CN1814127B - Shenbao preparation, its preparing method and quality control method - Google Patents

Abstract

The present invention relates to a Shenbao preparation, relating to the preparing method and quality control method thereof. It has advantages of good stability, no sugar, no antiseptic, convenient to take and carry, suitable to industrialized production, low production cost, etc, and great marketing potential, and the quality control method can identify Chinese Angelica, Rhizoma Chuanxiong, Common Cnidium Fruit, prepared Tuber Fleeceflower Root, Epimedium Herb, and Malaytea Scurfpea Fruit in the Shenbao preparation, and making content determination on active substance of prepared Tuber Fleeceflower Root-2, 3, 5, 4'-tetrahydroxy diphenylethylene -2-O-b-D- glucoside, active substance of Epimedium Herb-Icariin, active substance of Common Cnidium Fruit-Osthole, and active substance of Chinese Magnoliavine Fruit- Chinese Magnoliavine Fruit methanol.

Description

Shenbao preparation and preparation method thereof method of quality control

Technical field

The present invention relates to a kind of Chinese medicine preparation, particularly relate to Shenbao preparation, relate to preparation method's method of quality control of said preparation simultaneously.

Background technology

The impotence that causes by suffering from a deficiency of the kidney, seminal emission, soreness of waist pain, chilly is afraid of cold, night pollakiuria, and diseases such as spiritlessness and weakness, alopecia tinnitus, women leukorrhea, menorrhalgia are common clinically common frdquently encountered disease diseases, the precious prescription of kidney is rigorous, and compatibility is thorough, has YIN and YANG balance regulating, temperature male wind-supplying kidney, the controlling nocturnal emission with astringent drugs of calming the nerves, the merit of strengthening the body resistance to consolidate the constitution, the product of regulating the flow of vital energy with yin-nourishing because of Fang Zhongzuo, oiliness so mend, temperature and not dry, the good medicine of nourishing for the four seasons.Clinically be widely used in above-mentioned illness, evident in efficacy.

Shenbao preparation is the Chinese medicine preparation of being processed into through extraction by 22 flavor Chinese medicines such as barrenwort, prepared fleece flower root, existing SHENBAO HEJI, the precious syrup of kidney on the market, its prescription and method of quality control all record for the standard laid down by the ministries or commissions of the Central Government, both quality standard basically identicals, the quality standard that shows the precious syrup of kidney that records with the standard laid down by the ministries or commissions of the Central Government is:

[prescription] frutus cnidii 28g Ligusticum wallichii 28.3g seed of Chinese dodder 66g

Psoralea corylifolia 28.5g Poria cocos 30g red ginseng 20g

Fennel seeds 14.4g fruit of Chinese magnoliavine 36g fruit of Cherokee rose 94.6g

Bighead atractylodes rhizome 14.2g Radix Angelicae Sinensis 46.8g raspberry 32.9g

Prepared fleece flower root 74.4g plantain seed 16.5g prepared rhizome of rehmannia 94g

Fruit of Chinese wolfberry 66g Chinese yam 46.3g barrenwort 94.6g

Fenugreek 94g Radix Astragali 51.4g saline cistanche 47.3g

Honey-fried licorice root 14.2g

[method for making] above 22 flavors, frutus cnidii, barrenwort, Radix Angelicae Sinensis, Ligusticum wallichii, fennel seeds are ground into meal, according to the percolation under liquid extract and the extract item (appendix IO), make solvent with 70% ethanol, flood after 48 hours, with the speed of per minute 1～3ml diacolation slowly, collect percolate 1500ml, reclaim ethanol, being evaporated to relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed), filter filtrate for later use; Red ginseng powder is broken into meal, adds 20% alcohol dipping after 8 hours, the refluxing extraction secondary, and each 2 hours, merge extract, filter, filtrate recycling ethanol is concentrated into relative density and is the clear cream of 1.10 (80 ℃ of heat are surveyed), and is standby; Ten Six-elements such as all the other raspberries, after suitably cataclasm, with the dregs of a decoction of red ginseng through alcohol extracting, boiling secondary, each 2 hours, collecting decoction filters, and filtrate is concentrated into the clear cream that relative density is 1.16 (80 ℃ of heat are surveyed), adding ethanol makes and contains alcohol amount and reach 65%, left standstill 48 hours, and got supernatant and reclaim ethanol, being concentrated into relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed).The alcohol extract that adds extract such as above-mentioned frutus cnidii and red ginseng, mixing adds the about 700ml of simple syrup, boils 10 minutes, and it is an amount of to add Sodium Benzoate, adds water to 1000ml, filtration, promptly.

[proterties] this product is tan thick liquid; Sweet, little hardship of distinguishing the flavor of.

This product 40ml is got in [discriminating] (1), puts in the separating funnel, adds hydrochloric acid 2ml and ether 40ml, and jolting is extracted, and divides and gets ether layer, and evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, and 2～3 of dropping sodium test solutions promptly show red.

(2) get this product 10ml, put in the separating funnel, add sherwood oil (30～60 ℃), 10ml, jolting is extracted, petroleum ether extract is put in the evaporating dish, volatilizes, and adds methyl alcohol 1ml and makes dissolving, add 2%3,2 of the methyl alcohol saturated solutions of 2～3 of the methanol solutions of 5-dinitrobenzoic acid and potassium hydroxide, heating promptly shows reddish violet.

(3) get this product 10ml, by H type strongly acidic cation-exchange post (the internal diameter 1cm that has handled well, storng-acid cation exchange resin 2g), wash with water to effluent colourless, discard water lotion, use (2mol/L) ammonia solution 50ml wash-out again, discard colourless first washing lotion, collect continuous washing lotion 30ml, put evaporate to dryness in the water-bath, residue adds 3% hydrochloric acid solution 3ml makes dissolving, filters, and filtrate is placed in 3 test tubes, add the silico-tungstic acid test solution respectively, each 2 of bismuth potassium iodide test solution and test solution of mercuric potassium iodide promptly generate the light brown precipitation respectively, red-brown precipitation and brown precipitate.

[inspection] relative density should be not less than 1.26.

The pH value should be 4.2～5.5.

Other should meet every regulation relevant under the syrup item.

[function with cure mainly] YIN and YANG balance regulating, temperature male wind-supplying kidney, the controlling nocturnal emission with astringent drugs of calming the nerves is strengthened the body resistance to consolidate the constitution.Be used for impotence, seminal emission, the waist and leg ache, lassitude, night pollakiuria, chilly is afraid of cold; Women's menorrhalgia, all diseases of clear and thin leucorrhea.

[usage and consumption] is oral, a 10～20ml, 3 times on the one.

[attention] cold, fever phase withdraws.

[specification] every 10ml, every bottle of 100ml, 200ml

Shady and cool dry place is put in [storage] sealing.

The subject matter that prior art exists is: (1) liquid preparation instability, put easy generation precipitation for a long time; (2) production cost height, and carry transportation inconvenience; (3) existing formulation all contains antiseptic; (4) all there is the high deficiency of sugar content in existing formulation, is unsuitable for diabetes, hepatopath etc. and takes; (5) quality control standard of existing preparation is low, one of discriminating three physicochemical identification is only arranged, and specificity is poor, and does not have assay, and difficulty is really controlled product quality.

The invention provides a kind of Shenbao preparation, overcome the weak point of above-mentioned preparation aspect, have good stability, do not contain antiseptic, take easy to carry, be suitable for advantages such as suitability for industrialized production, production cost be low.And owing to do not contain sugar, not only be suitable for general patient and take, and be suitable for comprising the diabetic all be unsuitable for taking the patient who contains sugared medicine and take.A kind of method of quality control of Shenbao preparation is provided simultaneously, promptly can differentiate the Radix Angelicae Sinensis in the Shenbao preparation and Ligusticum wallichii, frutus cnidii, prepared fleece flower root, barrenwort with thin-layered chromatography, with high-efficient liquid phase technique the Psoralea corylifolia in the Shenbao preparation is differentiated, measure in the Shenbao preparation 2 with high-efficient liquid phase technique, 3,5, the content of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside, icariin, Osthole, schizandrin.Shenbao preparation must be controlled product quality if can adopt above-mentioned discriminating and assay project to carry out Quality Control wholly or in part, guarantees the safe and effective of medication, improves the quality of product.

Summary of the invention

1. Shenbao preparation, this Chinese medicine is the solid preparation of traditional Chinese medicine of being made by following Chinese medicinal raw materials, following proportioning can change in proportion,

The frutus cnidii 28 weight portion Ligusticum wallichiis 28.3 weight portion seed of Chinese dodders 66 weight portions

Psoralea corylifolia 28.5 weight portion Poria cocos 30 weight portion red ginsengs 20 weight portions

The fennel seeds 14.4 weight portion fruit ofs Chinese magnoliavine 36 weight portion fruits of Cherokee rose 94.6 weight portions

The bighead atractylodes rhizome 14.2 weight portion Radix Angelicae Sinensis 46.8 weight portion raspberries 32.9 weight portions

Prepared fleece flower root 74.4 weight portion plantain seeds 16.5 weight portion prepared rhizomes of rehmannia 94 weight portions

The fruit of Chinese wolfberry 66 weight portion Chinese yams 46.3 weight portion barrenwort 94.6 weight portions

The fenugreek 94 weight portion Radixs Astragali 51.4 weight portion saline cistanches 47.3 weight portions

Honey-fried licorice root 14.2 weight portions

Its preparation method process following steps:

(1) ethanol percolation or the refluxing extraction of frutus cnidii, barrenwort, Radix Angelicae Sinensis, Ligusticum wallichii, fennel seeds usefulness 50-80% in the prescription;

(2) ethanol percolation or the refluxing extraction of red ginseng usefulness 10-50% in the prescription, the dregs of a decoction are standby;

(3) all the other ten Six-elements such as raspberry are with the above-mentioned red ginseng dregs of a decoction in the prescription, and water is carried or water extract-alcohol precipitation;

(4) extract of above-mentioned (1), (2), (3) gained is concentrated in right amount, mixing after mixing/adding appropriate amount of auxiliary materials, vacuum drying/spray drying/freeze drying;

(5) dried cream powder of above-mentioned (4) gained is made various solid pharmaceutical preparations according to a conventional method.

Described solid pharmaceutical preparation, the various solid dosage forms that comprise the kidney treasured are as tablet, sugar coated tablet, film coated tablet, chewable tablet, effervescent tablet, Disket, capsule, hard capsule, soft capsule, pellt capsule, suck agent, electuary, granule, effervescent, pill, pill, micropill preparation.

2. the method for quality control of a Shenbao preparation, contain in following assay and the qualitative identification one or more:

(1) assay a. measures in the Shenbao preparation 2,3,5 with high-efficient liquid phase technique, the content of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside; B. measure content Determination of Icariin in the Shenbao preparation with high-efficient liquid phase technique; C. measure the content of Osthole in the Shenbao preparation with high-efficient liquid phase technique; D. measure the content of schizandrin in the Shenbao preparation with high-efficient liquid phase technique.

(2) qualitative identification a. differentiates Radix Angelicae Sinensis and Ligusticum wallichii in the Shenbao preparation with thin-layered chromatography; B. differentiate frutus cnidii in the Shenbao preparation with thin-layered chromatography; C. differentiate prepared fleece flower root in the Shenbao preparation with thin-layered chromatography; D. differentiate barrenwort in the Shenbao preparation with thin-layered chromatography; E. differentiate Psoralea corylifolia in the Shenbao preparation with high-efficient liquid phase technique.

3. the method for quality control of a Shenbao preparation, contain in following assay and the qualitative identification one or more:

(1) assay

A. high-efficient liquid phase technique measures 2,3,5, the content of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Second eyeball-water (5-70: 95-30) be moving phase; UV-detector or diode array detector.Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates should be not less than 2000.

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside reference substance adds ethanol or Diluted Alcohol or other The suitable solvent and makes the solution of determining concentration, product solution in contrast.

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, with the methanol/ethanol extraction of suitable concentration, as need testing solution.

Accurate respectively reference substance solution and an amount of volume of need testing solution drawn of determination method injects liquid chromatograph, measures, promptly.

B. high-efficient liquid phase technique is measured icariin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Second eyeball-water (5-70: 95-30) be moving phase; UV-detector or diode array detector.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol or other The suitable solvent and makes the solution of determining concentration, product solution in contrast.

