GC-MS (gas chromatography-mass spectrometer)-based method for quantifying eleven types of short-chain fatty acids in intestinal contents and fecal samples

A short-chain fatty acid, GC-MS technology, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problem of inaccurate quantitative short-chain fatty acid results.

Inactive Publication Date: 2016-06-08
CHINA PHARM UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, during the pretreatment process of this method, due to the effect of the introduction of acid, the extracted short-cha...

Method used

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  • GC-MS (gas chromatography-mass spectrometer)-based method for quantifying eleven types of short-chain fatty acids in intestinal contents and fecal samples
  • GC-MS (gas chromatography-mass spectrometer)-based method for quantifying eleven types of short-chain fatty acids in intestinal contents and fecal samples
  • GC-MS (gas chromatography-mass spectrometer)-based method for quantifying eleven types of short-chain fatty acids in intestinal contents and fecal samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Determination of the content of short-chain fatty acids in the mouse cecum content sample

[0018] (1) Sample pretreatment: collect mouse cecum contents, freeze-dry them into powder, weigh 150mg freeze-dried powder, add 150uL 1M hydrochloric acid saturated sodium chloride solution first, then add 1.5mL ethyl acetate (containing IS5ug / mL) , put the sample on a homogenizer, grind and mix well, take 220uL supernatant after centrifugation at 10000rpm for 10min, dry it with 100mg magnesium sulfate, centrifuge at 18000rpm for 10min, then take 150uL supernatant and dry it with 50mg magnesium sulfate, centrifuge at 18000rpm for 10min , take 72uL of the supernatant and put it in a sealed gas-phase vial, add 18uL of derivatization reagent MTBSTFA, tighten the bottle cap, mix well and place it in a water bath at 80°C for 20min, then place the sample at room temperature for 8h to be derivatized for analysis;

[0019] (2) GC-MS analysis:

[0020] Get the sample obtained ...

Embodiment 2

[0036] Embodiment 2: Measuring the content of short-chain fatty acids in mouse feces samples

[0037] (1) Sample pretreatment: collect mouse feces samples, lyophilize into powder, and all the other processing steps and GC-MS analysis steps are the same as in Example 1. The content of short-chain fatty acids in the measured blank mice is as follows: figure 2 -B.

[0038] Due to the low content of isocaproic acid and caproic acid in the intestinal tract, no isocaproic acid and caproic acid were detected in the intestinal contents and feces samples of mice by this method. However, the established method for the determination of 11 short-chain fatty acids in intestinal contents and stool samples can be extended to other tissues or liquids with high content of isocaproic acid and caproic acid, such as caproic acid bacteria fermentation broth, and the method should be verified , so the establishment of this method is still meaningful.

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Abstract

The invention belongs to the technical field of endogenous substance detection and discloses a GC-MS (gas chromatography-mass spectrometer)-based method for quantifying eleven types of short-chain fatty acids in intestinal contents and fecal samples. By adoption of a hydrochloric acid solution prepared from saturated salt water for acidification, high-stability ethyl acetate for extraction and N-tert-butyldimethylsilyl-N-methyl trifluoroacetamide MTBSTFA for derivatization, the eleven short-chain fatty acids in mouse intestinal contents and fecal samples are quantified by a gas chromatography-mass spectrometer. The method is high in accuracy and precision and low in detection limit, can be used for quantitative detection of eleven types of short-chain fatty acids and can meet detection requirements on determination of major short-chain fatty acids in the mouse intestinal contents and the fecal samples. Moreover, the method is conducive to analysis and comparison of metabolic disposition laws of the short-chain fatty acids under normal and enteritis conditions and observation whether changes of the short-chain fatty acids can represent dynamic conditions of enteritis or not, thereby providing significant bases for potential biomarkers under the enteritis condition.

Description

technical field [0001] The invention relates to a detection method for rapidly quantifying 11 kinds of short-chain fatty acids in intestinal contents and feces samples simultaneously by using gas chromatography-mass spectrometry, which belongs to the detection technology method of endogenous substances. Background technique [0002] Short-chain fatty acids are mainly produced by the fermentation of undigested and absorbed carbohydrates in the diet by intestinal flora, mainly including acetic acid, propionic acid and butyric acid. Under physiological conditions, short-chain fatty acids can provide intestinal epithelial cells with energy for their metabolism, and play an important role in intestinal mucosal cell proliferation, differentiation, maintenance of colon integrity and mucosal immune response. In a pathological state, such as enteritis, short-chain fatty acids can promote the proliferation of intestinal mucosal cells, maintain their normal shape, protect the intestina...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/02G01N30/88
CPCG01N30/06G01N30/02G01N30/88G01N2030/025G01N2030/884
Inventor 郝海平曹丽娟邓珊珊王广基
Owner CHINA PHARM UNIV
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