CN104597197B - A kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears - Google Patents
A kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears Download PDFInfo
- Publication number
- CN104597197B CN104597197B CN201510035405.3A CN201510035405A CN104597197B CN 104597197 B CN104597197 B CN 104597197B CN 201510035405 A CN201510035405 A CN 201510035405A CN 104597197 B CN104597197 B CN 104597197B
- Authority
- CN
- China
- Prior art keywords
- solution
- ethanol
- rhizoma belamcandae
- product
- long
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicinal Preparation (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of detection method of pharmaceutical preparation of the long-pending defaecation that disappears, this detection method do as one likes shape, discriminating, inspection, extract and assay composition, wherein differentiate it is that Qualitive test is carried out to Rhizoma Belamcandae, be contrast with Rhizoma Belamcandae control medicinal material, be developping agent with methenyl choloride-butanone-formic acid=1-5:0.5-2:0.5-2, differentiate Rhizoma Belamcandae according to thin-layered chromatography; Assay measures belamcandin in Rhizoma Belamcandae according to Chinese Pharmacopoeia version in 2010 annex VD high performance liquid chromatography; Compared with prior art, the present invention establishes the detection method of the pharmaceutical preparation of the long-pending defaecation that disappears, and the methodological science adopted is reasonable, accuracy is high, favorable reproducibility, the quality of the long-pending Tongbian capsule that can control fully and effectively to disappear, can better ensure that the Clinical efficacy and safety of said preparation.
Description
Technical field
The present invention relates to the detection method of medicine, particularly relate to a kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears, belong to the technical field of medicine.
Background technology
Constipation is common clinical symptoms, the many discomforts of patient can be caused, as difficult defecation, ight soil is dry and hard, a couple of days, even within 1 week, just defecation was once, can have the spastic pain of left abdomen and bearing down during defecation, some patients tells bitter taste, anorexia, abdominal distension, lower abdomen is uncomfortable, exhaust is many or have dizziness, headache, the neurosis such as tired.The long-pending Tongbian capsule that disappears is ground into by Rhizoma Belamcandae that fine powder is encapsulated to be formed.Rhizoma Belamcandae is the dry rhizome of irides iris IristectorumMaxim., and gas is micro-, and taste is sweet, bitter.The effect of Rhizoma Belamcandae is intactly summarised as by " Chinese medicine voluminous dictionary ": " disappear long-pending, the broken stasis of blood, row water, removing toxic substances, desinsection.Cure mainly dyspepsia turgor, note of the ancient Chinese lump in the abdomen, tympanites, abscess of throat, hemorrhoid complicated by anal fistula, pyogenic infections, fall injure swollen, roundworm suffer from abdominal pain.”
The long-pending Tongbian capsule that disappears records in national standard for traditional Chinese medicines compilation internal medicine taste fascicle, and standard No. is WS-11214 (ZD-1214)-2002, has effect of purgating heat and bowels, is mainly used in the constipation caused by Gastrointestinal excess heat, abdominal distention, poor appetite.In existing standard, when carrying out Qualitive test to medicine, it is not accurate enough to the description of crystal under microscope, Rhizoma Belamcandae is carried out TLC differentiate be, test sample preparation and the selection of developping agent not ideal enough, cause spot less, spot is unintelligible, is separated the phenomenons such as bad; Lack in primary standard in addition and assay is carried out to Rhizoma Belamcandae, therefore cause the quality control of medicine comprehensive and accurate not, thus cause a hidden trouble.
Chinese Pharmacopoeia version one in 2010 discloses the detection method of Rhizoma Belamcandae, utilizes methyl alcohol-0.05mol/L potassium dihydrogen phosphate (32: 68) to carry out assay as mobile phase to belamcandin in Rhizoma Belamcandae; Zou Kong Qiang etc. measure in HPLC method determination study (the traditional Chinese medicine Leader that Qian Bei introduces a fine variety belamcandin in blackberry lily, 2007,13 (9): 81 ~ 82.), in a literary composition, disclose employing acetonitrile-0.05mol/L potassium dihydrogen phosphate (19: 81) for mobile phase belamcandin and carry out assay; Guo Zhihui measures content (the Pharmaceutical Analysis magazine of belamcandin in blackberry lily granule in HPLC method, 2009,29 (8): 1375 ~ 1378.), in a literary composition, disclose employing acetonitrile-0.2% phosphoric acid solution (78:22) for mobile phase belamcandin and carry out assay.Although above document reports the content assaying method of Rhizoma Belamcandae, operate according to the method described above, occur or degree of separation not good, or analysis time is longer, and also have the phenomenons such as other chromatographic peak interference, the detection therefore offseting long-pending Tongbian capsule can not meet the demands.