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, adds the suitable quantity of water dissolving, extract with ethyl acetate or other The suitable solvent, and the extract evaporate to dryness, residue dissolves with The suitable solvent, as need testing solution.

Accurate respectively absorption reference substance solution of determination method and need testing solution are an amount of, inject liquid chromatograph, measure promptly.

C. high-efficient liquid phase technique is measured Osthole content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Methanol-water (40-85: 60-15) be moving phase; UV-detector or diode array detector.Number of theoretical plate calculates by the Osthole peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol or other The suitable solvent and makes the solution of determining concentration, product solution in contrast.

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, with the methanol/ethanol extraction of suitable concentration, as need testing solution.

Accurate respectively absorption reference substance solution of determination method and need testing solution are an amount of, inject liquid chromatograph, measure promptly.

D. high-efficient liquid phase technique is measured schizandrin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Methanol-water (5-30: 2-15) be moving phase; UV-detector or diode array detector.Number of theoretical plate calculates by the schizandrin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol or other The suitable solvent and makes the solution of determining concentration, product solution in contrast.

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, with the methanol/ethanol extraction of suitable concentration, as need testing solution.

Accurate respectively absorption reference substance solution of determination method and need testing solution are an amount of, inject liquid chromatograph, measure promptly.

(2) qualitative identification

A. thin-layered chromatography is differentiated Radix Angelicae Sinensis and Ligusticum wallichii in the Shenbao preparation

Get product or the product content thing is an amount of, add diethyl ether or other The suitable solvent is extracted, extract is as need testing solution, or extract steams/volatilize, and residue adds The suitable solvent makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis and the Ligusticum wallichii control medicinal material is an amount of, makes control medicinal material solution after the extraction.Each is an amount of to draw above-mentioned need testing solution and control medicinal material solution, puts respectively on same silica G or silica gel H thin layer plate, with normal hexane-ethyl acetate (3-20: 0.3-2) be developping agent, 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 fluorescence spot of same color.

B. thin-layered chromatography is differentiated frutus cnidii in the Shenbao preparation

Get product or the product content thing is an amount of, add diethyl ether or other The suitable solvent is extracted, extract is as need testing solution, or extract steams/volatilize, and residue adds The suitable solvent makes dissolving, as need testing solution.It is an amount of that other gets the Osthole reference substance, and solubilizer is made reference substance solution in right amount.Each is an amount of to draw above-mentioned need testing solution and reference substance solution, puts respectively on same silica G or silica gel H thin layer plate, with cyclohexane-ethyl acetate (3-20: 0.5-5) be developping agent, launch, take out, dry, put under the ultraviolet lamp and inspect.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

C. thin-layered chromatography is differentiated prepared fleece flower root in the Shenbao preparation

Get product or the product content thing is an amount of, adding ethyl acetate or other The suitable solvent extracts, extract extracts with the sodium carbonate liquor of suitable concentration, sodium carbonate extract hydrochloric acid adjust pH to 1～4, with ethyl acetate or the extraction of other The suitable solvent, extract is as need testing solution, or extract steams/volatilizes again, residue adds The suitable solvent makes dissolving, as need testing solution.

It is an amount of that other gets fleece-flower root control medicinal material, makes control medicinal material solution after the extraction.It is an amount of to get the archen reference substance again, and solubilizer is made reference substance solution in right amount.Each is an amount of to draw above-mentioned need testing solution, control medicinal material and reference substance solution, put respectively on same silica G or silica gel H thin layer plate, (5-25: 2-8: upper solution 0.4-2) is a developping agent with sherwood oil (30～60 ℃)-ethyl formate-formic acid, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot.Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness.

D. thin-layered chromatography is differentiated barrenwort in the Shenbao preparation

Get product or the product content thing is an amount of, add normal butyl alcohol or other The suitable solvent and extract, extract washs with ammonia solution, discards ammonia solution, and extract is as need testing solution, or extract steams/volatilize, and residue adds The suitable solvent makes dissolving, as need testing solution.Other gets the icariin reference substance, and solubilizer is made reference substance solution in right amount.Each is an amount of to draw above-mentioned two kinds of solution, put respectively on same silica G or silica gel H thin layer plate, with ethyl acetate-butanone-formic acid-water (3-30: 0.4-4: 0.4-4: 0.4-4) be developping agent, launch, take out, dry, spray is heated to colour developing with 10% ethanol solution of sulfuric acid, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

E. high-efficient liquid phase technique is differentiated Psoralea corylifolia in the Shenbao preparation

Get product or the product content thing is an amount of, add methyl alcohol or other The suitable solvent and extract, extract is as need testing solution.Other gets psoralen and the Isopsoralen reference substance is an amount of, and solubilizer is made reference substance solution in right amount.With the octadecylsilane chemically bonded silica is filling agent; Methanol-water (20-80: 80-20) be moving phase, UV-detector or diode array detector.Accurate respectively reference substance solution and each an amount of volume of need testing solution drawn injects liquid chromatograph, measures.The retention time of main peak should be consistent with reference substance in the test sample chromatogram.

Described extraction can be adopted conventional methods such as sonicated, refluxing extraction, Microwave Extraction, vibration extraction;

2,3,5, the moving phase of the assay of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside can also be methanol acetonitrile-water (2-10: 0.5-2: 4-20), methanol-water (5-70: 95-30) or other suitable moving phase;

The moving phase that content Determination of Icariin is measured can also be methanol-water-glacial acetic acid (40-80: 20-60: 0.2-1), methanol-water (5-70: 95-30) or other suitable moving phase;

The moving phase of the assay of Osthole can also be methanol-water-tetrahydrofuran (30-75: 20-60: 2-10), acetonitrile-water (5-70: 95-30) or other suitable moving phase;

The moving phase of schizandrin assay can also be acetonitrile-water (5-70: 95-30) or other suitable moving phase;

The developping agent that the thin-layer chromatography of Radix Angelicae Sinensis and Ligusticum wallichii is differentiated can also be petroleum ether-ethyl acetate (5-25: 0.5-5), cyclohexane-ethyl acetate (5-25: 0.5-5) or other suitable developping agent;

The developping agent that the thin-layer chromatography of frutus cnidii is differentiated can also be benzene-ethyl acetate (10-50: 0.3-2), normal hexane-ethyl acetate (4-20: 0.4-4) or other suitable developping agent;

The developping agent that the thin-layer chromatography of prepared fleece flower root is differentiated can also be chloroform-methanol (5-15: 0.5-5), sherwood oil (30-60 ℃)-ethyl acetate-glacial acetic acid (50-150: 5-50: 0.5-5) or other suitable developping agent;

Both can adopt reference substance among qualitative identification a, b, c, the d, and also can adopt control medicinal material to compare, or both all adopt; Both can adopt two kinds of reference substances among the qualitative identification e, also can only adopt wherein a kind of reference substance in contrast.

4. the method for quality control of a Shenbao preparation, contain in following assay and the qualitative identification one or more:

(1) assay

A. high-efficient liquid phase technique measures 2,3,5, the content of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (25: 75) is moving phase; The detection wavelength is 320nm.Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates should be not less than 2000.

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside reference substance adds Diluted Alcohol and makes the solution that every 1ml contains 20ug, promptly.

Product is got in the preparation of need testing solution or the product content thing is an amount of, puts in the conical flask, and the accurate Diluted Alcohol 50ml that adds claims decide weight, and reflux 30 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and filtration is got subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

B. high-efficient liquid phase technique is measured icariin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (30: 70) is moving phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 40ug, promptly.

Product is got in the preparation of need testing solution or the product content thing is an amount of, adds water 10ml, puts in the tool plug separating funnel, uses ethyl acetate extraction 4 times, each 10ml, combined ethyl acetate layer, evaporate to dryness, residue add dissolve with methanol and are transferred in the 25m1 measuring bottle, add methyl alcohol to scale, shake up, promptly.

Accurate respectively reference substance solution and the need testing solution 20ml of drawing of determination method injects liquid chromatograph, measures promptly.

C. high-efficient liquid phase technique is measured Osthole content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (67: 33) is a moving phase; The detection wavelength is 249nm.Number of theoretical plate calculates by the Osthole peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution that every 1ml contains 50ug, promptly.

Product is got in the preparation of need testing solution or the product content thing is an amount of, puts in the tool plug conical flask, the accurate ethanol 50ml that adds, and close plug claims to decide weight, and sonicated 1 hour claims to decide weight again, supplies the weight that subtracts mistake with ethanol, shakes up, and filters, and gets subsequent filtrate, promptly.

Accurate respectively reference substance solution and the need testing solution 20ml of drawing of determination method injects liquid chromatograph, measures, promptly.

D. high-efficient liquid phase technique is measured schizandrin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (13: 7) is a moving phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schizandrin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 30ug, promptly.

Product is got in the preparation of need testing solution or the product content thing is an amount of, puts in the tool plug conical flask, the accurate methyl alcohol 50ml that adds, and close plug claims to decide weight, and sonicated 20 minutes claims to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and gets subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

(2) qualitative identification

A. thin-layered chromatography is differentiated Radix Angelicae Sinensis and Ligusticum wallichii in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis and each 0.5g of Ligusticum wallichii control medicinal material, shines medicinal material solution in pairs with legal system.Drawing above-mentioned need testing solution and control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-ethyl acetate (9: 1), launches, and takes out, and dries, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

B. thin-layered chromatography is differentiated frutus cnidii in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Get the Osthole reference substance, add ethanol and make the solution that every 1ml contains 1mg, in contrast product solution.Drawing each 1 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (7: 2), launches, and takes out, and dries, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

C. thin-layered chromatography is differentiated prepared fleece flower root in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Add ethyl acetate 30ml, reflux 1 hour is put cold, filter, filtrate is extracted 3 times with 5% sodium carbonate liquor jolting, and each 20ml merges the sodium carbonate extract, with hydrochloric acid adjust pH to 2～3, extract 2 times each 30ml, combined ethyl acetate extract again with the ethyl acetate jolting, be concentrated into 1ml, as need testing solution.Other gets fleece-flower root control medicinal material 0.3g, shines medicinal material solution in pairs with legal system.Get the archen reference substance again, add methyl alcohol and make the solution that every 1ml contains 0.2mg, in contrast product solution.Draw above-mentioned need testing solution 10 μ l, each 2 μ l of control medicinal material and reference substance solution, put respectively on same silica gel g thin-layer plate, upper solution with sherwood oil (30～60 ℃)-ethyl formate-formic acid (15: 5: 1) is a developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot.Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness.

D. thin-layered chromatography is differentiated barrenwort in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Add normal butyl alcohol 30ml, ultrasonic Extraction 30 minutes filters, and filtrate adds ammonia solution washing 2 times, and each 30ml discards ammonia solution, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, 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, (10: 1: 1: 1) be developping agent, expansion was taken out with ethyl acetate-butanone-formic acid-water, dry, spray is put 105 ℃ of bakings several minutes, in the test sample chromatogram with 10% ethanol solution of sulfuric acid, with the corresponding position of reference substance chromatogram on, show the spot of same color.

E. high-efficient liquid phase technique is differentiated Psoralea corylifolia in the Shenbao preparation

Get product or the product content thing is an amount of, add methyl alcohol 40ml, ultrasonic Extraction 30 minutes is put coldly, filters, and filtrate is as need testing solution.Other gets psoralen and the Isopsoralen reference substance is an amount of, adds methyl alcohol and makes the mixed solution that every 1ml contains 4 μ g, in contrast product solution.Measure according to high performance liquid chromatography (appendix ID of Chinese Pharmacopoeia version in 2005).With the octadecylsilane chemically bonded silica is filling agent; Methanol-water (40: 60) is a moving phase, and the detection wavelength is 246nm.Accurate respectively reference substance solution and each 20ul of need testing solution of drawing injects liquid chromatograph, measures.The retention time of main peak should be consistent with reference substance in the test sample chromatogram.

The amount of described extraction sample or dissolving extract solvent for use, ultrasonic or time, constant volume or dissolving back volume, point sample amount or the sample size of refluxing extraction, the concentration of reference substance can be that benchmark changes with described occurrence, or change in proportion.

Both can adopt reference substance among described qualitative identification a, b, c, the d, and also can adopt control medicinal material to compare, or both all adopt, both can adopt two kinds of reference substances among the qualitative identification e, also can only adopt wherein a kind of reference substance in contrast.

Described sonicated also can adopt conventional extracting method such as refluxing extraction, Microwave Extraction, vibration extraction, and described refluxing extraction also can adopt conventional extracting method such as sonicated, Microwave Extraction, vibration extraction.

