A method for metabolomics extraction of porcine gastrointestinal contents

Abstract

The invention discloses a method for extracting metabolomics of pig gastrointestinal contents. The method is as follows: weigh 1 g of thawed gastrointestinal contents, use 1M NaOH solution and 1M HCl solution to adjust the pH value to 8.0, and then weigh 0.1 g mixed content, add 1mL of 0.1M phosphate buffer (pH 8.0), after liquid nitrogen quick freezing, ultrasonically thaw in a water bath, place in a tissue disruptor (20Hz, 30s) for crushing, and freeze and thaw repeatedly , ultrasonication and crushing treatment for 3 cycles. Finally, centrifuge at 5 000 rpm for 10 min at 4°C, and take 0.6 mL of the supernatant and add it to a 5 mm NMR tube for later use. 51 metabolites can be detected, including 20 amino acids, 7 short-chain fatty acids, 5 tricarboxylic acid cycle metabolites, and 19 other metabolites. The whole extraction process only takes 3 hours. The peak shift is little affected by pH. Therefore, this method not only improves the extraction rate of metabolites, improves the quality of NMR spectra, but also simplifies the extraction process.

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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