CN101433582A - Quality control method of capsule for treating prostatitis - Google Patents
Quality control method of capsule for treating prostatitis Download PDFInfo
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Abstract
The invention provides a quality control method for Qianlietai capsule. The method comprises the content mensuration of motherwort as raw material medicine, a Qianlietai capsule midbody and stachydrine hydrochloride in the Qianlietai capsule. Compared with the prior art, the quality control method is reasonable and has a stable mensuration result; and the quality control method is adopted to effectively control the quality of the Qianlietai capsule, thereby ensuring the clinical curative effect of the Qianlietai capsule.
Description
Technical field
The present invention relates to a kind of method of quality control for the treatment of prostatitic Chinese medicine preparation, particularly relate to the method for quality control of the capsule of this Chinese medicine preparation.
Background technology
Along with improving constantly and the senescence of population of socioeconomic continuous development, people's living standard, prostatitic sickness rate improves constantly, and the trend of rejuvenation is arranged, and the drug research of treatment prostatosis becomes the focus of new drug research.Prostatitis is male's commonly encountered diseases, and the overwhelming majority occurs in person between twenty and fifty, can be divided into acute and chronic two kinds clinically.Acute prostatitis is more rare clinically, and chronic prostatitis sickness rate in the crowd that grows up is higher, accounts for Urologic Surgery Clinic patient's about 1/5, because of chronic prostatitis many with seminal vesiculitis, so be called prostatovesticulitis again.
Cause of disease acute prostatitis: morbidity many tired, catch a cold, for a long time by bike, excessive drinking, sexual life excessively, when damage, per urethra instrumentation, whole body or local resistance weaken, pathogenic bacterium enter prostate by the focus menses fortune or the per urethra at other position of health, and topmost pathogenic bacterium are escherichia coli, staphylococcus, Bacillus proteus and streptococcus etc.
Chronic prostatitis: the cause of disease is comparatively complicated, and minority is failed thoroughly to cure to delay by acute prostatitis, and most humans had not then lived through clear and definite acute phase.The pathogenic microorganism that causes chronic prostatitis mainly is an antibacterial, and next has virus, mycoplasma, chlamydia and other sensitinogen etc.Libido is prosperous excessively, congestion of prostate, lower urinary tract obstruction, perineal position compressing, damage, the adjacent organs inflammatory disorders involves prostate and the whole body resistance descends or the like, all may be one of reason that causes chronic prostatitis, even patient's the mental status also be a factor that influences the symptom weight.In a word, chronic prostatitis cause of disease complexity exists the different causes of disease at different times probably, or in the same period has more than one paathogenic factor.
Treatment acute prostatitis: lie up, polydipsia water and relieving constipation etc. are general handles.The irritation sign of bladder severe patient can give analgesia spasmolytic medicine and hot water hip-bath with relief of symptoms.Antibacterials can be selected penicillin, streptomycin, ampicillin, cephalosporin and zinacef etc. for use.Acute prostatitis is behind general anti symptom treatment and anti-inflammatory treatment, and symptom often disappears in 1～2 week.Do not feel any better or increase the weight of on the contrary as symptom, the more swelling and fluctuation is arranged of prostate anus digital examination palpation, the visible abscess of ultrasound diagnosis forms, and extracts the pus person out through the perineum puncture, should be through the capable abscess incision and drainage of perineal position.
Chronic prostatitis: the treatment of chronic prostatitis generally will be treated in conjunction with means such as general treatment, massage of prostate, drug infusion, urethral dilatation, prostate periphery seal and antibacterials.But general antibacterials are difficult for entering prostata tissue, and this also is to treat one of comparatively difficult reason clinically.
Chinese medicine is in prevention and carry out dialectical executing when treating above-mentioned disease and controls, especially be the compound Chinese medicinal preparation made of primary raw material with the Herba Leonuri as: QIANLIETAI JIAONANG, QIANLIETAI PIAN etc. all has definite curative effect.
QIANLIETAI JIAONANG is the present patent application people obtains State Food and Drug Administration's medicine registration official written reply in August, 2005 a kind (drug standard numbering: YBZ15942005).QIANLIETAI JIAONANG is made up of Herba Leonuri, Herba Polygoni Avicularis, Flos Carthami, Brassica campestris L pollen, the Rhizoma Anemarrhenae (salt stir-fry), Cortex Phellodendri medical materials such as (salt stir-frys).The QIANLIETAI JIAONANG intermediates preparation of being mentioned among the present invention is: pulverize with high speed disintegrator behind the Brassica campestris L pollen breaking cellular wall, cross 100 mesh sieves, fine powder is standby; Herba Leonuri, Herba Polygoni Avicularis, Flos Carthami, the Rhizoma Anemarrhenae (salt stir-fry) and Cortex Phellodendri (salt stir-fry) are decocted with water secondary, each 2 hours, add 12 times of water gagings for the first time, add for the second time 10 times of water gagings, filter collecting decoction, filtrate decompression is concentrated into the thick paste that relative density is 1.33～1.37 (50 ℃ of heat are surveyed), drying under reduced pressure is pulverized, and adding Brassica campestris L pollen and starch are an amount of, mix homogeneously, ethanol with 85% (containing 3% soybean oil) is granulated, and 50～60 ℃ of dryings promptly make the QIANLIETAI JIAONANG intermediate; Intermediate is incapsulated, promptly make the QIANLIETAI JIAONANG finished product.