5. above-mentioned 2,3,4 described methods, can be used to detect any formulation of the Shenbao preparation that makes by above-mentioned 1 described preparation method or make by other preparation method, as tablet, sugar coated tablet, film coated tablet, chewable tablet, effervescent tablet, Disket, capsule, hard capsule, soft capsule, pellt capsule, oral liquid, syrup, suck agent, electuary, granule, effervescent, pill, pill, micropill preparation.

6. in the Shenbao preparation 2,3,5, the content limit of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside is: contain prepared fleece flower root with 2,3,5 in the once minimum dose, 4 '-tetrahydroxystilbene-2-O-b-D-glucoside (C ₂₀H ₂₂O ₉) meter, must not be less than 1.3mg; The content Determination of Icariin limit is: contain barrenwort with icariin (C in the once minimum dose ₃₃H ₄₀O ₁₅) meter, must not be less than 1.5mg; The content limit of Osthole is: contain frutus cnidii with Osthole (C in the once minimum dose ₁₅H ₁₆O ₃) meter, must not be less than 0.8mg; The content limit of schizandrin is: contain the fruit of Chinese magnoliavine with schizandrin (C in the once minimum dose ₂₄H ₃₂O ₇) meter, must not be less than 0.4mg; Above-mentioned content limit is the minimum content limit, can improve content limit according to the concrete condition of producing.

Experimental example 1: Shenbao preparation Study on Preparation

(1) red ginseng extraction process

With dried cream yield and ginsenoside Re+Rg ₁Extracted amount is an evaluation index, and the red ginseng reflux technique is investigated, and gets three parts of red ginseng meal 100g, adds 20% alcohol dipping after 8 hours, add 6 times, 9 times, 12 times solvents respectively, the refluxing extraction secondary each 2 hours, merges extract, filter filtrate recycling ethanol, concentrating under reduced pressure, vacuum drying.The result adds 6 times of amount solvents, and ginsenoside Re+Rg1 extracted amount, dried cream yield are respectively 766.37mg, 54.76%, adds 9 times of amount solvents and is respectively 957.22mg, 60.18%, adds 12 times of amount solvents and is respectively 960.38mg, 65.34%.So can think and can put forward most effective constituent when adding 9 times of amounts of solvent, increase extraction time and solvent amount again, can only increase the proposition of invalid components such as starch.From energy savings, reduce production costs and reduce dose and consider, preferred red ginseng refluxing extraction technology is: red ginseng powder is broken into meal, adds 9 times of amount 20% alcohol dipping after 8 hours, refluxing extraction secondary, each 2 hours.

(2) barrenwort, Radix Angelicae Sinensis, frutus cnidii, Ligusticum wallichii, fennel seeds five kinds of Chinese medicine diacolation technology are investigated

With dried cream yield, icariin extracted amount is evaluation index, and the diacolation technology of five kinds of Chinese medicine such as barrenwort is investigated, and takes by weighing three parts of five tastes 212.1g such as barrenwort by the amount of the precious syrup prescription of archinephridium, be ground into meal respectively, make solvent, flood after 48 hours with 70% ethanol, with the speed of per minute 1～3ml diacolation slowly, by testing program, collect percolate 1000ml (4.7 times of amounts), 1500ml (7 times of amounts), 2000ml (9.4 times of amounts) respectively, reclaim ethanol, concentrating under reduced pressure, vacuum drying.Collection percolate 1000ml icariin extracted amount, dried cream yield are respectively 598mg, 7.34%, and percolate 1500ml is respectively 650mg, 9.29%, collect percolate 2000ml and are respectively 667mg, 10.27%.So can think that percolate 1500ml can put forward most effective constituent, increase extraction time and solvent amount again, can only increase the proposition of invalid components such as starch.From energy savings, reduce production costs and reduce dose and consider, 5 flavor diacolation extraction processes such as preferred barrenwort are that ground spice such as barrenwort is broken into meal, make solvent with 70% ethanol, flood after 48 hours, with the speed of per minute 1～3ml diacolation slowly, collect 7 times of amounts of percolate.

(3) ten Six-element liquid medicine such as prepared fleece flower root are put forward technology

With dried cream yield, the Stibene-glucoside extracted amount is an evaluation index, ten Six-element liquid medicine such as prepared fleece flower root are put forward technology to be investigated, take by weighing 16 flavors such as prepared fleece flower root by 1/2 amount of the precious syrup prescription of archinephridium and add red ginseng through the dregs of a decoction of alcohol extracting three parts of 413.15g altogether, by testing program, the water that adds different amounts respectively, soaked 1 hour, and decocted secondary, each 2 hours, collecting decoction, filter, the filtrate filtrate decompression is concentrated into the clear cream that relative density is 1.16 (80 ℃ of heat are surveyed), adds ethanol and makes that to contain the alcohol amount be 65%, left standstill 48 hours, get the supernatant decompression recycling ethanol, concentrating under reduced pressure, vacuum drying.The result adds 8 times of water gaging Stibene-glucoside extracted amounts, dried cream yield is respectively 652mg, 12.46%, add 10 times of water gaging Stibene-glucoside extracted amounts, dried cream yield is respectively 725mg, 14.13%, add 12 times of water gaging Stibene-glucoside extracted amounts, dried cream yield is respectively 729mg, 15.62%, so can think when amount of water is 10 times, can put forward most effective constituent substantially, increase the solvent amount again, can only increase the proposition of invalid components such as starch, mucilaginous substance.From energy savings, reduce cost and reduce dose and consider, 16 flavor extraction processes such as preferred prepared fleece flower root are: 16 flavors such as prepared fleece flower root add the dregs of a decoction of red ginseng through alcohol extracting, adding 10 times of water gagings soaked 1 hour, decoct secondary, each 2 hours, collecting decoction, filter, the filtrate filtrate decompression is concentrated into relative density the be 1.16 clear cream of (80 ℃ of heat are surveyed), adds ethanol and makes that to contain the alcohol amount be 65%, leaves standstill 48 hours.

(4) drying process

The prescription Chinese crude drug is evaporated to thick paste through extracting, and when carrying out drying, is subjected to heat damage for a long time for preventing effective constituent, can adopt drying means such as vacuum drying, spray drying or freeze drying according to the situation of actual production equipment.With the contained active component with strong kidney effect 2 in contained active component icariin of barrenwort and the RADIX POLYGONI MULTIFLORI PREPARATA, 3,5, the extracted amount of 4-tetrahydroxystilbene-2-O-b-D-glucoside is an evaluation index, influence to vacuum drying centering pharmaceutically active ingredient is investigated, and finds that vacuum/spray drying front and back icariin extracted amount, Stibene-glucoside extracted amount all do not have significant difference.

(5) moulding process

The dried cream powder of gained is made various solid pharmaceutical preparations according to a conventional method, as tablet, sugar coated tablet, film coated tablet, chewable tablet, effervescent tablet, Disket, capsule, hard capsule, soft capsule, pellt capsule, suck agent, electuary, granule, effervescent, pill, pill, micropill preparation etc.Following Study on Forming is an example with tablet, chewable tablets, granule, capsule.

A. tablet

This product dried cream powder viscosity is stronger, test is the effect of 75% ethanol, 85 ethanol % and 95% alcohol granulation relatively, get medicinal extract powder 50g, add 15g starch, mixing, spray into above-mentioned wetting agent, phenomenon occurs cohering when 75% ethanol, 85% alcohol granulation as a result, 95% alcohol granulation effect is better, and preparation easily, grain color is even, and is easily dry.

Test intended is selected microcrystalline cellulose (MCC), L-HPC (HPC), pregelatinized starch (Pre-starch), superfine silica gel powder (SiO for use ₂), dolomol (Mg-st) is auxiliary material, test method: dried cream powder mixes with auxiliary material (except the dolomol), adds ethanol system particle, drying adds dolomol, mixing, compressing tablet, experimental design and the results are shown in Table 1.

Table 1: compression molding craft screening and result

By table 1 as seen, the 1 auxiliary material ratio of writing out a prescription accounts for 20%, and hardness is medium, because added auxiliary material space is little, test is found superfine silica gel powder to the hardness influence better, and helps preparation of granules, and is heavy for reducing sheet, and suitable compressing tablet and subsequent packet film-coating needs, select for use prescription 3 for determining prescription, granulate easily, the tablet appearance is bright and clean, the hardness height, disintegration is fast.

B. chewable tablets

In the precious extract of kidney heavier bitter taste is arranged, only add flavourings such as sucrose, aspartame, Stevioside, mannitol and be difficult to shelter its bitter taste, so attempt adopting in wet medicinal extract and add an amount of betadex, dry behind the mixing, add sweetener etc. again and carry out the method for flavoring, the result shows that this method is respond well.The amount of the betadex that adds, the amount that adds aspartame are carried out preferably, when the result adds 30% betadex, 1% aspartame, taste good.

Because dried cream powder viscosity is stronger, so during granulation, do not need to add in addition again bonding agent.Being wetting agent with 95% ethanol, 90% ethanol, 80% ethanol respectively, granulating, is evaluation index with granulation difficulty or ease, fine powder amount, and wetting agent is investigated, and the result is better with 90% alcohol granulation effect, is wetting agent so adopt 90% ethanol.

C. granule

A. wet granulation is got dried cream powder 200g, adds an amount of 90% ethanol, mixes, and the system softwood is crossed 14 mesh sieves and granulated.60 ℃ of dryings of wet granular, particle is with the whole grain of 14 mesh sieves, and fine grained and fine powder go with 60 mesh sieves sieve, promptly.

B. dry granulation adopts the dry granulation machine to granulate, and control workshop relative humidity is below 50%.

Two kinds of methods all can be granulated smoothly, and water cut, melting are all qualified, but dry granulation does not need guiding humid medium (ethanol), does not need heat drying, and technological process is short, and it is low to consume energy, and equipment is few, so dry granulation is better.

D. capsule

This product dried cream powder flowability is poor slightly, improve its flowability so add an amount of superfine silica gel powder, experiment shows that the superfine silica gel powder of adding 1% can effectively improve its flowability, control workshop relative humidity is below 50%, carry out the capsule trial assembly, add the dried cream powder can smoothly of 1% superfine silica gel powder, the capsule of being adorned is carried out inspections such as content uniformity, moisture, the result is all up to specification.

Experimental example 2: Shenbao preparation assay research

(1) in the Shenbao preparation 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glycoside assay

A. the chromatographic condition octadecylsilane chemically bonded silica is a filling agent; Second eyeball-water (25: 75) is moving phase; The detection wavelength is 320nm.Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates should be not less than 2000.

B. the preparation precision of the preparation reference substance solution of reference substance, test sample, negative control product solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside reference substance adds Diluted Alcohol and makes the solution that every 1ml contains 20ug, promptly.

The precious agreement that contracts a film or TV play to an actor or actress 0.15g of kidney is got in the preparation of need testing solution respectively, the kidney treasured is chewed agreement that contracts a film or TV play to an actor or actress 0.3g, the about 1.5g of granule for strengthening kidney, the about 0.15g of kidney capsule content, and accurate the title decides, and puts in the conical flask respectively, the accurate Diluted Alcohol 50ml that adds, claim to decide weight, reflux 30 minutes is put cold, claim to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.

Scarce prepared fleece flower root negative control 0.15g is got in the preparation that lacks the prepared fleece flower root negative control solution, operates with method according to " preparation of need testing solution ", promptly gets to lack the prepared fleece flower root negative control solution.

C. measure accurate respectively reference substance solution, need testing solution, scarce each 20ml of prepared fleece flower root negative control solution of drawing, inject liquid chromatograph, measure, promptly.

As seen in the test sample 2,3,5, degree of separation＞1.5 of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside, 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside reaches better with other component and separates, and feminine gender does not have interference, and interference component is not measured.Theoretical cam curve is with 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates and is not less than 2000.Moving phase employing methyl alcohol-acetonitrile-water (5: 1: 9) should.

D. this method is carried out methodological study, as a result 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside becomes the good linear relation in 15 μ g/ml～120 μ g/ml scopes, and the recovery is 98.99%, and precision, repeatability are all good.

(2) icariin Determination on content in the Shenbao preparation

A. the chromatographic condition octadecylsilane chemically bonded silica is a filling agent; Second eyeball-water (30: 70) is moving phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

B. the preparation of reference substance, test sample, negative control product solution

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 40ug, promptly.

