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Biological sample pretreatment method of vasopressin and analogues thereof

A technology for biological samples and analogues, applied in the field of analysis and detection, can solve problems such as reducing the accuracy of chromatographic detection and analysis

Inactive Publication Date: 2020-10-23
苏州旭辉检测有限公司
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a biological sample pretreatment method of vasopressin and its analogues with fast processing speed, low cost, simple method and easy promotion, which can effectively solve the problem of detection of vasopressin and its analogues. The problem of reducing the accuracy of chromatographic detection and analysis due to excessive impurities in the process

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  • Biological sample pretreatment method of vasopressin and analogues thereof

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Embodiment 1

[0021] Example 1 Biological sample pretreatment

[0022] (1) After the biological sample containing terlipressin was melted in an ice bath and protected from light, vortexed and mixed;

[0023] (2) Draw 500 μL of sample into a 1.5mL centrifuge tube, and the sample is a calibration standard, a quality control sample or a sample to be tested;

[0024] (3) Add 625 μL of 0.1% ammonia water to the sample, vortex for 2 minutes, then add 15 μL terlipressin internal standard, vortex evenly, when the sample is a double blank sample, use 15 μL of 50% methanol Aqueous solution instead of terlipressin internal standard;

[0025] (4) Activate the WCX SPE 96-well plate with 500 μL of 100% methanol and 500 μL of pure water in sequence, then transfer the aforementioned samples to the WCX SPE 96-well plate, and then use 1 mL of 2.0% ammonia water, Rinse with 1 mL of 100% methanol and 1 mL of 0.1% acetic acid methanol, and finally elute twice with 500 μL of 5.0% TFA methanol, and collect the ...

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Abstract

The invention provides a biological sample pretreatment method of vasopressin and analogues thereof. The method mainly comprises the following steps of melting a biological sample in an ice bath undera dark condition; vortex blending, sucking the sample into a centrifugal tube and then adding ammonia water into the sample; swirling, adding a terlipressin internal standard, swirling uniformly, activating a 96-well plate, transferring the sample into the 96-well plate, leaching with ammonia water, methanol and acetic acid methanol in sequence, eluting twice with TFA methanol, collecting eluenton a new 96-well plate, blow-drying the eluent, redissolving with a methanol aqueous solution, swirling, introducing a sample and analyzing. The method is high in treatment speed, low in cost, simpleand convenient, easy to popularize and capable of effectively solving a problem that in the detection process of the vasopressin and the analogues of the vasopressin, and chromatographic detection andanalysis accuracy is reduced due to too many impurities.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a pretreatment method for biological samples of vasopressin and its analogues. Background technique [0002] Vasopressin (AVP) belongs to the neurohypophysis hormone, also known as vasopressin or vasopressin, is a cyclic peptide hormone composed of nine amino acid residues produced by the optic nerve cells of the hypothalamus , and its theoretical molecular weight is 1084.24. Vasopressin has two receptors, V1 and V2. V1 is mainly distributed on the membrane of vascular smooth muscle cells, and acts through the receptor-G protein-second messenger pathway to constrict blood vessels and increase blood pressure. V2 is found in the epithelial cells of the distal tubule and collecting duct of the kidney, and physiological doses can promote the reabsorption of water by the distal tubule and collecting duct of the kidney, thereby exerting an antidiuretic effect. In view...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72
Inventor 李小川张枢石海兵李玉杰黄晓林
Owner 苏州旭辉检测有限公司
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