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Method for detecting content of asiaticoside-B and/or madecassoside by HPLC-CAD (High Performance Liquid Chromatography-Computer Aided Design) method

An HPLC-CAD, madecassoside technology, which is applied in the field of traditional Chinese medicine preparations, can solve the problems of contamination of chromatographic columns, increased column pressure, mixed isomeric volumes of madecassoside, etc., and achieves good effect and high accuracy Effect

Pending Publication Date: 2022-07-08
GUILIN SANJIN PHARMACEUTICALS CO LTD
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, with the progress of the research, it was found that the madecassoside detected by HPLC-ELSD method is actually mixed with the isomeric volume of madecassoside-B that is difficult to separate, which cannot truly reflect the quality attributes of the preparation
[0006] There have been related reports on the HPLC determination of related components in asiatica glycosides, all of which are determined under general HPLC conditions, and the mobile phase of the general methanol-water or acetonitrile-water system cannot effectively separate hydroxy asiatica Madecassoside and its isomer madecassoside B
In the existing literature, there is also a method of adding β-cyclodextrin to the mobile phase to improve the resolution, but the β-cyclodextrin mobile phase has significant defects, which can easily lead to abnormal increase in column pressure and contamination of the chromatographic column

Method used

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  • Method for detecting content of asiaticoside-B and/or madecassoside by HPLC-CAD (High Performance Liquid Chromatography-Computer Aided Design) method
  • Method for detecting content of asiaticoside-B and/or madecassoside by HPLC-CAD (High Performance Liquid Chromatography-Computer Aided Design) method
  • Method for detecting content of asiaticoside-B and/or madecassoside by HPLC-CAD (High Performance Liquid Chromatography-Computer Aided Design) method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] (1) Preparation of reference solution

[0062] Accurately weigh an appropriate amount of asiaticoside-B and madecassoside reference substance dried to constant weight, dissolve in methanol and dilute to 25ml and 10ml respectively, the prepared concentrations are 1.228mg / ml and 3.135mg / ml respectively The mixed standard solution of the above two components is prepared by 5, 10, 20, 25, 50 times dilution. Precisely draw 10 μl of each mixed standard solution, inject it into a liquid chromatograph, and measure.

[0063] (2) Preparation of the test solution:

[0064] Take 30 pieces (small pieces) or 20 pieces (large pieces) of three gold pieces, grind them into small pieces, take about 1.0 g, accurately weigh them, add 50 ml of methanol precisely, weigh them, and ultrasonically treat them for 45 minutes. The ultrasonic conditions are: power 250W, Frequency 40kHz; let cool, weigh again, make up the lost weight with methanol, shake well, and filter. Precisely measure 25ml o...

Embodiment 2

[0074] Embodiment 2 Optimization of test sample processing conditions

[0075] (1) Compare the contents of asiaticoside-B and madecassoside in the test solution prepared by different extraction methods and extraction time.

[0076] Ultrasonic extraction 45min and heating reflux extraction 2h were used as the comparison of extraction methods. In addition, three extraction times of ultrasonic extraction of 30min, 45min and 60min were compared. The results are shown in Table 3.

[0077] Table 3 Extraction method, determination result of extraction time selection

[0078]

[0079]

[0080] The results showed that there was no significant difference between ultrasonic extraction for 30 minutes and heating and reflux extraction for 2 hours. For the convenience of operation, ultrasonic extraction was selected as the extraction method, and it took 45 minutes to completely extract madecassoside, so the ultrasonic extraction time was set as 45 minutes.

[0081] (2) Selection of e...

Embodiment 3

[0092] Example 3 Methodological verification

[0093] (1) Linear

[0094]Accurately weigh an appropriate amount of asiaticoside-B and madecassoside reference substance dried to constant weight, dissolve in methanol and dilute to 25ml and 10ml respectively, the prepared concentrations are 1.228mg / ml and 3.135mg / ml respectively The mixed standard solution of the above two components is prepared by 5, 10, 20, 25, 50 times dilution. Precisely draw 10 μL of each mixed standard solution, inject it into a liquid chromatograph, and measure. Taking the amount of each reference substance as the abscissa (X), and the peak area of ​​the reference substance as the ordinate (Y), draw a standard curve, and calculate the regression equation. Asiaticoside-B is: Y=33.6543X-0.0340r=0.9993 ; Madecassoside is: Y=30.6748X+0.7686r=0.9984, the above test results are shown in Table 6. See working curve image 3 and Figure 4 .

[0095] Table 6 Linearity determination results

[0096]

[0097...

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Abstract

The invention discloses a method for detecting the content of asiaticoside-B and / or madecassoside by an HPLC-CAD (High Performance Liquid Chromatography-Computer Aided Design) method. The method comprises the following steps: (1) preparing a reference solution of the asiaticoside-B and / or madecassoside; (2) preparing a test solution, namely preparing a centella asiatica medicinal material, a centella asiatica medicinal material extract or a patent medicine preparation containing centella asiatica into the test solution; (3) detecting the content of asiaticoside-B and / or madecassoside in the reference substance and the test sample by adopting an HPLC-CAD (High Performance Liquid Chromatography-Computer Aided Design) method, and taking triacontylsilane chemically bonded silica as a filling agent as an adopted chromatographic column. According to the method, an HPLC-CAD method is adopted, a chromatographic column takes triacontylsilane chemically bonded silica as a filler, two components which are isomeride to each other can be effectively separated and the content of asiaticoside-B and madecassoside in a sample can be accurately evaluated by using a mobile phase with simple components, and the method is simple, convenient, sensitive and high in accuracy and can be used for establishing a quality control standard of a Sanjin preparation.

Description

technical field [0001] The invention belongs to the field of traditional Chinese medicine preparations, and in particular relates to a method for detecting the content of asiaticoside-B and / or madecassoside by HPLC-CAD method. Background technique [0002] Centella asiatica is the dried whole plant of Centella asiatica (L.) Urb. of Umbelliferae. Centella asiatica is used as medicine. It is cold in nature, bitter in taste and acrid. For damp-heat jaundice, carbuncle swollen poison, bruises, arsenic poisoning, lead poisoning, summer heat, infectious hepatitis, epidemic cerebrospinal meningitis, etc., but also anti-inflammatory, enhance memory, promote collagen synthesis and anti-cancer, etc. a variety of physiological activities. [0003] It is currently believed that triterpenoid saponins are the active components of Centella asiatica, mainly including three saponins, Asiaticoside, Madecassoside, and Asiaticoside-B. Compounds and their aglycones, their structural formulas a...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/64G01N30/08
CPCG01N30/88G01N30/64G01N30/08G01N2030/884Y02A50/30
Inventor 邹节明周艳林闵建国
Owner GUILIN SANJIN PHARMACEUTICALS CO LTD
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