The precious agreement that contracts a film or TV play to an actor or actress 0.25g of kidney is got in the preparation of need testing solution respectively, the kidney treasured is chewed agreement that contracts a film or TV play to an actor or actress 0.3g, the about 1.5g of granule for strengthening kidney, the about 0.25g of kidney capsule content, the accurate title, decide, and adds water 10ml respectively, puts in the tool plug separating funnel, with ethyl acetate extraction 4 times, each 10ml, combined ethyl acetate layer, evaporate to dryness, residue adds dissolve with methanol and is transferred in the 25m1 measuring bottle, add methyl alcohol to scale, shake up, promptly.

Scarce barrenwort negative control 0.18g is got in the preparation that lacks the barrenwort negative control solution, operates with method according to " preparation of need testing solution ", promptly gets to lack the negative negative control solution of barrenwort.

C. measure: get reference substance solution, need testing solution, lack barrenwort negative control solution and each 20 μ l of blank solution (moving phase), inject liquid chromatograph, measure.

As seen the degree of separation of icariin＞1.5 in the test sample, icariin reaches better with other component and separates, and feminine gender does not have interference, interference component is measured.Theoretical cam curve is calculated greater than 2000 with the icariin peak.Moving phase adopts methanol-water-glacial acetic acid (60: 40: 0.5) also can.

D. this method is carried out methodological study, icariin becomes the good linear relation in 20 μ g/ml～80 μ g/ml scopes as a result, and the recovery is 99.98%, and precision, repeatability are all good.Show that this law can be used for measuring the content Determination of Icariin of Shenbao preparation.

(3) Osthole Determination on content in the Shenbao preparation

A. the test of chromatographic condition and system suitability is a filling agent with octadecylsilane chemically bonded silica; Methanol-water (67: 33) is a moving phase; The detection wavelength is 249nm.Number of theoretical plate calculates by the Osthole peak should be not less than 2000.

B. the preparation of reference substance, test sample, negative control product solution

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution that every 1ml contains 50ug, promptly.

The precious agreement that contracts a film or TV play to an actor or actress 0.25g of kidney is got in the preparation of need testing solution respectively, the kidney treasured is chewed agreement that contracts a film or TV play to an actor or actress 0.3g, the about 1.5g of granule for strengthening kidney, the about 0.25g of kidney capsule content, the accurate title, decide, and puts respectively in the tool plug conical flask, the accurate ethanol 50ml that adds, close plug, claim to decide weight, sonicated 1 hour claims to decide weight, supply the weight that subtracts mistake with ethanol, shake up, filter, get subsequent filtrate promptly.

Scarce frutus cnidii negative control 0.25g is got in the preparation that lacks the frutus cnidii negative control solution, operates with method according to " preparation of need testing solution ", promptly gets to lack the negative negative control solution of frutus cnidii.

C. measure accurate respectively draw reference substance solution and need testing solution 20ml, inject liquid chromatograph, measure, promptly.

As seen the degree of separation of Osthole＞1.5 in the test sample, Osthole reaches better with other component and separates, and feminine gender does not have interference, interference component is measured.Theoretical cam curve is calculated greater than 2000 with the Osthole peak.Moving phase adopts methanol-water (67: 33), methanol-water-tetrahydrofuran (53: 42: 5) also can.

D. this method is carried out methodological study, Osthole becomes the good linear relation in 10 μ g/ml～100 μ g/ml scopes as a result, and the recovery is 99.35%, and precision, repeatability are all good.Show that this law can be used for measuring the content of the Osthole of Shenbao preparation.

(4) schizandrin Determination on content in the Shenbao preparation

A. the chromatographic condition octadecylsilane chemically bonded silica is a filling agent; Methanol-water (13: 7) is a moving phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schizandrin peak should be not less than 2000.

B. the preparation of reference substance, test sample, negative control product solution

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 30ug.

The precious agreement that contracts a film or TV play to an actor or actress 0.3g of kidney is got in the preparation of need testing solution respectively, the kidney treasured is chewed agreement that contracts a film or TV play to an actor or actress 0.5g, the about 2.0g of granule for strengthening kidney, the about 0.3g of kidney capsule content, and accurate the title decides, and puts respectively in the tool plug conical flask, the accurate methyl alcohol 50ml that adds, close plug claims to decide weight, sonicated 20 minutes, claim to decide weight again, supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate, promptly.

Scarce fruit of Chinese magnoliavine negative control 0.25g is got in the preparation that lacks fruit of Chinese magnoliavine negative control solution, operates with method according to " preparation of need testing solution ", promptly gets to lack the negative negative control solution of the fruit of Chinese magnoliavine.

C. measure accurate respectively draw reference substance solution and need testing solution 20ml, inject liquid chromatograph, measure, promptly.

As seen the degree of separation of schizandrin＞1.5 in the test sample, schizandrin reaches better with other component and separates, and feminine gender does not have interference, interference component is measured.Theoretical cam curve is calculated greater than 2000 with the schizandrin peak.Moving phase adopts methanol-water (13: 7) also can.

D. this method is carried out methodological study, schizandrin becomes the good linear relation in 10 μ g/ml～80 μ g/ml scopes as a result, and the recovery is 99.52%, and precision, repeatability are all good.Show that this law can be used for measuring the content of the Osthole of Shenbao preparation.

Experimental example 3: Shenbao preparation qualitative identification research

(1) thin-layer chromatography of Radix Angelicae Sinensis and Ligusticum wallichii is differentiated, with reference to " discriminating of Ligusticum wallichii under item of Chinese pharmacopoeia version in 2000. because of the spot basically identical of Radix Angelicae Sinensis and Ligusticum wallichii control medicinal material, so this experiment is contrast with Radix Angelicae Sinensis and two kinds of control medicinal materials of Ligusticum wallichii, negative sample also adopts and lacks Radix Angelicae Sinensis, Ligusticum wallichii two flavors simultaneously.With normal hexane-ethyl acetate (9: 1) is developping agent, and the Rf value is suitable as a result, and clear spot lacks Radix Angelicae Sinensis, the Ligusticum wallichii sample is noiseless.Developping agent adopts sherwood oil (60-90 ℃)-ethyl acetate (9: 2), cyclohexane-ethyl acetate (17: 3) also can.

(2) thin-layer chromatography of frutus cnidii is differentiated, this experiment is to be reference substance with the Osthole, and cyclohexane-ethyl acetate (7: 2) is developping agent, and the Rf value is suitable as a result, clear spot, and it is noiseless to lack the frutus cnidii sample.Developping agent adopts normal hexane-ethyl acetate (85: 15), benzene-ethyl acetate (30: 1) also can.

(3) thin-layer chromatography of prepared fleece flower root is differentiated, ethyl acetate extraction is used earlier in the test sample preparation, extract with 5% sodium carbonate liquor again, composition such as archen is dissolved in the alkali lye, again the acidifying ethyl acetate extraction, compare medicinal material with the fleece-flower root, archen compares product, and sherwood oil (30～60 ℃)-ethyl formate-formic acid (15: 5: 1) is developping agent, as a result the chromatogram clear spot, the Rf value is suitable, and it is noiseless to lack the prepared fleece flower root sample.Developping agent adopts chloroform-methanol (8: 2), sherwood oil (30～60 ℃)-ethyl acetate-glacial acetic acid (9: 1: 0.1) also can.

(4) be the thin-layer chromatography discriminating of barrenwort, with reference to the method under the black positive bushing syrup of National Drug Administration of People's Republic of China (PRC) standard (trying) [WS-10585 (ZD-0585) 2002] the discriminating item.Because of the prescription in the barrenwort amount more, after test adopts normal butyl alcohol to extract, use the need testing solution preparation method of alkali liquid washing again, with the icariin is reference substance, (10: 1: 1: 1) be developping agent, be developer with 10% ethanol solution of sulfuric acid to ethyl acetate-butanone-formic acid-water, as a result the chromatogram clear spot, the Rf value is suitable, and it is noiseless to lack the barrenwort sample.

(5) be the liquid chromatography discriminating of Psoralea corylifolia.With reference to the method under National Drug Administration of People's Republic of China (PRC) standard (trying) tonifying kidneys and nourishing brain ball [WS-10555 (ZD-0555) 2002] the discriminating item.The mixing reference substance solution of getting psoralen and Isopsoralen earlier is in 200-400nm place scanning ultra-violet absorption spectrum, and the result is presented at 246nm wavelength place absorption maximum, so select 246nm for measuring wavelength.Three batch samples sample introduction after methyl alcohol is ultrasonic, in the test sample chromatogram, with mix the corresponding position of reference substance chromatographic peak all arranged, lack the Psoralea corylifolia negative sample with the method preparation, in the negative sample chromatogram,, illustrate that negative sample is noiseless to it not having chromatographic peak with the corresponding position of reference substance.Moving phase adopts methanol-water (60: 40) also can.

Embodiment 4: the comparison of active constituent content in the Shenbao preparation (is example with SHENBAO HEJI, the precious sheet of kidney, the precious chewable tablets of kidney, granule for strengthening kidney)

(1) 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside content relatively

Relatively in the Shenbao preparation 2,3,5, the content of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside, the result is as follows: SHENBAO HEJI (mg/10ml) 1.3,2.1,6.4; The precious sheet of kidney (mg/ sheet) 5.07,5.11,5.09; The precious chewable tablets of kidney (mg/ sheet) 7.26,7.32,7.21; Granule for strengthening kidney (mg/ bag) 6.39,6.51,6.61; Kidney capsule (mg/ grain) 3.82,3.87,3.91.The amount that 3 of the precious sheets of SHENBAO HEJI 10ml and kidney, 2 of the precious chewable tablets of kidney, granule for strengthening kidney 1 bag, kidney capsule are 4 is suitable.Find in the SHENBAO GUTI ZHIJI 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside content is higher, shows that SHENBAO GUTI ZHIJI has bigger advantage at aspects such as preparation stabilities.

(2) icariin content relatively

Compare content Determination of Icariin in the Shenbao preparation, the result is as follows: SHENBAO HEJI (mg/10ml) 1.0,4.0,3.6; The precious sheet of kidney (mg/ sheet) 2.02,2.09,2.06; The precious chewable tablets of kidney (mg/ sheet) 3.31,3.17,3.10; Granule for strengthening kidney (mg/ bag) 4.52,4.12,4.37; Kidney capsule (mg/ grain) 1.55,1.58,1.62.The amount that 3 of the precious sheets of SHENBAO HEJI 10ml and kidney, 2 of the precious chewable tablets of kidney, granule for strengthening kidney 1 bag, kidney capsule are 4 is suitable.Find that icariin content is higher in the SHENBAO GUTI ZHIJI, show that SHENBAO GUTI ZHIJI has bigger advantage at aspects such as preparation stabilities.

(3) Osthole content relatively

Compare the content of Osthole in the Shenbao preparation, the result is as follows: SHENBAO HEJI (mg/10ml) 0.8,2.4,1.9; The precious sheet of kidney (mg/ sheet) 0.92,1.05,1.06; The precious chewable tablets of kidney (mg/ sheet) 1.51,1.44,1.60; Granule for strengthening kidney (mg/ bag) 3.14,3.26,3.10; Kidney capsule (mg/ grain) 0.86,0.92,0.95.The amount that 3 of the precious sheets of SHENBAO HEJI 10ml and kidney, 2 of the precious chewable tablets of kidney, granule for strengthening kidney 1 bag, kidney capsule are 4 is suitable.Find that Osthole content is higher in the SHENBAO GUTI ZHIJI, show that SHENBAO GUTI ZHIJI has bigger advantage at aspects such as preparation stabilities.

(4) schizandrin content relatively

Compare the content of schizandrin in the Shenbao preparation, the result is as follows: SHENBAO HEJI (mg/10ml) 0.64,0.45,1.32; The precious sheet of kidney (mg/ sheet) 0.78,0.62,0.83; The precious chewable tablets of kidney (mg/ sheet) 1.07,1.02,0.94; Granule for strengthening kidney (mg/ bag) 2.01,2.14,2.07; Kidney capsule (mg/ grain) 0.47,0.41,0.55.The amount that 3 of the precious sheets of SHENBAO HEJI 10ml and kidney, 2 of the precious chewable tablets of kidney, granule for strengthening kidney 1 bag, kidney capsule are 4 is suitable.Find that schizandrin content is higher in the SHENBAO GUTI ZHIJI, show that SHENBAO GUTI ZHIJI has bigger advantage at aspects such as preparation stabilities.

