Method of detecting amino acids in animal body fluid or tissue samples by ultrahigh efficient liquid chromatography-tandem quadrupole mass spectrometry

A tandem quadrupole, ultra-high performance liquid phase technology, applied in the field of animal food testing, can solve the problems of undiscovered amino acids, research blanks, etc., and achieve the effects of improving selectivity and sensitivity, simple processing method, and improved sensitivity

Inactive Publication Date: 2019-04-16
江苏澳华生物科技研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research report on the determination of amino acids in animal body fluids or tissue samples by LC-MS, and the current relevant research is still blank

Method used

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  • Method of detecting amino acids in animal body fluid or tissue samples by ultrahigh efficient liquid chromatography-tandem quadrupole mass spectrometry
  • Method of detecting amino acids in animal body fluid or tissue samples by ultrahigh efficient liquid chromatography-tandem quadrupole mass spectrometry
  • Method of detecting amino acids in animal body fluid or tissue samples by ultrahigh efficient liquid chromatography-tandem quadrupole mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1: Determination of Amino Acids in Animal Plasma

[0107] Animal plasma samples with batch number 2018091415 were taken out from the -80°C refrigerator and thawed on ice. Take 200 μL plasma and add 800 μL extract solution (methanol: acetonitrile volume ratio 1:1). After vortexing, the samples were centrifuged at 14500 rpm for 10 min at 4°C. The supernatant was collected and spin-dried in a vacuum centrifugal concentrator (Concentrator plus type, Eppendorf, Germany). The residue was reconstituted by adding 200 μL of methanol and water at a volume ratio of 1:1, vortexed and centrifuged again at 14500 rpm for 10 min at 4°C. The supernatant was transferred to the injection vial for detection by ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry.

[0108] The detection conditions are:

[0109] The chromatographic conditions of ultra-high performance liquid chromatography are:

[0110]Chromatographic column: Waters Atlantis T3 chroma...

Embodiment 2

[0127] Embodiment 2: the determination of amino acid in animal muscle

[0128] Take 15 mg of animal muscle sample with batch number 2018091515 and grind it in liquid nitrogen, and add 500 μL of extraction solution (methanol:water volume ratio is 8:2). After vortexing, the samples were placed on dry ice for 4 h, and centrifuged at 14500 rpm for 10 min at 4 °C. Collect the supernatant and add 400 μL of methanol extract to the residue, and vortex for 1 min. After standing on dry ice for 30 min, the sample was centrifuged again, and the supernatant was taken and mixed with the supernatant of the first extraction, and the supernatant was spin-dried in vacuum. The sample was reconstituted with 100 μL of methanol-water mixed solution with a volume ratio of 1:1, vortexed and centrifuged again at 14500 rpm for 10 min at 4 °C. The supernatant was transferred to the injection vial for detection by ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry.

[012...

Embodiment 3

[0130] Embodiment 3:: Determination of Amino Acids in Animal Liver

[0131] Take 15 mg of animal liver sample with batch number 2018101112 and grind it in liquid nitrogen, and add 500 μL of extraction solution (methanol:water volume ratio is 8:2). After vortexing, the samples were placed on dry ice for 4 h, and centrifuged at 14500 rpm for 10 min at 4 °C. Collect the supernatant and add 400 μL of methanol extract to the residue, and vortex for 1 min. After standing on dry ice for 30 min, the sample was centrifuged again, and the supernatant was taken and mixed with the supernatant of the first extraction, and the supernatant was spin-dried in vacuum. The sample was reconstituted with 100 μL of methanol-water mixed solution with a volume ratio of 1:1, vortexed and centrifuged again at 14500 rpm for 10 min at 4 °C. The supernatant was transferred to the injection vial for detection by ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry.

[0132] T...

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Abstract

The invention discloses a method of detecting amino acids in animal body fluid or tissue samples by ultrahigh efficient liquid chromatography-tandem quadrupole mass spectrometry. In particular, afterthe animal body fluid or the tissue sample is extracted and processed, the sample is detected by ultrahigh efficient liquid chromatography-tandem quadrupole mass spectrometry, and the amino acids is analyzed qualitatively and quantitatively. The method of detecting amino acids in animal body fluid or tissue samples by ultrahigh efficient liquid chromatography-tandem quadrupole mass spectrometry can simultaneously detect 26 kinds of amino acids in samples such as the animal body fluid and the tissue. Multi-reaction monitoring mass spectrometry is used for qualitative and quantitative analysis.The instrumental detection time of each sample is only 25 minutes, which is about 25% of the time of the existing method. The preprocessing method is simple and does not need derivation. Every amino acid can be separated well by extracting selective ions.

Description

technical field [0001] The invention belongs to the technical field of animal food detection, relates to a method for detecting amino acids in animal body fluids or tissue samples, in particular to a method for detecting amino acids in animal body fluids or tissue samples by ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry. Background technique [0002] As the main source of nitrogen required for animal growth, amino acids are not only substrates for protein synthesis but also signal molecules that play a vital role in animal growth and muscle protein deposition. Therefore, the accurate quantitative determination of amino acids in animal plasma, muscle, tissue and other samples provides strong technical support for evaluating the nutritional requirements of animals and finding the metabolic rules of amino acid synthesis and decomposition in animals. [0003] For most amino acids and their analogs, they are highly hydrophilic and difficult to r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 贺平丽孟成真许丹丹赵伟
Owner 江苏澳华生物科技研究院有限公司
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