572 ± 2,694 536.4 ± 156.7 137.2 ± 42.6 58.6 ± 16.7
ANLIETONG PIAN group 12893 ± 3,488 548.7 ± 124.1 185.7 ± 36.7 78.4 ± 12.1
Capsules group 13254 of the present invention ± 1,548 567.2 ± 134.8 159.6 ± 59.8 68.5 ± 13.4
Dripping pill group 12687 of the present invention ± 2,362 539.1 ± 112.7 165.2 ± 89.1 75.1 ± 19.5
As seen from the above table, after using ANLIETONG PIAN, capsule of the present invention, dripping pill treatment 45d, relatively each organizes the therapeutic action that all has in various degree with model control group, body of gland area average, body of gland maximum gauge average etc. are obviously raise, and the effect of product of the present invention is better than commercially available ANLIETONG PIAN.
Experimental example 5: the research of detection method
5.1 the thin-layer chromatography discrimination method of Berberine hydrochloride, golden cypress in the capsule
The purpose of this experiment is the feature for outstanding golden cypress, and for getting rid of in the preparation interference with other composition like the contained constituent classes such as Berberine hydrochloride of golden cypress, the experimenter compares test respectively to sample extraction, development system, coloration method etc.; Through screening, determined the best approach and condition: it is an amount of to get the prostatitis-treating preparation, the 20ml that adds diethyl ether, and sonicated 20 minutes discards ether solution, and the dregs of a decoction volatilize, and add methyl alcohol 20ml, and sonicated 20 minutes filters, and filtrate is concentrated into 2ml, as need testing solution; Get the golden cypress medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Berberine hydrochloride reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate-methyl alcohol-isopropyl alcohol-strong ammonia solution=4: 3: 2: be developping agent at 1.5: 0.5, launches, take out, dry, put under the uviol lamp 365nm and inspect, in the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.The expansion effect of several development systems relatively sees the following form in the experiment:
The thin-layer chromatography discrimination method of Berberine hydrochloride, golden cypress in the capsule
Developping agent Thin layer plate Effect
Benzene-ethyl acetate-ethanol-formic acid (8: 6: 1: 1) Silica G Difference: sample separation is unintelligible
Benzene-butyl acetate-methyl alcohol (8: 4: 3) Silica G Difference: feminine gender has interference, the spot hangover
Benzene-ethyl acetate-methyl alcohol-strong ammonia solution (4: 3: 3: 0.5) Silica gel H Preferable: it is clear that sample and reference substance all separate, negative noiseless
Benzene-ethyl acetate-methyl alcohol-isopropyl alcohol-strong ammonia solution (4: 3: 2: 1.5: 0.5) Silica G Best: it is clear, negative noiseless that sample and reference substance all separate
5.2 the thin-layer chromatography discrimination method of the radix paeoniae rubrathe, Paeoniflorin in the pill
The purpose of this experiment is the feature for the outstanding radix paeoniae rubrathe, and for getting rid of in the preparation interference with other composition like the contained constituent classes such as Paeoniflorin of the radix paeoniae rubrathe, the experimenter compares test respectively to sample extraction, development system, coloration method etc.; Through screening, determined the best approach and condition: it is an amount of to get the prostatitis-treating preparation, gets content, adds ethanol 25ml, and sonicated 20 minutes filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin-layered chromatography test, draw each 5 μ l of above-mentioned solution, respectively and on the same silica gel g thin-layer plate, with chloroform-ethyl acetate-methyl alcohol-formic acid=40: 5: 12: 0.2 was developping agent, launches, and takes out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ dry by the fire to spot colour developing clear; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
The thin-layer chromatography discrimination method of the radix paeoniae rubrathe, Paeoniflorin in the pill
Developping agent Thin layer plate Effect
Benzene-ethyl acetate-ethanol-formic acid (8: 4: 5: 1) Silica G Difference: sample separation is unintelligible
Ethyl acetate-methyl alcohol-formic acid (5: 12: 1) Silica gel H Difference: feminine gender has interference
Chloroform-ethyl formate-ethanol-acetate (20: 5: 8: 1) Silica G F 254 Preferable: it is clear that sample and reference substance all separate, negative noiseless
Chloroform-ethyl acetate-methyl alcohol-formic acid (40: 5: 12: 0.2) Silica G Best: it is clear, negative noiseless that sample and reference substance all separate
5.3 the thin-layer chromatography discrimination method of the red sage root, danshensu, protocatechualdehyde in the granule
The purpose of this experiment is the feature for the outstanding red sage root, and for getting rid of in the preparation interference of other composition like the constituent classes such as the danshensu contained with the red sage root, protocatechualdehyde, the experimenter compares test respectively to sample extraction, development system, coloration method etc.; Through screening, determined the best approach and condition: it is an amount of to get the prostatitis-treating preparation, adds water 20ml, ultrasonic 30 minutes, centrifugal, supernatant is regulated pH value to 2 with watery hydrochloric acid, extracts with ethyl acetate 20ml jolting, extract evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets red sage root control medicinal material 0.5g, and it is an amount of to add water, decocts 30 minutes, adds water to 20ml, shines medicinal material solution in pairs with legal system; Get danshensu or its sodium salt, protocatechualdehyde, add methyl alcohol and make reference substance solution; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with chloroform-acetone-formic acid=25: 10: 4 was developping agent, launched, and took out, dry, spray ferric trichloride ethanolic solution, place about 30 minutes to clear spot with 2%; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
The thin-layer chromatography discrimination method of the red sage root, danshensu, protocatechualdehyde in the granule
Developping agent Thin layer plate Effect
Benzene-ethyl acetate-ethanol (8: 4: 5) Silica G Difference: sample separation is unintelligible
Ethyl acetate-methyl alcohol-formic acid (5: 5: 1) Silica G F 254 Difference: feminine gender has interference
Methylene chloride-acetone-formic acid (5: 2: 4) Silica gel H Preferable: it is clear that sample and reference substance all separate, negative noiseless
Chloroform-acetone-formic acid (25: 10: 4) Silica G Best: it is clear, negative noiseless that sample and reference substance all separate
5.4 the thin-layer chromatography discrimination method of the root of Dahurain angelica in the capsule
The purpose of this experiment is the feature for the outstanding root of Dahurain angelica, and for getting rid of in the preparation interference with other composition like the contained constituent class of the root of Dahurain angelica, the experimenter compares test respectively to sample extraction, development system, coloration method etc.; Through screening, determined the best approach and condition: it is an amount of to get the prostatitis-treating preparation, and content adds water 20ml, ultrasonic 30 minutes, centrifugal, supernatant was regulated pH value to 12 with sodium hydroxide test solution, extract with chloroform 20ml jolting, extract is concentrated into 1ml, as need testing solution; Other gets root of Dahurain angelica control medicinal material 0.5g, adds chloroform 10ml, and ultrasonic 30 minutes, filter, filtrate is concentrated into 1ml, in contrast medicinal material solution; According to thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with chloroform-methanol=9: 1, put in the vapour-saturated expansion cylinder of ammonia, launch, taking-up is dried, and puts that 254nm inspects under the ultraviolet lamp; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
The thin-layer chromatography discrimination method of the root of Dahurain angelica in the capsule
Developping agent Thin layer plate Effect
Benzene-ethyl acetate-ethanol (8: 4: 1) Silica G Difference: sample separation is unintelligible
Ethyl acetate-ethanol-formic acid (5: 2: 1) Silica gel H Difference: feminine gender has interference
Chloroform-ethanol (4: 1) Silica gel H Preferable: it is clear that sample and reference substance all separate, negative noiseless
Chloroform-methanol (9: 1) Silica G Best: it is clear, negative noiseless that sample and reference substance all separate
5.5 the high-efficient liquid phase chromatogram process measuring method of capsule Central Plains catechu aldehyde
Test is reference substance with the protocatechualdehyde, with Alltech P426 high performance liquid chromatograph, has established the best approach of the assay of protocatechualdehyde in the prostatitis-treating capsule:
It is an amount of to get the prostatitis-treating preparation, and accurate the title decides, and puts in the tool plug conical flask, precision adds water 50ml, and weight, power 250W decided in accurate title, sonicated is 30 minutes under the frequency 33KHz, take out, put to room temperature, weight decided in accurate title, add water and supply the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 10ml that draws, with 10% salt acid for adjusting pH value to 2, extract 3 times each 10ml with the ether jolting, merge ether extracted liquid, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and quantitatively is transferred in the 10ml measuring bottle, adds methyl alcohol to scale, shake up, filter with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, and accurate the title decides, and puts in the brown measuring bottle, adds methyl alcohol and makes the solution that contains 12 μ g among every 1ml, in contrast product solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-1% glacial acetic acid aqueous solution=13: 87 is a moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by former a little tea aldehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: every of this product contains protocatechualdehyde, must not be lower than 18.0 μ g.
