WO2009062368A1

WO2009062368A1 - A process of quality control for a capsule of treatment for prostatitis

Info

Publication number: WO2009062368A1
Application number: PCT/CN2008/000737
Authority: WO
Inventors: Xin Zhou; Wen Xia; Huaguo Chen; Xing Li; Chao Zhao; Xiaojian Gong
Original assignee: Guizhou Bailing Enterprise Group Pharmaceutical Co., Inc.
Priority date: 2007-11-12
Filing date: 2008-04-10
Publication date: 2009-05-22
Also published as: CN101433582A; CN101433582B

Abstract

A process of quality control for a capsule of treatment for prostatitis comprising assaying of stachydrine hydrochloride in Herba Leonuri Japonici as a raw material, in the intermediates of the capsule and in the capsule content.

Description

Quality control method for capsules for treating prostatitis

The invention relates to a quality control method for a traditional Chinese medicine preparation for treating prostatitis, in particular to a quality control method for a capsule preparation of the traditional Chinese medicine preparation.

Background technique

With the continuous development of social economy, the improvement of people's living standards and the aging of the population, the incidence of prostatitis has been increasing, and there is a trend of rejuvenation. Drug research for treating prostate diseases has become a hot spot for new drug research. Prostatitis is a common disease in men. The vast majority occur in young and middle-aged patients. Clinically, it can be divided into acute and chronic. Acute prostatitis is rare in clinical practice. Chronic prostatitis has a high incidence in the adult population, accounting for about one-fifth of the urological outpatients. Because chronic prostatitis is accompanied by seminal vesiculitis, it is also called prostatic vesiculitis. .

Causes Acute prostatitis: When the onset is tired, cold, prolonged cycling, alcohol abuse, excessive sexual life, injury, transurethral device operation, weakened systemic or local resistance, pathogens are transported by lesions in other parts of the body. Or enter the prostate through the urethra, the main pathogens are Escherichia coli, Staphylococcus: Proteus and Streptococcus.

Chronic prostatitis: The cause is more complicated, and a small number of patients with acute prostatitis have not been completely cured. Most patients have not experienced a clear acute phase. The pathogenic microorganisms causing chronic prostatitis are mainly bacteria, followed by viruses, mycoplasma, chlamydia and other allergens. Excessive libido, prostate congestion, lower urinary tract obstruction, perineal compression, injury, inflammation of adjacent organs and prostate, and decreased systemic resistance may be one of the causes of chronic prostatitis, even the mental state of the patient. A factor that affects the severity of symptoms. In short, the cause of chronic prostatitis is complex, and it is likely that there are different causes at different times, or more than one pathogenic factor in the same period.

Treatment of acute prostatitis: general treatment of bed rest, drinking more water and laxatives. If the bladder irritation is severe, you can give an analgesic antispasmodic drug and hot water to relieve the symptoms. Antibacterial drugs can be selected from penicillin, streptomycin, ampicillin, cephalosporin, and xilixin. After acute symptomatic treatment and anti-inflammatory treatment of acute prostatitis, the symptoms often resolve within 1 to 2 weeks. If the symptoms do not improve or worse, the prostatic anal examination is more swollen and fluctuating. B-ultrasound can be seen in the formation of abscesses. If the pus is removed by perineal puncture, the perineal abscess should be opened and drained. Chronic prostatitis: The treatment of chronic prostatitis should generally be combined with general treatment, prostate massage, drug perfusion, urethral dilation, peri-prosthetic closure and antibiotics. However, general antibacterial drugs do not easily enter prostate tissue, which is one of the reasons for clinical difficulties.

Traditional Chinese medicine is used for dialectical treatment in the prevention and treatment of the above-mentioned diseases. In particular, traditional Chinese medicine preparations made of motherwort as main raw materials such as: Qianlietai Capsule and Qianlietai Tablet have exact curative effects.

Qianlietai Capsule is the variety that the applicant of the invention obtained the approval of the drug registration of the State Food and Drug Administration in August 2005 (drug standard number: YBZ15942005). Qianlietai Capsules are composed of medicinal herbs such as motherwort, saffron, safflower, rape bee pollen, Zhimu (salt fried) and cork (salt). The preparation method of the prostaglandin capsule intermediate mentioned in the present invention is as follows: the rapeseed bee pollen is broken by a high-speed pulverizer after being broken, passed through a 100-mesh sieve, and the fine powder is reserved; the motherwort, the essay, the safflower, the anemarrhena ( Salt fried) and Phellodendron (salted with salt) Add water to cook twice, each time for 2 hours, add 12 times the amount of water for the first time, add 10 times the amount of water for the second time, filter, combine the decoction, and concentrate the filtrate under reduced pressure. The relative density is 1. 33~1. 37 (50 °C thermal test) thick paste, dry under reduced pressure, pulverized, add rapeseed bee pollen and starch, mix well, use 85% ethanol (containing 3% soybean oil Granulation, 50~6CTC drying, that is, the preparation of Qianlietai capsule intermediate; the intermediate is filled into capsules, which is made into Qianlietai capsule

P Chinese medicine is mostly made up of a variety of raw materials, its composition is more complicated, and the quality control of medicine is also a big problem. Qianlietai Capsule is also prepared by compounding a variety of raw materials. In clinical application, there are defects of low quality standards and difficult to control. At present, there is no quality control method for more mature Qianlietai capsules. The quality control methods of the preparations were discussed and studied.

Summary of the invention

The technical problem to be solved by the present invention is to provide a quality control method for a prostaglandin capsule, which comprises a method for determining the content of stachysine hydrochloride in a raw material medicine motherwort, a prostaglandin capsule intermediate and a prostaglandin capsule, and improves the quality control standard. .

In order to solve the above technical problem, the present invention adopts the following technical solutions:

A method for quality control of Qianlietai capsule, comprising a method for determining the content of stachysine hydrochloride in the intermediates of the raw materials, motherwort, Qianlietai capsule and Qianlietai capsule.

