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Method for detecting aconitine in-vitro metabolic product by matrix assisted laser desorption mass spectrum

A matrix-assisted laser and mass spectrometry detection technology, which is applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of high toxicity

Active Publication Date: 2010-05-19
CHANGZHOU INST OF ENERGY STORAGE MATERIALS &DEVICES
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Problems solved by technology

(Zhao Yufeng et al., Chemical Journal of Chinese Universities: Vol.28(11), 2007, 2051-2055) Modern pharmacological studies have proved that diester-type alkaloids are more active, but also more toxic

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  • Method for detecting aconitine in-vitro metabolic product by matrix assisted laser desorption mass spectrum

Examples

Experimental program
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Effect test

Embodiment 1

[0010] Example 1: Put 0.5 mg of aconitine into 10 ml of common anaerobic culture medium (GAM medium) containing 0.2 g of healthy rat feces, culture it under anaerobic conditions at 37°C for 4 days, and take 100 microliters for culture The solution was extracted with 1 ml of dichloromethane, and the dichloromethane extract was centrifuged at a rate of 3000 rpm. After the supernatant was filtered through a 0.45 micron microporous membrane, the supernatant was mixed with the matrix, and the volume of the supernatant and the matrix was The ratio was 1:0.5, and the obtained mixture was analyzed by matrix-assisted laser desorption mass spectrometry.

[0011] The matrix was: 2 mg of α-cyano-4-hydroxycinnamic acid (CHCA), was added to 200 microliters of acetonitrile and water mixed solvent with a volume ratio of 3:7, and finally 8 microliters of trifluoroacetic acid was added.

[0012] Eighteen in vitro metabolites of aconitine were detected by mass spectrometry, including: monoester ...

Embodiment 2

[0013] Example 2: Put 1.5 mg of aconitine into 10 ml of common anaerobic culture medium (GAM medium) containing 0.2 g of healthy rat feces, cultivate it under anaerobic conditions at 37°C for 7 days, and take 200 microliters for culture The solution was extracted with 1 ml of dichloromethane, and the dichloromethane extract was centrifuged at a rate of 5000 rpm. After the supernatant was filtered through a 0.45 micron microporous membrane, the supernatant was mixed with the matrix, and the volume of the supernatant and the matrix was The ratio was 1:2, and the obtained mixture was analyzed by matrix-assisted laser desorption mass spectrometry.

[0014] The matrix was: 3 mg of α-cyano-4-hydroxycinnamic acid (CHCA), was added to 300 microliters of acetonitrile and water mixed solvent with a volume ratio of 7:3, and finally 10 microliters of trifluoroacetic acid was added.

[0015] Eighteen in vitro metabolites of aconitine were obtained through mass spectrometry detection (same ...

Embodiment 3

[0016] Example 3: Put 1 milligram of aconitine into 10 milliliters of common anaerobic culture medium (GAM culture medium) containing 0.2 grams of healthy rat feces, culture it under anaerobic conditions at 37°C for 5 days, and take 150 microliters for culture The solution was extracted with 1 ml of dichloromethane, and the dichloromethane extract was centrifuged at a rate of 3500 rpm. After the supernatant was filtered through a 0.45 micron microporous membrane, the supernatant was mixed with the matrix, and the volume of the supernatant and the matrix was The ratio was 1:1, and the obtained mixture was analyzed by matrix-assisted laser desorption mass spectrometry.

[0017] The matrix was: 2.5 mg of α-cyano-4-hydroxycinnamic acid (CHCA), was added to 250 microliters of acetonitrile and water mixed solvent with a volume ratio of 1:1, and finally 9 microliters of trifluoroacetic acid was added.

[0018] Eighteen in vitro metabolites of aconitine were obtained through mass spec...

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Abstract

The invention provides a method for detecting an aconitine in-vitro metabolic product by a matrix assisted laser desorption mass spectrum, which comprises the following steps of: adding aconitine into a common anaerobic bacterium culture medium containing the fresh excrement of a healthy rat to culture for a week to obtain culture liquid; extracting the culture liquid with dichloromethane; centrifuging the dichloromethane extract liquid, filtering supernatant liquid by a micropore filtering film and analyzing the aconitine in-vitro metabolic product by the matrix assisted laser desorption mass spectrum. The method analyzes the in-vitro metabolic products of 18 aconitines and is simple, easy, rapid and efficient.

Description

technical field [0001] The invention relates to a method for detecting metabolites of aconitine in vitro by matrix-assisted laser desorption mass spectrometry. technical background [0002] Ranunculaceae Aconitum, Aconitum aconitum, Aconitum aconitum and Aconitum aconitum are commonly used Chinese herbal medicines, which have various pharmacological activities such as strengthening the heart, relieving pain and reducing inflammation, and also have certain toxicity. Aconitine is one of the main active ingredients and toxic ingredients. Since traditional Chinese medicine is generally taken orally, its active ingredients will be affected by the intestinal flora in the intestinal tract before reaching the target organ for absorption. Aconitine can be metabolized into monoester alkaloids, diester alkaloids and lipid alkaloids under the action of intestinal flora. (Zhao Yufeng et al., Chemical Journal of Chinese Universities: Vol.28(11), 2007, 2051-2055) Modern pharmacological s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/64
Inventor 刘淑莹王曦烨宋凤瑞刘志强
Owner CHANGZHOU INST OF ENERGY STORAGE MATERIALS &DEVICES
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