Summary of the invention
Technical matters to be solved by this invention is the detection method of the pharmaceutical preparation providing a kind of long-pending defaecation that disappears, and comprises proterties, discriminating, inspection and assay; This detection method can control to disappear the quality of long-pending Tongbian capsule fully and effectively, thus guarantees clinical efficacy and the drug safety of this medicine.
In order to solve the problems of the technologies described above, the present invention adopts following technical scheme: the detection method of the pharmaceutical preparation of the long-pending defaecation that disappears, and described pharmaceutical preparation is capsule, this capsule is prepared like this: Rhizoma Belamcandae 350g, gets Rhizoma Belamcandae, is ground into fine powder, incapsulate, make 1000; This detection method do as one likes shape, discriminating, inspection, extract and assay composition, wherein differentiating is the Qualitive test to product, the TLC to Rhizoma Belamcandae differentiates, and assay is that photograph Chinese Pharmacopoeia version in 2010 annex VD high performance liquid chromatography measures belamcandin in Rhizoma Belamcandae.
The detection method of the pharmaceutical preparation of the above-mentioned long-pending defaecation that disappears, specifically such:
Proterties: this product is capsule, content is the light yellow powder to brown color; Gas is micro-, and taste is sweet, bitter;
Differentiate: Qualitive test is carried out to Rhizoma Belamcandae; With Rhizoma Belamcandae control medicinal material be contrast, with methenyl choloride-butanone-formic acid=1-5: 0.5-2: 0.5-2 for developping agent, according to thin-layered chromatography differentiate Rhizoma Belamcandae;
Check: every regulation relevant under meeting Chinese Pharmacopoeia version in 2010 annex IL capsule item;
Extract: the hot dipping measured under item according to Chinese Pharmacopoeia version in 2010 annex XA ethanol soluble extractives measures, and with ethanol as solvent, must not be less than 20.0%;
Assay: take tectoridin reference substance as contrast, according to the belamcandin content in Chinese Pharmacopoeia version in 2010 annex VD high effective liquid chromatography for measuring capsule.
The detection method of the capsule preparations of the aforesaid long-pending defaecation that disappears, wherein differentiate to be made up of following:
(1) get this product, put basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 20-40 minute, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=1-5: 0.5-2: 0.5-2 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color.
Further, described discrimination method is as follows:
(1) get this product, put basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 30 minutes, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=3: 1: 1 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color.
The detection method of the capsule preparations of the aforesaid long-pending defaecation that disappears, wherein content assaying method is as follows:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, carries out gradient elution; Flow velocity 0.5-2.0ml/min, column temperature 25-30 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 65-75% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 65-75% ethanol 25ml, ultrasonic process 50-70min, let cool, more weighed weight, the weight of less loss is supplied with 65-75% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 65-75% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product;
This product every contains Rhizoma Belamcandae with belamcandin (C
22h
24o
11) meter, must not 13.0mg be less than.
Further, described content assaying method is as follows:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, and according to the form below carries out gradient elution; Flow velocity 1.0ml/min, column temperature 25 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
Table 1
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 70% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 70% ethanol 25ml, ultrasonic process 60min, let cool, more weighed weight, the weight of less loss is supplied with 70% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 70% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product.
In order to study the detection method of the capsule preparations of the long-pending defaecation that disappears, applicant carried out a large amount of experiments to screen preferred plan, specific as follows:
1, Rhizoma Belamcandae Qualitive test
The long-pending Tongbian capsule that disappears is ground into by Rhizoma Belamcandae that fine powder is encapsulated to be formed, and Rhizoma Belamcandae is the dry rhizome of irides iris IristectorumMaxim..That examines under a microscope this product in primary standard is described as: cylindrulite is numerous, how disconnected broken, diameter 17 ~ 50 μm, long 50 ~ 80 ~ 380 μm.This description is stated not accurate enough to the length of crystal column and size, qualification operator and estimator is easily made to state unclear, therefore according to 2010 editions pharmacopeia to the character description of Rhizoma Belamcandae, in conjunction with the actual microscopy result of 10 batch sample, confirm that this product puts basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82.