Chinese medicine mostly is to be formed by plurality of raw materials recurrence due to taking drug side, and its composition more complicated also is an a great problem for the quality control of medicine.QIANLIETAI JIAONANG also is to be prepared from through compound recipe by the plurality of raw materials medicine, in clinical practice, exist quality standard low, uppity defective, also do not have at present the method for quality control of more sophisticated QIANLIETAI JIAONANG, the present patent application people inquires into and studies the method for quality control of this Chinese medicine preparation.
Summary of the invention
Technical problem to be solved by this invention provides a kind of method of quality control of QIANLIETAI JIAONANG, and this method comprises Determination of Contents of Hydrochloric Stachydrine assay method in crude drug Herba Leonuri, QIANLIETAI JIAONANG intermediate and the QIANLIETAI JIAONANG, has improved quality control standard.
In order to solve the problems of the technologies described above, the present invention adopts following technical scheme:
A kind of method of quality control of QIANLIETAI JIAONANG comprises Determination of Contents of Hydrochloric Stachydrine assay method in crude drug Herba Leonuri, QIANLIETAI JIAONANG intermediate and the QIANLIETAI JIAONANG.
The Determination of Contents of Hydrochloric Stachydrine assay method is in the above-mentioned raw materials medicine Herba Leonuri:
Chromatographic condition chromatographic column: Z0RBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 250～270nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;
The Herba Leonuri medical material is got in the preparation of need testing solution, pulverize, cross 40 mesh sieves, precision takes by weighing 1g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put coldly, claims to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shake up, filter, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filters, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, put in the ice bath and placed 0.5 hour, filter, with frozen water washing container and precipitation, discard filtrate, with 15ml acetone solution precipitation, the silver sulfate solution of dropping 0.5% produces to no longer including precipitation in acetone soln, places, filter, precipitation merges washing liquid and filtrate with 70% ethanol 15ml gradation washing, puts in 90 ℃ of water-baths and is concentrated into 2ml, put cold, 1% barium chloride solution of adding and silver sulfate equivalent, the potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shakes up, filter, promptly get need testing solution;
The algoscopy precision is measured need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/ml, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
Determination of Contents of Hydrochloric Stachydrine is not less than 0.50% in the crude drug Herba Leonuri.
The Determination of Contents of Hydrochloric Stachydrine assay method is in the aforementioned QIANLIETAI JIAONANG intermediate:
Chromatographic condition chromatographic column: ZORBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;
The QIANLIETAI JIAONANG intermediate is got in the preparation of need testing solution, pulverize, cross 40 mesh sieves, precision takes by weighing 1.5g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put cold, claim to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shakes up, and filters, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filter, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, puts in the ice bath and places 0.5 hour, filters, discard filtrate, with frozen water washing container and precipitation, discard filtrate, precipitate with the 15ml acetone solution, dripping 0.5% silver sulfate solution in acetone soln produces to no longer including precipitation, place, filter, with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put in 90 ℃ of water-baths and be concentrated into 2ml, put coldly, add 1% barium chloride solution with the silver sulfate equivalent, potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shake up, filter, promptly get need testing solution;
The algoscopy precision is measured need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/l, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
Determination of Contents of Hydrochloric Stachydrine is not less than 0.60% in the QIANLIETAI JIAONANG intermediate.
The Determination of Contents of Hydrochloric Stachydrine assay method is in the aforementioned QIANLIETAI JIAONANG:
Chromatographic condition chromatographic column: ZORBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;
20 of QIANLIETAI JIAONANG are got in the preparation of need testing solution, incline and get content, porphyrize, precision takes by weighing 2g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put coldly, claims to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shake up, filter, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filters, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, put in the ice bath and placed 0.5 hour, filter, with frozen water gradation washing container and precipitation, discard filtrate, with 15ml acetone solution precipitation, the silver sulfate solution of dropping 0.5% produces to no longer including precipitation in acetone soln, places, filter, with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put in 90 ℃ of water-baths and be concentrated into 2ml, put cold, 1% barium chloride solution of adding and silver sulfate equivalent, the potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shakes up, filter, promptly get need testing solution;
The algoscopy precision is measured above-mentioned need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/ml, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
Determination of Contents of Hydrochloric Stachydrine is not less than 0.50% in the QIANLIETAI JIAONANG.
In order to verify the reasonability of method of quality control of the present invention, the applicant has carried out experimental study and screening to this method, and is specific as follows:
1, the preparation of reference substance solution
Precision takes by weighing that to be dried to the stachydrine hydrochloride reference substance of constant weight through phosphorus pentoxide an amount of, makes the solution of the hydrochloric stachydrine 1mg of every 1ml with 0.01% potassium hydroxide solution, promptly.
2, the selection of mobile phase
Reference literature data chromatographic condition is tested, and selected mobile phase is the methanol-water of volume ratio 60:40.
3 detect the selection of wavelength
Get stachydrine hydrochloride reference substance derivative solution 10 μ l, inject chromatograph of liquid, carry out all-wave scanning in 190～400 scopes, the result has absorption maximum at the 260nm place, so select 260nm as detecting wavelength.