The method for determining the content of stachydrine hydrochloride in the above-mentioned raw material drug motherwort is:

Chromatographic conditions: ZORBAX SB-C ₁₈ column, 5 μ πι, 4. 6 X 250 let; mobile phase: methanol-water with a volume ratio of 6:4; temperature: 25~35 ° C; detection wavelength: 250~ 270nm; flow rate: lml/min; detector: MD detector, 190~400 full-wave scan; The preparation of the test solution is taken from the mother bean material, crushed, passed through a 40 mesh sieve, accurately weighed lg, placed in a 100 ml stoppered flask, and ultrasonically extracted with 50 ml of a solution of 1% ammonia in chloroform for 30 minutes, filtered. Discard the chloroform solution, add the residue to the chloroform: 50 ml of the mixed solution of methanol in a volume ratio of 7:3, weigh the weight, sonicate for 20 minutes, let cool, weigh the weight, use chloroform : Mixing solution with methanol volume ratio of 7:3 to make up the lost weight, shake well, filter, carefully extract 35ml of filtrate, place in evaporating dish, dry in 90°C water bath, dissolve residue in 20ml of 1% hydrochloric acid solution, Filtration, the filtrate was added to the newly prepared 2% ammonium hydrogen sulphate solution lOrnl, placed in an ice bath for 0.5 hours, filtered, washed with ice water and the precipitate, the filtrate was discarded, and the precipitate was dissolved in 15 ml of acetone. Add 0.5% silver sulfate solution to the acetone solution until no more precipitation occurs, place, filter, precipitate with 10% ethanol in 15 ml, wash the mixture and filtrate, and concentrate on a water bath at 90 °C to 2 ml. , let cool, add an equivalent of 1% barium chloride solution with silver sulfate With 0.01% potassium hydroxide solution was quantitatively transferred to a 10ml volumetric flask and dilute to volume, shake, filtration, to obtain the test solution;

The acetonitrile solution of p-bromo brominated acetophenone 1. 5ml and 0.05 mg / ml, the acetonitrile solution of p-bromo bromide acetophenone was added in an amount of 1. 5 ml and 0. 05 mg / ml The 18-crown ether-6 acetonitrile solution 1. 5ml, mixed, sealed, heated in a water bath at 80 ° C for 20 minutes, taken out, cooled to room temperature, add acetonitrile to 10ml, shake well, use 0 before injection. The 45 μm microporous membrane was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The content of the stachysine hydrochloride in the raw material drug motherwort is not less than 0.50%.

The method for determining the content of stachydrine hydrochloride in the aforementioned prostaglandin capsule intermediate is - chromatographic chromatography column: ZORBAX SB-C _lfi column, 5 μ ηι, 4. 6 X 250 painting; mobile phase: volume ratio of 6:4 Methanol-water; Temperature: 25 to 35. C; Detection wavelength: 260 nm _; Flow rate: lml/min; Detector: DAD detector, 190~400 full-wave scan;

The preparation of the test solution was taken from the middle of the capsule of the Qianlietai capsule, crushed, passed through a 40 mesh sieve, and accurately weighed 1. 5 g, placed in a 100 ml stoppered flask, and ultrasonically extracted with 50 ml of a solution of 1% ammonia in a solution of trichloromethane. Minutes, filtered, discard the chloroform solution, and add the chloroform after the residue is evaporated. The volume ratio of methanol is 50 ml of the mixed solution of 7·3, weighed, sonicated for 20 minutes, let cool, weigh Set the weight, use chloroform: methanol volume ratio of 7: 3 mixed solution to make up the lost weight, shake, filter, precision draw 35ml of filtrate, view the evaporating dish, 90 Ό water bath to dry, residue plus 1% hydrochloric acid The solution was dissolved in 20 ml, filtered, and the filtrate was added with 10 ml of a freshly prepared 2% ammonium hydrogen sulphate solution. The mixture was placed in an ice bath for 0.5 hr, filtered, and the filtrate was discarded. The vessel was washed with ice water and the filtrate was discarded. The precipitate was dissolved in 15 ml of acetone, and 0.5% of silver sulfate solution was added dropwise to the acetone solution until no more precipitate was formed. The mixture was filtered, and the precipitate was washed with 15 ml of 70% ethanol, and the washing liquid and the filtrate were combined. Concentrate to 2 ml on a °C water bath, let cool, add an equivalent of 1 ° / with barium sulfate. The ruthenium chloride solution was quantitatively transferred to a 10 ml volumetric flask with a 0.01% potassium hydroxide solution and diluted to the mark, shaken, and filtered to obtain a test solution; the measurement method accurately measured the sample solution 4 ml. 5毫升和0. 05 mg / ml of 18-crown ether-6 in acetonitrile solution, placed in a 10 ml stoppered reaction flask, precisely added 0. 5 mg / 1 of bromo bromide acetophenone acetonitrile solution 1. 5ml and 0. 05 mg / ml of 18-crown ether-6 acetonitrile solution 1. 5ml, mixed, condensed, heated in a water bath at 80 ° C for 20 minutes, taken out, cooled to room temperature, acetonitrile was added to 10 ml, shaken, filtered through a microporous membrane of 0. 45 μ m before injection. The filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The content of the stachysine hydrochloride in the intermediate of the prostaglandin capsule is not less than 0.60%.

The method for determining the content of stachydrine hydrochloride in the aforementioned Qianlietai capsule is as follows:

Chromatographic conditions: ZORBAX SB-C ₁₈ column, 5 μ πι, 4. 6 X 250 draw; Mobile phase: Methanol-water in a volume ratio of 6:4; Temperature: 25~35 ° C; Detection wavelength: 260 nm ; Flow rate: lml/min; Detector: DAD detector, 190~400 full-wave scan;

Preparation of the test solution Take 20 capsules of Qianlietai Capsule, pour the contents, grind finely, accurately weigh 2g, place it in a 100ml stoppered flask, and ultrasonically extract it with 50ml of 1% ammonia solution in chloroform solution for 30 minutes. , filtered, discarded the chloroform solution, the residue is added to the chloroform after the evaporation: the volume ratio of methanol is 7:3, 50ml, weighed, sonicated for 20 minutes, let cool, weighed Use the mixed solution of trichloromethane:methanol in a volume ratio of 7:3 to make up the lost weight, shake it, filter it, carefully extract 35ml of the filtrate, place it in the evaporating dish, spin in a 90'C water bath, add 1% residue. 20 ml of hydrochloric acid solution was dissolved, filtered, and the filtrate was added with 10 ml of a freshly prepared 2% ammonium thiolumate solution, placed in an ice bath for 0.5 hours, filtered, ice-washed to wash the container and precipitate, and the filtrate was discarded to 15 ml. The precipitate was dissolved in acetone, and 0.5% of the silver sulfate solution was added dropwise to the acetone solution until no more precipitate was formed. The mixture was filtered, and the precipitate was washed with 15 ml of 70% ethanol, and the washing liquid and the filtrate were combined and placed at 90 ° C. Concentrate to 2ml on a water bath, let cool, add with sulfuric acid Silver equivalent of 1% cerium chloride solution, quantitatively transferred to a 10 ml volumetric flask with 0.01% potassium hydroxide solution and diluted to the mark, shaken, filtered, to obtain a test solution;