2, TLC differentiates
Prepared by need testing solution: in primary standard, and select glue to disappear long-pending Tongbian capsule content 2g, add methyl alcohol 20ml, ultrasonic process 30 minutes, filter, filtrate is concentrated into 1ml, as need testing solution.Through practical operation, find that content is too much, sonication treatment time is inadequate, easily causes spot unintelligible, is separated bad phenomenon.Therefore need testing solution preparation made into: select to disappear long-pending Tongbian capsule content 1g, add methyl alcohol 10ml, ultrasonic process 30 minutes, filter, filtrate is concentrated into 1ml, as need testing solution.
The screening of development system: select developping agent 1---methenyl choloride-methyl alcohol-formic acid=10: 1: 0.1 (primary standard); Developping agent 2---methenyl choloride-butanone-formic acid=3: 1: 1; Developping agent 3---methenyl choloride-butanone-glacial acetic acid=4: 2: 1; Developping agent 4---methenyl choloride-acetone-glacial acetic acid=5: 0.5: 1; Developping agent 4---methenyl choloride-acetone-formic acid=4: 1: 2.Result shows, and except developping agent 2, all the other system results display dots are unintelligible, spot is few, or spot and R f value is lower.
Developping agent 2 is further investigated, finds that within the scope of methenyl choloride-butanone-formic acid=1-5: 0.5-2: 0.5-2, spot degree of separation is better, more clear, wherein methenyl choloride-butanone-formic acid=3: 1: 1 spot degree of separation is best, develop the color clear, and spot is more.
Compared by said method, with methenyl choloride-butanone-formic acid=3: 1: 1 for developping agent; Selection disappears long-pending Tongbian capsule content 1g, adds methyl alcohol 10ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into 1ml, as need testing solution.Method is simple, cost-saving, easy point sample, clear spot, therefore selects.
3, assay research
3.1 instruments and reagent
Instrument: Shimadzu LC-20AT high performance liquid chromatograph, Thermo-C185 μm of 4.6mm × 250mm post number: 25305-254630, Sai Duolisi CPA225D electronic balance (100,000/).
Reagent and reagent: tectoridin reference substance: Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 111632-200602; Sample: disappear long-pending Tongbian capsule, and lot number, is produced by Guizhou Shenqi Pharmaceutical Co., Ltd.; Acetonitrile is chromatographically pure, and phosphoric acid, ethanol are pure for analyzing.
The selection of 3.2 chromatographic conditions
Adopt C
184.6 × 250mm chromatographic column, be mobile phase (A) through overtesting with acetonitrile, 0.2% phosphoric acid solution is mobile phase (B), gradient elution: 0 ~ 30 minute, 17%A, 83%B; 31 ~ 40 minutes, 65%A, 35%B; 41 ~ 50 minutes, 17%A, 83%B.Flow velocity 1.0ml/min, determined wavelength is 265nm, and column temperature is 25 DEG C.
3.2.1, the selection of mobile phase:
Mobile phase: methyl alcohol-0.05mol/L potassium dihydrogen phosphate (32: 68), acetonitrile-0.05mol/L potassium dihydrogen phosphate (19: 81), methyl alcohol-0.2% phosphoric acid solution (53:47), acetonitrile-0.2% phosphoric acid solution (78:22), acetonitrile-0.2% phosphoric acid solution (gradient elution), methyl alcohol-glacial acetic acid (18:82), acetonitrile-water (20:80).
