CN111351866A

CN111351866A - Method for detecting hyodeoxycholic acid in Xihuang capsules

Info

Publication number: CN111351866A
Application number: CN201811567159.6A
Authority: CN
Inventors: 谢秋芳
Original assignee: Hebei Wanbang Folon Pharmaceutical Co ltd
Current assignee: Hebei Wanbang Folon Pharmaceutical Co ltd
Priority date: 2018-12-20
Filing date: 2018-12-20
Publication date: 2020-06-30

Abstract

The invention provides a method for detecting hyodeoxycholic acid in Xihuang capsules, thereby effectively controlling the product quality.

Description

Method for detecting hyodeoxycholic acid in Xihuang capsules

Technical Field

The invention relates to the technical field of drug detection, in particular to a method for detecting hyodeoxycholic acid in Xihuang capsules by using high performance liquid chromatography.

Background

In the state of the food and drug administration, the West yellow capsule is collected in the national drug Standard WS₃And each particle is 0.25 g-B-3210-98-2016, under the condition of the original system in the standard, the separation degree problem exists, the number of interference peaks is large, the result judgment is influenced, and the detection method is formulated for better controlling the product quality.

Disclosure of Invention

The technical problem to be solved by the invention is to design a detection method for detecting hyodeoxycholic acid in Xihuang capsules, so that the product quality is effectively controlled.

The invention provides a detection method for hyodeoxycholic acid in Xihuang capsules.

The object of the invention can be achieved by the following scheme:

instrument, reagent and test article

An Agilent 1260 high performance liquid chromatograph; shimadzu Inertsil ODS-3C18 column (4.6 μm 250mm, 5 μm)

The reagent acetonitrile is chromatographic grade reagent, and other reagents are analytical pure reagents. The water is purified water

Control hyodeoxycholic acid 111845-

Xihuang capsule 18061271807105180713318081011808110 for test

Chromatographic conditions and system applicability test:

high performance liquid chromatograph Agilent 1260

Chromatographic Shimadzu Inertsil ODS-3C18 column (4.6 μm 250mm, 5 μm)

Mobile phase acetonitrile-0.5% formic acid solution (35: 65)

Flow rate 0.8 ml/min^-1

Column temperature 30 deg.C

The spraying temperature of the evaporation light scattering detector is 35 ℃, and the temperature of the drift tube is 80 DEG C

The theoretical plate number is not lower than 4000 calculated according to the peak of hyodeoxycholic acid.

Second, test methods and results

(1) Preparation of test solution about 2g of the content of the product under the condition of different loading amount is taken, precisely weighed, added with 40ml of methanol, ultrasonically treated for 40 minutes, cooled, filtered, the filtrate is evaporated to dryness, the residue is added with a proper amount of methanol to dissolve and is transferred to a 10ml measuring flask, added with methanol to scale, shaken evenly and filtered, and the subsequent filtrate is taken as the test solution.

(2) Preparation of control solution A hyodeoxycholic acid control solution was prepared by adding methanol to obtain a solution containing 0.2mg per 1ml as a control solution.

(3) The determination method comprises precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into high performance liquid chromatograph, and determining.

And thirdly, in the chromatogram of the technical standard test sample, a chromatographic peak which is the same as the retention time of the hyodeoxycholic acid control sample cannot be presented.

Fourthly, the feasibility of the method can be illustrated by the following tests

Investigation of linear relationships

Precisely sucking 1, 2, 3, 4 and 5 mu l of hyodeoxycholic acid control solution (0.2064 mg/ml) respectively, injecting the solution into a high performance liquid chromatograph, measuring, and performing linear regression on peak area (Y) by using mass concentration (x), wherein the regression equation is Y =122.7x-1.938, R²The result of =0.9978 shows that the linear relationship is good. See fig. 1, 2.

Precision survey

Precisely sucking 10 μ l of hyodeoxycholic acid control solution (0.2064 mg/ml), and repeating the sample injection for 5 times, wherein the peak area RSD is 2.66%, and the precision is good. The results are shown in FIG. 3.

Specialization inspection

Sample adding test solution preparation: sucking 5ml of each of the sample solution and the reference solution, mixing in a 10ml measuring flask, filtering, and collecting the filtrate.

