CN109932434A - The detection method of cholic acid content in a kind of Xihuang capsules - Google Patents
The detection method of cholic acid content in a kind of Xihuang capsules Download PDFInfo
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- CN109932434A CN109932434A CN201711363968.0A CN201711363968A CN109932434A CN 109932434 A CN109932434 A CN 109932434A CN 201711363968 A CN201711363968 A CN 201711363968A CN 109932434 A CN109932434 A CN 109932434A
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Abstract
The present invention provides a kind of detection methods of cholic acid content in Xihuang capsules, and the specificity of this method is strong, and the linear of method, stability, reproducibility and sample recovery rate reach preferable technical requirements.The method of the present invention determines cholic acid detection method in Xihuang capsules, can more efficiently monitor the product quality of Xihuang capsules.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicines.More particularly to a kind of detection method of cholic acid content in Xihuang capsules.
Background technique
Xihuang capsules are recorded in state food pharmaceuticals administration general bureau national drug standards WS3-B-3210-98-2016,
There was only bilirubin, the detection of Determination of Muscone item, no cholic acid assay detection in proper mass standard.And the detection side of cholic acid
There was only thin layer chromatography scanning, no high performance liquid chromatography in method national standard.In order to preferably control product quality, now using efficient
Liquid chromatograph-ELSD carries out assay to cholic acid in Xihuang capsules, has formulated this detection method.
Summary of the invention
Technical problem to be solved by the present invention lies in design the side that cholic acid content detection is carried out in a kind of pair of Xihuang capsules
Method, to efficiently control product quality.
The present invention provides a kind of detection methods of cholic acid content in Xihuang capsules.
The object of the invention can be realized by following proposal:
One, instrument, reagent and test sample
Instrument: Agilent1260 type high performance liquid chromatograph
ELSD 2000ES evaporative light scattering detector
Reagent and reagent:
Cholic acid reference substance: National Institute for Food and Drugs Control, lot number: 100078-201415 are derived from
Acetonitrile is chromatography pure level reagent, and other reagents are analytical grade reagent.
Test sample: Xihuang capsules 1708103,1708104,1708105
Chromatographic condition and system suitability:
Chromatographic column: using octadecylsilane chemically bonded silica as filler；Mobile phase: with acetonitrile: 0.5% formic acid solution (38:62) stream
Speed: 1ml/min column temperature: 30 DEG C.
Evaporative light scattering detector.Number of theoretical plate presses cholic acid (C24H40O5) calculate should be not less than 4000.
Two, test method and result
(1) preparation of reference substance solution: taking cholic acid reference substance appropriate, and accurately weighed 10mg is set in 50ml measuring bottle, adds methanol to quarter
Degree, be made solution of every 1ml containing 0.2mg to get.
(2) preparation of test solution
This product content about 1.0g is taken, is set in stuffed conical flask, it is accurately weighed, add methanol 25ml, weighed weight, 40 points of ultrasound
Clock is let cool, and weighed weight adds methanol to supply less loss weight, shakes up, be centrifuged, filtration, take subsequent filtrate to get.
(3) measuring method
It is accurate respectively to draw 5 μ l of reference substance, 10 μ l and test solution 5-10 μ l, high performance liquid chromatograph is injected, is measured, is used
External standard two-point method logarithmic equation calculate to get.
Three, the technical standard detected
This product every containing In vitro cultured Calculus Bovis with cholic acid (C24H40O5) meter, 0.5mg must not be less than.
Four, it can illustrate the feasibility of this detection method by following tests
1. the investigation of linear relationship
1.1 precisions weigh cholic acid reference substance, add methanol that the solution of 0.2mg/ml is made.It takes the solution appropriate, takes respectively above-mentioned molten
1 μ l of liquid, 2 μ l, 5 μ l, 10 μ l, 20 μ l sample introductions are measured according to the chromatographic condition worked out, record sampling volume and peak area, with into
The amount logarithm of the reference substance of sample is abscissa, using peak area logarithm as ordinate, draws standard curve.Regression equation be y=
1.355x+1.099 R2At good linear relationship, linear relationship between=0.999, the range of linearity 0.20037-4.00743 μ g
Investigate the result is shown in Figure 1, standard curve See Figure 2.