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1 ml of 18-crown ether-6 acetonitrile solution, mix well, conceal, heat in a water bath at 80 ° C for 20 minutes, remove, cool to room temperature, add acetonitrile to 10 ml, shake well, use 0 before injection The 45 μ ra microporous membrane was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The content of the stachysine hydrochloride in the Qianlietai capsule is not less than 0.50%.

In order to verify the rationality of the quality control method of the present invention, the applicant conducted an experimental study on the method. And screening, as follows:

1. Preparation of reference solution

Accurately weigh the amount of reference substance of hydrochloric acid stachysine dried to constant weight with phosphorus pentoxide, and make a solution of lmg of stachysine hydrochloride per ml of 0.01% potassium hydroxide solution.

2. Choice of mobile phase

The test was carried out according to the chromatographic conditions of the literature, and the mobile phase was selected to be methanol-water having a volume ratio of 60:40. 3 Detection wavelength selection

A solution of the staphylillin hydrochloride reference derivative 10 μl was injected into the liquid chromatograph and subjected to full-wave scanning in the range of 190 to 400. The result was maximum absorption at 260 nm, so 260 nm was selected as the detection wavelength.

4 system suitability test

4. 1 Confirmation of the reference product and the chromatographic peak of the sample - Prepare 0. 01% K0H solution, 0.5 nig/nil of bromophenyl bromide in acetonitrile and

0. 05mg/ml 18-crown ether-6 acetonitrile solution, according to the steps listed in Table 1, to obtain 4 different solutions a~d: Table 1 Preparation process of 4 different solutions

The four solutions prepared according to Table 1 were sonicated for 10 minutes, heated in a 80 Torr water bath for 20 minutes, taken out, cooled to room temperature, and injected into a liquid chromatograph for detection. From the results of the test, it is known that the chromatographic peak generated by the catalyst and the derivatizing reagent added to the derivatization reaction does not interfere with the chromatographic peak of the stachysine hydrochloride derivative, and the hydrosalazine hydrochloride derivative on the chromatogram is easily confirmed.

4. 2 Negative sample interference test

Taking the negative samples of the prostaglandin and the motherwort, the test sample was prepared according to the method 4 of the fifth method. It was found that after adding 2% ammonium chromate solution, the negative sample produced no precipitate, and the former column Thai capsule sample There is a clear precipitate. The test results show that: the negative sample has no interference.

4. 3 Resolution and number of plates:

The sample solution and the reference solution were separately injected, and the data report showed that: the resolution was greater than 1.5, completely separated from other impurity peaks; and the number of theoretical plates was greater than 4000, so the test The number of theoretical plates should be no less than 4,000 based on the chromatographic peak of the stachysine hydrochloride derivative.

5. Preparation of test solution

5. 1 Selection of extraction methods

Method 1 Take 2g of the content of Qianlietai Capsule, place it in a 100ml stoppered flask, and ultrasonically extract it with 50ml of chloroform (containing 1% ammonia) solution for 20 minutes, filter it, discard the solution of trichloromethane, and drain the residue. , precisely add chloroformic methanol (in a volume ratio of 7:3) 50ml mixed solution, weighed, sonicated for 15 minutes, let cool, weighed, and used the above mixture of trichloromethane and methanol to make up the lost weight. Shake well, filter, take 25ml of filtrate, drain in a water bath, the residue is quantitatively transferred to a 25ml volumetric flask with 0.1% potassium hydroxide solution and diluted to the mark. Shake well, then the test solution is obtained. Become a negative sample solution.

Method 2 Take 2g of the content of Qianlietai Capsule, place it in a 100ml round bottom bottle, and extract it with 50ml of ethyl acetate solution for 30 minutes. Filter it, discard the ethyl acetate solution, and drain the residue. Add 50ml of absolute ethanol. The sulphuric acid is oxidized by a water bath. The residue is oxidized by a water bath. The residue is oxidized with 0.01% by weight. The potassium solution was quantitatively transferred to a 25 ml volumetric flask and diluted to the mark. Shake well to obtain the test solution 2. The same method was used to prepare a negative sample solution.

Method 3 Take 2g of the content of Qianlietai Capsule, put it into a 100ml Soxhlet extractor, use about 50ml of ethyl acetate solution, extract it by heating and reflux for 2 hours, filter it, discard the ethyl acetate solution, discard the residue, add methanol solution. 40ml, heated and refluxed for 2 hours, the extract was made up to 50ml with methanol, shaken, accurately extract the extract 25ml, drained in a water bath, the residue was quantitatively transferred to a 25ml volumetric flask and diluted with 0.01% potassium hydroxide solution. To the scale, shake well, that is, the test solution is obtained, and the negative sample solution is prepared by the same method.

5重量/毫升和18-, respectively, the above-mentioned sample solution, the first, second, third and the negative sample solution, respectively, 1 ml, respectively, placed in a 10 ml stoppered reaction bottle, precision addition of acetophenone bromide acetophenone 0. 5mg / ml and 18- Crown ether - 6 0. 05 mg / ml of acetonitrile solution 3ml, mix, tightly packed, heated in 80 ° water bath for 20 minutes, taken out, cooled to room temperature, add acetonitrile to a volume of 10ml, shake, to microporous membrane (0. 45 μ ιη) Filtered to obtain the test solution and the corresponding negative control solution, respectively.

According to the above chromatographic conditions (column: ZORBAX SB-C ₁₈ column, 5 μ πι, 4. 6 X 250 coffee; mobile phase: methanol-water with a volume ratio of 60:40; temperature: 25 to 35 ° C; detection wavelength : 260 nm _; flow rate: lml/min; Detector: DAD detector, 190~400 full-wave scan), inject the sample solution of the test solution one, two, three and its negative sample into the liquid chromatograph, determine the content, the results are shown in Table 2.