This method has been passed through following 1～11 methodological study test respectively:
1 detects the selection of wavelength
It is an amount of that precision takes by weighing the protocatechualdehyde reference substance, adds methyl alcohol respectively and make the solution that every 1ml contains 13 μ g, respectively in the interscan of 200～400nm wavelength coverage.The result shows that protocatechualdehyde has absorption maximum at the 281nm place, therefore selects 281nm as the detection wavelength of measuring prostatitis-treating capsule Central Plains catechu aldehyde.
The selection of 2 extraction times
Get 40 of this product, get content, mixing, therefrom precision takes by weighing about 6g (totally 4 minutes), splits in the tool plug conical flask, and precision adds water 50ml, weight decided in accurate title, respectively sonicated (power 250W, frequency 33KHz) 10,20,30,40 minutes, take out, put to room temperature, weight decided in accurate title, add water and supply the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 10ml that draws with 10% salt acid for adjusting pH value to 2, extracts 3 times with the ether jolting, each 10ml merges ether extracted liquid, puts evaporate to dryness in the water-bath, residue adds dissolve with methanol and quantitatively is transferred in the 10ml measuring bottle, add methyl alcohol to scale, shake up, filter with miillpore filter (0.45 μ m), get subsequent filtrate, promptly.See the following form.
Extraction time
Extraction time (min) Protocatechualdehyde (μ g/ grain)
10 21.47
20 22.46
30 23.69
40 23.72
The result shows that sonicated 30min can extract fully, so extraction time is decided to be 30 minutes.
3 chromatographic conditions
Chromatograph: Alltech P426;
Chromatographic column: Diamonsil (diamond) C 18(250 * 4.6mm, 5 μ m);
Moving phase: methyl alcohol-1% glacial acetic acid aqueous solution (13: 87);
Detect wavelength: 281nm;
Column temperature: 30 ℃;
Flow velocity: 1ml/min;
Sample size: 10 μ l.
Obtain protocatechualdehyde, test sample chromatogram according to above-mentioned condition, its number of theoretical plate is pressed the protocatechualdehyde peak and is calculated greater than 2000.In the sample protocatechualdehyde chromatographic peak separate with close peak clear fully, degree of separation is all greater than 1.5.
The test of 4 negative ELIMINATION OF ITS INTERFERENCE is for investigating the mensuration whether other medicinal material disturbs protocatechualdehyde, except that golden cypress, takes by weighing other medicinal material and auxiliary material is made negative control product solution and mensuration with method in the prescription ratio.The result shows that negative sample is noiseless to the assay of protocatechualdehyde.
5 reference substance purity tests provide protocatechualdehyde by Nat'l Pharmaceutical ﹠ Biological Products Control Institute, are respectively 99.1%, 99.2% through high effective liquid chromatography for measuring purity, meet assay reference substance requirement.
The investigation precision of 6 linear relationships is measured protocatechualdehyde reference substance solution 0.2ml, 0.6m, 1.0ml, 1.4ml, 1.8ml, splits in the 25ml measuring bottle, and it is fixed to scale to add methyl alcohol, shake up, the therefrom accurate respectively 10 μ l that draw inject liquid chromatograph, according to high effective liquid chromatography for measuring.Amount with protocatechualdehyde is a horizontal ordinate, and peak area is that ordinate is figure, the drawing standard curve.See the following form
The protocatechualdehyde linear relationship
Numbering Protocatechualdehyde amount (μ g) Peak area
1 0.034368 84202
2 0.103104 250682
3 0.171840 414574
4 0.240576 583623
5 0.309312 752806
Regression equation: Y=2429802.43X-359.85
Related coefficient: γ=0.9999
The result shows that protocatechualdehyde linear relationship within 0.034368 μ g～0.309312 μ g scope is good.
Through calculating, the protocatechualdehyde typical curve is one to cross the straight line of initial point, therefore selects one point external standard method to measure the content of protocatechualdehyde in the prostatitis-treating capsule.
The accurate protocatechualdehyde reference substance solution 10 μ l that draw of 7 precision test inject liquid chromatograph, and replication 5 times is investigated reference substance solution precision, and measurement result sees the following form.
The precision test
Test number (TN) 1 2 3 4 5 Mean value ?RSD(％)
Peak area 305123 310712 308924 307201 302658 306924 1.03
The result shows that reference substance solution precision is good.
8 stability tests
8.1 the accurate protocatechualdehyde reference substance solution 10 μ l that draw of reference substance stability test inject liquid chromatograph, measure at 0,2,6,10,24 hour sample introduction respectively, measurement result sees the following form.
Reference substance solution stability test result
Time (h) 0 2 6 10 24 Mean value ?RSD(％)
Peak area 305123 310712 308924 307201 302658 306924 1.03
The result shows that reference substance solution is good at 24 hours internal stabilities.
8.2 the accurate need testing solution 10 μ l that draw of need testing solution stability test inject liquid chromatograph, measure at 0,2,6,10,24 hour sample introduction respectively, measurement result sees the following form.
Need testing solution stability test result
Time (h) 0 2 6 10 24 Mean value ?RSD(％)
Content (μ g/ grain) 24.12 23.85 23.76 23.91 23.85 23.90 0.57
The result shows that need testing solution is good at 24 hours internal stabilities.
9 replica tests are got this product content, and mixing is therefrom got about 6.0g, and parallel 5 parts, precision claims fixed, by operating under preparation of text need testing solution and the mensuration item.The results are shown in following table.
Replica test
Time (h) 0 2 6 10 24 Mean value ?RSD(％)
Content (μ g/ grain) 23.39 23.27 22.58 23.61 23.35 23.24 1.68
The result shows that repeatability is good.
The test of 10 average recoveries
Get this product content, mixing, therefrom precision takes by weighing about 3g, parallel 6 parts, splits in the tool plug conical flask; Precision takes by weighing protocatechualdehyde 15.84mg, puts in the 100ml measuring bottle, adds water and makes dissolving and fixed to scale in right amount, shake up, precision is measured 0.8ml, 1.0ml, 1.2ml each 2 parts, put altogether in the above-mentioned tool plug conical flask, precision adds water to 50ml, and weight, sonicated 30 minutes decided in accurate title, take out, put to room temperature, the accurate title, decided weight, adds water and supply the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 10ml that draws is with 10% salt acid for adjusting pH value to 2, extract 3 times with the ether jolting, each 10ml merges ether extracted liquid, puts evaporate to dryness in the water-bath, residue adds dissolve with methanol and quantitatively is transferred in the 10ml measuring bottle, add methyl alcohol to scale, shake up, filter with miillpore filter, get subsequent filtrate, promptly.Measurement result sees the following form.
The test of protocatechualdehyde average recovery
Numbering Test sample weighing (g) Pure product amount (μ g) in the test sample Protocatechualdehyde addition (μ g) The amount of recording (μ g) The recovery (%)
1 2.94873 156.21 126.72 277.64 95.83
2 3.12065 165.32 126.72 286.23 95.42
3 3.04721 161.43 158.40 319.24 99.63
4 3.07638 162.97 158.40 320.51 99.46
5 3.04296 161.20 190.08 347.61 98.06
6 3.12553 165.58 190.08 351.74 97.94
Protocatechualdehyde average recovery rate=97.72%, RSD=1.81%.
11 sample sizes are measured the preparation and the operation down of determination method item of pressing the text need testing solution, measure ten batch samples, the results are shown in following table.
Ten batch sample assay results
Lot number Protocatechualdehyde (μ g/ grain)
1 23.12
2 20.97
3 24.56
4 43.87
5 20.64
6 23.12
7 22.48
8 21.79
9 22.03
10 20.58
Conclusion: tentative according to 10 batch sample assay results, every of this product contains protocatechualdehyde, must not be lower than 18.0 μ g.
Description of drawings:
Accompanying drawing 1 is a critical relative moisture curve map of the present invention.
Concrete embodiment:
Embodiments of the invention 1: golden cypress 200g, radix paeoniae rubrathe 200g, red sage root 100g, peach kernel 140g, Herba Lycopi 120g, root of three-nerved spicebush 120g, seed of cowherb 80g, root of Dahurain angelica 80g
Get golden cypress, the fine powder that was ground into 80 mesh sieves is standby; When the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb and the root of Dahurain angelica add 8 times of water and are heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction filtered, relative density was 1.05～1.10 thick paste when filtrate was condensed into 60 ℃, drying under reduced pressure is ground into fine powder, adds golden cypress fine powder and starch, talcum powder, amount of talc is 5%, mixing incapsulates, and should control relative humidity during packing below 65.0%, promptly get capsule, this product oral, a time 4～6,3 times on the one.