Experimental example 5: content limit determines in the Shenbao preparation assay

(1) 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside content limit is determined

Contain the about 0.744g of prepared fleece flower root medicinal material in the once minimum dose of Shenbao preparation, in a regulation of Chinese Pharmacopoeia version in 2005 the prepared fleece flower root medicinal material 2,3,5, the content of 4 '-tetrahydroxystilbene-2-O-b-D-glucoside must not be lower than 0.7%.With in the prepared fleece flower root medicinal material 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside content is 0.7%, when the rate of transform is 35%, contains 2,3,5 in the once minimum dose, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside is 1.82mg.In conjunction with in the SHENBAO HEJI 2,3,5, in 4 '-tetrahydroxystilbene-2-O-b-D-glucoside content and the precious sheet of the kidney of being developed, the precious chewable tablets of kidney, granule for strengthening kidney, the kidney capsule 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside the rate of transform is considered the factors such as loss in the big production link, with this product 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside content limit is decided to be: contain prepared fleece flower root with 2,3 in the once minimum dose, 5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside (C ₂₀H ₂₂O ₉) meter, must not be less than 1.3mg.

(2) the barrenwort content limit is determined

Contain the about 0.946g of epimedium herb in the once minimum dose of Shenbao preparation, content Determination of Icariin must not be lower than 0.5% in regulation epimedium herb of Chinese Pharmacopoeia version in 2005.In barrenwort content in the epimedium herb is 0.5%, and when the rate of transform was 35%, containing icariin in the once minimum dose was 1.6mg.In conjunction with the icariin rate of transform in the precious sheet of icariin content in the SHENBAO HEJI and the kidney of being developed, the precious chewable tablets of kidney, granule for strengthening kidney, the kidney capsule, consider the factors such as loss in the big production link, this product icariin content limit is decided to be: contain barrenwort with icariin (C in the once minimum dose ₃₃H ₄₀O ₁₅) meter, must not be less than 1.4mg.

(3) the Osthole content limit is determined

Contain the about 0.28g of frutus cnidii medicinal material in the once minimum dose of Shenbao preparation, the content of Osthole must not be lower than 1.0% in a regulation frutus cnidii of Chinese Pharmacopoeia version in 2005 medicinal material.In Osthole content in the frutus cnidii medicinal material is 1.0%, and when the rate of transform was 35%, containing Osthole in the once minimum dose was 0.98mg.In conjunction with the Osthole rate of transform in the precious sheet of Osthole content in the SHENBAO HEJI and the kidney of being developed, the precious chewable tablets of kidney, granule for strengthening kidney, the kidney capsule, consider the factors such as loss in the big production link, this product Osthole content limit is decided to be: contain frutus cnidii with Osthole (C in the once minimum dose ₁₅H ₁₆O ₃) meter, must not be less than 0.8mg.

(4) the schizandrin content limit is determined

Contain the about 0.36g of schisandra chinensis medicinal material in the once minimum dose of Shenbao preparation, the content of schizandrin must not be lower than 0.40% in 2005 editions regulation schisandra chinensis medicinal materials of Chinese Pharmacopoeia.In schizandrin content in the schisandra chinensis medicinal material is 0.40%, and when the rate of transform was 35%, containing schizandrin in the once minimum dose was 0.504mg.In conjunction with the schizandrin rate of transform in the precious sheet of schizandrin content in the SHENBAO HEJI and the kidney of being developed, the precious chewable tablets of kidney, granule for strengthening kidney, the kidney capsule, consider the factors such as loss in the big production link, this product schizandrin content limit is decided to be: contain the fruit of Chinese magnoliavine with schizandrin (C in the once minimum dose ₂₄H ₃₂O ₇) meter, must not be less than 0.4mg.

Embodiment

Further specify the present invention by the following examples, but do not limit range of application of the present invention therefrom.

Embodiment 1: the preparation of tablet

Frutus cnidii 93.3g Ligusticum wallichii 94.3g seed of Chinese dodder 220g

Psoralea corylifolia 95g Poria cocos 100g red ginseng 66.7g

Fennel seeds 48g fruit of Chinese magnoliavine 120g fruit of Cherokee rose 315.3g

Bighead atractylodes rhizome 47.3g Radix Angelicae Sinensis 156g raspberry 109.7g

Prepared fleece flower root 248g plantain seed 55g prepared rhizome of rehmannia 313.3g

Fruit of Chinese wolfberry 220g Chinese yam 154.3g barrenwort 315.3g

Fenugreek 313.3g Radix Astragali 171.3g saline cistanche 157.7g

Honey-fried licorice root 47.3g

More than 22 the flavor, frutus cnidii, barrenwort, Radix Angelicae Sinensis, Ligusticum wallichii, fennel seeds are ground into meal, make solvent with 70% ethanol, flood after 48 hours,, collect the about 5000ml of percolate with the speed of per minute 1～3ml diacolation slowly, reclaim ethanol, being evaporated to relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed), filters filtrate for later use; Red ginseng powder is broken into meal, adds 9 times of amount 20% alcohol dipping after 8 hours, the refluxing extraction secondary, and each 2 hours, merge extract, filter, filtrate recycling ethanol is concentrated into relative density and is the clear cream of 1.10 (80 ℃ of heat are surveyed), and is standby; Ten Six-elements such as all the other raspberries with the dregs of a decoction of red ginseng through alcohol extracting, add 10 times of water gagings and soaked 1 hour, decoct secondary, each 2 hours, collecting decoction, filter, filtrate is concentrated into the clear cream that relative concentration is 1.16 (80 ℃ of heat are surveyed), adds ethanol and makes that to contain the alcohol amount be 65%, left standstill 48 hours, and got supernatant and reclaim ethanol, being concentrated into relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed), add extract and red ginseng extracts such as above-mentioned frutus cnidii, mixing, vacuum drying, pulverize, add microcrystalline cellulose 51g, superfine silica gel powder 6g, mixing is made particle, dry, add dolomol 2.5g, mixing is pressed into 1000, dressing promptly gets the precious sheet of kidney.

Embodiment 2: the preparation of chewable tablet

Frutus cnidii 140g Ligusticum wallichii 141.5g seed of Chinese dodder 330g

Psoralea corylifolia 142.5g Poria cocos 150g red ginseng 100g

Fennel seeds 72g fruit of Chinese magnoliavine 180g fruit of Cherokee rose 473g

Bighead atractylodes rhizome 71g Radix Angelicae Sinensis 234g raspberry 164.5g

Prepared fleece flower root 372g plantain seed 82.5g prepared rhizome of rehmannia 470g

Fruit of Chinese wolfberry 330g Chinese yam 231.5g barrenwort 473g

Fenugreek 470g Radix Astragali 257g saline cistanche 236.5g

Honey-fried licorice root 71g

More than 22 the flavor, frutus cnidii, barrenwort, Radix Angelicae Sinensis, Ligusticum wallichii, fennel seeds are ground into meal, make solvent with 70% ethanol, flood after 48 hours,, collect the about 7500ml of percolate with the speed of per minute 1～3ml diacolation slowly, reclaim ethanol, being evaporated to relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed), filters filtrate for later use; Red ginseng powder is broken into meal, adds 9 times of amount 20% alcohol dipping after 8 hours, the refluxing extraction secondary, and each 2 hours, merge extract, filter, filtrate recycling ethanol is concentrated into relative density and is the clear cream of 1.10 (80 ℃ of heat are surveyed), and is standby; Ten Six-elements such as all the other raspberries with the dregs of a decoction of red ginseng through alcohol extracting, add 10 times of water gagings and soaked 1 hour, decoct secondary, each 2 hours, collecting decoction, filter, filtrate is concentrated into relative concentration the be 1.16 clear cream of (80 ℃ of heat are surveyed), adds ethanol and makes that to contain the alcohol amount be 65%, leaves standstill 48 hours, get supernatant and reclaim ethanol, being concentrated into relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed), adds extract and red ginseng extracts such as above-mentioned frutus cnidii, mixing, add betadex 318g, mixing, vacuum drying is pulverized, add aspartame 10g, mixing is made particle, drying in right amount with 90% ethanol; Add dolomol 3g, mixing is pressed into 1000, and the bag film-coating promptly gets the precious chewable tablets of kidney.

Embodiment 3: the preparation of granule

Frutus cnidii 93.3g Ligusticum wallichii 94.3g seed of Chinese dodder 220g

Psoralea corylifolia 95g Poria cocos 100g red ginseng 66.7g

The fennel seeds 48g 120g fruit of Cherokee rose 315.3g that distinguishes the flavor of

Bighead atractylodes rhizome 47.3g Radix Angelicae Sinensis 156g raspberry 109.7g

Prepared fleece flower root 248g plantain seed 55g prepared rhizome of rehmannia 313.3g

Fruit of Chinese wolfberry 220g Chinese yam 154.3g barrenwort 315.3g

Fenugreek 313.3g Radix Astragali 171.3g saline cistanche 157.7g

Honey-fried licorice root 47.3g

More than 22 the flavor, frutus cnidii, barrenwort, Radix Angelicae Sinensis, Ligusticum wallichii, fennel seeds are ground into meal, make solvent with 70% ethanol, flood after 48 hours,, collect the about 7000ml of percolate with the speed of per minute 1～3ml diacolation slowly, reclaim ethanol, being evaporated to relative density is the clear cream of 1.05 (heat is surveyed), filters filtrate for later use; Red ginseng powder is broken into meal, adds 9 times of amount 20% alcohol dipping after 8 hours, the refluxing extraction secondary, and each 2 hours, merge extract, filter, filtrate recycling ethanol is concentrated into relative density and is the clear cream of 1.05 (heat is surveyed), and is standby; Ten Six-elements such as all the other raspberries, with the dregs of a decoction of red ginseng through alcohol extracting, adding 10 times of water gagings soaked 1 hour, decoct secondary, each 2 hours, collecting decoction, filter, filtrate is concentrated into relative concentration the be 1.10 clear cream of (heat is surveyed), adds ethanol and makes and contain the alcohol amount and reach 65%, leaves standstill 48 hours, get supernatant and reclaim ethanol, being concentrated into relative density is the clear cream of 1.05 (heat is surveyed), adds extract and red ginseng extracts such as above-mentioned frutus cnidii, mixing, add dextrin 500g, spray drying, dry granulation is made particle 1000 grams altogether.

Embodiment 4: the preparation of capsule

Frutus cnidii 70g Ligusticum wallichii 70.75g seed of Chinese dodder 165g

Psoralea corylifolia 71.25g Poria cocos 75g red ginseng 50g

Fennel seeds 36g fruit of Chinese magnoliavine 90g fruit of Cherokee rose 236.5g

Bighead atractylodes rhizome 35.5g Radix Angelicae Sinensis 117g raspberry 82.25g

Prepared fleece flower root 186g plantain seed 41.25g prepared rhizome of rehmannia 235g

Fruit of Chinese wolfberry 165g Chinese yam 115.75g barrenwort 236.5g

Fenugreek 235g Radix Astragali 128.5g saline cistanche 118.25g

Honey-fried licorice root 35.5g

More than 22 the flavor, frutus cnidii, barrenwort, Radix Angelicae Sinensis, Ligusticum wallichii, fennel seeds are ground into meal, make solvent with 70% ethanol, flood after 48 hours,, collect the about 3750ml of percolate with the speed of per minute 1～3ml diacolation slowly, reclaim ethanol, being evaporated to relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed), filters filtrate for later use; Red ginseng powder is broken into meal, adds 9 times of amount 20% alcohol dipping after 8 hours, the refluxing extraction secondary, and each 2 hours, merge extract, filter, filtrate recycling ethanol is concentrated into relative density and is the clear cream of 1.10 (80 ℃ of heat are surveyed), and is standby; Ten Six-elements such as all the other raspberries with the dregs of a decoction of red ginseng through alcohol extracting, add 10 times of water gagings and soaked 1 hour, decoct secondary, each 2 hours, collecting decoction filtered, filtrate is concentrated into the clear cream that relative concentration is 1.16 (80 ℃ of heat are surveyed), add ethanol and make that to contain alcohol amount be 65%, left standstill 48 hours, get supernatant and reclaim ethanol, being concentrated into relative density is the clear cream of 1.10 (80 ℃ of heat are surveyed), add extract and red ginseng extracts such as above-mentioned frutus cnidii, mixing, vacuum drying, pulverize, add superfine silica gel powder 4g, mixing incapsulates, make 1000, promptly.