1.2 precisions weigh cholic acid reference substance, add methanol that the solution of 0.4mg/ml is made.Take the solution appropriate, with methanol system
For at 0.2mg/ml, 0.08mg/ml, 0.04mg/ml, 0.02mg/ml solution, above five are taken respectively according to the chromatographic condition worked out
A each 10 μ l of various concentration cholic acid reference substance solution, injection high performance liquid chromatograph measurement, records sampling volume and peak area, with
The amount logarithm of the reference substance of sample introduction is abscissa, using peak area logarithm as ordinate, draws standard curve.Regression equation is
Y=1.352x+1.221 R2At good linear relationship, linear relationship between=0.998, the range of linearity 0.2000-3.9797 μ g
It investigates result and sees that Fig. 3, standard curve are shown in Fig. 4.
2. solution stability testing
Same test sample solution is taken, is measured respectively in 0,2,6,8,12 hour sample introduction.
Test sample peak area logarithm (LgA): RSD is 1.2% (n=5), illustrates that cholic acid is stablized at least in 12 hours, surely
Qualitative test result is shown in Fig. 5.
3. precision -- repeatability
It takes with the uniform test sample of a batch, weighs 6 parts, each 1g, according to scheduled processing method, it is molten that 6 parts of different test samples are made
Liquid, precision draw each 10 μ l of test solution, inject high performance liquid chromatograph, measure peak area, calculate content.The result shows that
Test sample average content is 3.18mg/g, RSD=3.2%(n=6) the result shows that accuracy is good.Repetitive test result is shown in figure
6。
4. sample recovery rate is tested
Take sample-adding absorption method test.Reference substance concentration be 0.2006mg/ml, take known content Xihuang capsules (lot number:
1708105, content: 3.18mg/g) content, it is uniformly mixed, takes about 0.5g, it is accurately weighed, it is 9 parts of nominal, each successively to add respectively
Enter 50%, 100%, 120% a certain amount of cholic acid reference substance, sample-adding test solution is made according to sample solution preparation method, surveys
It is fixed.Test result show 50% rate of recovery be 100.4%, RSD=3.5%, 100% rate of recovery be 90.8%, RSD=3.9%, 120%
The rate of recovery is the % of 97.0%, RSD=3.2, illustrates that this measuring method measurement result is accurate.Sample recovery rate test result is shown in Fig. 7.
5. specificity is tested
It is accurate respectively to draw reference substance solution and each 10ul of test solution under the chromatographic condition drafted, it is injected separately into efficiently
Liquid chromatograph, measurement.The result shows that test sample separating degree is preferable, retention time is moderate, the retention time and peak of reference substance with
Cholic acid is consistent in test sample.
6, sample measures
Three batches of Xihuang capsules are taken (lot number: 1708103,1708104,1708105), to be made according to above-mentioned solution manufacturing method for examination
Product solution and reference substance solution.Under the chromatographic condition drafted, precision draws 5 μ l of reference substance solution, 10 μ l, test solution 10
μ l, injects high performance liquid chromatograph, and measurement is calculated, three batches of assay results of Xihuang capsules with external standard two-point method logarithmic equation
Respectively 0.81 mg/, 0.83 mg/, 0.80 mg/.
This measuring method specificity is strong, and the linear of method, stability, reproducibility, sample recovery rate reach preferable technology
It is required that.This method can the product quality to Xihuang capsules effectively controlled.
Detailed description of the invention
Fig. 1: linear relationship investigates result (0.2mg cholic acid/ml methanol)；Fig. 2: standard curve (0.2mg cholic acid/ml first
Alcohol)；
Fig. 3: linear relationship investigates result (0.4 mg cholic acid/ml methanol)；Fig. 4: standard curve (0.4mg cholic acid/ml methanol)；Figure
5: stability test result；Fig. 6: repetitive test result；Fig. 7: sample recovery rate test result.
Specific embodiment
One, instrument, reagent and test sample
Instrument: Agilent1260 type high performance liquid chromatograph,
ELSD 2000ES evaporative light scattering detector.
Reagent and reagent:
Cholic acid reference substance: National Institute for Food and Drugs Control, lot number: 100078-201415 are derived from
Acetonitrile is chromatography pure level reagent, and other reagents are analytical grade reagent.
Test sample: Xihuang capsules 1708103,1708104,1708105
Chromatographic condition and system suitability:
Chromatographic column: using octadecylsilane chemically bonded silica as filler；
Mobile phase: with acetonitrile: 0.5% formic acid solution (38:62)
Flow velocity: 1ml/min
Column temperature: 30 DEG C.