Table 2 Various extraction methods 'Test results table

NOTE - "+" indicates absorption at the corresponding retention time of the chromatographic peak of the stachysine hydrochloride derivative; "-" indicates no absorption at the corresponding retention time of the chromatographic peak of the stachysine hydrochloride derivative.

It can be seen from the above table that the samples processed according to the extraction methods one, two, and three have interferences in the negative samples, and therefore, the three extraction methods are not selected.

Cause analysis and solution for causing interference: The cause of the interference may be due to the fact that the negative sample contains a substance capable of reacting with the derivatizing reagent, and its molecular structure is similar to that of salicylate hydrochloride, which is reacted with the derivatizing reagent. The molecular weight of the reaction product is relatively large, and is close to the molecular weight of the reaction product of the stachysine hydrochloride derivatization reaction. Therefore, it is difficult to separate the two on the chromatographic column, resulting in a false positive, thereby interfering with the determination of the content. We have adjusted by adjusting the mobile phase and changing the pretreatment method, and the results have not been significantly improved.

Therefore, according to the nature of salicin hydrochloride as an amphoteric alkaloid, the precipitation by the precipitation method of Rayleigh salt is carried out, and the principle is as follows:

B ⁺ +NH„ [Cr (SCN) ₄ (NH3) ₂ ] — B [Cr (SCN) , (NH _;i ) ] I +NH

2B [Cr (SCN) , (NH ) ₂ ] I +Ag S0 ₄ →2Ag [Cr (SCN) , (NH ₃ ) ₂ ] I +B ₂ S0,

B ₂ S0 ₄ +BaCl ― 2BCl+BaS0 ₄ I

Wherein B = quaternary ammonium alkaloid cation

Test as follows:

Method 4: Take 3g of the content of Qianlietai Capsule, place it in a 100ml stoppered flask, ultrasonically extract it with 50ml of chloroform (containing 1% ammonia) solution for 20 minutes, filter it, discard the chloroform solution, and drain the residue. Precisely add 50ml of mixed solution of trichloromethane methanol (in a volume ratio of 7:3), weigh the weight, sonicate for 15 minutes, let cool, weigh the weight, make up the lost weight with the above chloroform methanol mixed solution, shake Uniform, filtered, take 25ml of filtrate, drain in a water bath, add 20% hydrochloric acid solution to the residue, dissolve in portions, filter, add new filtrate 10 ml of a 2% ammonium hydrogen sulphate solution was prepared, placed in an ice bath for 0.5 hours, filtered, and the container was washed with ice and water, and the filtrate was discarded. The precipitate was dissolved in 15 ml of acetone and added dropwise in acetone. 0. 5% silver sulfate solution until no more precipitation occurs, set, filter, wash the precipitate with 15ml of 70% ethanol, combine the washing liquid and filtrate, concentrate on water bath to about 2ml, let cool, add with silver sulfate An equivalent amount of 1% cesium chloride solution was quantitatively transferred to a 10 ml volumetric flask with a 0.01% potassium hydroxide solution and diluted to the mark. Shake well to obtain a test solution 4, and a negative sample solution was prepared in the same manner.

Method 5 Take the contents of this product 3. 5g, placed in a 100ml round bottom bottle, with a solution of trichloromethane (containing 1% ammonia) 50RAL reflux extraction for 1.5 hours, filtered, discarded chloroform solution The residue is evaporated, and the chloroformic methanol (volume ratio of 7:3) is precisely added to 50 ml of the mixed solution, and the weight is weighed, heated under reflux for 2 hours, allowed to cool, and weighed. The above chloroform methanol mixed solution is used to make up the loss. The weight, shake the hook, filter, take the filtrate 25ml, water bath to dry, the residue and 1% hydrochloric acid solution 20ral dissolved, filtered, the filtrate was added to freshly prepared 2% ammonium bisulfate solution 10ml, set the ice bath The mixture was placed for 0.5 hours, filtered, and the container was washed with ice and water, and the filtrate was discarded. The precipitate was dissolved in 15 ml of acetone, and 0.5% of silver sulfate solution was added dropwise to the acetone solution until no precipitation occurred. 0%, with a 1% cesium chloride solution equivalent to silver sulphate, 0. 01% Quantitative transfer of potassium hydroxide solution to 10m l Dilute to the mark in the measuring flask, shake it well, then get the test solution five, the same method to make a negative sample solution.

Method 6 Take 4g of the content of this product, put it in a 100ml Soxhlet extractor, extract it with a suitable amount of trichloromethane (containing 1% ammonia water) for 2 hours, filter it, discard the solution of trichloromethane, and slag Dry, add chloroform methanol (the volume ratio is 7:3), mix the appropriate amount of the solution, and heat to reflux for 2 hours. The extract is dissolved in 50 ml with the above chloroform methanol mixed solution, shake well, and accurately extract the extract 25 ml. The water was evaporated to dryness, and the residue was added with a 1% hydrochloric acid solution in 20 ml portions. The filtrate was filtered, and the filtrate was added to a freshly prepared solution of 2% ammonium hydrogen sulphate in 10 ml, placed in an ice bath for 0.5 hr. The washing container and the precipitate were washed, the filtrate was discarded, and the precipitate was dissolved in 15 ml of acetone. The 5% silver sulfate solution was added dropwise to the acetone solution until no more precipitate was formed, and the mixture was naturally filtered, and the precipitate was washed with 15 ml of 70% ethanol. The washing liquid and the filtrate were combined, concentrated to about 2 ml in a water bath, allowed to cool, and an equivalent of 1% cerium chloride solution with silver sulfate was added, and quantitatively transferred to a 10 ml volumetric flask with a 0.01% potassium hydroxide solution and diluted. To the scale, shake it, that is For the test solution, the same method is used to make a negative sample solution.