Embodiments of the invention 2: golden cypress 200g, radix paeoniae rubrathe 200g, red sage root 100g, peach kernel 140g, Herba Lycopi 120g, root of three-nerved spicebush 120g, seed of cowherb 80g, root of Dahurain angelica 80g
Get golden cypress, the fine powder that was ground into 80 mesh sieves is standby; The radix paeoniae rubrathe, the red sage root, the Herba Lycopi, the root of three-nerved spicebush, when the seed of cowherb and the root of Dahurain angelica add 8 times of water and are heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction, filter, relative density was 1.05～1.10 thick paste when filtrate was condensed into 60 ℃, added the golden cypress fine powder, was matrix with the Macrogol 4000, according to medicine: the part by weight of matrix=1: 2 adds Macrogol 4000, mixing, the employing internal diameter is 3.0mm, external diameter is the dropper of 4.0mm, drips 80 ℃ of system temperature, dripping speed is 20～30d/min, dripping distance is 5cm, splash in the long cooling stud of 130cm, be liquid coolant again with the methyl-silicone oil, pill promptly gets pill.
Embodiments of the invention 3: golden cypress 200g, radix paeoniae rubrathe 200g, red sage root 100g, peach kernel 140g, Herba Lycopi 120g, root of three-nerved spicebush 120g, seed of cowherb 80g, root of Dahurain angelica 80g
It is standby that golden cypress is ground into fine powder; When the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb and the root of Dahurain angelica add water and are heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction, filter, filtrate is condensed into thick paste, adds distilled water, simple syrup, mixes and promptly gets oral liquid.
Embodiments of the invention 4: golden cypress 200g, radix paeoniae rubrathe 200g, red sage root 100g, peach kernel 140g, Herba Lycopi 120g, root of three-nerved spicebush 120g, seed of cowherb 80g, root of Dahurain angelica 80g
It is standby that golden cypress is ground into fine powder; When the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb and the root of Dahurain angelica add water and be heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction, filter, filtrate is condensed into thick paste, adds carbomer, makes gel.
Embodiments of the invention 5: golden cypress 200g, radix paeoniae rubrathe 200g, red sage root 100g, peach kernel 140g, Herba Lycopi 120g, root of three-nerved spicebush 120g, seed of cowherb 80g, root of Dahurain angelica 80g
It is standby that golden cypress is ground into fine powder; When the radix paeoniae rubrathe, the red sage root, Herba Lycopi, the root of three-nerved spicebush, the seed of cowherb and the root of Dahurain angelica add water and be heated to 80 ℃, add peach kernel again, continue heating, decoct three times, 2 hours for the first time, 1.5 hours for the second time, 1 hour for the third time, collecting decoction filtered, filtrate is condensed into thick paste, and drying under reduced pressure is pulverized, add sodium carboxymethyl starch, crospolyvinylpyrrolidone, calcium monohydrogen phosphate, mix, compressing tablet is made dispersing tablet.
Embodiments of the invention 6: the thin-layer chromatography discrimination method of golden cypress in the tablet:
It is an amount of to get the prostatitis-treating tablet, adds Petroleum ether extraction, discards extract, and the dregs of a decoction volatilize, and adds alcohol reflux again, filters, and filtrate concentrates, as need testing solution; Get the golden cypress medicinal material, shine medicinal material solution in pairs with legal system; Test according to thin-layered chromatography, draw each 30 μ l of above-mentioned solution, put respectively on same silica gel H thin layer plate, with benzene-ethyl acetate-methyl alcohol-isopropyl alcohol-strong ammonia solution=1: 1: 3: be developping agent at 3: 1, takes out, and dries, put under the uviol lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Embodiments of the invention 7: the thin-layer chromatography discrimination method of Berberine hydrochloride, golden cypress in the granule
It is an amount of to get the prostatitis-treating preparation, and the extraction that adds diethyl ether discards extract, and the dregs of a decoction volatilize, and add methanol extraction again, filters, and filtrate concentrates, as need testing solution; Get the golden cypress medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Berberine hydrochloride reference substance, adds methyl alcohol or ethanol reference substance solution; Test according to thin-layered chromatography, draw each 1 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with dimethylbenzene-ethyl formate-methyl alcohol-dense ammoniacal liquor=8: 5: 3: 0.1 was developping agent, takes out, and dries, put under the uviol lamp and inspect, in the test sample chromatogram, with reference substance chromatogram, the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Embodiments of the invention 8: the thin-layer chromatography discrimination method of the radix paeoniae rubrathe, Paeoniflorin in the capsule
It is an amount of to get the prostatitis-treating capsule content, adds methyl alcohol 25ml, and sonicated 20 minutes filters, and filtrate evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Get radix paeoniae rubrathe control medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Paeoniflorin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin-layered chromatography test, draw each 5 μ l of above-mentioned solution, respectively and on the same silica gel g thin-layer plate, with chloroform-ethyl acetate-methyl alcohol-acetate=10: 1: 12: 2 was developping agent, launches, and takes out, dry, spray is with 1% vanillic aldehyde sulfuric acid solution, 105 ℃ dry by the fire to spot colour developing clear; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 9: the thin-layer chromatography discrimination method of Paeoniflorin in the pill
It is an amount of to get the prostatitis-treating preparation, gets content, adds ethanol or methanol extraction, filters, and filtrate concentrates, as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol or methyl alcohol is made reference substance solution; According to the thin-layered chromatography test, draw each 1 μ l of above-mentioned two kinds of solution, put in same silica G F respectively 254On the thin layer plate, be developping agent with chloroform-sour acid butyl ester-methyl alcohol=40: 1: 12, launch, take out, dry that spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiments of the invention 10: the thin-layer chromatography discrimination method of the red sage root, Sodium Danshensu in the capsule
It is an amount of to get the prostatitis-treating capsule content, adds water temperature and soaks, and filters, and supernatant is regulated pH value to 1 with watery hydrochloric acid, extracts with the chloroform jolting, and extract evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; Get Sodium Danshensu, add methyl alcohol and make reference substance solution; According to thin-layered chromatography test, draw each 1 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with chloroform-acetone-formic acid=5: 10: 4, launch, take out, dry, spray ferric trichloride ethanolic solution with 2%; In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiments of the invention 11: the thin-layer chromatography discrimination method of danshensu, protocatechualdehyde in the granule
It is an amount of to get the prostatitis-treating particle, adds the water ultrasonic Extraction, and extract is regulated pH to 4 with acid solution, extracts with the ethyl acetate jolting, and extract evaporate to dryness, residue add ethanol makes dissolving, as need testing solution; Get danshensu, protocatechualdehyde, add ethanol and make reference substance solution; According to the thin-layered chromatography test, draw each 30 μ l of above-mentioned two kinds of solution, put in same silica G F respectively 254On the thin layer plate, be developping agent, launch, take out that dry, spray is inspected under the visible light with the mixed solution of the ferric trichloride and the potassium ferricyanide with benzene-ethyl acetate-formic acid=8: 6: 1; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiments of the invention 12: the thin-layer chromatography discrimination method of the red sage root, root of red-rooted salvia phenolic acid B in the tablet
It is an amount of to get the prostatitis-treating tablet, adds methanol eddy and extracts, and extract concentrates, as need testing solution; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; Get root of red-rooted salvia phenolic acid B, add methyl alcohol and make reference substance solution; According to the thin-layered chromatography test, draw each 1 μ l of above-mentioned three kinds of solution, put in same silica G F respectively 254On the thin layer plate, with toluene-chloroform-ethyl acetate-methyl alcohol-formic acid=2: 0.5: 0.5: be developping agent at 0.5: 2, launches, and takes out, and dries, and 254nm inspects under the uviol lamp; In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiments of the invention 13: the thin-layer chromatography discrimination method of the red sage root in the pill
It is an amount of to get the prostatitis-treating dripping pill, extracts with 50% ethanol jolting, and extract concentrates, as need testing solution; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; According to thin-layered chromatography test, draw each 30 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with benzene-chloroform-ethyl formate-acetate=0.2: 0.5: 3: 1 developping agent, launch, take out, dry, spray is with ethanol solution of sulfuric acid, and heating is inspected under the visible light; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Embodiments of the invention 14: the thin-layer chromatography discrimination method of the root of Dahurain angelica in the capsule
It is an amount of to get the prostatitis-treating preparation, and it is ultrasonic that content adds water, centrifugal, and supernatant is regulated pH value to 14 with sodium hydroxide test solution, extracts with chloroform 20ml jolting, and extract concentrates, as need testing solution; Other gets root of Dahurain angelica control medicinal material and shines medicinal material solution in pairs with legal system; According to thin-layered chromatography test, draw each 1 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, be developping agent with chloroform-methanol=4: 1, launch, taking-up is dried, and puts that 254nm inspects under the ultraviolet lamp; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Embodiments of the invention 15: the thin-layer chromatography discrimination method of the root of Dahurain angelica in the tablet
