CN101874852A - Quality detecting method of restorex plus extract capsule - Google Patents
Quality detecting method of restorex plus extract capsule Download PDFInfo
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Abstract
The invention discloses a quality detecting method of a restorex plus extract capsule, wherein the restorex plus extract capsule comprises the following raw material drugs in parts by weight: 273 parts of leonurus, 68 parts of campanumaea pilosula, 137 parts of prepared rehmannia root, 68 parts of frying bighead atractylodes rhizome, 68 parts of poria cocos, 137 parts of Chinese angelica, 68 parts of wine white paeony root, 68 parts of szechuan lovage rhizome and 34 parts of liquorice. The method comprises thin-layer chromatography authentication of Chinese angelica, szechuan lovage rhizome and leonurus, as well as the measurement of content of paeoniflorin in the wine white paeony root. The quality detecting method can reflect the quality of the restorex plus extract capsule from multiple aspects, thereby ensuring the pharmacy safety and the clinical effects. The invention also discloses a preparation process of the restorex plus extract capsule.
Description
Technical field
The present invention relates to the detection method of the detection method of compound Chinese medicinal preparation, particularly BAZHEN YIMU JIAONANG.
Background technology
Bazhen yimu pills, bazhen yimu wan comes from Jing-Yue Complete Works, prescription consists of Herba Leonuri, Radix Codonopsis, Radix Rehmanniae Preparata, Rhizoma Atractylodis Macrocephalae (parched), Poria, Radix Angelicae Sinensis, Radix Paeoniae Alba, Rhizoma Chuanxiong and Radix Glycyrrhizae, have and fill blood, transfer the effect of menstruation, the clinical plain body of women that is used for loses after empty or the serious disease prolonged illness or Qi and blood deficiency, irregular menstruation due to the severe loss of blood, lose the first-selected prescription of menoxenia for QI and blood is two.But honeyed pill is directly beaten powder with whole crude drug and is used as medicine, and dose is big, takes inconvenience; Therefore, the pharmaceuticals researcher develops and makes things convenient for the patient to take and BAZHEN YIMU JIAONANG agent that curative effect increases, and studied the method for quality control of system, set up stachydrine hydrochloride thin layer chromatography discriminating in Radix Angelicae Sinensis in the preparation, Rhizoma Chuanxiong and the Herba Leonuri, formulate in the preparation paeoniflorin content assay method in the Radix Paeoniae Alba, thereby guaranteed drug safety.
Summary of the invention
The quality determining method that the purpose of this invention is to provide BAZHEN YIMU JIAONANG.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is:
The crude drug of BAZHEN YIMU JIAONANG of the present invention is composed as follows:
Herba Leonuri 273 weight portion Radix Codonopsis 68 weight portion Radix Rehmanniae Preparata 137 weight portions
Rhizoma Atractylodis Macrocephalae (parched) 68 weight portion Poria 68 weight portion Radix Angelicae Sinensis 137 weight portions
Radix Paeoniae Alba 68 weight portion Rhizoma Chuanxiongs 68 weight portion Radix Glycyrrhizaes 34 weight portions
Capsular preparation technology of the present invention is:
Get the Poria of 1/3rd weight portions and the Radix Paeoniae Alba of described weight portion and be ground into coarse powder; The Radix Angelicae Sinensis of described weight portion, Rhizoma Chuanxiong, Rhizoma Atractylodis Macrocephalae (parched) distillating extracting oil, the aqueous solution after the distillation device are in addition collected the Radix Codonopsis of medicinal residues and described weight portion, Radix Rehmanniae Preparata, Herba Leonuri, Radix Glycyrrhizae and the Poria that remains 2/3rds weight portions decoct with water secondary, 2 hours for the first time, 1.5 hours for the second time, decocting liquid filtered, filtrate merges, and with the aqueous solution merging after the distillation, relative density is 1.25～1.30 extractum when being concentrated into 60 ℃, add described coarse powder, stir evenly, 80～90 ℃ of oven dry are pulverized, add starch, sieve, mixing is with 90% ethanol system granule, dry, spray into described volatile oil, sealing incapsulates, every dress 0.28g, promptly.
Capsular preparation technology of the present invention, during the volatile oil of distillation extraction Radix Angelicae Sinensis, Rhizoma Chuanxiong, Rhizoma Atractylodis Macrocephalae (parched), the preferred distillation extraction time is 3 hours; When decocting with water, preferred total amount of water is 19 times of gross weight of Radix Angelicae Sinensis, Rhizoma Chuanxiong, Rhizoma Atractylodis Macrocephalae (parched), Radix Codonopsis, Radix Rehmanniae Preparata, Herba Leonuri, Radix Glycyrrhizae and 2/3rds weight portion Poria of described weight portion, adds 10 times of water gagings for the first time, adds 9 times of water gagings for the second time.
Described BAZHEN YIMU JIAONANG is oral, one time 3,3 times on the one, the described BAZHEN YIMU JIAONANG content of every 1g is converted to the raw material dose, is equivalent to 3.29g, the content of just per 1 described BAZHEN YIMU JIAONANG is converted to the raw material dose, is equivalent to 0.921g.