Number of theoretical plate presses cholic acid (C24H40O5) calculate should be not less than 4000.
Two, test method and result
(1) preparation of reference substance solution: taking cholic acid reference substance appropriate, and accurately weighed 10mg is set in 50ml measuring bottle, adds methanol to quarter
Degree, be made solution of every 1ml containing 0.2mg to get.
(2) preparation of test solution
This product content about 1.0g is taken, is set in stuffed conical flask, it is accurately weighed, add methanol 25ml, weighed weight, 40 points of ultrasound
Clock is let cool, and weighed weight adds methanol to supply less loss weight, shakes up, be centrifuged, filtration, take subsequent filtrate to get.
(3) measuring method
It is accurate respectively to draw 5 μ l of reference substance, 10 μ l and test solution 5-10 μ l, high performance liquid chromatograph is injected, is measured, is used
External standard two-point method logarithmic equation calculate to get.
Three, the technical standard detected
This product every containing In vitro cultured Calculus Bovis with cholic acid (C24H40O5) meter, 0.5mg must not be less than.
Claims (1)
1. the detection method of cholic acid content in a kind of Xihuang capsules: it is characterized in that this method includes the following steps:
(1) chromatographic condition and system suitability:
Chromatographic condition and system suitability: using octadecylsilane chemically bonded silica as filler；It is molten with -0.5% formic acid of acetonitrile
Liquid (38:62) is mobile phase；Column temperature: 30 DEG C；Flow velocity: 1ml/min；Evaporative light scattering detector；Number of theoretical plate presses cholic acid (C
H40O5) calculate should be not less than 4000；
(2) preparation of reference substance solution: taking cholic acid reference substance appropriate, and accurately weighed 10mg is set in 50ml measuring bottle, adds methanol to quarter
Degree, be made solution of every 1ml containing 0.2mg to get；
(3) preparation of test solution
This product content about 1.0g is taken, is set in stuffed conical flask, it is accurately weighed, add methanol 25ml, weighed weight, 40 points of ultrasound
Clock is let cool, and weighed weight adds methanol to supply less loss weight, shakes up, be centrifuged, filtration, take subsequent filtrate to get；
(4) measuring method: it is accurate respectively to draw 5 μ l of reference substance, 10 μ l and test solution 5-10 μ l, inject high performance liquid chromatography
Instrument, measurement, with external standard two-point method logarithmic equation calculate to get；
This product every containing In vitro cultured Calculus Bovis with cholic acid (CH40O5) meter, 0.5mg must not be less than.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115575542A (en) * | 2022-10-19 | 2023-01-06 | 陕西科技大学 | Fingerprint construction method and fingerprint of Xihuang capsule |
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| CN101332287A (en) * | 2008-06-25 | 2008-12-31 | 浙江永宁药业股份有限公司 | Quality control method of Danle capsule |
| CN101829266A (en) * | 2010-04-28 | 2010-09-15 | 江西南昌桑海制药厂 | Method for detecting quality of bezoar snake bile bulbus fritilariae liquid |
| CN102879495A (en) * | 2012-09-27 | 2013-01-16 | 山东阿如拉药物研究开发有限公司 | Antidotal capsule of Tibetan medicine compound and quality detection method of preparation of antidotal capsule |
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2017
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Patent Citations (3)
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|---|---|---|---|---|
| CN101332287A (en) * | 2008-06-25 | 2008-12-31 | 浙江永宁药业股份有限公司 | Quality control method of Danle capsule |
| CN101829266A (en) * | 2010-04-28 | 2010-09-15 | 江西南昌桑海制药厂 | Method for detecting quality of bezoar snake bile bulbus fritilariae liquid |
| CN102879495A (en) * | 2012-09-27 | 2013-01-16 | 山东阿如拉药物研究开发有限公司 | Antidotal capsule of Tibetan medicine compound and quality detection method of preparation of antidotal capsule |
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| 金芬: "西黄软胶囊的药学研究", 《中国优秀硕士论文全文数据库 医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115575542A (en) * | 2022-10-19 | 2023-01-06 | 陕西科技大学 | Fingerprint construction method and fingerprint of Xihuang capsule |
| CN115575542B (en) * | 2022-10-19 | 2024-11-19 | 陕西科技大学 | A fingerprint map construction method and fingerprint map of Xihuang capsule |
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Application publication date: 20190625 |