Method for rapidly detecting content of B vitamins in pig intestinal contents/excrement

A technology of B vitamins and content, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as lack, and achieve the effect of rapid detection and simple sample pretreatment

Active Publication Date: 2020-12-25
ANIMAL SCI RES INST GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of methods for the efficient quantitative determination of multiple B vitamins in pig intestinal contents / feces

Method used

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  • Method for rapidly detecting content of B vitamins in pig intestinal contents/excrement
  • Method for rapidly detecting content of B vitamins in pig intestinal contents/excrement
  • Method for rapidly detecting content of B vitamins in pig intestinal contents/excrement

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] 1. Reagents and instruments

[0056] Formic acid (Sigma-Aldrich, 94318, USA), methanol (Fisher Chemical, A452-4, USA), acetonitrile (Sigma-Aldrich, 34851-4L, USA), isopropanol (Fisher Chemical, A461-4, USA), B vitamins standard products (both purchased from Sigma-Aldrich, the United States), and the test water is ultrapure water.

[0057] 5500QTRAP mass spectrometer (AB SCIEX, USA), ultra-high performance liquid chromatography system (Waters ACQUITY UPLCI-Class, USA), chromatographic column (HSS T3 2.5μm, 2.1mm×100mm column, USA), low-temperature high-speed centrifuge (Eppendorf, 5430R, the United States), high-speed homogenizer (T25, German IKA company), vortex oscillator (MS3 German IKA company).

[0058] 2. Instrument conditions

[0059] 2.1), ultra-high performance liquid chromatography conditions

[0060] Samples were separated using Waters ACQUITY UPLCI-Class system. Chromatographic column: Waters, HSS T3 (2.5μm, 2.1mm×100mm column); column temperature: 45°C; ...

Embodiment 2

[0087] 1. Sample collection and processing

[0088] 1.1), sample collection

[0089] In the experimental base of the Institute of Animal Science, Guangdong Academy of Agricultural Sciences, 35-day-old piglets were selected to collect fecal samples and cecal contents, and 63-day-old piglets were selected to collect cecal content samples and fecal samples. The specific information of the samples is shown in Table 4. In this experiment, all piglets were weaned at the age of 21 days, and all piglets were fed with the same diet in the pig farm.

[0090]Table 4 Basic information of samples

[0091]

[0092]

[0093] 1.2), sample pretreatment

[0094] Weigh about 200 mg of cecal contents / stool sample, 750 μL of methanol / acetonitrile / water (volume ratio 2:2:1) solution, extract homogeneously for 30 seconds, and vortex for 2 minutes.

[0095] 2. Sample testing

[0096] Centrifuge the pretreated sample solution at 14 000 r / min at 4°C for 15 minutes in an Eppendorf-5430R low-te...

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Abstract

The invention discloses a method for rapidly detecting the content of B vitamins in pig intestinal contents / excrement. The method comprises the following steps: weighing cecum contents / excrement of pigs as a sample, adding a methanol / acetonitrile / water solution, and performing homogeneous extraction to obtain a sample solution; and performing UPLC-MS / MS-MRM detection analysis on the sample solution to finally obtain the content of eight B vitamins in the sample, wherein the eight B vitamins are vitamin B1, vitamin B2, nicotinic acid, nicotinamide, pyridoxal, pyridoxine, vitamin B9 and vitaminB12. According to the method, an MRM technology is utilized, an ultra-high performance liquid chromatography system and a mass spectrometer are combined, and the content of a plurality of B vitamins in the samples such as the pig intestinal contents and the excrement can be rapidly detected based on targeted metabonomics of an MRM method.

Description

technical field [0001] The invention belongs to the field of animal husbandry and veterinary medicine, and in particular relates to a method for quickly detecting B vitamins in pig intestinal contents. Background technique [0002] B vitamins are components of various coenzymes and enzymes that participate in important metabolic activities in the body. The synthesis of intestinal microorganisms is one of the important sources of B vitamins for the host. B vitamins are important metabolites of intestinal microorganisms and play an important role in intestinal development, immune function and nutrient metabolism. They are widely used as feed additives Livestock and poultry farming. [0003] Accurately grasping the dynamic changes of B vitamins in intestinal contents is of great significance for studying the biological functions of B vitamins in the intestine. At present, the existing high-performance liquid chromatography (HLPC), high-performance capillary electrophoresis, m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/72G01N30/8634
Inventor 易宏波蒋宗勇王丽陈一波
Owner ANIMAL SCI RES INST GUANGDONG ACADEMY OF AGRI SCI
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