Above-mentioned mobile phase, compare through test, result shows: except the collocation of acetonitrile-0.2% phosphoric acid solution, all the other mobile phases or degree of separation are not good, or analysis time is longer, wherein arrange in pairs or groups with methyl alcohol or acetonitrile and 0.05mol/L potassium dihydrogen phosphate, belamcandin peak and other impurities peak degree of separation against regulation, analysis time is very long, also has chromatographic peak at 130 minutes.And adopting acetonitrile-0.2% phosphoric acid solution to carry out gradient elution as mobile phase, belamcandin is separated completely with other impurities peak, and peak shape is better.Through test of many times, according to the form below is selected to carry out gradient elution:
Table 2
3.2.2 the investigation of chromatographic column:
With ThermoC
185 μm of 4.6 × 250mm, Yi Lite C
185 μm of 4.6 × 2150mm measure, and belamcandin peak and other impurities peak reach baseline separation, and degree of separation is good, and appearance time is short; With enlightening horse C
185 μm of 4.6 × 250mm, enlightening horse C
185 μm of 4.6 × 150mm measure, and belamcandin appearance time is longer, belamcandin peak and other impurities peak degree of separation against regulation.Therefore recommendation ThermoC
185 μm of 4.6 × 250mm chromatographic columns measure.
3.2.3 determined wavelength is selected:
Get need testing solution and carry out uv scan at 190nm ~ 370nm, near 268nm, have absorption maximum.Get same reference substance solution and need testing solution measures at 268nm ± 5nm, content is respectively 47.80mg/g, 47.99mg/g, 48.46mg/g, illustrates that measuring wavelength does not make significant difference to assay thus.Determined wavelength in comprehensive 2010 editions pharmacopeia Rhizoma Belamcandae assays, therefore select 265nm to measure wavelength as this product.
The selection of 3.3 extracting modes:
Get the sample of same lot number, accurately weighed 0.2g, precision adds 70% ethanol 25ml with refluxing extraction and ultrasonic process 1 hour respectively, measure by content assaying method, belamcandin content is respectively 48.23mg/g, 47.94mg/g, and content without significant difference, therefore selects ultrasonic extraction.
3.4 ultrasonic times are investigated
Get the sample of same lot number, accurately weighed 0.2g, precision adds the 70% ethanol 25ml extraction effect of ultrasonic 30 minutes, 45 minutes, 60 minutes, 90 minutes respectively.Test findings is in table 3.
Table 3 ultrasonic time is on the impact of assay
Test findings shows: ultrasonic time does not make significant difference to assay.For guaranteeing to extract completely, therefore 60 minutes will be decided to be extraction time.
3.5 specificity tests
This product does not add auxiliary material in the formulation, does not have negative sample.Detect by above-mentioned chromatographic condition, solvent has no absorption peak in relevant position; The retention time at belamcandin peak in working sample is consistent with the retention time at belamcandin peak in reference substance solution.
3.6 system suitability
Measure the content of belamcandin in preparation by above-mentioned chromatographic condition, belamcandin peak tailing factor, degree of separation meet the requirements.Number of theoretical plate is not less than 3000.
The investigation of 3.7 linear relationships
Precision takes through the phosphorus pentoxide drying under reduced pressure tectoridin reference substance of more than 12 hours (111632-200602) 9.42mg, put in 50ml volumetric flask, the ethanol adding 70% dissolves and is diluted to scale, shakes up, and makes the reference substance storing solution containing 0.1884mg in every 1ml.Precision measures above-mentioned reference substance concentrated wiring liquid 1.0ml, put in 10ml measuring bottle, the ethanol ethanol adding 70% is diluted to scale, shake up, make the reference substance solution containing 0.01884mg in every 1ml, precision measures each 1 μ l of above-mentioned reference substance solution respectively, 3 μ l, 5 μ l, 10 μ l, 15 μ l, 18 μ l, 20 μ l injection liquid chromatographies, record chromatogram, with sample size X (μ g) for horizontal ordinate, peak area Y (mv) ordinate, mapping, obtain the straight-line equation substantially crossing initial point, regression equation Y=4624.8X-17303, correlation coefficient r=0.9999, measure gained peak area with reference substance and substitute into aforementioned regression equation calculation content, acquired results and real content mean deviation are less than 1%, therefore can think that typical curve was similar to initial point.Assay can adopt one point external standard method to calculate (see table 4) thus.
Test findings shows: belamcandin has good linear relationship within the scope of 18.84 μ g ~ 376.80 μ g.
Quantitative limit and detectability are respectively: 0.00336 μ g/ml and 0.00112 μ g/ml.
