A kind of assay method of ginsenosides in rat plasma

A ginsenoside and plasma technology, which is applied in the detection field of ginsenoside, can solve the problem of high instrument cost, and achieve the effects of high sensitivity, high accuracy and good recovery rate of standard addition.

Active Publication Date: 2020-09-15
康龙化成(北京)生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the prior art, the method for studying the pharmacokinetics of ginsenosides in plasma is mainly liquid chromatography-tandem mass spectrometry, which has relatively high instrument costs, and there is no research report on the above five ginsenosides by simply using gas chromatography

Method used

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  • A kind of assay method of ginsenosides in rat plasma

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Take 100 μL of rat plasma, put it in a 2 mL centrifuge tube, add 200 μL methanol, vortex for 1 min, and centrifuge. Take the supernatant, put it into another clean 2mL centrifuge tube, add 500μL of dichloromethane, vortex for 1min, and centrifuge. The above two centrifugation conditions are 1000g, 3min. The Oasis HLB column was activated sequentially with methanol, water, and dichloromethane, and the extracted supernatant was added, eluted with 3 mL of methanol, and the eluate was collected. The eluent was taken, blown with nitrogen gas to near dryness, and the residue was redissolved in methanol.

[0027] The chromatographic conditions are as follows: DB-35MS, 30m×0.25mm, 0.25μm; heating program: the initial temperature is 60°C, rising to 160°C at a rate of 50°C / min, and then rising to 220°C at a rate of 20°C; Temperature 250°C; carrier gas: nitrogen, purity ≥99.999%; flow rate 1.2mL / min; injection volume 1μL; splitless injection.

[0028] The detection conditions a...

Embodiment 2

[0031] standard curve drawing

[0032] Add different volumes of ginsenoside reference solution in the centrifuge tube, after drying in a water bath, add blank plasma to form plasma samples containing different concentrations of ginsenoside, analyze using the pretreatment-analysis method of Example 1, and obtain a chromatogram Figure, record the peak area, take the peak area as the ordinate, and the concentration as the abscissa, draw the standard curve. The regression curves of ginsenosides Ra1, Rb1, Rh1, Re and Ro are as follows: y=0.057x+0.027, y=0.007x+0.215, y=0.098x-0.125, y=0.105x-0.024, y=0.0127x+ 0.114.

[0033] Method recovery

[0034] In order to verify the recovery rate of the method, the sample recovery experiment was carried out at three levels of high, medium and low respectively. The added material was then extracted and quantified. Three repetitions were performed for each of the three levels of high, middle and low. The results showed that the recoveries ...

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Abstract

The invention relates to detection of drug metabolism in plasma, and in particular relates to a method for determining five ginsenosides in rat plasma, and the method is characterized in that: Ra1, Rb1, Rh1, Re and Ro in the plasma can be detected by a gas chromatography method by dichloromethane extraction after precipitating of proteins with methanol, and solid phase extraction column purifying.Compared with the prior art, the method has the advantages of high accuracy, high sensitivity and good repeatability, is suitable for simultaneous quantitative detection of the five ginsenosides in the plasma, and can be used for the study of ginsenoside metabolism pathways in the plasma.

Description

technical field [0001] The invention relates to a method for detecting ginsenosides in rat plasma, which includes simultaneous quantitative detection of five ginsenosides, Ra1, Rb1, Rh1, Re and Ro. Background technique [0002] Ginseng has a wide range of pharmacological activities on the central nervous system, cardiovascular, endocrine, and immune systems. Ginseng saponin and its hydrolyzate in the intestinal tract are considered to be the main pharmacologically active ingredients in Asian ginseng, American ginseng, and Panax notoginseng. However, their mechanism of action is still unclear. It is very clear that the evaluation of the exposure level of the body to the components of saponins after taking ginseng preparations will help to uncover the material basis of its medicinal effects. [0003] In recent years, the pharmacokinetics of traditional Chinese medicine has been widely used in the interpretation of the material basis and mechanism of action of traditional Chine...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 马梅伍
Owner 康龙化成(北京)生物技术有限公司
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