The 4 samples of the above three test solutions and their negative sample solutions are accurately taken separately, and the test solution is treated according to the method of treating the test solution one, two, three and the negative sample solution for the fourth, fifth, sixth and corresponding negative For the sample solution, the test solution and the corresponding negative control solution were obtained. According to the above chromatographic conditions (column: ZORBAX SB-CJ3⁄4, 5 μ πι, 4. 6 X 250 mm _; mobile phase: methanol-water with a volume ratio of 60:40; temperature: 25 to 35 ° C; detection wavelength: 260 nm _; Flow rate: Iml/min, detector: DAD detector, 190~400 full-wave scan), inject the derivative solution of the test solution four, five, six and its negative sample into the liquid chromatograph to determine the content, - The results are shown in Table 3 _:

Table 3 Test results of various extraction methods

Note: "+" indicates absorption at the corresponding retention time of the chromatographic peak of the stachysine hydrochloride derivative; "-" indicates no absorption at the corresponding retention time of the chromatographic peak of the stachysine hydrochloride derivative.

It can be seen from the above table that the samples processed in the fourth, fifth and sixth extraction methods have corresponding peaks at the retention time corresponding to the chromatographic peak of the stachysine hydrochloride derivative, and the negatives have no interference, so the extraction method is four or five. And six can be used as the extraction method of this product. Considering that extraction method 4 is simpler than method 5 and method 6, the extraction effect is not much different. Therefore, extraction method 4 is finally determined as the extraction method of this product.

5. 2 The number of extractions

According to the method 4 of item 5.1, take 3 parts of Qianlietai capsule (batch number: 20040301), each part is 3g, ultrasonic extraction, respectively, 1 time, 2 times, 3 times, each time 10 minutes, the results are shown in the table 4:

Number of extraction times

It can be seen from the above table that the extraction of stachydrine hydrochloride in the product can be completely extracted once.

5. 3 extraction time inspection

According to the method 4 in item 5.1, take 3 parts of Qianlietai Capsule (batch number: 20040301), and each part is 3g, ultrasonic extraction, the extraction time is 15 minutes, 20 minutes, 25 minutes respectively. The results are shown in Table 5:

It can be seen from the above table that the extraction of stachydrine hydrochloride in the product can be completely extracted in 20 minutes, so the extraction time is determined to be 20 minutes.

6. Selection of derivatization reaction conditions

6. 1 Reaction principle "Make (Talk - Male)

6. 2 Selection of derivatization reagents

The commonly used derivatization reagents brominated acetophenone and p-bromo acetophenone were investigated as follows: Take 3 10 mL stoppered reaction bottles, numbered A to C, and precisely add 1 mg/ml hydrochloric acid water. 5毫升/毫升和18-冠等-6 0. 05mg。 The lyobase reference solution 1mL (formed with 0. 01% K0H solution), A bottle of acetonitrile 3mL; B bottle containing brominated acetophenone 0. 5mg / ml and 18-crown ether - 6 0. 05mg 3 ml of acetonitrile solution of /ml; 3 ml of acetonitrile containing 0.5 mg/ml of p-bromophenyl acetophenone and 6 0. 05 mg/ml. A is a blank control, and B and C are experimental liquids. The A, B, and C solutions were heated in an 80-inch water bath, and the capillary was sampled at 2 μΐ at 10, 20, and 30 minutes, and spotted on the same silica gel G thin-layer plate, and examined at the position corresponding to the control. The presence or absence of stachysamine hydrochloride spots was used to judge the degree of reaction. The results are shown in Table 6:

Table 6 Comparison of Derivatization Results of Two Derivatization Reagents (η=3)

Note: "+" in the table indicates that stachydrine hydrochloride was detected; "-" indicates that stachydrine hydrochloride was not detected.

It can be seen from Table 6 that under the same conditions, the derivatization reaction rate of acetophenone bromide is faster, and the reaction is complete after 20 minutes, so it is selected as the derivatizing agent of the method.

6. 3 Selection of derivatization reaction conditions (pH, temperature)

5 mg/ml Each of the reaction flasks of the reaction flasks of the reaction volume of 1 to 5 was added to each well. 5毫升和0. 05mg/ml 18-crown ether-6 in acetonitrile solution 1. 5ral as a test solution; 6~10 reaction flasks each added acetonitrile 3raL (without derivatization reagent) as a control solution. After heating in a 80-inch water bath for 20 minutes, the mixture was cooled to room temperature, and the experimental solution and the control solution were accurately pipetted for 2 μM each, and each was spotted on the same silica gel G thin-layer plate to give n-butanol: hydrochloric acid: ethyl acetate (4 : 0. 5: 0. 5) is a developing agent, the cesium iodide test solution is a color developing agent, and is scanned by chromatography (λ s is 510 nm, λ R is 700 nm), and the test solution and the control solution hydrochloric acid water are measured. The integral value of the surface area of the sulphate base, the ratio of the integral value of the staphylline spot area of the test solution and the control solution, R (%), is used as the percentage of the completion of the derivatization reaction, thereby judging the extent of the derivatization reaction, and then 100%_R is derived. The percentage of unresolved reactions is shown in Table 7:

From Table 7 in combination with the previously determined heating time, the conditions for the derivatization reaction were selected: 0. 01% K0H solution, 80 ° C water bath, heating for 20 min.

7, linear relationship inspection

5毫升和毫升的18-冠ethers, and the solution of the bromo brominated acetophenone acetonitrile solution 1. 5ml and 0. 5mg / ml of 18-crown ether - 6 acetonitrile solution 1 · 5ml, mix, close the plug, heat in a water bath at 80 ° C for 20 minutes, remove, cool to room temperature, add acetonitrile to the mark, shake well, to microporous The filter (0.45 μm) was filtered to obtain a reference derivative solution.

The above reference substance derivative solutions 1μ 1 , 2μ1 , 4μ 1 , 6μ 1 , 8μ 1 , 10 μΐ were accurately injected into the high performance liquid chromatograph, and the peak area was determined according to the chromatographic conditions described in the technical scheme. The chromatogram was recorded. Table 8.

Study on linear relationship of solution of stachysine hydrochloride reference derivative

The standard curve is drawn with the peak area as the ordinate and the injection amount (yg) as the abscissa. The linear equation is - Y=1666.4X-37.2, the correlation coefficient Υ =0.9999, and the equation fitted to the origin is: y=1654x, The correlation coefficient is Y = 0.9999. Substituting the peak area of a sample (A=1339.7097) into the above two formulas, the relative deviation of the result is 0.99%. It can be considered that the intercept is zero. The content can be calculated by the external standard method. The results show that the reference substance of the stachysine hydrochloride is The linear relationship is good in the range of 0.4195 to 4.1950 μg.