The test of 4 system suitabilitys
4.1 the affirmation at reference substance, sample chromatogram peak:
Prepare the acetonitrile solution of 0.01%KOH solution, 0.5mg/ml respectively, press the listed step operation of table 1, obtain 4 kinds of different solutions a～d the hexaoxacyclooctadecane-6-6 of the acetonitrile solution of bromo bromination 1-Phenylethanone. and 0.05mg/ml:
The process for preparation of table 14 kind of different solutions
4 kinds of solution that will prepare by table 1, supersound process are after 10 minutes, and heating is 20 minutes in 80 ℃ of water-baths, after the taking-up, is cooled to room temperature, inject chromatograph of liquid and detect.From testing result as can be known, the chromatographic peak that the catalyst that adds in the derivative reaction, derivatization reagent produce does not disturb the chromatographic peak of stachydrine hydrochloride derivant, and the stachydrine hydrochloride derivant on the chromatogram is easy to confirm.
4.2 negative sample interference test
Get QIANLIETAI JIAONANG and do not contain the negative sample of Herba Leonuri,, found that after adding 2% sulfur hydracid chromium ammonium salt solution, the negative sample deposit-free produces, and the QIANLIETAI JIAONANG sample has tangible precipitation by 5.1 following methods four preparation test samples.Result of the test shows: negative sample is noiseless.
4.3 separating degree and theoretical cam curve:
To need testing solution and reference substance solution difference sample introduction, show on data report: its separating degree can be separated with other impurity peaks fully all greater than 1.5; And the theoretical tray number average is greater than 4000, so the theoretical cam curve of this test should be not less than 4000 in stachydrine hydrochloride derivant chromatographic peak.
5, the preparation of need testing solution
5.1 the selection of extracting method
Method one is got QIANLIETAI JIAONANG content 2g, put in the 100ml tool plug triangular flask,, filter with chloroform (containing 1% ammonia) solution 50ml supersound extraction 20 minutes, discard chloroform liquid, residue volatilizes, and accurate chloroform methanol (its volume ratio is 7:3) the mixed solution 50ml that adds claims to decide weight, supersound process 15 minutes, put coldly, claim to decide weight, supply the weight of loss with above-mentioned chloroform methanol mixed solution, shake up, filter, get filtrate 25ml, water-bath volatilizes, residue quantitatively is transferred in the 25ml measuring bottle with 0.01% potassium hydroxide solution and is diluted to scale, shake up, promptly get need testing solution one, make negative sample solution with method.
Method two is got QIANLIETAI JIAONANG content 2g, put in the 100ml tool plug round-bottomed bottle, use ethyl acetate solution 50ml reflux, extract, 30 minutes, filter, discard acetic acid ethyl fluid, residue volatilizes, and the accurate dehydrated alcohol 50ml that adds claims to decide weight, reflux 2 hours, put coldly, claim to decide weight, supply the weight of loss with ethanol solution, shake up, filter, get filtrate 25ml, water-bath volatilizes, residue quantitatively is transferred in the 25ml measuring bottle with 0.01% potassium hydroxide solution and is diluted to scale, shake up, promptly get need testing solution two, make negative sample solution with method.
Method three is got QIANLIETAI JIAONANG content 2g, put in the 100ml apparatus,Soxhlet's, with the about 50ml of ethyl acetate solution, heating and refluxing extraction 2 hours, filter, discard acetic acid ethyl fluid, residue volatilizes, add the about 40ml of methanol solution, heating and refluxing extraction 2 hours, extracting solution, shakes up to 50ml with methanol constant volume, the accurate extracting solution 25ml that draws, water-bath volatilizes, and residue quantitatively is transferred in the 25ml measuring bottle with 0.01% potassium hydroxide solution and is diluted to scale, shakes up, promptly get need testing solution three, make negative sample solution with method.
Accurate respectively above-mentioned need testing solution one, two, three and each 1ml of negative sample solution thereof of drawing, place 10ml tool plug reaction bulb respectively, the accurate acetonitrile solution 3ml that contains bromo bromination 1-Phenylethanone. 0.5mg/ml and hexaoxacyclooctadecane-6-60.05mg/ml, the mixing of adding, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, and is cooled to room temperature, add acetonitrile and be settled to 10ml, shake up, filter, obtain need testing solution and corresponding negative control solution respectively with microporous filter membrane (0.45 μ m).
According to aforementioned chromatographic condition (chromatographic column: ZORBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 60:40; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves), the derivative solution of need testing solution one, two, three and negative sample thereof is injected chromatograph of liquid, measure content, the results are shown in Table 2
The various extracting method result of the test of table 2 table
Annotate: "+" is illustrated in the corresponding retention time of stachydrine hydrochloride derivant chromatographic peak place absorption; "-" is illustrated in the corresponding retention time of stachydrine hydrochloride derivant chromatographic peak place does not have absorption.
As seen from the above table, press extracting method one, two, three handled samples, its negative sample has interference, therefore, does not select this three kinds of extracting method for use.