The quality determining method of BAZHEN YIMU JIAONANG of the present invention comprises the authentication method of following I, II:
I, get the content 6g of described BAZHEN YIMU JIAONANG, add normal hexane 25ml, supersound process 15 minutes filters, and filtrate volatilizes, and residue adds normal hexane 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1g of Rhizoma Chuanxiong control medicinal material, adds normal hexane 15ml respectively, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launches, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
II, get the content 8g of described BAZHEN YIMU JIAONANG, add an amount of kieselguhr, grind well, put in the apparatus,Soxhlet's, added alcohol reflux 4 hours, extracting solution filters, filtrate recycling ethanol, dissolve with 0.1mol/L hydrochloric acid solution 8ml gradation, acid solution filters, and merging filtrate adds the chromic thiocyanate ammonium salt saturated solution 12ml that faces with preparation, placing 1 hour below 10 ℃, filter with sintered glass funnel, precipitation adds acetone 5ml and makes dissolving after washing with low amounts of water, 0.5% silver sulfate solution to the precipitation that adds about 5ml is more no longer separated out, filter, precipitation is washed with small amount of acetone, and washing liquid and filtrate merge, be concentrated into about 2ml, add the about 2.5ml of 1% barium chloride solution, mixing, being added in internal diameter is 15mm, in adorn on the chromatographic column of 80～100 order neutral alumina 1.5g, with 70% ethanol 35ml eluting, collect eluent, be concentrated into dried, residue adds ethanol 1.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Main flavour of a drug in the Radix Paeoniae Alba side of being, and be used as medicine with former powder, content of paeoniflorin can directly reflect the grade quality and the inventory of Radix Paeoniae Alba, is significant so peoniflorin is carried out assay to the quality testing of compound Chinese medicinal preparation of the present invention.Therefore, quality determining method of the present invention also comprises the assay of following high performance liquid chromatography:
The test of chromatographic condition and system suitability: with the octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solution is a mobile phase with volume ratio, and detections wavelength is 230nm, and number of theoretical plate should be not less than 1500 by the calculating of peoniflorin peak;
The preparation of reference substance solution: get the peoniflorin reference substance, the accurate title, decide, and adds 50% methanol and make the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get the content 2.8g of described BAZHEN YIMU JIAONANG, mixing, porphyrize, get about 0.3g, the accurate title, decide, the accurate 50% methanol 50ml that adds, claim to decide weight, close plug, the supersound process of power 250W, frequency 25kHz 60 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with 50% methanol, shake up, filter, get subsequent filtrate, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
Every described BAZHEN YIMU JIAONANG contains Radix Paeoniae Alba with peoniflorin C
23H
28O
11Meter must not be less than 0.90mg.
In the above-mentioned high performance liquid chromatography,, during the preparation reference substance solution, preferably be dried to the peoniflorin reference substance of constant weight through phosphorus pentoxide for guaranteeing the accuracy of measurement result.
Preferred described discrimination method I of the quality determining method of BAZHEN YIMU JIAONANG of the present invention and II, and the combination of described content assaying method can reflect the quality of preparation from many aspects.
Below by the Study on Preparation of description of test BAZHEN YIMU JIAONANG of the present invention, and the foundation of quality determining method of the present invention.
1 eight gusts of capsular preparation technologies of Herba Leonuri of experimental example
The preparation of BAZHEN YIMU JIAONANG of the present invention relates to the volatile oil extraction, water soluble ingredient extracts and crude drug directly is used as medicine, and therefore, preparation technology investigates above-mentioned three aspects, to determine optimum process condition.
1.1 volatile oil extraction time
Radix Angelicae Sinensis, Rhizoma Chuanxiong, the Rhizoma Atractylodis Macrocephalae contain a large amount of volatile active matter, therefore, should separately volatile ingredient be extracted.Crude drug according to description of the present invention record is formed proportioning, takes by weighing each three parts of Radix Angelicae Sinensis, Rhizoma Chuanxiong and the Rhizoma Atractylodis Macrocephalaes of common 400g, and every part adds water 2000ml, and distillating extracting oil is that index has been investigated different extraction times with must measuring of volatile oil, the results are shown in Table 1.
The extraction time investigation of table 1 volatile oil
The result shows, generally extract 3 hours after, volatile oil extracts fully substantially, yield is about 0.12%.Therefore, the extraction time of determining volatile oil is 3 hours.
1.2 the optimum process condition that decocting boils
The extraction process that decocting boils, the influence that is decocted factors such as number of times, amount of water, decocting time influences factor in order to simplify, and is first-selected under the situation of fixedly amount of water and decocting time, investigates the decoction number of times; Secondly, decoct under the number of times, investigate amount of water and decocting time preferred.
1.2.1 decocting boils number of times
Form according to preferred crude drug of the present invention, take by weighing the crude drug that a certain amount of water decocts, total amount of water be 15 times under crude drug and decocting time are each 4.5 hours rigid condition, serve as to investigate index with the dry extract yield, investigate different decoction number of times, the results are shown in Table 2.
Table 2 decocting boils the investigation of number of times
The result shows, is adding 15 times of amounts of water, and each decocting time is under 4.5 hours the decocting condition, to decoct 2 dry extract yields of 3 ratios and only improve 1%, and from the production cost consideration, the number of times that decocting boils is defined as 2 times.
1.2.2 amount of water and decocting time
Forming according to preferred crude drug of the present invention, take by weighing the crude drug that a certain amount of water decocts, decocting under 2 times the condition, serves as to investigate index with dry extract yield and stachydrine hydrochloride content, has investigated different amount of water and decocting time, the results are shown in Table 3.
Table 3 amount of water and decocting time are investigated
The result shows, always adds 19 times of amounts of water---the 1st time 10 times, the 2nd time 9 times, extraction time is respectively---the 1st time 2 hours, under the 2nd time 1.5 hours the condition, dry extract yield and stachydrine hydrochloride content were all the highest; So determining this condition is best process conditions.
1.3 directly beating powder, Radix Paeoniae Alba, Poria be used as medicine
Radix Paeoniae Alba, Poria rich in starch and protein, color and luster is white, easily pulverizes; The two is beaten powder directly be used as medicine, can reduce moist, increase disintegration of drawing of preparation, be convenient to dry extract pulverizing, granulation, encapsulated and preservation.If but Poria all beat powder and be used as medicine, it is excessive then to press synthetic day dose volume of 8.3g crude drug folding, 9 No. 1 capsules also can not load.Adopt 1/3rd Poria to beat powder and be used as medicine, by synthetic day dose of 8.3g crude drug folding 9 No. 1 capsules of just packing into.Event determines that beating powder with the Poria of Radix Paeoniae Alba of total weight part and 1/3rd weight portions is used as medicine.