8. Precision test

' 8.1 Instrument Precision Test

Take 5 μl of the staphylillin hydrochloride reference derivative solution, inject it into the liquid chromatograph, repeat 6 times, the peak area, and record the chromatogram. The results are as follows:

Table 9 Precision test results table

The results showed that the relative deviation of the peak area of the reference substance of the stachysine hydrochloride was less than 2%, indicating that the precision of the test was good.

8. 2 Repeatability test Weigh accurately the contents of this product by 6 parts, according to the previous method, to obtain the test solution, the injection ΙΟ μ Ι test, determine the peak area, record the chromatogram, the results are as follows:

Table 10 Repeatability test results table

The results showed that the content of the reference substance of the stachysine hydrochloride was less than 2% relative to the standard, indicating that the test has good reproducibility.

8. 3 Intermediate Precision Experiments We examined the results of intermediate precision tests for formulations with different dates, different analysts, and different equipment.

The above results show that the intermediate precision of the content determination method is good.

9. Accuracy test

Using the sample recovery test to determine the same batch of samples with the known content (batch number: 20040301, the average content of stacyl hydrochloride is 0. 546%) 9 parts, each lg, precision weighing, according to the hydrochloric acid containing water in the sample Alkali conversion, 1~3 parts respectively added 1: 0.8 hydrochloric acid stachysine reference substance; 4~6 parts respectively added 1:1 hydrochloric acid stachysine reference substance; 7~9 parts respectively added 1: 1. 2 The reference substance of stachysamine hydrochloride. According to the preparation method of the test solution, the test solution is prepared. According to the above chromatographic conditions, the sample is injected and measured, the chromatogram is recorded, the content is calculated, and the relative standard deviation is obtained. The result shows that the method has a good sample recovery rate. The accuracy is good, and the results are shown in Table 12.

Table 12 Test sample recovery test results

10, durability test

10. 1 Stability of the tested solution Accurately weigh the appropriate amount of Qianlietai Capsule, prepare the test solution, and place the test solution at room temperature for 0, 2, 4, 6, 8, 24 hours, Pipette lOul to inject the peak area and record the chromatogram. The results are as follows:

Table 13 Solution Stability Test Results Table

The results showed that: 'The relative deviation of the peak area of the reference substance of the stachysine hydrochloride was less than 2%, indicating that the product has good stability within 24 hours at room temperature.

10. 2 Comparison of different columns We compared four different types of columns and the results are as follows:

Table 14 Table of results of different columns

10. 3 Comparison of different column temperatures We compared the effects of three different column temperatures on the determination results. The results are as follows:

10. 4 Comparison of different flow rates We compared the effects of three different flow rates on the results of the measurements. The results are as follows:

Different flow rate inspection results table

10. 5 Comparison of mobile phase composition ratio changes We compared the effects of three different flow relative measurement results. The results are as follows -

Methanol-water (65: 35) >5 2. 46 1. 7 Methanol-water (60: 40) >5 2. 45

10. 6 Comparison of different detection wavelengths We compared the effects of three different wavelengths on the measurement results. The measurement results are as follows - the results of different detection wavelengths

Compared with the prior art, the quality control method of the invention is reasonable, and the measurement result is stable. The quality control method of the invention can effectively control the quality of the Qianlietai capsule, thereby ensuring the clinical efficacy of the Qianlietai capsule.

detailed description

The quality control method of Qianlietai Capsule includes the determination methods of the content of the raw material medicine motherwort, Qianlietai capsule intermediate and the prostaglandine hydrochloride staphylline hydrochloride, as follows:

The method for determining the content of stachydrine hydrochloride in the raw material motherwort is - chromatographic conditions column: ZORBAX SB-C _l8 column, 5 μ ηι, 4. 6 X 250 painting; mobile phase: methanol-water with a volume ratio of 6:4 ; Temperature: 25~35°C; Detection wavelength: 250~270nm _; Flow rate: lml/min; Detector: DAD detector, 190~400 full-wave scan;

The preparation of the test solution is taken from the motherwort, crushed, passed through a 40 mesh sieve, accurately weighed lg, placed in a 100 ml stoppered flask, and ultrasonically extracted with 50 ml of a chloroform solution containing 1% ammonia in water for 30 minutes, filtered, discarded. After removing the residue, the residue is evaporated and then added with chloroform: 50 ml of a mixed solution of methanol in a volume ratio of 7:3, weighed, sonicated for 20 minutes, allowed to cool, weighed, and used trichloromethane.垸: Mixing solution with methanol volume ratio of 7:3 to make up the lost weight, shake well, filter, accurately draw 35ml of filtrate, place in evaporating dish, float in 90°C water bath, dissolve residue with 1% hydrochloric acid solution 20ml After filtration, the filtrate was added with 10 ml of a freshly prepared 2% ammonium hydrogen sulphate solution, placed in an ice bath for 0.5 hours, filtered, washed with ice water and precipitated, and the filtrate was discarded, and the precipitate was dissolved in 15 ml of acetone. Add 0.5% silver sulfate solution to the acetone solution until no more precipitation occurs, place, filter, precipitate with 10% ethanol in 15 ml, wash the mixture and filtrate, and concentrate on a 90 ° C water bath. 2ml, let cool, add an equivalent of 1% barium chloride solution with silver sulfate, 0.01% potassium hydroxide solution Quantitative transfer to a 10ml volumetric flask and dilute to the mark, shake well, filter, then get the test solution;

The acetonitrile solution of bromo brominated acetophenone 1. 5ml and 0. 05 mg / ml, the acetonitrile solution of bromo brominated acetophenone was added in an amount of 0.5 ml / ml. 18-crown ether _6 acetonitrile solution 1. 5ml, mixed, densely packed, heated in a water bath at 80 ° C for 20 minutes, taken out, cooled to room temperature, add acetonitrile to 10ml, shake well, use 0 before injection. The 45 μm microporous membrane was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The determination method of the content of stacyl hydrochloride in the intermediate of Qianlietai capsule is - chromatographic conditions column: ZORBAX SB-C ₁₈ column, 5 μ πι, 4. 6 X 250; mobile phase: methanol with a volume ratio of 6:4 - water; temperature: 25~35 ° C; detection wavelength: 260 nm _; flow rate: lml/min; detector: DAD detector, 190~400 full-wave scan;