Precisely sucking 10 mul of a control solution and 10 mul, 15 mul, 20 mul and 30 mul of a test solution into a high performance liquid chromatograph respectively by a measuring method, and measuring a result: the sample does not have chromatographic peak at the same retention time as the hyodeoxycholic acid control, and the labeled sample has peak at the same retention time as the control, thus proving good specificity. The results are shown in FIG. 4.

Durability test

Preparation of test solution: weighing 1808109 batches of 2.0079g of Xihuang capsules, 1808110 batches of 2.0163g of Xihuang capsules, 1807133 batches of 2.0099g of Xihuang capsules and 1806127 batches of 2.0234g of Xihuang capsules, and preparing a test solution according to a preparation method of the test solution.

Using a Watts Waters symmetry purified RP18 type C18 (No. LC-016) (4.6X 250), and Shimadzu Inertsil ODS-3 type C18 (No. LC-008) (4.6X 250) chromatographic columns, 10. mu.l of the sample was precisely injected under the above chromatographic conditions, and detection was carried out. As a result, 2 chromatographic columns of different brands are well separated, and no interference peak exists near a target peak in the chromatogram of the test sample. The results are shown in FIG. 5.

Drawings

The determination method has strong specificity, and the linearity, the precision and the durability of the method all meet better technical requirements. The method can effectively control the quality of the Xihuang capsule product.

FIG. 1 is a graph of results of a linear relationship investigation; FIG. 2 is a linear trend graph; FIG. 3 is a graph showing the results of precision examination; FIG. 4 is a diagram of results of a specificity study; FIG. 5 is a graph showing the result of durability test.

Detailed Description

Instrument, reagent and test article

Control hyodeoxycholic acid 111845-

Xihuang capsule 18071051806127180713318081011808110 for test

Test method and results

High performance liquid chromatography is adopted.

Chromatographic conditions and system applicability test:

high performance liquid chromatograph Agilent 1260

Chromatographic Shimadzu Inertsil ODS-3C18 column (4.6 μm 250mm, 5 μm)

Mobile phase acetonitrile-0.5% formic acid solution (35: 65)

Flow rate 0.8 ml/min^-1

Column temperature 30 deg.C

The chromatogram of the test sample does not show a chromatographic peak with the same retention time as that of the hyodeoxycholic acid control.

Claims

1. A method for detecting hyodeoxycholic acid in Xihuang capsules is characterized by comprising the following steps:

preparing a test solution, precisely weighing about 2g of the content of the product under the condition of different loading amounts, adding 40ml of methanol, performing ultrasonic treatment for 40 minutes, cooling, filtering, evaporating filtrate, dissolving residue in a proper amount of methanol, transferring to a 10ml measuring flask, adding methanol to scale, shaking up, filtering, and taking a subsequent filtrate as the test solution;

preparing a control solution by adding methanol into hyodeoxycholic acid control to obtain a solution containing 0.2mg per 1ml as the control solution;

the liquid chromatography condition and system applicability test is tested by high performance liquid chromatography, and octadecylsilane chemically bonded silica is used as a filler; acetonitrile-0.5% formic acid solution (35: 65) is used as a mobile phase; flow rate: 0.8 ml/min^-1(ii) a Column temperature: 30 ℃; evaporating a light scattering detector, wherein the spraying temperature is 35 ℃, and the drift tube temperature is 80 ℃;

the number of theoretical plates is not less than 4000 calculated according to the peak of hyodeoxycholic acid;

the determination method comprises precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into high performance liquid chromatograph, and determining.

2. The detection method according to claim 1, characterized in that chromatographic conditions and system suitability tests of the method;

octadecyl bonded silica gel is used as a filling agent; acetonitrile-0.5% formic acid solution (35: 65) as mobile phase; flow rate: 0.8 ml/min^-1(ii) a Column temperature: 30 ℃; evaporating a light scattering detector, wherein the spraying temperature is 35 ℃, and the drift tube temperature is 80 ℃;

the theoretical plate number is not lower than 4000 calculated according to the peak of hyodeoxycholic acid;

3. The test method according to claim 1, characterized in that the test result of the method is that chromatographic peaks of the sample of the Xihuang capsule cannot be shown in the sample chromatogram of the sample, and the chromatographic peaks can not be shown in the sample chromatogram of the sample of the Hehyodeoxycholic acid.