The pharmacodynamics of experimental example 2 BAZHEN YIMU JIAONANG and bazhen yimu pills, bazhen yimu wan of the prior art relatively
The raw material of bazhen yimu pills, bazhen yimu wan consists of:
Herba Leonuri 200g Radix Codonopsis 50g Radix Rehmanniae Preparata 100g Rhizoma Atractylodis Macrocephalae (parched) 50g Poria 50g Radix Angelicae Sinensis 100g Radix Paeoniae Alba 50g Rhizoma Chuanxiong 50g Radix Glycyrrhizae 25g
Prepare by following method:
More than nine flavors pulverize, be crude drug according to weight ratio: refined honey=100: 45, add refined honey and an amount of water pill, drying is made water-honeyed pill.
The described bazhen yimu pills, bazhen yimu wan of every 1g is converted to the raw material dose, is equivalent to 0.69g.
Find that by pharmacology test two kinds of dosage forms all can promote rat uterus to grow, improve estradiol content and reduction progesterone content in the blood; The movable nothing detail of normal isolated rat uterine smooth muscle is influenced, can obviously increase the excitation of oxytocin to the isolated rat uterus, increase its contraction frequency and energy, the inhibitory action of Progesterone be can resist, its contraction frequency, amplitude and energy increased the excited isolated rat uterine smooth muscle of oxytocin; Can improve mice carbon clearance speed, serum hemolysin level, peripheral blood lymphocyte conversion ratio; Can prolong mice swimming, hypoxia endurance time; Can promote the improvement and the body weight gain of losing blood property blood deficiency rat symptom and sign, accelerate blood RBC, Hb and recover; In addition, still can alleviate rat paw edema and granuloma hypertrophy due to the agar.Capsule of the present invention is compared with described bazhen yimu pills, bazhen yimu wan, promote the rat uterus growth, improving estradiol content in the blood, strengthen the excitation of oxytocin to the isolated rat uterus, improve mice carbon clearance speed, all there is enhancing in various degree aspects such as prolongation mice swimming time.Explanation is compared with pill of the prior art, and capsule of the present invention can improve curative effect to a certain extent.
The specificity test that experimental example 3 Radix Angelicae Sinensis, Rhizoma Chuanxiong thin layer chromatography are differentiated
Discrimination method I is that the thin layer chromatography of Radix Angelicae Sinensis, Rhizoma Chuanxiong is differentiated.Radix Angelicae Sinensis, Rhizoma Chuanxiong crude drug derive from samphire, and both chemical constituents are seemingly closer, and under suitable thin layer chromatography condition, the thin layer chromatography of the two also has to a certain degree similar; Therefore, by a thin layer chromatography, just can while qualitative identification Radix Angelicae Sinensis and Rhizoma Chuanxiong.
Get the preparation method that other medical material of not containing Radix Angelicae Sinensis, Rhizoma Chuanxiong in the prescription puts down in writing to specifications and make negative sample; get content (hereinafter to be referred as the test sample) 6g of BAZHEN YIMU JIAONANG of the present invention; negative sample 6g; Radix Angelicae Sinensis control medicinal material 1g (Nat'l Pharmaceutical ﹠ Biological Products Control Institute; lot number: 120927-200613); (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 120918-200501) the described method of by specification is mixed with need testing solution, negative sample solution, control medicinal material solution to Rhizoma Chuanxiong control medicinal material 1g.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution, negative sample solution, each 3 μ l of control medicinal material solution, putting respectively on same silica gel g thin-layer plate, is that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launches, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color; In the negative sample chromatograph with the corresponding position of reference substance chromatograph on no corresponding speckle.Illustrate negative noiseless.(see figure 1)
The specificity test that experimental example 4 Herba Leonuri thin layer chromatographys are differentiated
Discrimination method II is that the thin layer chromatography of Herba Leonuri is differentiated.
Get the preparation method that other medical material of not containing Herba Leonuri in the prescription puts down in writing to specifications and make negative sample, get test sample 8g, negative sample 8g, (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 110712-200508) the described method of by specification is mixed with need testing solution, negative sample solution, reference substance solution to the stachydrine hydrochloride reference substance.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 10 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launch, take out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; In the negative sample chromatograph with the corresponding position of reference substance chromatograph on no corresponding speckle.Illustrate negative noiseless.(see figure 2)
Experimental example 5 paeoniflorin contents are measured
1, instrument and reagent
SHIMADZU LC-2010AHT high performance liquid chromatograph, SHIMADZU LC-2010AHT chem workstation, VP-ODS C18 chromatographic column (150mm * 4.6mm, 5 μ m), the variable detector of ultraviolet.
Water is ultra-pure water, and acetonitrile is a chromatographically pure, and other reagent is analytical pure; Peoniflorin reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 110736-200526).
2, system suitability test
With octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solutions are mobile phase with volume ratio, and the detection wavelength is 230nm, and theoretical cam curve is calculated by the peoniflorin peak should be not less than 1500.
3, wavelength is selected
It is an amount of to get the peoniflorin reference substance, makes suitable concentration with 50% methanol, is blank with 50% methanol, carries out spectral scan in 400～200nm wave-length coverage, and the result has the absorption maximum (see figure 3) at 230nm wavelength place, so determine that detecting wavelength is 230nm.
4, the selection of mobile phase
In the prior art, mobile phase is that volume ratio is acetonitrile-0.05mol/L potassium dihydrogen phosphate of 13: 87, but the peoniflorin peak shape is bad, number of theoretical plate is low, after to change volume ratio into be 13: 87 acetonitrile-0.1% phosphoric acid solution, the peoniflorin peak shape was good, the high (see figure 4) of number of theoretical plate.
5, the preparation of need testing solution
5.1 extract the investigation of solvent
Get four parts of the contents of BAZHEN YIMU JIAONANG of the present invention, each 0.3g, accurate claim fixed, investigate with 50% methanol, ethanol, 50% ethanol, four kinds of solvents of methanol respectively, adopt supersound process (power 250W, frequency 25kHz) extracting method to extract, measurement result sees Table 8.
Table 8 different solvents extraction and determination result
Determine that 50% methanol is the optimum extraction solvent.
5.2 the investigation of extraction time
Get three parts of the capsular contents of eight gusts of Herba Leonuri of the present invention, each 0.3g, the accurate title, decide, and supersound process (power 250W, frequency 25kHz) is 30 minutes, 60 minutes, 120 minutes respectively, the results are shown in Table 9.
