Metabonomics extraction method of pig gastrointestinal tract content

Abstract

The invention discloses a metabonomics extraction method of a pig gastrointestinal tract content. The method comprises: weighting 1g of the unfrozen gastrointestinal tract content; adjusting the pH value to 8.0 by adopting a NaOH solution with the concentration being 1M and a HCl solution with the concentration being 1M; weighting 0.1g of the uniformly-mixed content; adding 1mL of a phosphate buffer with the concentration being 0.1 M (and pH value being 8.0); performing fast liquid-nitrogen quick freezing; performing supersonic treatment and unfreezing in a water-bath pot; crushing the obtained product in a tissue crusher (20Hz, 30s); repeatedly performing circulation of freezing, supersonic treatment, and crushing for three times; finally, performing centrifugation with the rotational speed being 5000rpm for 10 min at 4 DEG C; and adding 0.6 mL of the supernatant liquid into an NMR tube of 5mm for later use. According to the method, 51 kinds of metabolites can be detected, wherein the metabolites comprise 20 kinds of amino acids, 7 kinds of short-chain fatty acids, 5 kinds of tricarboxylic acid circulation metabolites, and 19 kinds of other metabolites, and the whole extraction process needs only 3h. The peak shift is less susceptible to the pH value. Therefore, the extraction efficiency of metabolites is increased; the quality of an NMR Spectra is improved; and the extraction process is simplified.

Description

technical field [0001] The invention belongs to the technical field of metabolomics. Specifically relates to a method for extracting metabolomics of pig gastrointestinal tract contents. Background technique [0002] Intestinal health problems in piglets caused by factors such as stress and weaning syndrome are important causes of piglet mortality and reduced pig production efficiency. Indigestion in piglets can cause diarrhea and bacterial infections, and cause decreased immunity, increased morbidity, and even death. Since the feed will first pass through the intestinal flora after reaching the intestinal tract, the intestinal flora plays an important role in the digestion, absorption and metabolism of the feed, and the metabolites of the flora play an important role in the metabolism of the host. Intestinal flora, as a special community colonized in the intestine, participates in the metabolism of dietary carbohydrates, amino acids, lipids and other nutrients together wit...

AI Technical Summary

Problems solved by technology

The disadvantage is that the extraction process takes a long time

Although the concentrations of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid can be determined simultaneously, amino acids and other compounds cannot be detected at the same time because only short-chain fatty acids are targeted.

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