Cause the interferential analysis of causes and solution: cause that interferential reason may be owing to contain the material that can react with derivatization reagent in the negative sample, its molecular structure is similar to stachydrine hydrochloride, after it and derivatization reagent react, the molecular weight of gained product is bigger, and it is approaching with the molecular weight of stachydrine hydrochloride derivative reaction product, therefore on chromatographic column, be difficult for the two is separated, cause false positive, thereby disturb assay.We once adjusted by regulating means such as mobile phase, change pre-treating method, and the result does not all have obviously improvement.
Therefore, be this character of both sexes alkaloid according to stachydrine hydrochloride, adopt the Reinecke salt sedimentation method to carry out remove impurity, its principle is as follows:
B
++NH
4[Cr(SCN)
4(NH3)
2]→B[Cr(SCN)
4(NH3)
2]↓+NH
4 +
2B[Cr(SCN)
4(NH
3)
2]↓+Ag
2SO
4→2Ag[Cr(SCN)
4(NH
3)
2]↓+B
2SO
4
B
2SO
4+BaCl→2BCl+BaSO
4↓
Wherein, B=quartermary ammonium alkaloids cation
Test as follows:
Method four is got QIANLIETAI JIAONANG content 3g, put in the 100ml tool plug triangular flask, with chloroform (containing 1% ammonia) solution 50ml supersound extraction 20 minutes, filter, discard chloroform liquid, residue volatilizes, and accurate chloroform methanol (its volume ratio is 7:3) the mixed solution 50ml that adds claims to decide weight, supersound process 15 minutes, put coldly, claim to decide weight, supply the weight of loss with above-mentioned chloroform methanol mixed solution, shake up, filter, get filtrate 25ml, water-bath volatilizes, residue adds 1% hydrochloric acid solution 20ml gradation dissolving, filter, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, puts in the ice bath and places 0.5 hour, filter, with frozen water gradation washing container and precipitation, discard filtrate, precipitation is with the 15ml acetone solution, dripping 0.5% silver sulfate solution in acetone soln produces to no longer including precipitation, place, filter, with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put and be concentrated into about 2ml in the water-bath, put coldly, add 1% barium chloride solution with the silver sulfate equivalent, potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shake up, promptly get need testing solution four, make negative sample solution with method.
Method five is got this product content 3.5g, put in the 100ml tool plug round-bottomed bottle, with chloroform (containing 1% ammonia) solution 50ml reflux, extract, 1.5 hours, filter, discard chloroform liquid, residue volatilizes, and accurate chloroform methanol (its volume ratio is 7:3) the mixed solution 50ml that adds claims to decide weight, reflux 2 hours, put coldly, claim to decide weight, supply the weight of loss with above-mentioned chloroform methanol mixed solution, shake up, filter, get filtrate 25ml, water-bath volatilizes, residue adds 1% hydrochloric acid solution 20ml gradation dissolving, filter, filtrate adds freshly prepared 2% sulfur hydracid chromium ammonium salt solution 10ml, puts in the ice bath and places 0.5 hour, filter, with frozen water gradation washing container and precipitation, discard filtrate, precipitation is with the 15ml acetone solution, dripping 0.5% silver sulfate solution in acetone soln produces to no longer including precipitation, place, natural filtration is with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put and be concentrated into about 2ml in the water-bath, put coldly, add 1% barium chloride solution with the silver sulfate equivalent, potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shake up, promptly get need testing solution five, make negative sample solution with method.
Method six is got this product content 4g, put in the 100ml apparatus,Soxhlet's, with an amount of heating and refluxing extraction of chloroform (containing 1% ammonia) solution 2 hours, filter, discard chloroform liquid, residue volatilizes, it is an amount of to add chloroform methanol (its volume ratio is 7:3) mixed solution, heating and refluxing extraction 2 hours, extracting solution is fixed molten to 50ml with above-mentioned chloroform methanol mixed solution, shake up, the accurate extracting solution 25ml that draws, water-bath volatilizes, residue adds 1% hydrochloric acid solution 20ml gradation dissolving, filter, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, puts in the ice bath and places 0.5 hour, filter, with frozen water gradation washing container and precipitation, discard filtrate, precipitate with the 15ml acetone solution, dripping 0.5% silver sulfate solution at acetone soln produces to no longer including precipitation, place, natural filtration is with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put and be concentrated into about 2ml in the water-bath, put coldly, add 1% barium chloride solution with the silver sulfate equivalent, potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shake up, promptly get need testing solution six, make negative sample solution with method.
Accurate respectively above-mentioned three kinds of need testing solutions and each 4ml of negative sample solution thereof of drawing, handle need testing solution four, five, six and corresponding negative sample solution according to the method for handling need testing solution one, two, three and negative sample solution thereof, get need testing solution and corresponding negative control solution.
According to aforementioned chromatographic condition (chromatographic column: ZORBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 60:40; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves), the derivative solution of need testing solution four, five, six and negative sample thereof is injected chromatograph of liquid, measures content, the results are shown in Table 3:
The various extracting method result of the test of table 3 table
Annotate: "+" is illustrated in the corresponding retention time of stachydrine hydrochloride derivant chromatographic peak place absorption; "-" is illustrated in the corresponding retention time of stachydrine hydrochloride derivant chromatographic peak place does not have absorption.