Table 4 belamcandin linear relationship is investigated
3.8 precision test
Precision measures same tectoridin reference substance solution (0.01884mg/ml) 10 μ l, continuous sample introduction 6 times, record chromatogram, peak area relative standard deviation (RSD) is 0.14%, and test findings shows precision good (see table 5).
Table 5 belamcandin Precision test result
3.9 stability test
Get same need testing solution, respectively at 0h, 2h, 4h, 6h, 8h, 12h sample introduction 6 times, record peak area, peak area relative standard deviation (RSD) is 0.30%.Test findings shows, need testing solution was at 12 hours internal stabilities good (see table 6).
Table 6 belamcandin stability test result
3.10 replica test
Get 6 parts, the sample of same lot number, every part of about 0.2g, accurately weighed, measure by the method under assay item, record chromatogram, calculate water belamcandin content: average content is 46.43mg/g, RSD is 0.85%; Test findings shows, repeatability good (see table 7).
Table 7 belamcandin replica test result
3.11 average recovery is tested
Get 9 parts, the sample (48.43mg/g) of known content, every part of accurately weighed 0.1g, put in tool plug conical flask, the accurate reference substance solution 25ml adding 0.2166mg/ml, 0.2640mg/ml, 0.3242mg/ml respectively, measure by the method under assay item, record chromatogram, calculates the recovery.Test findings shows, the recovery good (see table 8).
Table 8 belamcandin average recovery test findings
3.12 sample size measures
Adopt 10 lot number assay results (see table 9) that this law measures.
Table 9 belamcandin assay result
The formulation of 3.13 sample size limits:
2010 editions pharmacopeia Rhizoma Belamcandae content must not be and are less than 3.6%, calculate by theoretical amount, and every must not be less than 12.6mg containing belamcandin; Above 10 batches of content mean values are 18.14mg/ grain, and due to the place of production and different harvest time difference, medicinal material content difference to some extent, calculating every content by 80% is 14.51mg.
Therefore, content limit is decided to be every and contains belamcandin and be not less than 13.0mg by comprehensive above factor.
Embodiment
Embodiment 1
Disappear the detection method of pharmaceutical preparation of long-pending defaecation, and step is as follows:
Proterties: this product is capsule, content is the light yellow powder to brown color; Gas is micro-, and taste is sweet, bitter;
Differentiate: (1) gets this product, puts basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 30 minutes, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=3: 1: 1 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color;
Check: every regulation relevant under meeting Chinese Pharmacopoeia version in 2010 annex IL capsule item;
Extract: the hot dipping measured under item according to Chinese Pharmacopoeia version in 2010 annex XA ethanol soluble extractives measures, and with ethanol as solvent, must not be less than 20.0%;
Assay:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, gradient elution: 0 ~ 30 minute, 17%A, 83%B; 31 ~ 40 minutes, 65%A, 35%B; 41 ~ 50 minutes, 17%A, 83%B; Flow velocity 1.0ml/min, column temperature 25 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 70% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 70% ethanol 25ml, ultrasonic process 60min, let cool, more weighed weight, the weight of less loss is supplied with 70% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 70% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product.
Embodiment 2
Disappear the detection method of pharmaceutical preparation of long-pending defaecation, and step is as follows:
Proterties: this product is capsule, content is the light yellow powder to brown color; Gas is micro-, and taste is sweet, bitter;
Differentiate: (1) gets this product, puts basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 20 minutes, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=1: 0.5: 0.5 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color;
Check: every regulation relevant under meeting Chinese Pharmacopoeia version in 2010 annex IL capsule item;
Extract: the hot dipping measured under item according to Chinese Pharmacopoeia version in 2010 annex XA ethanol soluble extractives measures, and with ethanol as solvent, must not be less than 20.0%;
Assay:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, carries out gradient elution; Flow velocity 0.5ml/min, column temperature 25 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 65% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 65% ethanol 25ml, ultrasonic process 50min, let cool, more weighed weight, the weight of less loss is supplied with 65% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 65% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product.