Embodiment 5: the discriminating of the precious sheet of kidney and assay (can comprise following all or part of)

1. differentiate

(1) get 6 of this product, porphyrize, the 20ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis and each 0.5g of Ligusticum wallichii control medicinal material, shines medicinal material solution in pairs with legal system.Drawing above-mentioned need testing solution and control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-ethyl acetate (9: 1), launches, and takes out, and dries, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

(2) get the Osthole reference substance, add ethanol and make the solution that every 1ml contains 1mg, in contrast product solution.Drawing above-mentioned reference substance solution and [discriminating] (1) item each 1 μ l of need testing solution down, put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (7: 2), launches, and taking-up is dried, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

(3) get 12 of this product, porphyrize adds ethyl acetate 30ml, reflux 1 hour is put coldly, filters, filtrate is extracted 3 times with 5% sodium carbonate liquor jolting, and each 20ml merges the sodium carbonate extract, with hydrochloric acid adjust pH to 2～3, extract 2 times each 30ml, combined ethyl acetate extract again with the ethyl acetate jolting, be concentrated into 1ml, as need testing solution.Other gets fleece-flower root control medicinal material 0.3g, shines medicinal material solution in pairs with legal system.Get the archen reference substance again, add methyl alcohol and make the solution that every 1ml contains 0.2mg, in contrast product solution.Draw above-mentioned need testing solution 10 μ l, each 2 μ l of control medicinal material and reference substance solution, put respectively on same silica gel g thin-layer plate, upper solution with sherwood oil (30～60 ℃)-ethyl formate-formic acid (15: 5: 1) is a developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot.Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness.

(4) get 10 of this product, porphyrize adds normal butyl alcohol 30ml, and sonicated 30 minutes filters, and filtrate adds ammonia solution washing 2 times, and each 30ml discards ammonia solution, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, 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, (10: 1: 1: 1) be developping agent, expansion was taken out with ethyl acetate-butanone-formic acid-water, dry, spray is put 105 ℃ of bakings several minutes, in the test sample chromatogram with 10% ethanol solution of sulfuric acid, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(5) get 1 of this product, porphyrize adds methyl alcohol 20ml, and ultrasonic 30 minutes, put coldly, filter, filtrate is as need testing solution.Other gets psoralen and the Isopsoralen reference substance is an amount of, adds methyl alcohol and makes the mixed solution that every 1ml contains 4 μ g, in contrast product solution.With the octadecylsilane chemically bonded silica is filling agent; Methanol-water (40: 60) is a moving phase, and the detection wavelength is 246nm.Accurate respectively reference substance solution and each 20ul of need testing solution of drawing injects liquid chromatograph, measures.The retention time of main peak should be consistent with reference substance in the test sample chromatogram.

2. assay

(1) 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside lucifuge operation.

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (25: 75) is moving phase; The detection wavelength is 320nm.Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates should be not less than 2000.

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside reference substance adds Diluted Alcohol and makes the solution that every 1ml contains 20ug, promptly.

This product under the weight differential item, porphyrize, mixing are got in the preparation of need testing solution, get about 0.25g, the accurate title, decide, and puts in the conical flask, the accurate Diluted Alcohol 50ml that adds claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains prepared fleece flower root with 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside (C ₂₀H ₂₂O ₉) meter, must not be less than 0.6mg.

(2) icariin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (30: 70) is moving phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 40ug, promptly.

This product under the weight differential item, porphyrize, mixing are got in the preparation of need testing solution, get about 0.25g, the accurate title, decide, and adds water 10ml, put in the tool plug separating funnel, use ethyl acetate extraction 4 times, each 10ml, the combined ethyl acetate layer, evaporate to dryness, residue add dissolve with methanol and are transferred in the 25m1 measuring bottle, add methyl alcohol to scale, shake up, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains barrenwort with icariin (C ₃₃H ₄₀O ₁₅) meter, must not be less than 0.5mg.

(3) Osthole

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (67: 33) is a moving phase; The detection wavelength is 249nm.Number of theoretical plate calculates by the Osthole peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution that every 1ml contains 50ug, promptly.

The precious sheet of kidney is got in the preparation of need testing solution, porphyrize, and mixing is got about 0.25g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate ethanol 50ml that adds, close plug claims to decide weight, and sonicated 1 hour claims to decide weight again, supply the weight that subtracts mistake with ethanol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and the need testing solution 20ml of drawing of determination method injects liquid chromatograph, measures, promptly.

Every of this product contains frutus cnidii with Osthole (C ₁₅H ₁₆O ₃) meter, must not be less than 0.25mg;

(4) schizandrin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (13: 7) is a moving phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schizandrin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 30ug, promptly.

The precious sheet of kidney is got in the preparation of need testing solution, porphyrize, and mixing is got about 0.3g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol 50ml that adds, close plug claims to decide weight, and sonicated 20 minutes claims to decide weight again, supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains the fruit of Chinese magnoliavine with schizandrin (C ₂₄H ₃₂O ₇) meter, must not be less than 0.15mg;

Embodiment 6: the discriminating of the precious chewable tablets of kidney and assay (can comprise following all or part of)

1. differentiate

(1) get 4 of this product, porphyrize, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis and each 0.5g of Ligusticum wallichii control medicinal material, shines medicinal material solution in pairs with legal system.Drawing above-mentioned need testing solution and control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-ethyl acetate (9: 1), launches, and takes out, and dries, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

(2) get the Osthole reference substance, add ethanol and make the solution that every 1ml contains 1mg, in contrast product solution.Drawing above-mentioned reference substance solution and [discriminating] (1) item each 1 μ l of need testing solution down, put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (7: 2), launches, and taking-up is dried, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

(3) get 8 of this product, porphyrize adds ethyl acetate 30ml, reflux 1 hour is put coldly, filters, filtrate is extracted 3 times with 5% sodium carbonate liquor jolting, and each 20ml merges the sodium carbonate extract, with hydrochloric acid adjust pH to 2～3, extract 2 times each 30ml, combined ethyl acetate extract again with the ethyl acetate jolting, be concentrated into 1ml, as need testing solution.Other gets fleece-flower root control medicinal material 0.3g, shines medicinal material solution in pairs with legal system.Get the archen reference substance again, add methyl alcohol and make the solution that every 1ml contains 0.2mg, in contrast product solution.Draw above-mentioned need testing solution 10 μ l, each 2 μ l of control medicinal material and reference substance solution, put respectively on same silica gel g thin-layer plate, upper solution with sherwood oil (30～60 ℃)-ethyl formate-formic acid (15: 5: 1) is a developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot.Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness.

(4) get 5 of this product, porphyrize adds normal butyl alcohol 30ml, and sonicated 30 minutes filters, and filtrate adds ammonia solution washing 2 times, and each 30ml discards ammonia solution, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, 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, (10: 1: 1: 1) be developping agent, expansion was taken out with ethyl acetate-butanone-formic acid-water, dry, spray is put 105 ℃ of bakings several minutes, in the test sample chromatogram with 10% ethanol solution of sulfuric acid, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(5) get 1 of this product, porphyrize adds methyl alcohol 40ml, and ultrasonic 30 minutes, put coldly, filter, filtrate is as need testing solution.Other gets psoralen and the Isopsoralen reference substance is an amount of, adds methyl alcohol and makes the mixed solution that every 1ml contains 4 μ g, in contrast product solution.With the octadecylsilane chemically bonded silica is filling agent; Methanol-water (40: 60) is a moving phase, and the detection wavelength is 246nm.Accurate respectively reference substance solution and each 20ul of need testing solution of drawing injects liquid chromatograph, measures.The retention time of main peak should be consistent with reference substance in the test sample chromatogram.

2. assay

(1) 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside lucifuge operation.Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version appendix in 2005 VID).

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (25: 75) is moving phase; The detection wavelength is 320nm.Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates should be not less than 2000.

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside reference substance adds Diluted Alcohol and makes the solution that every 1ml contains 20ug, promptly.

This product under the weight differential item, porphyrize, mixing are got in the preparation of need testing solution, get about 0.3g, the accurate title, decide, and puts in the conical flask, the accurate Diluted Alcohol 50ml that adds claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains prepared fleece flower root with 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside (C ₂₀H ₂₂O ₉) meter, must not be less than 0.9mg.

(2) icariin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (30: 70) is moving phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 40ug, promptly.

This product under the weight differential item, porphyrize, mixing are got in the preparation of need testing solution, get about 0.3g, the accurate title, decide, and dissolves with suitable quantity of water, put in the tool plug separating funnel, use ethyl acetate extraction 4 times, each 10ml, the combined ethyl acetate layer, evaporate to dryness, residue add dissolve with methanol and are transferred in the 25m1 measuring bottle, add methyl alcohol to scale, shake up, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains barrenwort with icariin (C ₃₃H ₄₀O ₁₅) meter, must not be less than 0.75mg.

(3) Osthole

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (67: 33) is a moving phase; The detection wavelength is 249nm.Number of theoretical plate calculates by the Osthole peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution that every 1ml contains 50ug, promptly.

The precious chewable tablets of kidney is got in the preparation of need testing solution, porphyrize, and mixing is got about 0.3g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate ethanol 50ml that adds, close plug claims to decide weight, and sonicated 1 hour claims to decide weight again, supply the weight that subtracts mistake with ethanol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and the need testing solution 20ml of drawing of determination method injects liquid chromatograph, measures, promptly.

Every of this product contains frutus cnidii with Osthole (C ₁₅H ₁₆O ₃) meter, must not be less than 0.4mg.

(4) schizandrin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (13: 7) is a moving phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schizandrin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 30ug, promptly.

The precious chewable tablets of kidney is got in the preparation of need testing solution, porphyrize, and mixing is got about 0.5g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol 50ml that adds, close plug claims to decide weight, and sonicated 20 minutes claims to decide weight again, supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains the fruit of Chinese magnoliavine with schizandrin (C ₂₄H ₃₂O ₇) meter, must not be less than 0.2mg;

Embodiment 7: the discriminating of granule for strengthening kidney and assay (can comprise following all or part of)

1. differentiate

(1) get this product 6g, porphyrize, the 30ml that adds diethyl ether soaked 15 minutes, and sonicated is 15 minutes again, puts coldly, filters, and filtrate volatilizes solvent, and residue adds ethanol 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis and each 0.2g of Ligusticum wallichii control medicinal material, shines medicinal material solution in pairs with legal system.Draw above-mentioned need testing solution 15 μ l, control medicinal material solution 2 μ l put respectively on same silica gel g thin-layer plate, are developping agent with normal hexane-ethyl acetate (9: 1), launch, and take out, and dry, and put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

(2) get the Osthole reference substance, add ethanol and make the solution that every 1ml contains 1mg, in contrast product solution.Drawing each the 1 μ l of need testing solution under the item of above-mentioned reference substance solution and [discriminating] (1), put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (7: 2), launches, and takes out, and dries, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

(3) get this product 6g, porphyrize adds ethyl acetate 30ml, reflux 1 hour is put coldly, filters, filtrate is extracted 3 times with 5% sodium carbonate liquor jolting, and each 20ml merges the sodium carbonate extract, with hydrochloric acid adjust pH to 2～3, extract 2 times with the ethyl acetate jolting, each 30ml merges the ethyl acetate extract again, be concentrated into 1ml, as need testing solution.Other gets fleece-flower root control medicinal material 0.3g, shines medicinal material solution in pairs with legal system.Get the archen reference substance again, add methyl alcohol and make the solution that every 1ml contains 0.2mg, in contrast product solution.Draw above-mentioned need testing solution 10 μ l, each 2 μ l of control medicinal material and reference substance solution, put respectively on same silica gel g thin-layer plate, upper solution with sherwood oil (30～60 ℃)-ethyl formate-formic acid (15: 5: 1) is a developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot.Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness.