As seen from the above table, extracting method four, five, six handled samples are having chromatographic peak corresponding with it with the corresponding retention time of stachydrine hydrochloride derivant chromatographic peak place, and negative noiseless, so extracting method four, five, six all can be used as the extracting method of this product.Consider that extracting method four is more easy than method five and method six, extraction effect is more or less the same, and therefore, finally determines the extracting method of extracting method four as this product.
5.2 extraction time is investigated
By 5.1 following methods four, get 3 parts of QIANLIETAI JIAONANG contents (lot number: 20040301), every part of 3g carries out supersound extraction, extracts respectively 1 time, 2 times, 3 times, each 10 minutes, the results are shown in Table 4:
Table 4 extraction time is investigated table as a result
As seen from the above table, extract the stachydrine hydrochloride that just can extract fully in this product for 1 time.
5.3 extraction time is investigated
By 5.1 following methods four, get 3 parts of QIANLIETAI JIAONANG contents (lot number: 20040301), every part of 3g carries out supersound extraction, and extraction time was respectively 15 minutes, 20 minutes, 25 minutes, the results are shown in Table 5:
Table 5 extraction time investigation is table as a result
As seen from the above table, extract and to extract stachydrine hydrochloride in this product fully in 20 minutes, so extraction time is defined as 20 minutes.
6, the selection of derivative reaction condition
6.1 reaction principle
6.2 the selection of derivatization reagent
To using derivatization reagent bromination 1-Phenylethanone. always, bromo bromination 1-Phenylethanone. being investigated, its method is as follows: get 3 10mL tool plug reaction bulbs, numbering A～C, each accurate stachydrine hydrochloride reference substance solution 1mL (the KOH solution preparation with 0.01%) that adds 1mg/ml adds acetonitrile 3mL in the A bottle; Add the acetonitrile solution 3mL that contains bromination 1-Phenylethanone. 0.5mg/ml and hexaoxacyclooctadecane-6-60.05mg/ml in the B bottle; Add the acetonitrile 3mL that contains bromo bromination 1-Phenylethanone. 0.5mg/ml and hexaoxacyclooctadecane-6-60.05mg/ml in the C bottle.A is a blank, and B, C are experiment liquid.A, B, C solution are put into 80 ℃ of water-baths to be heated, the capillary tube 2 μ l that sample during respectively at 10,20 and 30 minutes, point sample launches on same silica gel g thin-layer plate, with the reference substance correspondence position on check and have or not the stachydrine hydrochloride speckle, judge the extent of reaction with this, the results are shown in Table 6:
Two kinds of derivatization reagent derivative reactions of table 6 result is (n=3) relatively
Annotate: "+" expression detects stachydrine hydrochloride in the table; "-" expression does not detect stachydrine hydrochloride.
As shown in Table 6, under the same conditions, use comparatively fast to bromo bromination 1-Phenylethanone. derivative reaction speed, 20min promptly reacts completely, and is the derivatization agent of this method so select it.
6.3 the selection of derivative reaction condition (pH value, temperature)
Get 10 of 10mL tool plug reaction bulbs, numbering 1～10, among each accurate KOH solution 1mL that adds hydrochloric stachydrine reference substance 1mg/ml, 1～No. 5 reaction bulb each accurate add 0.5mg/ml to the acetonitrile solution 1.5ml of the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml as experiment liquid; Respectively add acetonitrile 3mL (not containing derivatization reagent) liquid in contrast in 6～No. 10 reaction bulbs.In 80 ℃ of water-baths behind the reacting by heating 20min, be cooled to room temperature, the regularly accurate experiment liquid of drawing, each 2 μ l of contrast liquid, point sample is on same silica gel g thin-layer plate respectively, with n-butyl alcohol: hydrochloric acid: ethyl acetate (4:0.5:0.5) is developing solvent, rare bismuth potassium iodide test solution is a developer, scan by chromatography that (λ s is 510nm, λ R is 700nm), experiments of measuring liquid and contrast liquid stachydrine hydrochloride speck area integrated value, to test the percentage rate that liquid and contrast liquid stachydrine speck area integrated value ratio R (%) are finished as derivative reaction, judge the degree that derivative reaction carries out with this, then 100%-R is as the uncompleted percentage rate of derivative reaction, and data see Table 7:
The uncompleted percentage rate of derivative reaction (n=5) under the different basicity conditions of table 7
By table 7 and heat time heating time of determining in conjunction with the front, select the condition of derivative reaction to be: 0.01% KOH solution, 80 ℃ of water-baths, heating 20min.
7, linear relationship is investigated
Precision is measured reference substance solution 1ml, place 5ml tool plug reaction bulb, the accurate hexaoxacyclooctadecane-6-6 acetonitrile solution 1.5ml that adds 0.5mg/ml, mixing to bromo bromination 1-Phenylethanone. acetonitrile solution 1.5ml and 0.5mg/ml, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, and is cooled to room temperature, add acetonitrile to scale, shake up, filter, obtain the reference substance derivative solution with microporous filter membrane (0.45 μ m).
The above-mentioned reference substance derivative solution 1 μ l of accurate respectively absorption, 2 μ l, 4 μ l, 6 μ l, 8 μ l, 10 μ l inject high performance liquid chromatograph, measure peak area by the described chromatographic condition of technical scheme, the record chromatogram, and measurement result sees Table 8.
