Pretreatment method for determining 18 amino acids in C57 mouse brain tissues and blood plasma

A brain tissue and amino acid technology, applied in the preparation of test samples, measurement devices, color/spectral characteristics measurement, etc., can solve problems such as difficulties, interference measurement, and instrument contamination

Pending Publication Date: 2020-10-27
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
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Problems solved by technology

Based on the existing technology, there are still some difficulties in the simultaneous extraction of 18 kinds of amino acids by solid phase extraction.
In the existing pretreatment for the determination of amino acids in biological samples, the protein precipitation method is mainly used to examine the pros and cons of the pretreatment method through the removal rate of protein and the extraction rate of amino acids, but the influence of lipids and other substances on the determination is not considered , Lipids will interfere with the determination and pollute the instrument when the analyte is determined

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  • Pretreatment method for determining 18 amino acids in C57 mouse brain tissues and blood plasma
  • Pretreatment method for determining 18 amino acids in C57 mouse brain tissues and blood plasma
  • Pretreatment method for determining 18 amino acids in C57 mouse brain tissues and blood plasma

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Embodiment Construction

[0030] The present invention will be further described below in conjunction with the accompanying drawings and embodiments, but the scope of the present invention is not limited thereto.

[0031] Case 1: Brain tissue homogenate and plasma sample pretreatment

[0032] Take 200 μL of sample homogenate, add 200 μL of methyl tert-butyl ether, vortex shake for 30 s, centrifuge at 15000 g for 15 min at 4 °C, and discard the upper layer to remove most of the lipids in the sample.

[0033] Take 200 μL of the above-mentioned lipid-free sample, add 400 μL of acetonitrile, vortex shake for 30 s, centrifuge at 15000 g at 4 °C for 15 min, and take the supernatant to remove the protein in the sample; take 200 μL of the above-mentioned lipid-free sample, add 400 μL of 0.4mol·L -1 The perchloric acid solution was vortexed for 30s, centrifuged at 15000g for 15min at 4°C, and the supernatant was taken to remove the protein in the sample.

[0034] Case 2: Evaluation of pre-processing methods ...

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Abstract

The invention discloses a pretreatment method for determination of 18 amino acids in C57 mouse brain tissue and plasma and belongs to the field of pretreatment of endogenous substances in biological samples. According to the method, an ultraviolet spectrophotometer and a high performance liquid chromatograph are used, a method for treating brain tissue homogenate and plasma is investigated by combining liquid-liquid extraction with a protein precipitation method, and the method mainly investigates the influence of a sample pretreatment method on the removal amount of proteins, the removal amount of total lipids and the extraction amount of 18 amino acids in a sample, and further optimizes to obtain the optimal pretreatment method for determining 18 amino acids in C57 rat brain tissues andplasma.

Description

technical field [0001] The invention belongs to the field of pretreatment of endogenous substances of biological samples, and in particular relates to a pretreatment method for determining 18 kinds of amino acids in brain tissue and plasma of C57 mice. Background technique [0002] Quantitative determination of various endogenous markers in different biological samples is playing an increasingly important role in the study of disease pathogenesis. Changes in the content of amino acids that play the role of neurotransmitters in brain tissue are hot spots in the research of diseases such as depression and cerebral ischemia. The biological samples involved are mainly animal brain tissue and plasma. Before the quantitative determination of endogenous amino acids, biological samples need to be pretreated with special pretreatment methods to remove endogenous interfering substances and maximize the retention of endogenous amino acids to be determined. Brain tissue and plasma cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N21/31G01N30/06G01N30/74
CPCG01N1/28G01N21/3103G01N30/06G01N30/74
Inventor 蒋心惠李建沙张静龚涛
Owner CHONGQING MEDICAL UNIVERSITY
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