(4) get this product 6g, porphyrize adds normal butyl alcohol 30ml, and sonicated 30 minutes filters, and filtrate adds ammonia solution washing 2 times, and each 30ml discards ammonia solution, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, 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, (10: 1: 1: 1) be developping agent, expansion was taken out with ethyl acetate-butanone-formic acid-water, dry, spray is put 105 ℃ of bakings several minutes, in the test sample chromatogram with 10% ethanol solution of sulfuric acid, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(5) get this product 1g, porphyrize adds methyl alcohol 20ml, and ultrasonic 30 minutes, put coldly, filter, filtrate is as need testing solution.Other gets psoralen and the Isopsoralen reference substance is an amount of, adds methyl alcohol and makes the mixed solution that every 1ml contains 3 μ g, in contrast product solution.With the octadecylsilane chemically bonded silica is filling agent; Methanol-water (40: 60) is a moving phase, and the detection wavelength is 246nm.Accurate respectively reference substance solution and each 10ul of need testing solution of drawing injects liquid chromatograph, measures.The retention time of main peak should be consistent with reference substance in the test sample chromatogram.

2. assay

(1) 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside lucifuge operation.

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (25: 75) is moving phase; The detection wavelength is 320nm.Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates should be not less than 2000.

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside reference substance adds Diluted Alcohol and makes the solution that every 1ml contains 60ug, promptly.

The about 1.5g of this product is got in the preparation of need testing solution, accurate claims surely, puts in the conical flask, and the accurate Diluted Alcohol 50ml that adds claims decide weight, and reflux 30 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and gets the supernatant filtration, promptly.

Accurate respectively reference substance solution and each 10ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

This product contains 2,3,5 for every bag, 4 '-tetrahydroxystilbene-2-O-b-D-glucoside (C ₂₀H ₂₂O ₉) must not be less than 1.3mg.

(2) icariin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (30: 70) is moving phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 40ug, promptly.

The about 0.6g of this product is got in the preparation of need testing solution, and accurate the title decides, and with the suitable quantity of water dissolving, puts in the tool plug separating funnel, uses ethyl acetate extraction 4 times, and each 10ml merges the ethyl acetate layer.Evaporate to dryness, residue 10ml dissolve with methanol filters promptly.

Accurate respectively reference substance solution and each 10ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

This product contains icariin (C for every bag ₃₃H ₄₀O ₁₅) must not be less than 1.5mg.

(3) Osthole

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (67: 33) is a moving phase; The detection wavelength is 249nm.Number of theoretical plate calculates by the Osthole peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution that every 1ml contains 50ug, promptly.

Granule for strengthening kidney is got in the preparation of need testing solution, porphyrize, and mixing is got about 1.5g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate ethanol 50ml that adds, close plug claims to decide weight, and sonicated 1 hour claims to decide weight again, supply the weight that subtracts mistake with ethanol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and the need testing solution 20ml of drawing of determination method injects liquid chromatograph, measures, promptly.

This product contains Osthole (C for every bag ₁₅H ₁₆O ₃) must not be less than 0.8mg.

(4) schizandrin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (13: 7) is a moving phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schizandrin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 30ug, promptly.

Granule for strengthening kidney is got in the preparation of need testing solution, porphyrize, and mixing is got about 2.0g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol 50ml that adds, close plug claims to decide weight, and sonicated 20 minutes claims to decide weight again, supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

This product contains schizandrin (C for every bag ₂₄H ₃₂O ₇) must not be less than 0.4mg.

Embodiment 8: the discriminating of kidney capsule and assay

1. differentiate

(1) get 8 of this product, the 20ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Sinensis and each 0.5g of Ligusticum wallichii control medicinal material, shines medicinal material solution in pairs with legal system.Drawing above-mentioned need testing solution and control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate, is developping agent with normal hexane-ethyl acetate (9: 1), launches, and takes out, and dries, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color.

(2) get the Osthole reference substance, add ethanol and make the solution that every 1ml contains 1mg, in contrast product solution.Drawing above-mentioned reference substance solution and [discriminating] (1) item each 1 μ l of need testing solution down, put respectively on same silica gel g thin-layer plate, is developping agent with cyclohexane-ethyl acetate (7: 2), launches, and taking-up is dried, and puts under the ultraviolet lamp (365nm) and inspects.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.

(3) get 15 of this product, porphyrize adds ethyl acetate 30ml, reflux 1 hour is put coldly, filters, filtrate is extracted 3 times with 5% sodium carbonate liquor jolting, and each 20ml merges the sodium carbonate extract, with hydrochloric acid adjust pH to 2～3, extract 2 times each 30ml, combined ethyl acetate extract again with the ethyl acetate jolting, be concentrated into 1ml, as need testing solution.Other gets fleece-flower root control medicinal material 0.3g, shines medicinal material solution in pairs with legal system.Get the archen reference substance again, add methyl alcohol and make the solution that every 1ml contains 0.2mg, in contrast product solution.Draw above-mentioned need testing solution 10 μ l, each 2 μ l of control medicinal material and reference substance solution, put respectively on same silica gel g thin-layer plate, upper solution with sherwood oil (30～60 ℃)-ethyl formate-formic acid (15: 5: 1) is a developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect.In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot.Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness.

(4) get 12 of this product, porphyrize adds normal butyl alcohol 30ml, and sonicated 30 minutes filters, and filtrate adds ammonia solution washing 2 times, and each 30ml discards ammonia solution, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, 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, (10: 1: 1: 1) be developping agent, expansion was taken out with ethyl acetate-butanone-formic acid-water, dry, spray is put 105 ℃ of bakings several minutes, in the test sample chromatogram with 10% ethanol solution of sulfuric acid, with the corresponding position of reference substance chromatogram on, show the spot of same color.

(5) get 2 of this product, porphyrize adds methyl alcohol 20ml, and ultrasonic 30 minutes, put coldly, filter, filtrate is as need testing solution.Other gets psoralen and the Isopsoralen reference substance is an amount of, adds methyl alcohol and makes the mixed solution that every 1ml contains 4 μ g, in contrast product solution.With the octadecylsilane chemically bonded silica is filling agent; Methanol-water (40: 60) is a moving phase, and the detection wavelength is 246nm.Accurate respectively reference substance solution and each 20ul of need testing solution of drawing injects liquid chromatograph, measures.In the test sample chromatogram retention time of main peak should with reference substance-cause.

2. assay

(1) 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside lucifuge operation.

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (25: 75) is moving phase; The detection wavelength is 320nm.Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside peak calculates should be not less than 2000.

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-b-D-glucoside reference substance adds Diluted Alcohol and makes the solution that every 1ml contains 20ug, promptly.

This product under the weight differential item, porphyrize, mixing are got in the preparation of need testing solution, get about 0.25g, the accurate title, decide, and puts in the conical flask, the accurate Diluted Alcohol 50ml that adds claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains prepared fleece flower root with 2,3,5,4 '-tetrahydroxystilbene-2-O-b-D-glucoside (C ₂₀H ₂₂O ₉) meter, must not be less than 0.4mg.

(2) icariin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Second eyeball-water (30: 70) is moving phase; The detection wavelength is 270nm.Number of theoretical plate calculates by the icariin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 40ug, promptly.

This product under the weight differential item, porphyrize, mixing are got in the preparation of need testing solution, get about 0.25g, the accurate title, decide, and adds water 10ml, put in the tool plug separating funnel, use ethyl acetate extraction 4 times, each 10ml, the combined ethyl acetate layer, evaporate to dryness, residue add dissolve with methanol and are transferred in the 25ml measuring bottle, add methyl alcohol to scale, shake up, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains barrenwort with icariin (C ₃₃H ₄₀O ₁₅) meter, must not be less than 0.35mg

(3) Osthole

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (67: 33) is a moving phase; The detection wavelength is 249nm.Number of theoretical plate calculates by the Osthole peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution that every 1ml contains 50ug, promptly.

This product content is got in the preparation of need testing solution, porphyrize, and mixing is got about 0.25g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate ethanol 50ml that adds, close plug claims to decide weight, and sonicated 1 hour claims to decide weight again, supply the weight that subtracts mistake with ethanol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and the need testing solution 20ml of drawing of determination method injects liquid chromatograph, measures, promptly.

Every of this product contains frutus cnidii with Osthole (C ₁₅H ₁₆O ₃) meter, must not be less than 0.2mg

(4) schizandrin

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Methanol-water (13: 7) is a moving phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schizandrin peak should be not less than 2000.

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 30ug, promptly.

This product content is got in the preparation of need testing solution, porphyrize, and mixing is got about 0.3g, and accurate the title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol 50ml that adds, close plug claims to decide weight, and sonicated 20 minutes claims to decide weight again, supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each 20ml of need testing solution of drawing of determination method injects liquid chromatograph, measures promptly.

Every of this product contains the fruit of Chinese magnoliavine with schizandrin (C ₂₄H ₃₂O ₇) meter, must not be less than 0.1mg.

Claims (4)

1. the detection method of a Shenbao preparation, be used to detect the quality of the Shenbao preparation of making by following Chinese medicinal raw materials, frutus cnidii 28 weight portions, Ligusticum wallichii 28.3 weight portions, the seed of Chinese dodder 66 weight portions, Psoralea corylifolia 28.5 weight portions, Poria cocos 30 weight portions, red ginseng 20 weight portions, fennel seeds 14.4 weight portions, the fruit of Chinese magnoliavine 36 weight portions, the fruit of Cherokee rose 94.6 weight portions, the bighead atractylodes rhizome 14.2 weight portions, Radix Angelicae Sinensis 46.8 weight portions, raspberry 32.9 weight portions, prepared fleece flower root 74.4 weight portions, plantain seed 16.5 weight portions, prepared rhizome of rehmannia 94 weight portions, the fruit of Chinese wolfberry 66 weight portions, Chinese yam 46.3 weight portions, barrenwort 94.6 weight portions, fenugreek 94 weight portions, the Radix Astragali 51.4 weight portions, saline cistanche 47.3 weight portions, honey-fried licorice root 14.2 weight portions, said ratio can change in proportion, it is characterized in that, contain following assay and qualitative identification:

(1) assay

A. high-efficient liquid phase technique measures in the Shenbao preparation 2,3,5, the content of 4 '-tetrahydroxystilbene-2-O-β-D-glucoside

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Second eyeball 5-70 part: water 95-30 part is a moving phase; UV-detector or diode array detector; Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-β-D-glucoside peak calculates should be not less than 2000;

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-β-D-glucoside reference substance adds ethanol or Diluted Alcohol and makes the solution of determining concentration, product solution in contrast;

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, with the methanol/ethanol extraction of suitable concentration, as need testing solution;

Accurate respectively reference substance solution and an amount of volume of need testing solution drawn of determination method injects liquid chromatograph, measures, promptly;

B. high-efficient liquid phase technique is measured icariin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Second eyeball 5-70 part: water 95-30 part is a moving phase; UV-detector or diode array detector; Number of theoretical plate calculates by the icariin peak should be not less than 2000;

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution of determining concentration, product solution in contrast;

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, adds the suitable quantity of water dissolving, uses ethyl acetate extraction, the extract evaporate to dryness, and residue dissolves with The suitable solvent, as need testing solution;

Accurate respectively reference substance solution and an amount of volume of need testing solution drawn of determination method injects liquid chromatograph, measures, promptly;

C. high-efficient liquid phase technique is measured Osthole content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Methyl alcohol 40-85 part: water 60-15 part is a moving phase; UV-detector or diode array detector; Number of theoretical plate calculates by the Osthole peak should be not less than 2000;

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution of determining concentration, product solution in contrast;

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, with the methanol/ethanol extraction of suitable concentration, as need testing solution;

Accurate respectively reference substance solution and an amount of volume of need testing solution drawn of determination method injects liquid chromatograph, measures, promptly;

D. high-efficient liquid phase technique is measured schizandrin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels; Methyl alcohol 5-30 part: water 2-15 part is a moving phase; UV-detector or diode array detector; Number of theoretical plate calculates by the schizandrin peak should be not less than 2000;

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution of determining concentration, product solution in contrast;

The preparation precision of need testing solution takes by weighing product or the product content thing is an amount of, with the methanol/ethanol extraction of suitable concentration, as need testing solution;

Accurate respectively reference substance solution and an amount of volume of need testing solution drawn of determination method injects liquid chromatograph, measures, promptly;

(2) qualitative identification

A. thin-layered chromatography is differentiated Radix Angelicae Sinensis and Ligusticum wallichii in the Shenbao preparation

Get product or the product content thing is an amount of, the extraction that adds diethyl ether, extract is as need testing solution, or extract steams/volatilize, and residue adds The suitable solvent makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis and the Ligusticum wallichii control medicinal material is an amount of, makes control medicinal material solution after the extraction; Each is an amount of to draw above-mentioned need testing solution and control medicinal material solution, puts respectively on same silica G or silica gel H thin layer plate, and with normal hexane 3-30 part: ethyl acetate 0.3-3 part is a developping agent, launches, and takes out, and dries, and puts under the ultraviolet lamp and inspects; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