It is an amount of to get the prostatitis-treating preparation, adds the water temperature lixiviate and gets, and extract is regulated pH value to 9 with the potassium hydroxide test solution, extracts with the chloroform jolting, and extract concentrates, as need testing solution; Other gets root of Dahurain angelica control medicinal material, adds ethyl acetate extraction, extracts to concentrate, in contrast medicinal material solution; According to thin-layered chromatography test, draws each 30 μ l of above-mentioned kind of solution, put respectively on same silica gel H thin layer plate, be developping agent with ethyl acetate-methyl alcohol=9: 1, expansion, taking-up is dried, and puts that 365nm inspects under the ultraviolet lamp; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Embodiments of the invention 16: the thin-layer chromatography discrimination method of peach kernel, amygdalin in the granule
It is an amount of to get the prostatitis-treating particle, adds methyl alcohol 20ml sonicated 20min, filters, filtrate evaporate to dryness, residue add water 5ml low-grade fever makes dissolving, puts cold, D101 type macroporous adsorptive resins by internal diameter 1.5cm, long 8cm adds water 20m prewashing once, last sample, discard water liquid,, discard ammoniacal liquor with ammonia solution 30ml wash-out, water 10ml wash-out discards water liquid again, continues with 20% ethanol 30ml wash-out, collect eluent, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving as need testing solution; Get the peach kernel medicinal material, the 20ml that adds diethyl ether refluxes, and discards ether solution, adds methyl alcohol 20ml again and refluxes, and filters, and filtrate concentrates, in contrast medicinal material solution; Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution; Draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate,, spray immediately, at 105 ℃ of about 10min of baking with 10% sulfuric acid ethanol so that chloroform-methanol-water=the lower floor's solution in placement below 10 ℃ was developping agent in 13: 7: 2, launched, and took out; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 17: one or both thin-layer chromatography discrimination method in Herba Lycopi, the ursolic acid in the pill
It is an amount of to get the prostatitis-treating preparation, adds ethanol 30ml, and sonicated 10 minutes is put coldly, filter, and the filtrate evaporate to dryness, residue soaks with sherwood oil 10ml, and the sherwood oil that inclines, residue add absolute ethyl alcohol 1ml makes dissolving, as need testing solution; Other gets Herba Lycopi's control medicinal material, shines medicinal material solution in pairs with legal system; Get the ursolic acid reference substance again, add absolute ethyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 1 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, with cyclohexane-methylene chloride-ethyl acetate-acetate=10: 5: 1: 1 was developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 18: the high performance liquid chromatography discrimination method of protocatechualdehyde in the capsule
It is an amount of to get the prostatitis-treating capsule content, adds the water sonicated, filters, and subsequent filtrate is with 10% salt acid for adjusting pH value to 1, extract with the ethyl acetate jolting, evaporate to dryness in the extract water-bath, residue adds dissolve with methanol, filter with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, puts in the brown measuring bottle, adds methyl alcohol and makes reference substance solution; With eight alkyl silane bonded silica gels is filling agent; Methanol-water-phosphoric acid=50: 50: 0.01 is a moving phase; The detection wavelength is 361nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 19: the high performance liquid chromatography discrimination method of protocatechualdehyde in the tablet
It is an amount of to get the prostatitis-treating tablet, adds water temperature and soaks processing, takes out, and puts to room temperature, filter, subsequent filtrate is regulated pH value to 3 with watery hydrochloric acid, extracts the extract evaporate to dryness with the chloroform jolting, residue adds dissolve with ethanol and constant volume, filters with miillpore filter, gets subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, adds ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Acetonitrile-water-acetate=10: 95: 5 is moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 30 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 20: the high performance liquid chromatography discrimination method of tanshin polyphenolic acid B in the pill
It is an amount of to get the prostatitis-treating dripping pill, adds 80% methyl alcohol sonicated, filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 80% methyl alcohol and makes reference substance solution; With the dialkyl silane bonded silica gel is filling agent; Methyl alcohol-formic acid-water=30: 1: 59 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 21: the high performance liquid chromatography discrimination method of tanshin polyphenolic acid B in the capsule
It is an amount of to get the prostatitis-treating capsule content, adds 95% ethanol sonicated, takes out, and puts to room temperature, filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 95% ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-phosphoric acid-water=10: 10: 0.4: 70 is moving phase; The detection wavelength is 245nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 22: the high performance liquid chromatography discrimination method of Paeoniflorin in the particle
It is an amount of to get the prostatitis-treating particle, adds the methyl alcohol sonicated, filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance, adds methyl alcohol and makes reference substance solution; With eight alkyl silane bonded silica gels is filling agent; Acetonitrile-water-potassium dihydrogen phosphate=5: 95: 5 is a moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 23: the high performance liquid chromatography discrimination method of Paeoniflorin in the tablet
It is an amount of to get the prostatitis-treating tablet, adds ethanol ultrasonic extraction, filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance, adds ethanol and makes reference substance solution; With eight alkyl silane bonded silica gels is filling agent; Methanol-water-phosphoric acid=50: 50: 0.5 is a moving phase; The detection wavelength is 360nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 30 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 24: the high performance liquid chromatography discrimination method of Berberine hydrochloride in the pill
It is an amount of to get the prostatitis-treating pill, and the mixed solution sonicated that adds hydrochloric acid-methyl alcohol=5: 100 is taken out, and puts to room temperature, filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With octadecylsilane chemically bonded silica is filling agent; With water-acetate-acetonitrile=20: 5: 20 was moving phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 25: the high performance liquid chromatography discrimination method of Berberine hydrochloride in the capsule
It is an amount of to get the prostatitis-treating capsule content, adds the ethanol sonicated, and extract is a need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With the dialkyl silane bonded silica gel is filling agent; With water-acetate-acetonitrile=60: 1: 5 was moving phase; The detection wavelength is 370nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 1 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 26: the high-efficient liquid phase chromatogram process measuring method of dripping pill Central Plains catechu aldehyde
It is an amount of to get the prostatitis-treating dripping pill, and accurate the title decides, and puts in the tool plug conical flask, precision adds water 50ml, and weight, sonicated 30 minutes decided in accurate title, take out, put to room temperature, weight decided in accurate title, add water and supply the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 10ml that draws, regulate pH value to 2 with watery hydrochloric acid, extract 3 times, each 10ml with the ethyl acetate jolting, the combined ethyl acetate extract, put evaporate to dryness in the water-bath, residue adds dissolve with ethanol and quantitatively is transferred in the 10ml measuring bottle, adds ethanol to scale, shake up, filter with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, and accurate the title decides, and puts in the brown measuring bottle, adds ethanol and makes the solution that contains 10 μ g among every 1ml, in contrast product solution; With eight alkyl silane bonded silica gels is filling agent; Acetonitrile-water-phosphoric acid=13: 87: 0.3 is moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 216 μ g with containing the red sage root in protocatechualdehyde in the prostatitis-treating dripping pill on 1st.
Embodiments of the invention 27: the high-efficient liquid phase chromatogram process measuring method of particle Central Plains catechu aldehyde
It is an amount of to get the prostatitis-treating preparation, and accurate the title decides, and adds the water sonicated, add water and supply the weight that subtracts mistake, shake up, filter, subsequent filtrate is regulated pH value to 3 with acid solution, extract with the chloroform jolting, merge extract, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and constant volume, filter with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, puts in the brown measuring bottle, adds methyl alcohol and is made as reference substance solution; With the dialkyl silane bonded silica gel is filling agent; Methanol-water-acetate=30: 95: 2 is moving phase; The detection wavelength is 200nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method or calibration curve method; Content limit should be: must not be lower than 216 μ g with containing the red sage root in protocatechualdehyde in the prostatitis-treating dripping pill on 1st.
Embodiments of the invention 28: the high-efficient liquid phase chromatogram process measuring method of content of danshinolic acid B in the dripping pill
It is an amount of to get the prostatitis-treating dripping pill, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, and accurate the title decides, and adds methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-acetate-water=10: 30: 1: 60 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 21.6mg with containing the red sage root in tanshin polyphenolic acid B in the prostatitis-treating preparation on 1st.
Embodiments of the invention 29: the high-efficient liquid phase chromatogram process measuring method of content of danshinolic acid B in the tablet
It is an amount of to get the prostatitis-treating tablet, and accurate the title decides, and adds 50% ethanol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds 50% ethanol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, and accurate the title decides, and adds 50% ethanol and makes reference substance solution; With the dialkyl silane bonded silica gel is filling agent; Acetonitrile-phosphoric acid-water=30: 0.3: 60 is moving phase; The detection wavelength is 376nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 21.6mg with containing the red sage root in tanshin polyphenolic acid B in the prostatitis-treating preparation on 1st.