The preparation of the test solution was taken from the middle of the capsule of the Qianlietai capsule, crushed, passed through a 40 mesh sieve, and accurately weighed 1. 5 g, placed in a 100 ml stoppered flask, and ultrasonically extracted with 50 ml of a solution of 1% ammonia in a solution of trichloromethane. Minutes, filtered, discarded the chloroform solution, the residue was evaporated and then added with chloroform: 50 ml of a mixed solution of methanol in a volume ratio of 7:3, weighed, sonicated for 20 minutes, allowed to cool, weighed Weight, use chloroform: methanol volume ratio of 7: 3 mixed solution to make up the lost weight, shake, filter, precision draw 35ml of filtrate, placed in the evaporating dish, 9 (TC water bath, the residue plus 1% 20 ml of hydrochloric acid solution was dissolved, filtered, and the filtrate was added to a freshly prepared solution of 2% ammonium hydrogen sulphate 10 ml, placed in an ice bath for 0.5 hr, filtered, the filtrate was discarded, the container was washed with ice water and precipitated, discarded. The filtrate was dissolved in 15 ml of acetone, and 0.5% silver sulfate solution was added dropwise to the acetone solution until no more precipitate was formed. The mixture was filtered, and the precipitate was washed with 15 ml of 70% ethanol, and the washing liquid and the filtrate were combined. Concentrate to 2ml on a 90 °C water bath, let cool Adding an equivalent of 1% cerium chloride solution with silver sulfate, quantitatively transferring to a 10 ml volumetric flask with 0.01% potassium hydroxide solution and diluting to the mark, shaking the hook, filtering, and obtaining the test solution; The acetonitrile solution of p-bromo brominated acetophenone 1. 5 ml and 0. 05 mg / ml. The 18-crown ether _6 acetonitrile solution 1. 5ml, mixed hook, dense plug, heated in a water bath at 80 ° C for 20 minutes, taken out, cooled to room temperature, add acetonitrile to 10ml, shake well, use 0 before injection. The 45 m microporous membrane was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The determination method of stachydrine hydrochloride in Qianlietai capsule is - chromatographic conditions column: ZORBAX SB- C ₁₈ column, 5 m, 4. 6 X 250 诵; mobile phase: volume The ratio is 6··4 of methanol-water; Temperature: 25~35°C; Detection wavelength: 260 nm _; Flow rate: lml/mi _n; Detector: DAD detector, 190~400 full-wave scan;

Preparation of the test solution: Take 20 capsules of Qianlietai capsule, pour the contents, grind finely, accurately weigh 2g, place in a 100ml stoppered flask, and ultrasonically extract with 50ml of trichloromethane solution containing i% ammonia for 30 minutes. , filtered, discard the chloroform solution, and add the chloroform after the residue is evaporated. The volume ratio of methanol is 50 ml of the mixed solution of 7:3, weighed, sonicated for 20 minutes, allowed to cool, weighed, Use a mixed solution of chloroform:methanol in a volume ratio of 7:3 to make up the lost weight. Shake well, filter, finely and carefully take 35 ml of the filtrate, place in an evaporating dish, spin in a 90 °C water bath, add 1 residue. 20 ml of % hydrochloric acid solution was dissolved, filtered, and the filtrate was added with 10 ml of a freshly prepared 2% ammonium hydrogen sulphate solution, placed in an ice bath for 0, 5 hours, filtered, and the container was washed with ice and water, and the filtrate was discarded. The precipitate was dissolved in 15 ml of acetone, and 0.5% of a silver sulfate solution was added dropwise to the acetone solution until no more precipitate was formed. The mixture was filtered, and the precipitate was washed with 70 ml of alcohol in 15 ml portions, and the washing liquid and the filtrate were combined. Concentrate to 2 ral on a 90 ° C water bath, let cool, add Silver sulfate with an equivalent amount of 1% barium chloride solution and quantitated by 0.01% potassium hydroxide solution was transferred to a lOinl volumetric flask and dilute to volume, shake, filtration, to obtain the test solution;

The acetonitrile solution of p-bromo brominated acetophenone 1. 5ml and 0.05 mg / 0.25 mg / ml of acetonitrile bromide acetophenone acetonitrile solution 1. 5ml and 0. 05 mg / 1 ml of 18-crown ether-6 acetonitrile solution, mix well, conceal, heat in a water bath at 80 ° C for 20 minutes, remove, cool to room temperature, add acetonitrile to 10 ml, shake well, use 0 before injection The 45 μm microporous membrane was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

Claims

Claim

[Claim 1] A method for quality control of a prostaglandin capsule, characterized in that: the quality control method comprises a method for determining a content of stachysine hydrochloride in a raw material medicine motherwort, a prostaglandin capsule intermediate, and a prostaglandin capsule.

[Claim 2] The method for controlling quality of the prostaglandin capsule according to claim 1, wherein: the method for determining the content of stachydrine hydrochloride in the raw material drug motherwort is:

Chromatographic conditions: ZORBAX SB-C18 column, 5 μ ιη, 4. 6X 250 mm _; mobile phase: methanol-water in a volume ratio of 6:4; temperature: 25 to 35 ° C; detection wavelength: 250 to 270 nm; : lml/min; detector: DAD detector, 190~400 full-wave scan;

The preparation of the test solution is taken from the mother-of-pearl material, crushed, passed through a 40-mesh sieve, accurately weighed lg, placed in a 100 ml stoppered flask, and ultrasonically extracted with a solution of 1% ammonia in a solution of 50 ml for 30 minutes, filtered. Discard the chloroform solution, add the residue to the chloroform: 50 ml of the mixed solution of methanol in a volume ratio of 7:3, weigh the weight, sonicate for 20 minutes, let cool, weigh the weight, use trichloromethane.垸: The volume ratio of methanol is 7:3 to make up the lost weight. Shake well, filter, carefully extract 35ml of filtrate, place in evaporating dish, dry in 90Ό water bath, dissolve residue in 1% hydrochloric acid solution, dissolve in 20ml, filter After the filtrate was added to a freshly prepared 2% sulphuric acid chrome hinge solution 10 ml, placed in an ice bath for 0.5 hours, filtered, washed with ice water and precipitated, the filtrate was discarded, and the precipitate was dissolved in acetone (15 ml) in acetone. The solution was added dropwise 0.5% silver sulfate solution until no more precipitates were formed, and the mixture was filtered, and the precipitate was washed with 15 ml of 70% ethanol, and the washing liquid and the filtrate were combined, and concentrated to 2 ml in a water bath at 90 ° C. Let cool, add with silver sulfate Equivalent of 1% barium chloride solution, 0.01% potassium hydroxide solution was quantitatively transferred to a 10ml volumetric flask and dilute to volume, shake, filtration, to obtain the test solution;