B. thin-layered chromatography is differentiated frutus cnidii in the Shenbao preparation

Get product or the product content thing is an amount of, the extraction that adds diethyl ether, extract is as need testing solution, or extract steams/volatilize, and residue adds The suitable solvent makes dissolving, as need testing solution; It is an amount of that other gets the Osthole reference substance, and solubilizer is made reference substance solution in right amount; Each is an amount of to draw above-mentioned need testing solution and reference substance solution, puts respectively on same silica G or silica gel H thin layer plate, and with benzene 10-50 part: ethyl acetate 0.3-2 part is a developping agent, launches, and takes out, and dries, and puts under the ultraviolet lamp and inspects; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

C. thin-layered chromatography is differentiated prepared fleece flower root in the Shenbao preparation

Get product or the product content thing is an amount of, add ethyl acetate extraction, extract extracts with the sodium carbonate liquor of suitable concentration, sodium carbonate extract hydrochloric acid adjust pH to 1～4, use ethyl acetate extraction again, extract is as need testing solution, or extract steams/volatilizes, residue adds The suitable solvent makes dissolving, as need testing solution; It is an amount of that other gets fleece-flower root control medicinal material, makes control medicinal material solution after the extraction; It is an amount of to get the archen reference substance again, and solubilizer is made reference substance solution in right amount; Each is an amount of to draw above-mentioned need testing solution, control medicinal material and reference substance solution, put respectively on same silica G or silica gel H thin layer plate, with 30～60 ℃ of sherwood oil 5-25 parts: ethyl formate 2-8 part: the upper solution of formic acid 0.4-2 part is a developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot; Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness;

D. thin-layered chromatography is differentiated barrenwort in the Shenbao preparation

Get product or the product content thing is an amount of, add normal butyl alcohol and extract, extract washs with ammonia solution, discards ammonia solution, and extract is as need testing solution, or extract steams/volatilize, and residue adds The suitable solvent makes dissolving, as need testing solution; Other gets the icariin reference substance, and solubilizer is made reference substance solution in right amount; Each is an amount of to draw above-mentioned two kinds of solution, put respectively on same silica G or silica gel H thin layer plate, with ethyl acetate 3-30 part: butanone 0.4-4 part: formic acid 0.4-4 part: water 0.4-4 part is a developping agent, launches, and takes out, dry, spray is heated to colour developing with 10% ethanol solution of sulfuric acid, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

E. high-efficient liquid phase technique is differentiated Psoralea corylifolia in the Shenbao preparation

Get product or the product content thing is an amount of, add methanol extraction, extract is as need testing solution; Other gets psoralen and the Isopsoralen reference substance is an amount of, and solubilizer is made reference substance solution in right amount; With the octadecylsilane chemically bonded silica is filling agent; Methyl alcohol 20-80 part: water 80-20 part is a moving phase, UV-detector or diode array detector; Accurate respectively reference substance solution and each an amount of volume of need testing solution drawn injects liquid chromatograph, measures; The retention time of main peak should be consistent with reference substance in the test sample chromatogram.

2. the detection method of the described Shenbao preparation of claim 1 is characterized in that:

(1) described extraction can adopt sonicated, refluxing extraction, Microwave Extraction, vibration to extract conventional method;

(2) 2,3,5, the moving phase of the assay of 4 '-tetrahydroxystilbene-2-O-β-D-glucoside can also be methyl alcohol 2-10 part: acetonitrile 0.5-2 part: water 4-20 part, methyl alcohol 5-70 part: water 95-30 part;

(3) moving phase of content Determination of Icariin mensuration can also be methyl alcohol 40-80 part: water 20-60 part: glacial acetic acid 0.2-1 part, methyl alcohol 5-70 part: water 95-30 part;

(4) moving phase of the assay of Osthole can also be methyl alcohol 30-75 part: water 20-60 part: tetrahydrofuran 2-10 part, acetonitrile 5-70 part: water 95-30 part;

(5) moving phase of the assay of schizandrin can also be acetonitrile 5-70 part: water 95-30 part;

(6) developping agent of the thin-layer chromatography of Radix Angelicae Sinensis and Ligusticum wallichii discriminating can also be 60-90 ℃ of sherwood oil 5-25 part: ethyl acetate 0.5-5 part, cyclohexane 5-25 part: ethyl acetate 0.5-5 part;

(7) developping agent of the thin-layer chromatography of frutus cnidii discriminating can also be cyclohexane 3-20 part: ethyl acetate 0.5-5 part, normal hexane 4-20 part: ethyl acetate 0.4-4 part;

(8) developping agent of the thin-layer chromatography of prepared fleece flower root discriminating can also be chloroform 5-15 part: methyl alcohol 0.5-5 part, 30-60 ℃ sherwood oil 50-150 part: ethyl acetate 5-50 part: glacial acetic acid 0.5-5 part;

(9) both can adopt reference substance among qualitative identification a, b, c, the d, and also can adopt control medicinal material to compare, or both all adopt; Both can adopt two kinds of reference substances among the qualitative identification e, also can only adopt wherein a kind of reference substance in contrast.

3. the detection method of the described a kind of Shenbao preparation of claim 1 is characterized in that, contains following assay and qualitative identification:

(1) assay

A. high-efficient liquid phase technique measures in the Shenbao preparation 2,3,5, the content of 4 '-tetrahydroxystilbene-2-O-β-D-glucoside

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; 25 parts in second eyeball: water is moving phase for 75 parts; The detection wavelength is 320nm; Number of theoretical plate is by 2,3,5, and 4 '-tetrahydroxystilbene-2-O-β-D-glucoside peak calculates should be not less than 2000;

The preparation precision of reference substance solution takes by weighing 2,3,5, and 4 '-tetrahydroxystilbene-2-O-β-D-glucoside reference substance adds Diluted Alcohol and makes the solution that every 1ml contains 20ug, promptly;

Product is got in the preparation of need testing solution or the product content thing is an amount of, puts in the conical flask, and the accurate Diluted Alcohol 50ml that adds claims decide weight, and reflux 30 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and filtration is got subsequent filtrate, promptly;

Accurate respectively reference substance solution and each the 20 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly;

B. high-efficient liquid phase technique is measured icariin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; 30 parts in second eyeball: water is moving phase for 70 parts; The detection wavelength is 270nm; Number of theoretical plate calculates by the icariin peak should be not less than 2000;

The preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 40ug, promptly;

Product is got in the preparation of need testing solution or the product content thing is an amount of, adds water 10ml, puts in the tool plug separating funnel, uses ethyl acetate extraction 4 times, each 10ml, combined ethyl acetate layer, evaporate to dryness, residue add dissolve with methanol and are transferred in the 25m1 measuring bottle, add methyl alcohol to scale, shake up, promptly;

Accurate respectively reference substance solution and the need testing solution 20 μ l of drawing of determination method inject liquid chromatograph, measure, promptly;

C. high-efficient liquid phase technique is measured Osthole content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; 67 parts of methyl alcohol: water is moving phase for 33 parts; The detection wavelength is 249nm; Number of theoretical plate calculates by the Osthole peak should be not less than 2000;

The preparation precision of reference substance solution takes by weighing the Osthole reference substance, adds methyl alcohol and makes the solution that every 1ml contains 50ug, promptly;

Product is got in the preparation of need testing solution or the product content thing is an amount of, puts in the tool plug conical flask, the accurate ethanol 50ml that adds, and close plug claims to decide weight, and sonicated 1 hour claims to decide weight again, supplies the weight that subtracts mistake with ethanol, shakes up, and filters, and gets subsequent filtrate, promptly;

Accurate respectively reference substance solution and the need testing solution 20 μ l of drawing of determination method inject liquid chromatograph, measure, promptly;

D. high-efficient liquid phase technique is measured schizandrin content in the Shenbao preparation

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; 13 parts of methyl alcohol: water is moving phase for 7 parts; The detection wavelength is 250nm; Number of theoretical plate calculates by the schizandrin peak should be not less than 2000;

The preparation precision of reference substance solution takes by weighing the schizandrin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 30ug, promptly;

Product is got in the preparation of need testing solution or the product content thing is an amount of, puts in the tool plug conical flask, the accurate methyl alcohol 50ml that adds, and close plug claims to decide weight, and sonicated 20 minutes claims to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filters, and gets subsequent filtrate, promptly;

Accurate respectively reference substance solution and each the 20 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly;

(2) qualitative identification

A. thin-layered chromatography is differentiated Radix Angelicae Sinensis and Ligusticum wallichii in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis and each 0.5g of Ligusticum wallichii control medicinal material, shines medicinal material solution in pairs with legal system; Draw above-mentioned need testing solution and control medicinal material solution 2 μ l, put respectively on same silica gel g thin-layer plate, with 9 parts of normal hexanes: ethyl acetate is developping agent for 1 part, launches, and takes out, and dries, and puts under the 365nm ultraviolet lamp and inspects; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the fluorescence spot of same color;

B. thin-layered chromatography is differentiated frutus cnidii in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Get the Osthole reference substance, add ethanol and make the solution that every 1ml contains 1mg, in contrast product solution; Draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 30 parts of benzene: 7 parts of 1 part in ethyl acetate or cyclohexanes: ethyl acetate be developping agent for 2 parts, launches, and taking-up is dried, and puts under the 365nm ultraviolet lamp and inspects; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;

C. thin-layered chromatography is differentiated prepared fleece flower root in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Add ethyl acetate 30ml, reflux 1 hour is put cold, filter, filtrate is extracted 3 times with 5% sodium carbonate liquor jolting, and each 20ml merges the sodium carbonate extract, with hydrochloric acid adjust pH to 2～3, extract 2 times each 30ml, combined ethyl acetate extract again with the ethyl acetate jolting, be concentrated into 1ml, as need testing solution; Other gets fleece-flower root control medicinal material 0.3g, shines medicinal material solution in pairs with legal system; Get the archen reference substance again, add methyl alcohol and make the solution that every 1ml contains 0.2mg, in contrast product solution; Draw above-mentioned need testing solution 10 μ l, each 2 μ l of control medicinal material and reference substance solution put respectively on same silica gel g thin-layer plate, with 15 parts of 30～60 ℃ of sherwood oils: 5 parts of ethyl formates: the upper solution that formic acid is 1 part is a developping agent, launches, and takes out, dry, put under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with control medicinal material and the corresponding position of reference substance chromatogram on, show identical orange-yellow fluorescence spot; Put in the ammonia steam smoked after, inspect under the daylight, spot becomes redness;

D. thin-layered chromatography is differentiated barrenwort in the Shenbao preparation

Get product or the product content thing is an amount of, the 30ml that adds diethyl ether, reflux 1 hour filters. and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Add normal butyl alcohol 30ml, ultrasonic Extraction 30 minutes filters, and filtrate adds ammonia solution washing 2 times, and each 30ml discards ammonia solution, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the icariin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, 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, with 10 parts in ethyl acetate: 1 part of butanone: 1 part in formic acid: water is developping agent for 1 part, launches, and takes out, dry, spray is put 105 ℃ of bakings several minutes, in the test sample chromatogram with 10% ethanol solution of sulfuric acid, with the corresponding position of reference substance chromatogram on, show the spot of same color;

E. high-efficient liquid phase technique is differentiated Psoralea corylifolia in the Shenbao preparation

Get product or the product content thing is an amount of, add methyl alcohol 40ml, ultrasonic Extraction 30 minutes is put coldly, filters, and filtrate is as need testing solution; Other gets psoralen and the Isopsoralen reference substance is an amount of, adds methyl alcohol and makes the mixed solution that every 1ml contains 4 μ g, in contrast product solution; According to high effective liquid chromatography for measuring; With the octadecylsilane chemically bonded silica is filling agent; 40 parts of methyl alcohol: water is moving phase for 60 parts, and the detection wavelength is 246nm; Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject liquid chromatograph, measure; The retention time of main peak should be consistent with reference substance in the test sample chromatogram.

4. claim 1,3 described methods, it is characterized in that, can be used to detect any formulation Shenbao preparation, contain tablet, sugar coated tablet, film coated tablet, chewable tablet, effervescent tablet, Disket, capsule, hard capsule, soft capsule, pellt capsule, oral liquid, syrup, suck agent, electuary, granule, effervescent, pill, pill, micropill preparation.