Table 9 measurement result of different extraction time
The result shows, supersound process (power 250W, frequency 25kHz) 60 minutes and 120 minutes assays be basically identical as a result, illustrates that supersound extraction can be effective in 60 minutes, so determine that the supersound process time is 60 minutes.
6, blank assay
Get other control sample that does not contain Radix Paeoniae Alba in the prescription, the need testing solution preparation method of by specification record is made negative sample, the need testing solution preparation method of middle record is made negative sample solution to specifications again, content assaying method test according to the description record, compare need testing solution chromatograph and peoniflorin reference substance chromatograph, other compositions are to peoniflorin peak noiseless (seeing Fig. 4,5,6) in the negative sample as a result, and peoniflorin has chromatographic isolation preferably.
7, the preparation of reference substance solution
Precision takes by weighing the described peoniflorin reference substance 12.59mg that is dried to constant weight through phosphorus pentoxide, puts in the 50ml measuring bottle, adds 50% dissolve with methanol and is diluted to scale, shakes up.Precision is measured 2ml and is put in the 25ml measuring bottle, adds 50% methanol to scale, shakes up, promptly.
8, linear relationship is investigated
Accurate above-mentioned reference substance solution each 5 μ l, 10 μ l, 20 μ l, 30 μ l, the 40 μ l of drawing inject chromatograph of liquid, and Ji Zai chromatographiccondition is measured peak area to specifications, the results are shown in Table 10.
Table 10 linear relationship measurement result
With the peak area integrated value is vertical coordinate, and the reference substance sample size is an abscissa, and the drawing standard curve is seen Fig. 7.Regression equation is:
Y＝1194141.298504X-1983.228659，r
2＝0.999993
Peoniflorin is good linear between 0.10072～0.80576 μ g.
9, precision test
9.1 reference substance precision test
The described reference substance solution 20 μ l of accurate absorption, the liquid phase chromatogram condition of by specification record repeats sample introduction 5 times, the results are shown in Table 11, and peoniflorin area relative standard deviation is 0.2%.
Table 11 reference substance Precision test result
9.2 test sample precision test
Get the content 0.3g of described BAZHEN YIMU JIAONANG, the accurate title, decide, Ji Zai method prepares need testing solution to specifications, the described need testing solution 20 μ l of accurate absorption, the liquid phase chromatogram condition of by specification record, repeat sample introduction 5 times, the results are shown in Table 12, peoniflorin area relative standard deviation is 0.2%.
Table 12 test sample Precision test result
10, stability test
Get the content 0.3g of described BAZHEN YIMU JIAONANG, the accurate title, decide, Ji Zai method is made need testing solution to specifications, the chromatographic condition of by specification record, accurate described need testing solution 20 μ l, the injection chromatograph of liquid drawn, sample introduction is measured once at regular intervals, in 24 hours, peak area is consistent substantially, RSD=1.6%.Show that peoniflorin is stable in 24 hours in described mobile phase solution, the results are shown in Table 13.
Table 13 stability test result
11, replica test
Get respectively with 5 parts of the contents of a collection of described BAZHEN YIMU JIAONANG, accurately claim surely, Ji Zai method is made need testing solution to specifications, under the chromatographic condition of description record, measure content of paeoniflorin, RSD=1.9%, show that this law repeatability is good, the results are shown in Table 14.
Table 14 replica test result
12, average recovery test
5 parts of the contents of the described BAZHEN YIMU JIAONANG that to get with a collection of known paeoniflorin content be 2.9113mg/g, every part of about 0.15g, the accurate title, decide, put in the tool plug conical flask, accurate respectively peoniflorin reference substance solution (the concentration 7.554 μ g/ml) 50ml that adds, Ji Zai method is made need testing solution to specifications, under the chromatographic condition of description record, measure content of paeoniflorin, calculate recovery rate, average recovery rate are 99.86%, RSD=0.9%, show that this law response rate is good, the results are shown in Table 15.
Table 15 recovery test result
13, sample determination
Get 6 batches of BAZHEN YIMU JIAONANG of the present invention, Ji Zai method is made need testing solution to specifications, under the chromatographic condition of description record, measures content of paeoniflorin, and measurement result sees Table 16.
Table 16 sample determination result
According to sample determination result in the table 16, draft every of BAZHEN YIMU JIAONANG of the present invention and contain Radix Paeoniae Alba with peoniflorin C
23H
28O
11Meter must not be less than 0.90mg.
Those skilled in the art are to be understood that, the quality determining method of BAZHEN YIMU JIAONANG of the present invention is equally applicable to crude drug to be formed and same or analogous other compound Chinese medicinal preparation of BAZHEN YIMU JIAONANG of the present invention, as pill, tablet, granule, oral liquid, concentrated pill etc.The day clothes dosage of different compound Chinese medicinal preparation is variant because of the difference of preparation process, but every day taking dose to be converted to crude drug be consistent, the amount that therefore can be converted to crude drug takes by weighing desired for the preparation that detects.
Description of drawings
Fig. 1 is that Radix Angelicae Sinensis and Rhizoma Chuanxiong thin layer are differentiated, 1 negative sample wherein, and 2 is the Radix Angelicae Sinensis control medicinal material, and 3 is the Rhizoma Chuanxiong control medicinal material, and 4,5,6 is three batch samples.
Fig. 2 is that the Herba Leonuri thin layer is differentiated, 1 negative sample wherein, and 2 is the stachydrine hydrochloride reference substance, 3,4,5 is three batch samples.
Fig. 3 is the ultra-violet absorption spectrum of peoniflorin reference substance
Fig. 4 is a peoniflorin reference substance solution HPLC collection of illustrative plates
Fig. 5 is described capsule need testing solution HPLC collection of illustrative plates
Fig. 6 is a negative sample Solution H PLC collection of illustrative plates
Fig. 7 is a peoniflorin standard solution canonical plotting
The specific embodiment
Following embodiment all can realize the effect of above-mentioned experimental example, but the present invention is not limited to described embodiment.