Table 8 stachydrine hydrochloride reference substance derivative solution linear relationship investigation table
| Sequence number | Sample size (μ g) | Area value (mv.s) |
| 1 | 0.4195 | 666.6107 |
| 2 | 0.8390 | 1339.7097 |
| 3 | 1.6780 | 2756.3310 |
| 4 | 2.5170 | 4193.0879 |
| 5 | 3.3560 | 5554.3076 |
| 6 | 4.1950 | 6937.2690 |
With the peak area is that vertical coordinate, sample size (μ g) they are abscissa drawing standard curve, and linear equation is: Y=1666.4X-37.2, and correlation coefficient γ=0.9999, the equation of crossing initial point through match is: y=1654x, correlation coefficient γ=0.9999.With two formulas in peak area (A=1339.7097) substitution of a sample, relative deviation is 0.99% as a result, can think that intercept is zero, and available one point external standard method calculates content, and the result shows that stachydrine hydrochloride reference substance derivant is 0.4195～4.1950
μ g scope internal linear relation is good.
8. precision test
8.1 instrument precision test
Get stachydrine hydrochloride reference substance derivative solution 5 μ l, inject chromatograph of liquid, repeat 6 times, measure peak area, the record chromatogram, the result is as follows:
Table 9 Precision test result table
The result shows: the peak area relative deviation of stachydrine hydrochloride reference substance derivant illustrates that less than 2% this test precision is good.
8.2 replica test precision respectively takes by weighing 6 parts of this product contents, handles by preceding method, makes need testing solution, sample introduction 10 μ l test, and measure peak area, the record chromatogram, and measurement result is as follows:
Table 10 replica test is table as a result
The result shows: the content relative standard of stachydrine hydrochloride reference substance derivant is less than normal in 2%, illustrates that this test repeatability is good.
8.3 middle precision is tested us and has been investigated the not assay result of same date, different analyst, distinct device, the results are shown in Table 11
Precision test result in the middle of table 11 preparation
The above results shows that the middle precision of this content assaying method is good.
9. accuracy test
Adopt the application of sample recovery test to measure, get 9 parts of the same batch samples (lot number: 20040301, the stachydrine hydrochloride average content is 0.546%) of known content, each 1g, the accurate title, decide, and presses contained stachydrine hydrochloride conversion in the sample, 1～3 part of stachydrine hydrochloride reference substance that adds 1:0.8 respectively; 4～6 parts of stachydrine hydrochloride reference substances that add 1:1 respectively; 7～9 parts of stachydrine hydrochloride reference substances that add 1:1.2 respectively.Be prepared into need testing solution by the need testing solution preparation method, according to above-mentioned chromatographic condition, sample introduction, mensuration, the record chromatogram calculates content, asks relative standard deviation, and the result shows that the method has average recovery preferably, and accuracy is good, the results are shown in Table 12.
Table 12 preparation application of sample recovery test result
10, serviceability test
It is an amount of that 10.1 the study on the stability precision of detected solution takes by weighing the QIANLIETAI JIAONANG content, the preparation need testing solution, with need testing solution put place 0,2,4,6,8,24 hour respectively under the room temperature after, draw the 10ul sample introduction, measure peak area, the record chromatogram, measurement result is as follows:
Table 13 solution stability testing is table as a result
The result shows: the peak area relative deviation of stachydrine hydrochloride reference substance derivant is less than 2%, illustrates that this product has good stable in following 24 hours of the room temperature.
10.2 we have compared the chromatographic column of four kinds of different models the comparison of different chromatographic columns, measurement result is as follows:
The different chromatographic columns of table 14 are investigated table as a result
10.3 we have compared the influence of three kinds of different column temperatures to measurement result different column temperature comparisons, measurement result is as follows:
The different column temperatures of table 15 are investigated table as a result
10.4 we have compared the influence of three kinds of different in flow rate to measurement result the different in flow rate comparison, measurement result is as follows:
Table 16 different in flow rate is investigated table as a result
| Flow velocity | Separating degree | The average content of measuring (mg/ grain) | RSD(％) |
| 0.8ml/min | >5 | 2.47 | |
| 1.0ml/min | >5 | 2.45 | 0.8 |
| 1.2ml/min | >5 | 2.46 |
10.5 we have compared three kinds of mobile relative determination results' of difference influence the comparison that the mobile phase proportion of composing changes, measurement result is as follows:
The different mobile phases of table 17 are investigated table as a result
| Mobile phase | Separating degree | The average content of measuring (mg/ grain) | RSD(％) |
| Methanol-water (70:30) | >5 | 2.48 | |
| Methanol-water (65:35) | >5 | 2.46 | 1.7 |
| Methanol-water (60:40) | >5 | 2.45 |
10.6 we have compared the influence of three kinds of different wave lengths to measurement result different comparisons that detect wavelength, measurement result is as follows:
The different wavelength that detect of table 18 are investigated table as a result
| Detect wavelength | Separating degree | The average content of measuring (mg/ grain) | RSD(％) |
| 255nm | >5 | 2.46 | |
| 260nm | >5 | 2.45 | 0.6 |
| 265nm | >5 | 2.46 |
Compared with prior art, method of quality control of the present invention is that reasonably measurement result is stable, adopts method of quality control of the present invention can effectively control the quality of QIANLIETAI JIAONANG, thereby guarantees the clinical efficacy of QIANLIETAI JIAONANG.