Embodiment 3
Disappear the detection method of pharmaceutical preparation of long-pending defaecation, and step is as follows:
Proterties: this product is capsule, content is the light yellow powder to brown color; Gas is micro-, and taste is sweet, bitter;
Differentiate: (1) gets this product, puts basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 40 minutes, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=5: 2: 2 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color;
Check: every regulation relevant under meeting Chinese Pharmacopoeia version in 2010 annex IL capsule item;
Extract: the hot dipping measured under item according to Chinese Pharmacopoeia version in 2010 annex XA ethanol soluble extractives measures, and with ethanol as solvent, must not be less than 20.0%;
Assay:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, carries out gradient elution; Flow velocity 2.0ml/min, column temperature 30 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 75% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 75% ethanol 25ml, ultrasonic process 70min, let cool, more weighed weight, the weight of less loss is supplied with 75% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 75% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product.
Embodiment 4
Get the sample that lot number is 140703, detect according to the method for embodiment 1-3, result is as following table 10.
Table 10 sample quality measurement result
Compared with prior art, the method applied in the present invention is scientific and reasonable, and accuracy is high, favorable reproducibility, is easy to the mensuration differentiated, the quality of the long-pending Tongbian capsule that can control fully and effectively to disappear, can better guarantee the Clinical efficacy and safety of said preparation.
Claims (4)
1. disappear the detection method of pharmaceutical preparation of long-pending defaecation, and described preparation is the long-pending Tongbian capsule that disappears, and this capsule is prepared like this: Rhizoma Belamcandae 350g, gets Rhizoma Belamcandae, is ground into fine powder, incapsulate, and makes 1000; This detection method do as one likes shape, discriminating, inspection, extract and assay composition, discriminating carries out Qualitive test to Rhizoma Belamcandae, is contrast with Rhizoma Belamcandae control medicinal material, take methenyl choloride-butanone-formic acid=1-5: 0.5-2: 0.5-2 as developping agent, differentiates Rhizoma Belamcandae according to thin-layered chromatography; Assay measures belamcandin in Rhizoma Belamcandae according to Chinese Pharmacopoeia version in 2010 annex VD high performance liquid chromatography; It is characterized in that: discrimination method is as follows:
(1) get this product, put basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 20-40 minute, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=1-5: 0.5-2: 0.5-2 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color;
Content assaying method is as follows:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, carries out gradient elution, and order is: 0 ~ 30 minute, 17%A, 83%B; 31 ~ 40 minutes, 65%A, 35%B; 41 ~ 50 minutes, 17%A, 83%B; Flow velocity 0.5-2.0ml/min, column temperature 25-30 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 65-75% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 65-75% ethanol 25ml, ultrasonic process 50-70min, let cool, more weighed weight, the weight of less loss is supplied with 65-75% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 65-75% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product;
This product every contains Rhizoma Belamcandae with belamcandin (C
22h
24o
11) meter, must not 13.0mg be less than.
2. the detection method of the pharmaceutical preparation of the long-pending defaecation that disappears according to claim 1, is characterized in that: discrimination method is as follows:
(1) get this product, put basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 30 minutes, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=3: 1: 1 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color.
3. the detection method of the pharmaceutical preparation of the long-pending defaecation that disappears according to claim 1, is characterized in that: content assaying method is as follows:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, gradient elution: 0 ~ 30 minute, 17%A, 83%B; 31 ~ 40 minutes, 65%A, 35%B; 41 ~ 50 minutes, 17%A, 83%B; Flow velocity 1.0ml/min, column temperature 25 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 70% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 70% ethanol 25ml, ultrasonic process 60min, let cool, more weighed weight, the weight of less loss is supplied with 70% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 70% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product.