Herba Leonuri 273g Radix Codonopsis 68g Radix Rehmanniae Preparata 137g Rhizoma Atractylodis Macrocephalae (parched) 68g Poria 68g Radix Angelicae Sinensis 137g Radix Paeoniae Alba 68g Rhizoma Chuanxiong 68g Radix Glycyrrhizae 34g
Poria and the 68g Radix Paeoniae Alba of getting 22.5g are ground into coarse powder; The Radix Angelicae Sinensis of described recipe quantity, Rhizoma Chuanxiong, the Rhizoma Atractylodis Macrocephalae extract volatile oil, and the aqueous solution after distillation device is in addition collected, and the Radix Codonopsis of medicinal residues and described recipe quantity, Radix Rehmanniae Preparata, Herba Leonuri, Radix Glycyrrhizae and remaining Poria decoct with water secondary, 2 hours for the first time, add 10 times of water gagings, 1.5 hours for the second time, add 9 times of water gagings; Collecting decoction filters, and the aqueous solution after filtrate and the distillation merges, and relative density is 1.25～1.30 extractum when being concentrated into 60 ℃, add described coarse powder, stir evenly, 80～90 ℃ of oven dry, pulverize, adding starch adjustment total amount is 280g, sieves, mixing is granulated drying with 90% alcohol, spray into above-mentioned volatile oil, sealing incapsulates, make 1000, every dress 0.28g, promptly.
Instructions of taking: oral, one time 3, one day three times.
The described BAZHEN YIMU JIAONANG content of every 1g is converted to the raw material dose, is equivalent to 3.29g.
Herba Leonuri 300g Radix Codonopsis 75g Radix Rehmanniae Preparata 120g Rhizoma Atractylodis Macrocephalae (parched) 60g Poria 72g Radix Angelicae Sinensis 150g Radix Paeoniae Alba 75g Rhizoma Chuanxiong 65g Radix Glycyrrhizae 35g
Poria and the 75g Radix Paeoniae Alba of getting 48g are ground into coarse powder; The Radix Angelicae Sinensis of described recipe quantity, Rhizoma Chuanxiong, the Rhizoma Atractylodis Macrocephalae extract volatile oil, and the aqueous solution after distillation device is in addition collected, and the Radix Codonopsis of medicinal residues and described recipe quantity, Radix Rehmanniae Preparata, Herba Leonuri, Radix Glycyrrhizae and remaining Poria decoct with water secondary, 2 hours for the first time, add 10 times of water gagings, 1.5 hours for the second time, add 9 times of water gagings; Collecting decoction filters, and the aqueous solution after filtrate and the distillation merges, relative density is 1.25～1.30 extractum when being concentrated into 60 ℃, adds described coarse powder, stirs evenly, 80～90 ℃ of oven dry are pulverized, and add conventional adjuvant, granulate, spray into the volatile oil that dilutes with an amount of dehydrated alcohol in advance, sealing, be pressed into tablet, make 1000, every 0.3g, promptly.
Instructions of taking: oral, one time 3, one day three times.
The described BAZHEN YIMU PIAN of every 1g is converted to the raw material dose, is equivalent to 3.17g.
Herba Leonuri 250g Radix Codonopsis 60g Radix Rehmanniae Preparata 150g Rhizoma Atractylodis Macrocephalae (parched) 70g Poria 66g Radix Angelicae Sinensis 120g Radix Paeoniae Alba 70g Rhizoma Chuanxiong 65g Radix Glycyrrhizae 30g
More than nine the flavor, prepare granule according to the conventional formulation method, every bag of 1.5g.
Instructions of taking: one time one bag, one day three times.
The described BAZHEN YIMU KELI of every 1g is converted to the raw material dose, is equivalent to 1.89g.
Herba Leonuri 200g Radix Codonopsis 50g Radix Rehmanniae Preparata 100g Rhizoma Atractylodis Macrocephalae (parched) 50g Poria 50g Radix Angelicae Sinensis 100g Radix Paeoniae Alba 50g Rhizoma Chuanxiong 50g Radix Glycyrrhizae 25g
More than nine flavors pulverize, be crude drug according to weight ratio: refined honey=100: 45, add refined honey and an amount of water pill, drying is made water-honeyed pill.
Instructions of taking: one time 1 ball, one day secondary.
The described bazhen yimu pills, bazhen yimu wan of every 1g is converted to the raw material dose, is equivalent to 0.69g.
Herba Leonuri 300g Radix Codonopsis 75g Radix Rehmanniae Preparata 150g Rhizoma Atractylodis Macrocephalae (parched) 70g Poria 75g Radix Angelicae Sinensis 150g Radix Paeoniae Alba 75g Rhizoma Chuanxiong 70g Radix Glycyrrhizae 35g
Poria and the 60g Radix Paeoniae Alba of getting 36g are ground into coarse powder; The Radix Angelicae Sinensis of described recipe quantity, Rhizoma Chuanxiong, the Rhizoma Atractylodis Macrocephalae extract volatile oil, and the aqueous solution after distillation device is in addition collected, and the Radix Codonopsis of medicinal residues and described recipe quantity, Radix Rehmanniae Preparata, Herba Leonuri, Radix Glycyrrhizae and remaining Poria decoct with water secondary, 2 hours for the first time, add 10 times of water gagings, 1.5 hours for the second time, add 9 times of water gagings; Collecting decoction filters, and the aqueous solution after filtrate and the distillation merges, and relative density is 1.25～1.30 extractum when being concentrated into 60 ℃, add described coarse powder, stir evenly, 80～90 ℃ of oven dry are pulverized, add conventional adjuvant, make 1000 concentrated pills, bottling, 100 every bottle.
Instructions of taking: each 8 balls, one day three times
The described BAZHEN YIMU NONGSUOWAN of every 1g is converted to the raw material dose, is equivalent to 2.78g.