Embodiments of the invention 30: the high-efficient liquid phase chromatogram process measuring method of paeoniflorin content in the tablet
It is an amount of to get the prostatitis-treating tablet, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance that is dried to constant weight, and accurate the title decides, and adds methyl alcohol and makes reference substance solution; With eight alkyl silane bonded silica gels is filling agent; Methanol-water-phosphoric acid=8: 60: 0.5 is a moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 26.4mg with containing the radix paeoniae rubrathe in Paeoniflorin in the prostatitis-treating preparation on 1st.
Embodiments of the invention 31: the high-efficient liquid phase chromatogram process measuring method of content of berberine hydrochloride in the particle
It is an amount of to get the prostatitis-treating particle, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, supplies the solvent of loss, shakes up, and filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; Water-potassium dihydrogen phosphate-acetonitrile=60: 5: 5 is a moving phase; The detection wavelength is 350nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 72mg with containing golden cypress in the prostatitis-treating preparation with Berberine hydrochloride on 1st.
Embodiments of the invention 32: the thin-layer chromatography discrimination method of Berberine hydrochloride, golden cypress in the capsule
It is an amount of to get the prostatitis-treating preparation, the 20ml that adds diethyl ether, and sonicated 20 minutes discards ether solution, and the dregs of a decoction volatilize, and add methyl alcohol 20ml, and sonicated 20 minutes filters, and filtrate is concentrated into 2ml, as need testing solution; Get the golden cypress medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Berberine hydrochloride reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate-methyl alcohol-isopropyl alcohol-strong ammonia solution=4: 3: 2: be developping agent at 1.5: 0.5, launches, take out, dry, put under the uviol lamp 365nm and inspect, in the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 33: the thin-layer chromatography discrimination method of the radix paeoniae rubrathe, Paeoniflorin in the dripping pill
It is an amount of to get the prostatitis-treating preparation, gets content, adds ethanol 25ml, and sonicated 20 minutes filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin-layered chromatography test, draw each 5 μ l of above-mentioned solution, respectively and on the same silica gel g thin-layer plate, with chloroform-ethyl acetate-methyl alcohol-formic acid=40: 5: 12: 0.2 was developping agent, launches, and takes out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ dry by the fire to spot colour developing clear; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 34: the thin-layer chromatography discrimination method of the red sage root, danshensu, protocatechualdehyde in the preparation
It is an amount of to get the prostatitis-treating preparation, adds water 20ml, and ultrasonic 30 minutes, centrifugal, supernatant was regulated pH value to 2 with watery hydrochloric acid, extracted with ethyl acetate 20ml jolting, and extract evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets red sage root control medicinal material 0.5g, and it is an amount of to add water, decocts 30 minutes, adds water to 20ml, shines medicinal material solution in pairs with legal system; Get danshensu, protocatechualdehyde, add methyl alcohol and make reference substance solution; According to the thin-layered chromatography test, draw above-mentioned solution 5 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-acetone-formic acid=25: 10: 4 was developping agent, launched, and took out, dry, spray ferric trichloride ethanolic solution, place about 30 minutes to clear spot with 2%; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 35: the thin-layer chromatography discrimination method of the red sage root, root of red-rooted salvia phenolic acid B in the tablet
It is an amount of to get the prostatitis-treating preparation, and 75% methyl alcohol jolting is extracted, and extract concentrates, as need testing solution; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; Root of red-rooted salvia phenolic acid B or its magnesium salts add 75% methyl alcohol and make reference substance solution; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned solution, put in same silica G F respectively 254On the thin layer plate, with toluene-chloroform-ethyl acetate-methyl alcohol-formic acid=2: 3: 4: be developping agent at 0.5: 2, launches, and takes out, and dries, and 254nm inspects under the uviol lamp; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 36: the thin-layer chromatography discrimination method of the root of Dahurain angelica in the particle
It is an amount of to get the prostatitis-treating preparation, and content adds water 20ml, and ultrasonic 30 minutes, centrifugal, supernatant was regulated pH value to 12 with sodium hydroxide test solution, extracted with chloroform 20ml jolting, and extract is concentrated into 1ml, as need testing solution; Other gets root of Dahurain angelica control medicinal material 0.5g, adds chloroform 10ml, and ultrasonic 30 minutes, filter, filtrate is concentrated into 1ml, in contrast medicinal material solution; According to thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with chloroform-methanol=9: 1, put in the vapour-saturated expansion cylinder of ammonia, launch, taking-up is dried, and puts that 254nm inspects under the ultraviolet lamp; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Embodiments of the invention 37: the thin-layer chromatography discrimination method of peach kernel, amygdalin in the capsule
It is an amount of to get the prostatitis-treating preparation, adds methyl alcohol 20ml sonicated 20min, filters, filtrate evaporate to dryness, residue add water 5ml low-grade fever makes dissolving, puts cold, D101 type macroporous adsorptive resins by internal diameter 1.5cm, long 8cm adds water 20m prewashing once, last sample, discard water liquid,, discard ammoniacal liquor with ammonia solution 30ml wash-out, water 10ml wash-out discards water liquid again, continues with 20% ethanol 30ml wash-out, collect eluent, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving as need testing solution; Get the peach kernel medicinal material, the 20ml that adds diethyl ether refluxes, and discards ether solution, adds methyl alcohol 20ml again and refluxes, and filters, and filtrate concentrates, in contrast medicinal material solution; Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution; Draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-methanol-water=15: 40: 22: 10 was developping agent at 5～10 ℃ of lower floor's solution of placing 12h, launch, take out, spray immediately, at 105 ℃ of about 10min of baking with the phosphomolybdic acid sulfuric acid solution; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.
Embodiments of the invention 38: the thin-layer chromatography discrimination method of Herba Lycopi, ursolic acid in the tablet
It is an amount of to get the prostatitis-treating preparation, adds acetone 30ml, and sonicated 10 minutes is put coldly, filter, and the filtrate evaporate to dryness, residue soaks with sherwood oil 10ml, and the sherwood oil that inclines, residue add absolute ethyl alcohol 1ml makes dissolving, as need testing solution; Other gets Herba Lycopi's control medicinal material, shines medicinal material solution in pairs with legal system; Get the ursolic acid reference substance again, add absolute ethyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, with cyclohexane-methenyl choloride-ethyl acetate-formic acid=20: 5: 8: 0.1 was developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiments of the invention 39: the high performance liquid chromatography discrimination method of protocatechualdehyde in the particle
It is an amount of to get the prostatitis-treating preparation, adds water 50ml, sonicated 30 minutes, take out, put, filter to room temperature, subsequent filtrate extracts 3 times with the ether jolting with 10% salt acid for adjusting pH value to 2, each 10ml, merge ether extracted liquid, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and constant volume, filters with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, puts in the brown measuring bottle, adds methyl alcohol and makes the solution that contains 12 μ g among every 1ml, in contrast product solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-1% glacial acetic acid aqueous solution=13: 87 is a moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 40: the high performance liquid chromatography discrimination method of tanshin polyphenolic acid B in the pill
It is an amount of to get the prostatitis-treating preparation, adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 75% methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-formic acid-water=30: 10: 1: 59 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 41: the high performance liquid chromatography discrimination method of Paeoniflorin in the tablet
It is an amount of to get the prostatitis-treating preparation, adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance that is dried to constant weight, adds methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.05mol/L potassium dihydrogen phosphate=40: 65 is moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 42: the high performance liquid chromatography discrimination method of Berberine hydrochloride in the capsule
It is an amount of to get the prostatitis-treating preparation, adds the mixed solution sonicated 30 minutes of hydrochloric acid-methyl alcohol=1: 100, takes out, and puts to room temperature, supplies the solvent of loss, shakes up, and filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With the 0.05mol/L sodium dihydrogen phosphate-acetonitrile of phosphorus acid for adjusting pH value to 3.0=75: 25 was moving phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
Embodiments of the invention 43: the high-efficient liquid phase chromatogram process measuring method of capsule Central Plains catechu aldehyde
It is an amount of to get the prostatitis-treating preparation, and accurate the title decides, and puts in the tool plug conical flask, precision adds water 50ml, and weight, sonicated 30 minutes decided in accurate title, take out, put to room temperature, weight decided in accurate title, add water and supply the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 10ml that draws, with 10% salt acid for adjusting pH value to 2, extract 3 times each 10ml with the ether jolting, merge ether extracted liquid, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and quantitatively is transferred in the 10ml measuring bottle, adds methyl alcohol to scale, shake up, filter with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, and accurate the title decides, and puts in the brown measuring bottle, adds methyl alcohol and makes the solution that contains 12 μ g among every 1ml, in contrast product solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-1% glacial acetic acid aqueous solution=13: 87 is a moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit is: be not less than 216 μ g with containing the red sage root in protocatechualdehyde in the prostatitis-treating preparation on 1st.
