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Extraction separation method of fat-soluble metabolite in faeces

A metabolite and separation method technology, applied in the field of extraction and separation of fat-soluble metabolites in feces, can solve the problems of inability to distinguish metabolites, etc., and achieve the effect of simple process and high extraction and separation efficiency

Active Publication Date: 2017-02-01
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Through the above-mentioned technical scheme, the present invention overcomes the disadvantage that HPLC-MS cannot distinguish metabolites of the same molecular weight in the existing research. At the same time, the extraction and separation efficiency of the present invention is high and the process is simple. While ensuring the completion of subsequent structure determination, it can also Obtain the extracted and separated metabolite components

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Study on the extraction and separation of metabolites from the feces of rats fed with polymethoxyflavonoids

[0023] The research object of this example is F344 rats fed with 5-hydroxylisotriment. They were raised in a metabolic cage and given standard food for AIN-76A rats (added with a total weight of 0.1% 5-hydroxy lichenpin). The food tank was replenished regularly every day and rat feces were collected within a week.

[0024] In order to conduct in-depth excavation and development of the metabolites in rat 5-hydroxyl chuanxifen feces, the present invention adopts the following extraction and separation methods:

[0025] (1) Crush 4.2g of rat feces samples, extract with 1000ml of organic solvent under reflux for 24 hours, concentrate under reduced pressure and evaporate the solvent; fecal residue is extracted again with the same organic solvent under reflux, and combined with the first extract to obtain the primary Extract the product. The primary extraction p...

Embodiment 2 2

[0030] Example 2 Study on the extraction and separation of metabolites from the feces of mice fed with dihydrochalcone (phlorizin)

[0031] The research object of this example is AJ mice fed epiphloridin. They were housed in a metabolic cage and given standard food for AIN-76A mice (with the addition of 0.1% phloridin in total weight). The food tank was replenished regularly every day and 3.6 g of all rat feces were collected within one week. The extraction and separation method of the present invention is used for the metabolites of phloridin in mouse feces:

[0032] (1) Crush 3.6 g of rat feces samples, and use the three organic solvents of n-hexane, ethyl acetate, and n-butanol to extract 1000ml of each under reflux for 24 hours, and concentrate under reduced pressure to evaporate the solvent; Solvent reflux extraction, combined with the first extract to obtain the primary extraction product. Concentrate, evaporate and weigh to obtain the weight of the primary extraction prod...

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Abstract

The invention discloses an extraction separation method of a fat-soluble metabolite in faeces. The extraction separation method comprises the following steps: extracting the metabolite in the faeces by an organic solvent, carrying out gradient chromatography by a petroleum ether / ethyl acetate elution system I and a dichloromethane / methyl alcohol elution system II, and separating metabolite monomers through high performance liquid chromatography. According to the extraction separation method, large-polarity and small-polarity impurities in a sample are removed through normal-phase silica gel column chromatography of gradient elution, the method plays a good role in enrichment and separation for the metabolite, the metabolite monomers can be obtained through extraction and separation, and the disadvantage that isomerides of the metabolite are difficultly identified by HPLC-MS is overcome; and moreover, while subsequent structure determination is guaranteed to be completed, an extracted and separated metabolite standard sample can be obtained.

Description

Technical field [0001] The present disclosure relates to a method for extracting and separating metabolites, in particular, to a method for extracting and separating fat-soluble metabolites in feces. Background technique [0002] Nutrients or drugs in food enter the body and are digested and absorbed through the mouth, esophagus, stomach, small intestine and other organs. Nutrients are absorbed from the small intestine into the blood circulation, and enter the liver through the hepatic portal vein. The liver is the main organ that metabolizes endogenous and exogenous substances. Through metabolism, these nutrients or drugs are broken down into other products. Metabolized products may have stronger biological activity than the parent body, so the study of metabolites in the body is of great significance to the pharmaceutical and food industries. [0003] At present, the research of metabolites mainly relies on HPLC-MS methods. However, HPLC-MS cannot separate and identify metaboli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/34B01D15/42
CPCB01D15/426G01N1/34G01N1/405
Inventor 郑金铠赵成英田桂芳肖航杨莹陈钰莹张晔
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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