The specific embodiment
The method of quality control of QIANLIETAI JIAONANG comprises Determination of Contents of Hydrochloric Stachydrine assay method in crude drug Herba Leonuri, QIANLIETAI JIAONANG intermediate and the QIANLIETAI JIAONANG, and is specific as follows:
The Determination of Contents of Hydrochloric Stachydrine assay method is in the crude drug Herba Leonuri:
Chromatographic condition chromatographic column: ZORBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 250～270nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;
The Herba Leonuri medical material is got in the preparation of need testing solution, pulverize, cross 40 mesh sieves, precision takes by weighing 1g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put coldly, claims to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shake up, filter, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filters, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, put in the ice bath and placed 0.5 hour, filter, with frozen water washing container and precipitation, discard filtrate, with 15ml acetone solution precipitation, the silver sulfate solution of dropping 0.5% produces to no longer including precipitation in acetone soln, places, filter, precipitation merges washing liquid and filtrate with 70% ethanol 15ml gradation washing, puts in 90 ℃ of water-baths and is concentrated into 2ml, put cold, 1% barium chloride solution of adding and silver sulfate equivalent, the potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shakes up, filter, promptly get need testing solution;
The algoscopy precision is measured need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/ml, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
Determination of Contents of Hydrochloric Stachydrine is not less than 0.50% in the crude drug Herba Leonuri.
The Determination of Contents of Hydrochloric Stachydrine assay method is in the QIANLIETAI JIAONANG intermediate:
Chromatographic condition chromatographic column: ZORBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;
The QIANLIETAI JIAONANG intermediate is got in the preparation of need testing solution, pulverize, cross 40 mesh sieves, precision takes by weighing 1.5g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put cold, claim to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shakes up, and filters, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filter, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, puts in the ice bath and places 0.5 hour, filters, discard filtrate, with frozen water washing container and precipitation, discard filtrate, precipitate with the 15ml acetone solution, dripping 0.5% silver sulfate solution in acetone soln produces to no longer including precipitation, place, filter, with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put in 90 ℃ of water-baths and be concentrated into 2ml, put coldly, add 1% barium chloride solution with the silver sulfate equivalent, potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shake up, filter, promptly get need testing solution;
The algoscopy precision is measured need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/l, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
Determination of Contents of Hydrochloric Stachydrine is not less than 0.60% in the QIANLIETAI JIAONANG intermediate.
The Determination of Contents of Hydrochloric Stachydrine assay method is in the QIANLIETAI JIAONANG:
Chromatographic condition chromatographic column: Z0RBAX SB-C
18Post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;
20 of QIANLIETAI JIAONANG are got in the preparation of need testing solution, incline and get content, porphyrize, precision takes by weighing 2g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put coldly, claims to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shake up, filter, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filters, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, put in the ice bath and placed 0.5 hour, filter, with frozen water gradation washing container and precipitation, discard filtrate, with 15ml acetone solution precipitation, the silver sulfate solution of dropping 0.5% produces to no longer including precipitation in acetone soln, places, filter, with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put in 90 ℃ of water-baths and be concentrated into 2ml, put cold, 1% barium chloride solution of adding and silver sulfate equivalent, the potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shakes up, filter, promptly get need testing solution;
The algoscopy precision is measured above-mentioned need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/ml, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
Determination of Contents of Hydrochloric Stachydrine is not less than 0.50% in the QIANLIETAI JIAONANG.
Claims (7)
- The method of quality control of [claim 1] a kind of QIANLIETAI JIAONANG is characterized in that: this method of quality control comprises Determination of Contents of Hydrochloric Stachydrine assay method in crude drug Herba Leonuri, QIANLIETAI JIAONANG intermediate and the QIANLIETAI JIAONANG.
- The method of quality control of [claim 2] QIANLIETAI JIAONANG according to claim 1 is characterized in that: the Determination of Contents of Hydrochloric Stachydrine assay method is in the described crude drug Herba Leonuri:Chromatographic condition chromatographic column: ZORBAX SB-C18 post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 250～270nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;The Herba Leonuri medical material is got in the preparation of need testing solution, pulverize, cross 40 mesh sieves, precision takes by weighing 1g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put coldly, claims to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shake up, filter, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filters, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, put in the ice bath and placed 0.5 hour, filter, with frozen water washing container and precipitation, discard filtrate, with 15ml acetone solution precipitation, the silver sulfate solution of dropping 0.5% produces to no longer including precipitation in acetone soln, places, filter, precipitation merges washing liquid and filtrate with 70% ethanol 15ml gradation washing, puts in 90 ℃ of water-baths and is concentrated into 2ml, put cold, 1% barium chloride solution of adding and silver sulfate equivalent, the potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shakes up, filter, promptly get need testing solution;The algoscopy precision is measured need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/ml, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
- [claim 3] is characterized in that according to the method for quality control of the described QIANLIETAI JIAONANG of claim 2: Determination of Contents of Hydrochloric Stachydrine is not less than 0.50% in the crude drug Herba Leonuri.
