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Method for measuring paeoniflorin metabolic products in liver microsome

A technology for liver microsomes and metabolites, which is applied in the field of medical detection technology-biochemical analysis, can solve the problems of long detection process, inability to detect and distinguish, large experimental errors, etc., and achieve rich structural feature information, high selectivity, High sensitivity and the effect of

Active Publication Date: 2017-07-21
LIAONING UNIVERSITY
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Problems solved by technology

[0004] The commonly used detection technology takes a long time, and the experimental error is relatively large. It is impossible to achieve high-throughput detection of a large number of metabolites at the same time. The structure of paeoniflorin metabolites is very similar, which cannot be detected and distinguished by conventional detection methods, and the specificity is poor.

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  • Method for measuring paeoniflorin metabolic products in liver microsome
  • Method for measuring paeoniflorin metabolic products in liver microsome
  • Method for measuring paeoniflorin metabolic products in liver microsome

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Embodiment

[0066] Example Method for Determination of Paeoniflorin Metabolites in Liver Microsomes

[0067] (1) Optimization of metabolic reaction conditions of paeoniflorin in liver microsomes and acquisition of metabolites

[0068] The NADPH regeneration system (Table 2) was used to conduct six groups of condition comparison experiments. HPLC was used to detect the content of paeoniflorin metabolites, and the metabolic rate was calculated and compared to determine the best conditions for the in vitro metabolic reaction of paeoniflorin in liver microsomes.

[0069] Method: Add appropriate amount of 0.05mol / L Tris-HCl buffer solution (pH 7.4) to 6 groups of culture flasks, and then add liver microsome solution, NADP solution, NADH solution, G-6-P solution, G- 6-P DH solution and MgCl 2 solution, shake well, put the 6 groups of culture bottles into the preheated, and keep the temperature at 37 ℃ in the full temperature shaker and incubate for 5min, then add appropriate amount of paeonifl...

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Abstract

The invention discloses a method for measuring paeoniflorin metabolic products in liver microsome and belongs to the technical field of medical detection. The method provided by the invention mainly comprises the following steps: transforming paeoniflorin liver microsome; performing sample pretreatment; and measuring the paeoniflorin metabolic products in the liver microsome by a liquid chromatography-mass spectrometry combination method. The method has the advantages of simplicity and convenience in operation, simplicity in sample treatment, high analysis speed, high specificity and high sensitivity. The method can be used for detecting more metabolic products of the paeoniflorin in the liver microsome; furthermore, by adoption of the analysis method, the structure of the metabolic products can be speculated more and accurately, technical support is provided for researching in vitro metabolic products of the paeoniflorin in the liver microsome, and important significance in deep research on active products and the action mechanism thereof is achieved.

Description

technical field [0001] The invention belongs to the field of medical detection technology-biochemical analysis, and specifically relates to a method for measuring paeoniflorin metabolites in liver microsomes. Background technique [0002] Radix Paeoniae Rubra is a traditional Chinese medicinal material in my country, which was first recorded in "The Classic". It has the effects of clearing away heat and cooling blood, dissipating blood stasis and relieving pain. Ulcers and other diseases. The chemical components in Radix Paeoniae Rubra are relatively complex, but the monoterpene glycosides in Radix Radix Radix Paeoniae Rubra are considered to be the main active components of Radix Paeoniae Rubra, mainly paeoniflorin and so on. Modern pharmacology shows that paeoniflorin has multiple effects such as dilating blood vessels, analgesic and sedative, anti-inflammatory and anti-ulcer, antipyretic and antispasmodic, and diuretic. [0003] Judging from the current domestic research...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8813
Inventor 刘玉峰朱丽君孙珊珊胡延喜徐亮卢晓丹
Owner LIAONING UNIVERSITY
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