4. the detection method of the pharmaceutical preparation of the long-pending defaecation that disappears according to any one of claim 1-3, is characterized in that: detection method is as follows:
Proterties: this product is capsule, content is the light yellow powder to brown color; Gas is micro-, and taste is sweet, bitter;
Differentiate: (1) gets this product, puts basis of microscopic observation: cylindrulite is more, how disconnected broken, diameter 16 ~ 52 μm, complete person long 15 ~ 82;
(2) get this product content 1g, add methyl alcohol 10ml, ultrasonic process 30 minutes, filter, filtrate is concentrated into 1ml, as need testing solution; Separately get Rhizoma Belamcandae control medicinal material 1g, be made in the same way of control medicinal material solution; According to the annex VI B thin-layered chromatography test of Chinese Pharmacopoeia version in 2010, draw each 1 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, with methenyl choloride-butanone-formic acid=3: 1: 1 for developping agent, launch, take out, dry, spray with aluminium choride test solution, inspect under putting 365nm ultraviolet lamp, in test sample chromatogram, with on control medicinal material chromatogram relevant position, show the fluorescence spot of same color;
Check: every regulation relevant under meeting Chinese Pharmacopoeia version in 2010 annex IL capsule item;
Extract: the hot dipping measured under item according to Chinese Pharmacopoeia version in 2010 annex XA ethanol soluble extractives measures, and with ethanol as solvent, must not be less than 20.0%;
Assay:
Chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, gradient elution: 0 ~ 30 minute, 17%A, 83%B; 31 ~ 40 minutes, 65%A, 35%B; 41 ~ 50 minutes, 17%A, 83%B; Flow velocity 1.0ml/min, column temperature 25 DEG C, determined wavelength is 265nm; Number of theoretical plate calculates should be not less than 3000 by belamcandin peak;
The preparation of reference substance solution: precision takes tectoridin reference substance, adds 70% ethanol and makes the solution of every 1ml containing 20 μ g, to obtain final product;
The preparation of need testing solution: get this product content, mixing, gets 0.2g, accurately weighed, put in conical flask, add 70% ethanol 25ml, ultrasonic process 60min, let cool, more weighed weight, the weight of less loss is supplied with 70% ethanol, shake up, filter, precision measures subsequent filtrate 1ml, put in 25ml measuring bottle, add 70% ethanol to scale, shake up, filter, get subsequent filtrate, to obtain final product;
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product;
This product every contains Rhizoma Belamcandae with belamcandin (C
22h
24o
11) meter, must not 13.0mg be less than.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510035405.3A CN104597197B (en) | 2015-01-23 | 2015-01-23 | A kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510035405.3A CN104597197B (en) | 2015-01-23 | 2015-01-23 | A kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104597197A CN104597197A (en) | 2015-05-06 |
| CN104597197B true CN104597197B (en) | 2016-04-13 |
Family
ID=53123116
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510035405.3A Active CN104597197B (en) | 2015-01-23 | 2015-01-23 | A kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104597197B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106442843A (en) * | 2016-06-30 | 2017-02-22 | 贵阳中医学院第二附属医院 | Quality check method of children's granules for clearing heat from throat |
| CN107748220B (en) * | 2017-09-26 | 2020-02-04 | 四川省中医药科学院 | Sichuan blackberry lily flavone capsule and detection method of Sichuan blackberry lily flavone extract |
| CN108344842A (en) * | 2018-01-23 | 2018-07-31 | 牟会玉 | A kind of medicament reaction checking device |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6172702A (en) * | 1984-09-18 | 1986-04-14 | Taisho Pharmaceut Co Ltd | Attracting and appetizing agent for periplaneta americana |
| CN101987146A (en) * | 2010-09-17 | 2011-03-23 | 四川逢春制药有限公司 | Preparation method of capsule for treating pharyngitis and infection of upper respiratory tract and detection method of capsule |
| CN102579861A (en) * | 2011-01-17 | 2012-07-18 | 山东阿如拉药物研究开发有限公司 | Method for detecting quality of An'erning granules |
| CN102603834A (en) * | 2012-01-17 | 2012-07-25 | 武汉回盛生物科技有限公司 | Method for extracting and separating tectoridin from iris tectorum |