The BAZHEN YIMU JIAONANG quality determining method of embodiment 6 embodiment 1 preparation
[discriminating] is according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin layer chromatography
I, get the content 6g of described BAZHEN YIMU JIAONANG, porphyrize adds normal hexane 25ml, and supersound process 15 minutes filters, and filtrate volatilizes, and residue adds normal hexane 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1g of Rhizoma Chuanxiong control medicinal material, adds normal hexane 15ml respectively, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launches, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
II, get the content 8g of described BAZHEN YIMU JIAONANG, add an amount of kieselguhr, grind well, put in the apparatus,Soxhlet's, added alcohol reflux 4 hours, extracting solution filters, filtrate recycling ethanol, dissolve with 0.1mol/L hydrochloric acid solution 8ml gradation, acid solution filters, and merging filtrate adds the chromic thiocyanate ammonium salt saturated solution 12ml that faces with preparation, placing 1 hour below 10 ℃, filter with sintered glass funnel, precipitation adds acetone 5ml and makes dissolving after washing with low amounts of water, 0.5% silver sulfate solution to the precipitation that adds about 5ml is more no longer separated out, filter, precipitation is washed with small amount of acetone, and washing liquid and filtrate merge, be concentrated into about 2ml, add the about 2.5ml of 1% barium chloride solution, mixing, being added in internal diameter is 15mm, in adorn on the chromatographic column of 80～100 order neutral alumina 1.5g, with 70% ethanol 35ml eluting, collect eluent, be concentrated into dried, residue adds ethanol 1.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[assay] measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D)
The test of chromatographic condition and system suitability: with the octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solution is a mobile phase with volume ratio, and detections wavelength is 230nm, and number of theoretical plate should be not less than 1500 by the calculating of peoniflorin peak;
The preparation of reference substance solution: the phosphorus pentoxide of learning from else's experience is dried to the peoniflorin reference substance of constant weight, and accurate the title decides, and adds 50% methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get the content 2.8g of described BAZHEN YIMU JIAONANG, mixing, porphyrize, get 0.3g, the accurate title, decide, the accurate 50% methanol 50ml that adds, claim to decide weight, close plug, power 250W, the supersound process of frequency 25kHz 60 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with 50% methanol, shake up, filter, get subsequent filtrate, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
Every described BAZHEN YIMU JIAONANG contains Radix Paeoniae Alba with peoniflorin C
23H
28O
11Meter must not be less than 0.90mg.
The quality determining method of the tablet of embodiment 7 embodiment 2 preparations
[discriminating] is according to an appendix VI of Chinese Pharmacopoeia version in 2000 B thin layer chromatography
I, get described tablet 6.3g, porphyrize adds normal hexane 25ml, and supersound process 15 minutes filters, and filtrate volatilizes, and residue adds normal hexane 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1g of Rhizoma Chuanxiong control medicinal material, adds normal hexane 15ml respectively, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
II, get described tablet 8.2g, add an amount of kieselguhr, grind well, put in the apparatus,Soxhlet's, added alcohol reflux 4 hours, extracting solution filters, filtrate recycling ethanol, dissolve with 0.1mol/L hydrochloric acid solution 8ml gradation, acid solution filters, and merging filtrate adds the chromic thiocyanate ammonium salt saturated solution 12ml that faces with preparation, placing 1 hour below 10 ℃, filter with sintered glass funnel, precipitation adds acetone 5ml and makes dissolving after washing with low amounts of water, 0.5% silver sulfate solution to the precipitation that adds about 5ml is more no longer separated out, filter, precipitation is washed with small amount of acetone, and washing liquid and filtrate merge, be concentrated into about 2ml, add the about 2.5ml of 1% barium chloride solution, mixing, being added in internal diameter is 15mm, in adorn on the chromatographic column of 80～100 order neutral alumina 1.5g, with 70% ethanol 35ml eluting, collect eluent, be concentrated into dried, residue adds ethanol 1.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[assay] measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D)
The test of chromatographic condition and system suitability: with the octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solution is a mobile phase with volume ratio, and detections wavelength is 230nm, and number of theoretical plate should be not less than 1500 by the calculating of peoniflorin peak;
The preparation of reference substance solution: the phosphorus pentoxide of learning from else's experience is dried to the peoniflorin reference substance of constant weight, and accurate the title decides, and adds 50% methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get described tablet 3g, porphyrize is got 0.3g, the accurate title, decide, and the accurate 50% methanol 50ml that adds claims to decide weight, power 250W, the supersound process of frequency 25kHz 60 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 50% methanol, shake up, filter, get subsequent filtrate, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
The described tablet that is equivalent to crude drug 0.92g contains Radix Paeoniae Alba with peoniflorin (C
23H
28O
11) meter, must not be less than 0.90mg.