The acetonitrile solution of p-bromo brominated acetophenone 1. 5ml and 0.05 mg / 0.25 mg / ml of acetonitrile bromide acetophenone acetonitrile solution 1. 5ml and 0. 05 mg / 1 ml of 18-crown ether-6 acetonitrile solution, mix well, conceal, heat in a water bath at 80 ° C for 20 minutes, remove, cool to room temperature, add acetonitrile to 10 ml, shake the hook, use 0 before injection The 45 μπι microporous membrane was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The content of the sulphate hydrochloride in the raw material drug motherwort is not less than 0.50%. The method of the present invention is as follows.

[Claim 4] The method for controlling quality of the prostaglandin capsule according to claim 1, wherein: the method for determining the content of stachydrine hydrochloride in the prostaglandin capsule intermediate is: Chromatographic conditions: ZORBAX SB-C18 column, 5 μ ιη, 4. 6 X 250 mm _; mobile phase: methanol-water in a volume ratio of 6:4; temperature: 25 to 35 ° C; detection wavelength: 260 nm; : lml/min; detector: DAD detector, 190~400 full-wave scan;

The preparation of the test solution was prepared by taking the intermediate of the prostaglandin capsule, crushed, passing through a 40 mesh sieve, and accurately weighed 1.5 g, placed in a 100 ml stoppered flask, and ultrasonically extracted with 50 ml of a chloroform solution containing 1% aqueous ammonia for 30 minutes. After filtering, discard the trichloromethane solution, and then add the chloroform after the residue is evaporated. The volume ratio of methanol is 50 ml of the mixed solution of 7:3, weigh the weight, sonicate for 20 minutes, let cool, weigh the weight. Use the mixed solution of trichloromethane:methanol with a ratio of 7:3 to make up the lost weight. Shake well, filter, carefully draw 35ml of filtrate, place in evaporating dish, dry in 90Ό water bath, add 1% hydrochloric acid to residue. The solution was dissolved in 20 ml, filtered, and the filtrate was added with 10 ml of a freshly prepared 2% ammonium sulphate solution. The mixture was placed in an ice bath for 0.5 hour, filtered, and the filtrate was discarded. The vessel was washed with ice water and the filtrate was discarded. The precipitate was dissolved in 15 ml of acetone, and 0.5% silver sulfate solution was added dropwise to the acetone solution until no more precipitate was formed. The mixture was filtered, and the precipitate was washed with 15 ml of 70% ethanol, and the washing liquid and the filtrate were combined. Concentrate to 2ml on a °C water bath, let cool And silver sulfate was added an equivalent amount of 1% barium chloride solution, quantitative 0.01% potassium hydroxide solution was transferred to a 10ml volumetric flask and dilute to volume, shake, filtration, to obtain the test solution;

The acetonitrile solution of bromo brominated acetophenone 1. 5 ml and 0. 05 mg / ml. The 18-crown ether-6 acetonitrile solution 1. 5ml, mix well, close the plug, heat in a water bath at 80 ° C for 20 minutes, remove, cool to room temperature, add acetonitrile to 10ml, shake well, use 0 before injection. The microporous membrane of 45 μm was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The content of the sulphate hydrochloride in the intermediate of the prostaglandin capsule is not less than 0.60%.

[Claim 6] The method for controlling quality of the prostaglandin capsule according to claim 1, wherein: the method for determining the content of stachydrine hydrochloride in the Qianlietai capsule is:

Chromatographic conditions: ZORBAX SB-C18 column, 5 μ πι, 4. 6 X 250 mm _; mobile phase: methanol to water in a volume ratio of 6:4; temperature: 25 to 35 ° C; detection wavelength: 260 nm; : lml/min; detector: DAD detector, 190~400 full-wave scan;

Preparation of the test solution Take 20 capsules of Qianlietai capsule, pour the contents, grind finely, accurately weigh 2g, place it in a 100ml stoppered flask, and ultrasonically extract it with 50ml of chloroform solution containing 1% ammonia water for 30 minutes. After filtering, discard the chloroform solution, and then add the chloroform after the residue is evaporated. The volume ratio of methanol is 50 ml of the mixed solution of 7:3, weighed, sonicated for 20 minutes, allowed to cool, weighed, used. Trichloromethane: methanol The volume ratio of 7:3 mixed solution to make up the lost weight, shake, filter, precision draw the filtrate 35ral, set in the evaporating dish, 90 Ό water bath to dry, the residue and 1% hydrochloric acid solution 20ml dissolved, filtered, the filtrate was added 10 ml of freshly prepared 2% ammonium hydrogen sulphate solution, placed in an ice bath for 0.5 hours, filtered, washed the container and precipitate with ice water, discarded the filtrate, dissolved in 15 ml of acetone, and dropped in acetone solution. Add 0.5% silver sulfate solution until no more precipitation occurs, place, filter, wash the precipitate with 15ml of 70% ethanol, combine the washing liquid and filtrate, concentrate on a 90Ό water bath to 2ml, let cool, add and An equivalent of 1 °/ of silver sulfate. The ruthenium chloride solution was quantitatively transferred to a 10 ml volumetric flask with a 0.01% potassium hydroxide solution and diluted to the mark, shaken, and filtered to obtain a test solution;

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1 ml of 18-crown ether _6 acetonitrile solution 1. 5ml, mixed, sealed, heated in 80 Ό water bath for 20 minutes, taken out, cooled to room temperature, acetonitrile to 10ml, shake well, before injection 0.55 The microporous membrane of m was filtered, and the filtrate was subjected to HPLC analysis under the above chromatographic conditions, and the content was calculated by an external standard method.

The content of the stalihydrin hydrochloride in the Qianlietai capsule is not less than 0.50%. The method of controlling the quality of the prostaglandin capsule according to claim 6.