- The method of quality control of [claim 4] QIANLIETAI JIAONANG according to claim 1 is characterized in that: the Determination of Contents of Hydrochloric Stachydrine assay method is in the described QIANLIETAI JIAONANG intermediate:Chromatographic condition chromatographic column: ZORBAX SB-C18 post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;The QIANLIETAI JIAONANG intermediate is got in the preparation of need testing solution, pulverize, cross 40 mesh sieves, precision takes by weighing 1.5g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put cold, claim to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shakes up, and filters, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filter, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, puts in the ice bath and places 0.5 hour, filters, discard filtrate, with frozen water washing container and precipitation, discard filtrate, precipitate with the 15ml acetone solution, dripping 0.5% silver sulfate solution in acetone soln produces to no longer including precipitation, place, filter, with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put in 90 ℃ of water-baths and be concentrated into 2ml, put coldly, add 1% barium chloride solution with the silver sulfate equivalent, potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shake up, filter, promptly get need testing solution;The algoscopy precision is measured need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/l, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
- [claim 5] is characterized in that according to the method for quality control of the described QIANLIETAI JIAONANG of claim 4: Determination of Contents of Hydrochloric Stachydrine is not less than 0.60% in the QIANLIETAI JIAONANG intermediate.
- The method of quality control of [claim 6] QIANLIETAI JIAONANG according to claim 1 is characterized in that: the Determination of Contents of Hydrochloric Stachydrine assay method is in the described QIANLIETAI JIAONANG:Chromatographic condition chromatographic column: ZORBAX SB-C18 post, 5 μ m, 4.6 * 250mm; Mobile phase: volume ratio is the methanol-water of 6:4; Temperature: 25～35 ℃; Detect wavelength: 260nm; Flow velocity: 1ml/min; Detector: DAD detector, the scanning of 190～400 all-waves;20 of QIANLIETAI JIAONANG are got in the preparation of need testing solution, incline and get content, porphyrize, precision takes by weighing 2g, put in the 100ml tool plug triangular flask, with the chloroform soln 50ml supersound extraction that contains 1% ammonia 30 minutes, filter, discard chloroform liquid, residue volatilizes the accurate chloroform that adds in back: the volume ratio of methanol is the mixed solution 50ml of 7:3, claim to decide weight, supersound process 20 minutes is put coldly, claims to decide weight, use chloroform: the volume ratio of methanol is the weight that the mixed solution of 7:3 is supplied loss, shake up, filter, the accurate filtrate 35ml that draws, put in the evaporating dish, 90 ℃ of water-baths volatilize, and residue adds 1% hydrochloric acid solution 20ml dissolving, filters, filtrate adds 2% sulfur hydracid chromium ammonium salt solution 10ml of new preparation, put in the ice bath and placed 0.5 hour, filter, with frozen water gradation washing container and precipitation, discard filtrate, with 15ml acetone solution precipitation, the silver sulfate solution of dropping 0.5% produces to no longer including precipitation in acetone soln, places, filter, with 70% ethanol 15ml gradation washing precipitation, merge washing liquid and filtrate, put in 90 ℃ of water-baths and be concentrated into 2ml, put cold, 1% barium chloride solution of adding and silver sulfate equivalent, the potassium hydroxide solution with 0.01% quantitatively is transferred in the 10ml measuring bottle and is diluted to scale, shakes up, filter, promptly get need testing solution;The algoscopy precision is measured above-mentioned need testing solution 4ml, places 10ml tool plug reaction bulb, the accurate acetonitrile solution 1.5ml to the hexaoxacyclooctadecane-6-6 of the acetonitrile solution 1.5ml of bromo bromination 1-Phenylethanone. and 0.05mg/ml that adds 0.5mg/ml, mixing, close plug, heating is 20 minutes in 80 ℃ of water-baths, takes out, be cooled to room temperature, add acetonitrile to 10ml, shake up, the microporous filter membrane with 0.45 μ m before the sample introduction filters, get filtrate and under above-mentioned chromatographic condition, carry out the HPLC analysis, calculate content with one point external standard method.
- [claim 7] is characterized in that according to the method for quality control of the described QIANLIETAI JIAONANG of claim 6: Determination of Contents of Hydrochloric Stachydrine is not less than 0.50% in the QIANLIETAI JIAONANG.
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Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1973897B (en) * | 2006-12-21 | 2010-04-14 | 贵州益佰制药股份有限公司 | Quality control method for puerperal blood clot dispersing tablet |
-
2007
- 2007-11-12 CN CN2007102024975A patent/CN101433582B/en active Active
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103245753A (en) * | 2013-04-25 | 2013-08-14 | 昆明中药厂有限公司 | Mass detection method for motherwort grains |
| CN104535707A (en) * | 2014-12-31 | 2015-04-22 | 贵州百灵企业集团制药股份有限公司 | Detection method for Qianlietai capsules |
| CN109655565A (en) * | 2019-01-07 | 2019-04-19 | 南京海昌中药集团有限公司 | The fingerprint atlas detection method of yimucao paste |
| CN109655565B (en) * | 2019-01-07 | 2020-10-30 | 南京海昌中药集团有限公司 | Fingerprint spectrum detection method of motherwort herb paste |
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| WO2009062368A1 (en) | 2009-05-22 |
| CN101433582B (en) | 2012-01-11 |
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