Embodiments of the invention 44: the high-efficient liquid phase chromatogram process measuring method of content of danshinolic acid B in the pill
It is an amount of to get the prostatitis-treating preparation, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, and accurate the title decides, and adds 75% methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-formic acid-water=30: 10: 1: 59 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit is: be not less than 21.6mg with containing the red sage root in tanshin polyphenolic acid B in the prostatitis-treating preparation on 1st.
Embodiments of the invention 45: the high-efficient liquid phase chromatogram process measuring method of paeoniflorin content in the granule
It is an amount of to get the prostatitis-treating preparation, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance that is dried to constant weight, and accurate the title decides,, add methyl alcohol and make reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.05mol/L potassium dihydrogen phosphate=40: 65 is moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit is: be not less than 26.4mg with containing the radix paeoniae rubrathe in Paeoniflorin in the prostatitis-treating preparation on 1st.
Embodiments of the invention 46: the high-efficient liquid phase chromatogram process measuring method of content of berberine hydrochloride in the tablet
It is an amount of to get the prostatitis-treating preparation, accurate claims surely, adds the mixed solution sonicated 30 minutes of hydrochloric acid-methyl alcohol=1: 100, takes out, and puts to room temperature, supplies the solvent of loss, shakes up, and filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With the 0.05mol/L sodium dihydrogen phosphate-acetonitrile of phosphorus acid for adjusting pH value to 3.0=75: 25 was moving phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit is: be not less than 72mg with containing golden cypress in the prostatitis-treating preparation with Berberine hydrochloride on 1st.

Claims (5)

  1. One kind the treatment prostatitic pharmaceutical preparation detection method, described medicine is made by golden cypress 200g, radix paeoniae rubrathe 200g, red sage root 100g, peach kernel 140g, Herba Lycopi 120g, root of three-nerved spicebush 120g, seed of cowherb 80g and root of Dahurain angelica 80g or with their extract of corresponding weight portion, and it is characterized in that: this method is:
    (1) the differential test method of Berberine hydrochloride, golden cypress, the radix paeoniae rubrathe, Paeoniflorin, the red sage root, danshensu or its sodium salt, protocatechualdehyde, tanshin polyphenolic acid B or its magnesium salts, the root of Dahurain angelica, peach kernel, amygdalin, Herba Lycopi, ursolic acid;
    (2) content test method of Berberine hydrochloride, Paeoniflorin, tanshin polyphenolic acid B or its magnesium salts, protocatechualdehyde;
  2. 2. according to the detection method of the prostatitic pharmaceutical preparation of the described treatment of claim 1, it is characterized in that: the discrimination method of described preparation is:
    A. one or both thin-layer chromatography discrimination method in Berberine hydrochloride, the golden cypress in the preparation: it is an amount of to get this preparation, add diethyl ether or sherwood oil or ethyl acetate or chloroform extraction, discard extract, the dregs of a decoction volatilize, add 20～100% methyl alcohol or alcohol extract again, the preparation need testing solution; Or it is an amount of to get this preparation, directly adds 20～100% methyl alcohol or alcohol extract, the preparation need testing solution; Get the golden cypress medicinal material, with reference to a kind of control medicinal material solution of making among the need testing solution preparation method; Other gets the Berberine hydrochloride reference substance, adds 20～100% methyl alcohol or ethanol and makes reference substance solution; Test according to thin-layered chromatography, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively on same silica G or silica gel H thin layer plate, with benzene or toluene or dimethylbenzene-ethyl acetate or ethyl formate or sour acid butyl ester-isopropyl alcohol or normal butyl alcohol-methyl alcohol or ethanol-dense ammoniacal liquor=1～12: be developping agent at 1～6: 0～3: 0～3: 0～1, take out, dry, put under the uviol lamp and inspect, in the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    B. one or both thin-layer chromatography discrimination method in the radix paeoniae rubrathe, the Paeoniflorin in the preparation:
    It is an amount of to get this preparation, adds 20～100% ethanol or methanol extraction, the preparation need testing solution; Get radix paeoniae rubrathe medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Paeoniflorin reference substance, adds 20～100% ethanol or methyl alcohol and makes reference substance solution; According to the thin-layered chromatography test, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with chloroform or methylene chloride-ethyl acetate or ethyl formate or sour acid butyl ester-methyl alcohol or ethanol-formic acid or acetate=10～40: be developping agent at 1～5: 2～12: 0～2, launches, take out, dry, spray is with vanillic aldehyde sulfuric acid or ethanol solution of sulfuric acid, and it is clear to be heated to spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    C. the thin-layer chromatography discrimination method of the red sage root, danshensu or its sodium salt, protocatechualdehyde in the preparation:
    It is an amount of to get this preparation, extracting in water, and extract is regulated pH1～4 with acid solution, and with ethyl acetate or chloroform extraction, extract evaporate to dryness, residue add methyl alcohol or ethanol makes dissolving, as need testing solution; Or it is an amount of to get this preparation, adds 20～100% ethanol or methanol extraction, the preparation need testing solution; Get red sage root control medicinal material,, make control medicinal material solution with reference to one of need testing solution preparation method; Get danshensu or its sodium salt, protocatechualdehyde, add 20～100% methyl alcohol or ethanol and make reference substance solution; According to the thin-layered chromatography test, draw one or more each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with chloroform or methylene chloride or benzene or ethyl acetate-acetone or butanone or ethyl acetate-formic acid or acetate=5～25: be developping agent at 1～10: 0～4, launch, take out, dry, spray is inspected under the visible light with the mixed solution of liquor ferri trichloridi or the ferric trichloride and the potassium ferricyanide, or with the ammonia steam smoked after, under uviol lamp, inspect; In the test sample chromatogram, with control medicinal material chromatogram, the corresponding position of reference substance chromatogram on, show the spot of same color;
    D. the thin-layer chromatography discrimination method of the red sage root, root of red-rooted salvia phenolic acid B or its magnesium salts in the preparation:
    It is an amount of to get this preparation, extracts the preparation need testing solution with 10～100% methyl alcohol or ethanol jolting; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; Root of red-rooted salvia phenolic acid B or its magnesium salts are made reference substance solution; According to the thin-layered chromatography test, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with toluene or benzene or dimethylbenzene-chloroform or methylene chloride-ethyl acetate or ethyl formate or sour acid butyl ester-methyl alcohol or ethanol-formic acid or acetate=0.2～2: be developping agent at 0.5～3: 0.5～4: 0～1: 0～2, launch, take out, dry, inspect under the uviol lamp, or spray is with ethanol solution of sulfuric acid, heating is inspected under the visible light; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    E. the thin-layer chromatography discrimination method of the root of Dahurain angelica in the preparation:
    It is an amount of to get this preparation, extracting in water, and extract concentrates with chloroform or ethyl acetate extraction, extract, as need testing solution; Or it is an amount of to get this preparation, extracts the preparation need testing solution with chloroform or ethyl acetate jolting; Other gets root of Dahurain angelica control medicinal material, with reference to one of need testing solution preparation method, makes control medicinal material solution; Test according to thin-layered chromatography, draw each 1～30 μ l of above-mentioned two kinds of solution, put respectively on same silica G or silica gel H thin layer plate, with chloroform or methylene chloride or ethyl acetate-methyl alcohol or ethanol or acetone=1～9: 0.2～3 is developping agent, put in the vapour-saturated expansion cylinder of ammonia, launch, take out, dry, put under the ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
    F. one or both thin-layer chromatography discrimination method in peach kernel, the amarogentin in the preparation:
    It is an amount of to get this preparation, adds 20～100% methyl alcohol or alcohol extract, passes through macroporous adsorptive resins after extract is handled again,, with 0～80% ethanol elution, collect the part eluent, evaporate to dryness, residue add 10～100% methyl alcohol or ethanol makes dissolving, as need testing solution; Get the peach kernel medicinal material, add diethyl ether or Petroleum ether extraction, discard extract, add 20～100% methyl alcohol or alcohol extract again, preparation control medicinal material solution; Other gets the amarogentin reference substance, adds 20～100% methyl alcohol or ethanol and makes reference substance solution; Draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform or methylene chloride-ethyl acetate or ethyl formate or sour acid butyl ester-methyl alcohol or alcohol-water=0～15: 0～40: 1～22: 1～10 lower floor's solution 0～10 ℃ of placement is developping agent, launch, take out, spray is with phosphomolybdic acid sulfuric acid solution or ethanol solution of sulfuric acid, and it is clear to be heated to colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    G. one or both thin-layer chromatography discrimination method in Herba Lycopi, the ursolic acid in the preparation:
    It is an amount of to get this preparation, adds diethyl ether or methenyl choloride or ethyl acetate or ethanol or methyl alcohol or acetone extraction the preparation need testing solution; Other gets Herba Lycopi's control medicinal material, shines medicinal material solution in pairs with legal system; Get the ursolic acid reference substance again, add methyl alcohol or methenyl choloride or ethyl acetate or ethanol and make dissolving, in contrast product solution; According to the thin-layered chromatography test, draw one or both each 1～30 μ l in above-mentioned need testing solution, control medicinal material or the reference substance solution, put respectively in same silica G or silica gel H or silica G F 254On the thin layer plate, with thiacyclohexane or normal hexane-methenyl choloride or the suitable cave セ centipede appearance Mei the moon of the cloudy セ Kuo of the suitable Mei of methylene chloride jujube? formic acid or acetate=0～20: 1～5: 1～8: 0～1 is developping agent, launches, take out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    H. the high performance liquid chromatography discrimination method of protocatechualdehyde in the preparation:
    It is an amount of to get this preparation, extracting in water, and water liquid prepares need testing solution with ether or ethyl acetate or chloroform jolting extraction; Or it is an amount of to get this preparation, adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the protocatechualdehyde reference substance, adds 20～100% methyl alcohol or ethanol and is made as reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-acetate or formic acid or phosphoric acid=10～50: 30～95: 0.01～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
    I. the high performance liquid chromatography discrimination method of tanshin polyphenolic acid B in the preparation:
    It is an amount of to get this preparation, adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 10～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or ethanol-acetonitrile-formic acid or acetate or phosphoric acid-water=10～30: 0～30: 0～5: 50～95 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
    J. the high performance liquid chromatography discrimination method of Paeoniflorin in the preparation:
    It is an amount of to get this preparation, adds 20～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the Paeoniflorin reference substance, adds 20～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate=5～50: 20～60: 0～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
    K. the high performance liquid chromatography discrimination method of Berberine hydrochloride in the preparation:
    This preparation is an amount of before getting, and adds hydrochloric acid-methyl alcohol or ethanol=0～5: 100 solution extracts, the preparation need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; With water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate-acetonitrile or methyl alcohol=0～60: be moving phase at 0～5: 5～50; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
  3. 3. according to the detection method of the prostatitic pharmaceutical preparation of the described treatment of claim 2, it is characterized in that: the discrimination method of described preparation is:
    A. one or both thin-layer chromatography discrimination method in Berberine hydrochloride, the golden cypress in the preparation:
    It is an amount of to get this preparation, the 20ml that adds diethyl ether, and sonicated 20 minutes discards ether solution, and the dregs of a decoction volatilize, and add methyl alcohol 20ml, and sonicated 20 minutes filters, and filtrate is concentrated into 2ml, as need testing solution; Get the golden cypress medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Berberine hydrochloride reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate-methyl alcohol-isopropyl alcohol-strong ammonia solution=4: 3: 2: be developping agent at 1.5: 0.5, launches, take out, dry, put under the uviol lamp 365nm and inspect, in the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    B. one or both thin-layer chromatography discrimination method in the radix paeoniae rubrathe, the Paeoniflorin in the preparation:
    It is an amount of to get this preparation, gets content, adds ethanol 25ml, and sonicated 20 minutes filters, and filtrate evaporate to dryness, residue add ethanol 2ml makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Test according to thin-layered chromatography, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, respectively with same silica gel g thin-layer plate on, with chloroform-ethyl acetate-methyl alcohol-formic acid=40: 5: 12: 0.2 was developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ dry by the fire to the spot colour developing clear; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    C. the thin-layer chromatography discrimination method of the red sage root, danshensu or its sodium salt, protocatechualdehyde in the preparation:
    It is an amount of to get this preparation, adds water 20ml, and ultrasonic 30 minutes, centrifugal, supernatant was regulated pH value to 2 with watery hydrochloric acid, extracted with ethyl acetate 20ml jolting, and extract evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets red sage root control medicinal material 0.5g, and it is an amount of to add water, decocts 30 minutes, adds water to 20ml, shines medicinal material solution in pairs with legal system; Get danshensu or its sodium salt, protocatechualdehyde, add methyl alcohol and make reference substance solution; Test according to thin-layered chromatography, draw one or more each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-acetone-formic acid=25: 10: 4 was developping agent, launch, take out, dry, spray is placed about 30 minutes to clear spot with 2% ferric trichloride ethanolic solution; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    D. the thin-layer chromatography discrimination method of the red sage root, root of red-rooted salvia phenolic acid B or its magnesium salts in the preparation:
    It is an amount of to get this preparation, and 75% methyl alcohol jolting is extracted, and extract concentrates, as need testing solution; Other gets red sage root control medicinal material, shines medicinal material solution in pairs with legal system; Root of red-rooted salvia phenolic acid B or its magnesium salts add 75% methyl alcohol and make reference substance solution; According to the thin-layered chromatography test, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put in same silica G F respectively 254On the thin layer plate, with toluene-chloroform-ethyl acetate-methyl alcohol-formic acid=2: 3: 4: be developping agent at 0.5: 2, launches, and takes out, and dries, and 254nm inspects under the uviol lamp; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    E. the thin-layer chromatography discrimination method of the root of Dahurain angelica in the preparation:
    It is an amount of to get this preparation, and content adds water 20ml, and ultrasonic 30 minutes, centrifugal, supernatant was regulated pH value to 12 with sodium hydroxide test solution, extracted with chloroform 20ml jolting, and extract is concentrated into 1ml, as need testing solution; Other gets root of Dahurain angelica control medicinal material 0.5g, adds chloroform 10ml, and ultrasonic 30 minutes, filter, filtrate is concentrated into 1ml, in contrast medicinal material solution; According to thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with chloroform-methanol=9: 1, put in the vapour-saturated expansion cylinder of ammonia, launch, taking-up is dried, and puts that 254nm inspects under the ultraviolet lamp; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
    F. one or both thin-layer chromatography discrimination method in peach kernel, the amarogentin in the preparation:
    It is an amount of to get this preparation, adds methyl alcohol 20ml sonicated 20min, filters, filtrate evaporate to dryness, residue add water 5ml low-grade fever makes dissolving, puts cold, by the D101 type macroporous adsorptive resins of the internal diameter 1.5cm of water 20m prewashing, long 8cm,, discard water liquid, with ammonia solution 30ml wash-out, discard ammoniacal liquor, again water 10ml wash-out, discard water liquid, continue, collect eluent with 20% ethanol 30ml wash-out, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving as need testing solution; Get the peach kernel medicinal material, the 20ml that adds diethyl ether refluxes, and discards ether solution, adds methyl alcohol 20ml again and refluxes, and filters, and filtrate concentrates, in contrast medicinal material solution; Other gets the amarogentin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution; Draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-methanol-water=15: 40: 22: 10 was developping agent at 5～10 ℃ of lower floor's solution of placing 12h, launch, take out, spray immediately with the phosphomolybdic acid sulfuric acid solution, at 105 ℃ of about 10min of baking; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color;
    G. one or both thin-layer chromatography discrimination method in Herba Lycopi, the ursolic acid in the preparation:
    It is an amount of to get this preparation, adds acetone 30ml, and sonicated 10 minutes is put coldly, filter, and the filtrate evaporate to dryness, residue soaks with sherwood oil 10ml, and the sherwood oil that inclines, residue add absolute ethyl alcohol 1ml makes dissolving, as need testing solution; Other gets Herba Lycopi's control medicinal material, shines medicinal material solution in pairs with legal system; Get the ursolic acid reference substance again, add absolute ethyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw one or both each the 5 μ l in above-mentioned need testing solution, reference substance or the control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, with thiacyclohexane-methenyl choloride jujube Yi Meiyin? formic acid=20: 5: 8: 0.1 is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the spot of same color;
    H. the high performance liquid chromatography discrimination method of protocatechualdehyde in the preparation:
    It is an amount of to get this preparation, adds water 50ml, sonicated 30 minutes, take out, put, filter to room temperature, subsequent filtrate extracts 3 times with the ether jolting with 10% salt acid for adjusting pH value to 2, each 10ml, merge ether extracted liquid, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and constant volume, filters with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, puts in the brown measuring bottle, adds methyl alcohol and makes the solution that contains 12 μ g among every 1ml, in contrast product solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-1% glacial acetic acid aqueous solution=13: 87 is a moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
    I. the high performance liquid chromatography discrimination method of tanshin polyphenolic acid B in the preparation:
    It is an amount of to get this preparation, adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 75% methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-formic acid-water=30: 10: 1: 59 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Draw each 10 μ l of reference substance solution and need testing solution respectively, inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
    J. the high performance liquid chromatography discrimination method of Paeoniflorin in the preparation:
    It is an amount of to get this preparation, adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance that is dried to constant weight, adds methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.05mol/L potassium dihydrogen phosphate=40: 65 is moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention;
    K. the high performance liquid chromatography discrimination method of Berberine hydrochloride in the preparation:
    It is an amount of to get this preparation, adds the mixed solution sonicated 30 minutes of hydrochloric acid-methyl alcohol=1: 100, takes out, and puts to room temperature, supplies the solvent of loss, shakes up, and filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With the 0.05mol/L sodium dihydrogen phosphate-acetonitrile of phosphorus acid for adjusting pH value to 3.0=75: 25 was moving phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; In the test sample chromatogram, have the chromatographic peak consistent with the reference substance chromatographic retention.