The quality determining method of the granule of embodiment 8 embodiment 3 preparations
[discriminating] is according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin layer chromatography
I, get described granule 10.6g, porphyrize adds normal hexane 25ml, and supersound process 15 minutes filters, and filtrate volatilizes, and residue adds normal hexane 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1g of Rhizoma Chuanxiong control medicinal material, adds normal hexane 15ml respectively, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
II, get described granule 13.8g, add an amount of kieselguhr, grind well, put in the apparatus,Soxhlet's, added alcohol reflux 4 hours, extracting solution filters, filtrate recycling ethanol, dissolve with 0.1mol/L hydrochloric acid solution 8ml gradation, acid solution filters, and merging filtrate adds the chromic thiocyanate ammonium salt saturated solution 12ml that faces with preparation, placing 1 hour below 10 ℃, filter with sintered glass funnel, precipitation adds acetone 5ml and makes dissolving after washing with low amounts of water, 0.5% silver sulfate solution to the precipitation that adds about 5ml is more no longer separated out, filter, precipitation is washed with small amount of acetone, and washing liquid and filtrate merge, be concentrated into about 2ml, add the about 2.5ml of 1% barium chloride solution, mixing, being added in internal diameter is 15mm, in adorn on the chromatographic column of 80～100 order neutral alumina 1.5g, with 70% ethanol 35ml eluting, collect eluent, be concentrated into dried, residue adds ethanol 1.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[assay] measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D)
The test of chromatographic condition and system suitability: with the octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solution is a mobile phase with volume ratio, and detections wavelength is 230nm, and number of theoretical plate should be not less than 1500 by the calculating of peoniflorin peak;
The preparation of reference substance solution: the phosphorus pentoxide of learning from else's experience is dried to the peoniflorin reference substance of constant weight, and accurate the title decides, and adds 50% methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get described granule 5g, porphyrize is got 0.5g, the accurate title, decide, and the accurate 50% methanol 50ml that adds claims to decide weight, power 250W, the supersound process of frequency 25kHz 60 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 50% methanol, shake up, filter, get subsequent filtrate, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
The quality determining method of the honey pill agent of embodiment 9 embodiment 4 preparations
[discriminating] is according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin layer chromatography
I, get described honey pill agent 28g, grind evenly, add normal hexane 25ml, supersound process 15 minutes filters, and filtrate volatilizes, and residue adds normal hexane 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1g of Rhizoma Chuanxiong control medicinal material, adds normal hexane 15ml respectively, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
II, get described honey pill agent 37.7g, add an amount of kieselguhr, grind well, put in the apparatus,Soxhlet's, added alcohol reflux 4 hours, extracting solution filters, filtrate recycling ethanol, dissolve with 0.1mol/L hydrochloric acid solution 8ml gradation, acid solution filters, and merging filtrate adds the chromic thiocyanate ammonium salt saturated solution 12ml that faces with preparation, placing 1 hour below 10 ℃, filter with sintered glass funnel, precipitation adds acetone 5ml and makes dissolving after washing with low amounts of water, 0.5% silver sulfate solution to the precipitation that adds about 5ml is more no longer separated out, filter, precipitation is washed with small amount of acetone, and washing liquid and filtrate merge, be concentrated into about 2ml, add the about 2.5ml of 1% barium chloride solution, mixing, being added in internal diameter is 15mm, in adorn on the chromatographic column of 80～100 order neutral alumina 1.5g, with 70% ethanol 35ml eluting, collect eluent, be concentrated into dried, residue adds ethanol 1.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[assay] measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D)
The test of chromatographic condition and system suitability: with the octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solution is a mobile phase with volume ratio, and detections wavelength is 230nm, and number of theoretical plate should be not less than 1500 by the calculating of peoniflorin peak;
The preparation of reference substance solution: the phosphorus pentoxide of learning from else's experience is dried to the peoniflorin reference substance of constant weight, and accurate the title decides, and adds 50% methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get described honey pill agent 13g, grind evenly, get 1.5g, the accurate title, decide, and the accurate 50% methanol 50ml that adds claims to decide weight, power 250W, the supersound process of frequency 25kHz 60 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 50% methanol, shake up, filter, get subsequent filtrate, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
The described honey pill agent that is equivalent to crude drug 0.92g contains Radix Paeoniae Alba with peoniflorin (C
23H
28O
11) meter, must not be less than 0.90mg.
The method of quality control of the concentrated pill of embodiment 10 embodiment 5 preparations
[discriminating] is according to an appendix VI of Chinese Pharmacopoeia version in 2005 B thin layer chromatography
I, get described concentrated pill 7.2g, grind evenly, add normal hexane 25ml, supersound process 15 minutes filters, and filtrate volatilizes, and residue adds normal hexane 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1g of Rhizoma Chuanxiong control medicinal material, adds normal hexane 15ml respectively, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
II, get described concentrated pill 9.4g, add an amount of kieselguhr, grind well, put in the apparatus,Soxhlet's, added alcohol reflux 4 hours, extracting solution filters, filtrate recycling ethanol, dissolve with 0.1mol/L hydrochloric acid solution 8ml gradation, acid solution filters, and merging filtrate adds the chromic thiocyanate ammonium salt saturated solution 12ml that faces with preparation, placing 1 hour below 10 ℃, filter with sintered glass funnel, precipitation adds acetone 5ml and makes dissolving after washing with low amounts of water, 0.5% silver sulfate solution to the precipitation that adds about 5ml is more no longer separated out, filter, precipitation is washed with small amount of acetone, and washing liquid and filtrate merge, be concentrated into about 2ml, add the about 2.5ml of 1% barium chloride solution, mixing, being added in internal diameter is 15mm, in adorn on the chromatographic column of 80～100 order neutral alumina 1.5g, with 70% ethanol 35ml eluting, collect eluent, be concentrated into dried, residue adds ethanol 1.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
[assay] measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D)
The test of chromatographic condition and system suitability: with the octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solution is a mobile phase with volume ratio, and detections wavelength is 230nm, and number of theoretical plate should be not less than 1500 by the calculating of peoniflorin peak;
The preparation of reference substance solution: the phosphorus pentoxide of learning from else's experience is dried to the peoniflorin reference substance of constant weight, and accurate the title decides, and adds 50% methanol and makes the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get described concentrated pill 3.3g, grind evenly, get 0.36g, the accurate title, decide, and the accurate 50% methanol 50ml that adds claims to decide weight, power 250W, the supersound process of frequency 25kHz 60 minutes is put cold, claim to decide weight again, supply the weight that subtracts mistake with 50% methanol, shake up, filter, get subsequent filtrate, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
The described compound Chinese medicinal preparation that is equivalent to crude drug 0.92g contains Radix Paeoniae Alba with peoniflorin (C
23H
28O
11) meter, must not be less than 0.90mg.