| CN103134897A (en) * | 2013-02-02 | 2013-06-05 | 四川逢春制药有限公司 | Rhiaoma iridis tectori capsule detecting method |
-
2015
- 2015-01-23 CN CN201510035405.3A patent/CN104597197B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6172702A (en) * | 1984-09-18 | 1986-04-14 | Taisho Pharmaceut Co Ltd | Attracting and appetizing agent for periplaneta americana |
| CN101987146A (en) * | 2010-09-17 | 2011-03-23 | 四川逢春制药有限公司 | Preparation method of capsule for treating pharyngitis and infection of upper respiratory tract and detection method of capsule |
| CN102579861A (en) * | 2011-01-17 | 2012-07-18 | 山东阿如拉药物研究开发有限公司 | Method for detecting quality of An'erning granules |
| CN102603834A (en) * | 2012-01-17 | 2012-07-25 | 武汉回盛生物科技有限公司 | Method for extracting and separating tectoridin from iris tectorum |
| CN103134897A (en) * | 2013-02-02 | 2013-06-05 | 四川逢春制药有限公司 | Rhiaoma iridis tectori capsule detecting method |
Non-Patent Citations (6)
| Title |
|---|
| Eun-Kyung PARK 等.Passive Cutaneous Anaphylaxis-Inhibitory Action of Tectorigenin,a Metabolite of Tectoridin by Intestinal Microfl * |
| ora.《Biol. Pharm. Bull.》.2004,第27卷(第7期),1099-1102. * |
| 利咽颗粒的定性定量方法研究;李士敏 等;《中国中药杂志》;20030831;第28卷(第8期);727-729 * |
| 射干与鸢尾的薄层色谱鉴别;吴颖等;《中医药信息》;19980228(第1期);第2.2节 * |
| 射干地龙颗粒质量标准;辛蕊华 等;《中国实验方剂学杂志》;20130731;第19卷(第13期);57-60 * |
| 高效液相色谱法测定射干利咽口服液中射干苷、次野鸢尾黄素的含量;曹岳华 等;《中南药学》;20100630;第8卷(第6期);431-434 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104597197A (en) | 2015-05-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103149320B (en) | Quality control method of loins-strengthening and kidney-invigorating medicine | |
| CN103954724B (en) | Method for detecting Jingfang granules | |
| CN106822203B (en) | Radix angelicae pubescentis granules and preparation method and quality control method thereof | |
| CN104597197B (en) | A kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears | |
| CN100437112C (en) | Method for inspecting Chinese medicinal preparation quality in treatment of old man eyes dieases | |
| CN102269752B (en) | Detection method for pharmaceutical composition preparation | |
| CN101028460B (en) | Method for inspecting throat-clearing Chinese medicinal pills | |
| CN101332287B (en) | Quality control method of Danle capsule | |
| CN102707006B (en) | Quality detection method of cudrania tricuspidata formula granules | |
| CN108490083A (en) | The quality determining method of SuHuang ZhiKe capsule | |
| CN104678004A (en) | Quality control method for kudzuvine root and hawthorn lipid-lowering particles | |
| CN102068549A (en) | Quality control method for Chinese medicinal preparation heat clearing and blood cooling pills | |
| CN106596777B (en) | The method of quality control of Dandengtongnao preparation | |
| CN113759057B (en) | Characteristic spectrum of allium macrostemon white water extract and preparation thereof and construction method thereof | |
| CN101028474B (en) | Method for inspecting the quality of Chinese preparation with Yang-and kidney tonifying functions | |
| CN105842381A (en) | Detection method of Qigu capsule | |
| CN110960617B (en) | Preparation method and quality analysis method of medicinal composition for joint protection | |
| CN100432670C (en) | Method for inspecting Chinese-medicinal preparation Kaiyinwan | |
| CN107064325A (en) | A kind of method of quality control of Qige granules | |
| CN102721782B (en) | Method for detecting quality of philippine flemingia root formula granules | |
| CN102068599B (en) | Detection method for phlegm eliminating and asthma relieving cough syrup | |
| CN105004833A (en) | Detection method for traditional Chinese medicine preparation for treating acute gouty arthritis and gout | |
| CN109917041A (en) | The measuring method of a variety of active ingredients content in xiaoshuan tongluo tablet | |
| CN103175938B (en) | Detecting method of drug for treating cough | |
| CN102370721A (en) | Method for detecting dredging particular Chinese medicinal preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221214 Address after: No. 177, Chuang Bei Road, Shawen Ecological Science and Technology Industrial Park, Guiyang National High tech Industrial Development Zone, Guiyang, Guizhou 550014 Patentee after: GUIZHOU SHENQI PHARMACEUTICAL Co.,Ltd. Patentee after: GUIZHOU SHENQI PHARMACEUTICAL Research Institute Address before: No. 1 Beijing Road, Guiyang, Guizhou Patentee before: GUIZHOU SHENQI PHARMACEUTICAL Research Institute |
|
| TR01 | Transfer of patent right |