  4. 4. according to the detection method of the prostatitic pharmaceutical preparation of the described treatment of claim 1, it is characterized in that: the content assaying method of described preparation is:
    A. the high-efficient liquid phase chromatogram process measuring method of preparation Central Plains catechu aldehyde:
    It is an amount of to get this preparation, and accurate the title decides, extracting in water, and water liquid prepares need testing solution with ether or ethyl acetate or chloroform jolting extraction; Or it is an amount of to get this preparation, adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the protocatechualdehyde reference substance, adds 10～100% methyl alcohol or ethanol and is made as reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-acetate or formic acid or phosphoric acid=10～50: 30～95: 0.01～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures, and calculates with one point external standard method or calibration curve method; Content limit should be: must not be lower than 108 μ g with containing the red sage root in this preparation in protocatechualdehyde on 1st;
    B. the high-efficient liquid phase chromatogram process measuring method of content of danshinolic acid B in the preparation:
    It is an amount of to get this preparation, and accurate the title decides, and adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, adds 10～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or ethanol-acetonitrile-formic acid or acetate or phosphoric acid-water=10～30: 0～30: 0～5: 50～95 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures, and calculates with one point external standard method or calibration curve method; Content limit should be: must not be lower than 10.8mg with containing the red sage root in this preparation in tanshin polyphenolic acid B on 1st;
    C. the high-efficient liquid phase chromatogram process measuring method of paeoniflorin content in the preparation:
    It is an amount of to get this preparation, and accurate the title decides, and adds 10～100% methyl alcohol or alcohol extract, the preparation need testing solution; It is an amount of to get the Paeoniflorin reference substance, adds 10～100% methyl alcohol or ethanol and makes reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; Methyl alcohol or acetonitrile-water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate=5～50: 20～60: 0～5 is moving phase; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method or calibration curve method; Content limit should be: must not be lower than 13.2mg with containing the radix paeoniae rubrathe in this preparation in Paeoniflorin on 1st;
    D. the high-efficient liquid phase chromatogram process measuring method of content of berberine hydrochloride in the preparation:
    It is an amount of to get this preparation, and accurate the title decides, and adds hydrochloric acid-methyl alcohol=0～5: 100 solution extracts, the preparation need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel is filling agent; With water-potassium dihydrogen phosphate or potassium dihydrogen phosphate or phosphoric acid or formic acid or acetate-acetonitrile or methyl alcohol=20～60: be moving phase at 0～5: 5～50; The detection wavelength is one or several among 190～410nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 1～30 μ l of need testing solution of drawing injects liquid chromatograph, measures, and calculates with one point external standard method or calibration curve method; Content limit should be: must not be lower than 36mg with containing golden cypress in this preparation in Berberine hydrochloride on 1st.
  5. 5. according to the detection method of the prostatitic pharmaceutical preparation of the described treatment of claim 4, it is characterized in that: the content assaying method of described preparation is:
    A. the high-efficient liquid phase chromatogram process measuring method of preparation Central Plains catechu aldehyde:
    It is an amount of to get this preparation, and accurate the title decides, and puts in the tool plug conical flask, precision adds water 50ml, and weight, sonicated 30 minutes decided in accurate title, take out, put to room temperature, weight decided in accurate title, add water and supply the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 10ml that draws, with 10% salt acid for adjusting pH value to 2, extract 3 times each 10ml with the ether jolting, merge ether extracted liquid, put evaporate to dryness in the water-bath, residue adds dissolve with methanol and quantitatively is transferred in the 10ml measuring bottle, adds methyl alcohol to scale, shake up, filter with miillpore filter, get subsequent filtrate, as need testing solution; It is an amount of to get the protocatechualdehyde reference substance, and accurate the title decides, and puts in the brown measuring bottle, adds methyl alcohol and makes the solution that contains 12 μ g among every 1ml, in contrast product solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-1% glacial acetic acid aqueous solution=13: 87 is a moving phase; The detection wavelength is 281nm; Number of theoretical plate calculates by protocatechualdehyde should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 216 μ g with containing the red sage root in this preparation in protocatechualdehyde on 1st;
    B. the high-efficient liquid phase chromatogram process measuring method of content of danshinolic acid B in the preparation:
    It is an amount of to get this preparation, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the tanshin polyphenolic acid B reference substance, and accurate the title decides, and adds 75% methyl alcohol and makes reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-acetonitrile-formic acid-water=30: 10: 1: 59 is moving phase; The detection wavelength is 286nm; Number of theoretical plate calculates by tanshin polyphenolic acid B should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 21.6mg with containing the red sage root in this preparation in tanshin polyphenolic acid B on 1st;
    C. the high-efficient liquid phase chromatogram process measuring method of paeoniflorin content in the preparation:
    It is an amount of to get this preparation, and accurate the title decides, and adds the methyl alcohol sonicated 30 minutes, takes out, and puts to room temperature, and accurate the title decided weight, adds methyl alcohol and supplies the weight that subtracts mistake, shakes up, and filters, as need testing solution; It is an amount of to get the Paeoniflorin reference substance that is dried to constant weight, and accurate the title decides,, add methyl alcohol and make reference substance solution; With octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.05mol/L potassium dihydrogen phosphate=40: 65 is moving phase; The detection wavelength is 230nm; Number of theoretical plate calculates by Paeoniflorin should be not less than 3000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 26.4mg with containing the radix paeoniae rubrathe in this preparation in Paeoniflorin on 1st;
    D. the high-efficient liquid phase chromatogram process measuring method of content of berberine hydrochloride in the preparation:
    It is an amount of to get this preparation, accurate claims surely, adds the mixed solution sonicated 30 minutes of hydrochloric acid-methyl alcohol=1: 100, takes out, and puts to room temperature, supplies the solvent of loss, shakes up, and filters, and gets subsequent filtrate, is need testing solution; Get the Berberine hydrochloride reference substance, add above-mentioned solvent and make reference substance solution; With the 0.05mol/L sodium dihydrogen phosphate-acetonitrile of phosphorus acid for adjusting pH value to 3.0=75: 25 was moving phase; The detection wavelength is 270nm; Number of theoretical plate calculates by the Berberine hydrochloride peak should be not less than 2000; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure, and calculate with one point external standard method, promptly; Content limit should be: must not be lower than 72mg with containing golden cypress in this preparation with Berberine hydrochloride on 1st.
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CN102818875A (en) * 2011-06-10 2012-12-12 天津同仁堂集团股份有限公司 Quality control method of vessel rehabilitation tablets
CN103048416A (en) * 2011-10-14 2013-04-17 西安千禾药业有限责任公司 Detection method of Chinese medicinal composition for treating acute and chronic prostatitis
CN103134896A (en) * 2011-11-25 2013-06-05 张金荣 Detection method for Chinese materia medica preparation for treating chronic prostatitis
CN102526305A (en) * 2012-02-14 2012-07-04 史志辉 Chinese medicinal capsules for treating prostatitis and prostatic hyperplasia and preparation method for Chinese medicinal capsules
CN105012994B (en) * 2015-08-25 2018-02-16 东莞市达庆医疗器械有限公司 A kind of medical bio uropoiesis hydrogel functional dressings and preparation method thereof
CN107782816A (en) * 2016-08-31 2018-03-09 天津中新药业研究中心 A kind of detection method of clearing heat and detoxicating cool blood stranguria-treating drug
CN106511631A (en) * 2016-10-25 2017-03-22 西安千禾药业股份有限公司 Qianlieping preparation used for treating prostatitis and preparation method thereof
CN107884508B (en) * 2017-04-16 2021-01-12 湖南安邦制药有限公司 Quality detection method of Yinhuang lung-clearing capsule
CN109406707B (en) * 2018-11-29 2020-11-27 四川新绿色药业科技发展有限公司 Thin-layer chromatography identification method for fried peach kernels and preparation thereof
CN113834895B (en) * 2021-08-28 2023-02-24 海南葫芦娃药业集团股份有限公司 Quality control method of amygdalin in infantile lung heat cough and asthma granules

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