Claims (3)
1. the quality determining method of crude drug BAZHEN YIMU JIAONANG composed as follows,
Herba Leonuri 273 weight portions, Radix Codonopsis 68 weight portions, Radix Rehmanniae Preparata 137 weight portions, Rhizoma Atractylodis Macrocephalae (parched) 68 weight portions, Poria 68 weight portions, Radix Angelicae Sinensis 137 weight portions, Radix Paeoniae Alba 68 weight portions, Rhizoma Chuanxiong 68 weight portions, Radix Glycyrrhizae 34 weight portions;
Described capsule prepares by the following method:
Get the Poria of 1/3rd weight portions and the Radix Paeoniae Alba of described weight portion and be ground into coarse powder; The Radix Angelicae Sinensis of described weight portion, Rhizoma Chuanxiong, Rhizoma Atractylodis Macrocephalae distillating extracting oil, the aqueous solution after the distillation device are in addition collected the Radix Codonopsis of medicinal residues and described weight portion, Radix Rehmanniae Preparata, Herba Leonuri, Radix Glycyrrhizae and the Poria that remains 2/3rds weight portions decoct with water secondary, 2 hours for the first time, 1.5 hours for the second time, decocting liquid filtered, filtrate merges, and with the aqueous solution merging after the distillation, relative density is 1.25～1.30 extractum when being concentrated into 60 ℃, add described coarse powder, stir evenly, 80～90 ℃ of oven dry are pulverized, add starch, sieve, mixing is used 90% alcohol granulation, dry, spray into described volatile oil, sealing incapsulates, every dress 0.28g, promptly;
It is characterized in that described quality determining method comprises the authentication method of following I, II:
I, get the content 6g of described BAZHEN YIMU JIAONANG, add normal hexane 25ml, supersound process 15 minutes filters, and filtrate volatilizes, and residue adds normal hexane 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1g of Rhizoma Chuanxiong control medicinal material, adds normal hexane 15ml respectively, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that 60～90 ℃ of petroleum ether-ethyl acetates of 10: 0.4 are developing solvent with volume ratio, launches, take out, dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
II, get the content 8g of described BAZHEN YIMU JIAONANG, add an amount of kieselguhr, grind well, put in the apparatus,Soxhlet's, added alcohol reflux 4 hours, extracting solution filters, filtrate recycling ethanol, dissolve with 0.1mol/L hydrochloric acid solution 8ml gradation, acid solution filters, and merging filtrate adds the chromic thiocyanate ammonium salt saturated solution 12ml that faces with preparation, placing 1 hour below 10 ℃, filter with sintered glass funnel, precipitation adds acetone 5ml and makes dissolving after washing with low amounts of water, 0.5% silver sulfate solution to the precipitation that adds about 5ml is more no longer separated out, filter, precipitation is washed with small amount of acetone, and washing liquid and filtrate merge, be concentrated into about 2ml, add the about 2.5ml of 1% barium chloride solution, mixing, being added in internal diameter is 15mm, in adorn on the chromatographic column of 80～100 order neutral alumina 1.5g, with 70% ethanol 35ml eluting, collect eluent, be concentrated into dried, residue adds ethanol 1.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that n-butyl alcohol-ethyl acetate-hydrochloric acid of 8: 1: 3 is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
2. the quality determining method of BAZHEN YIMU JIAONANG according to claim 1 is characterized in that described quality determining method also comprises the assay of following photograph high performance liquid chromatography:
The test of chromatographic condition and system suitability: with the octadecylsilane chemically bonded silica is filler, is that 13: 87 acetonitrile-0.1% phosphoric acid solution is a mobile phase with volume ratio, and detections wavelength is 230nm, and number of theoretical plate should be not less than 1500 by the calculating of peoniflorin peak;
The preparation of reference substance solution: get the peoniflorin reference substance, the accurate title, decide, and adds 50% methanol and make the solution that every 1ml contains 20 μ g, promptly;
The preparation of need testing solution: get the content 2.8g of described BAZHEN YIMU JIAONANG, mixing, porphyrize, get 0.3g, the accurate title, decide, the accurate 50% methanol 50ml that adds, claim to decide weight, close plug, the supersound process of power 250W, frequency 25kHz 60 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with 50% methanol, shake up, filter, get subsequent filtrate, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly; Every described BAZHEN YIMU JIAONANG contains Radix Paeoniae Alba with peoniflorin C
23H
28O
11Meter must not be less than 0.90mg.
3. quality determining method according to claim 1 and 2 is characterized in that Radix Angelicae Sinensis, the Rhizoma Chuanxiong of described weight portion, the time of Rhizoma Atractylodis Macrocephalae distillating extracting oil are 3 hours; The total amount of water that decocts with water is 19 times of gross weight of Radix Angelicae Sinensis, Rhizoma Chuanxiong, the Rhizoma Atractylodis Macrocephalae, Radix Codonopsis, Radix Rehmanniae Preparata, Herba Leonuri, Radix Glycyrrhizae and 2/3rds weight portion Poria of described weight portion, 10 times for the first time, and 9 times for the second time.
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| CN102824457A (en) * | 2012-06-04 | 2012-12-19 | 江西南昌桑海制药厂 | Eight-treasure motherwort active site and preparation process of capsules |
| CN104198615A (en) * | 2014-09-17 | 2014-12-10 | 上海海虹实业(集团)巢湖今辰药业有限公司 | Method for determining content of paeoniflorin in flavored motherwort cream |
| CN107412513A (en) * | 2017-07-21 | 2017-12-01 | 刘道鹏 | A kind of irregular menstruation tea and preparation method thereof |
| CN108159373A (en) * | 2018-01-06 | 2018-06-15 | 明光市千里浓酒业有限公司 | Corydalis tuber wine and preparation method thereof |
| CN110333304A (en) * | 2019-06-30 | 2019-10-15 | 广西万寿堂药业有限公司 | The detection method of content of puerperal blood clot dispersing pharmaceutical preparation |
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| CN1456206A (en) * | 2003-01-03 | 2003-11-19 | 江西南昌桑海制药厂 | Qi-blood tonifying capsule |
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| CN102579669A (en) * | 2012-03-05 | 2012-07-18 | 宁夏紫荆花制药有限公司 | Chinese medicinal composition for nourishing qi and blood and conditioning menstruation and preparation method thereof |
| CN102824457A (en) * | 2012-06-04 | 2012-12-19 | 江西南昌桑海制药厂 | Eight-treasure motherwort active site and preparation process of capsules |
| CN104198615A (en) * | 2014-09-17 | 2014-12-10 | 上海海虹实业(集团)巢湖今辰药业有限公司 | Method for determining content of paeoniflorin in flavored motherwort cream |
| CN107412513A (en) * | 2017-07-21 | 2017-12-01 | 刘道鹏 | A kind of irregular menstruation tea and preparation method thereof |
| CN108159373A (en) * | 2018-01-06 | 2018-06-15 | 明光市千里浓酒业有限公司 | Corydalis tuber wine and preparation method thereof |
| CN110333304A (en) * | 2019-06-30 | 2019-10-15 | 广西万寿堂药业有限公司 | The detection method of content of puerperal blood clot dispersing pharmaceutical preparation |
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