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Kit for determining trimethylamine oxide as well as preparation method and application of kit

A trimethylamine oxide and kit technology, applied in the field of detection of endogenous biomarkers, can solve the inconsistency between batches of mixed matrix, the difficulty of applying in vitro diagnostic kits to industrialization, the source of mixed human blood matrix and ethical restrictions and other issues to achieve the effect of guaranteeing precision and accuracy and excellent acceleration stability

Active Publication Date: 2022-04-19
长沙都正医学检验有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this approach ensures the “original taste” of the matrix, it ignores the fact that there are significant differences in TMAO content among samples, and the fact that mixed matrixes also have batch-to-batch inconsistencies, coupled with the fact that the source of mixed human blood matrix and ethics are restricted. Many limitations, so it is difficult to apply to the industrialization of the development of in vitro diagnostic kits (IVD)

Method used

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  • Kit for determining trimethylamine oxide as well as preparation method and application of kit
  • Kit for determining trimethylamine oxide as well as preparation method and application of kit
  • Kit for determining trimethylamine oxide as well as preparation method and application of kit

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preparation example Construction

[0063] The preparation of the above kits provided by the present invention may include the following steps:

[0064] The known concentration of the trimethylamine oxide standard solution is diluted with a matrix solution to obtain the series of oxidized trimethylamine oxide calibrator working liquid having a concentration gradient; wherein the concentration of the known concentration of the trimethylamine oxide standard solution may, for example, be about 1mg / mL, subject to the actual weighing preparation; the known concentration of the oxidized trimethylamine standard solution may also be referred to as the trimethylamine oxide calibration reserve solution;

[0065] The deuterated trimethylamine standard solution of the known concentration of deuterium oxide trimethylamine is diluted with a matrix solution to obtain the predetermined concentration of the deuterated trimethylamine oxide standard working liquid; wherein the concentration of the known concentration of the deuterat...

Embodiment 1

[0093] 1.1. Prepare the kit as follows:

[0094] 1) Weighing: Weigh TMAO-d9 standards, BSA, TMAO standards respectively.

[0095] 2) Preparation of stock solution: TMAO standards were prepared with 50% (volume) aqueous methanol solutions into a concentration of 1mg / mL of trimethylamine oxide calibrator stock solution, a concentration of 1mg / mL of trimethylamine oxide quality control liquid, and TMAO-d9 standards were prepared with a 50% (volume) aqueous methanol solution with a concentration of 1mg / mL of deuterated oxidation trimethylamine reserve liquid.

[0096] 3) Matrix aqueous solution: The matrix BSA is formulated with 10mM pH 7.4 PBS to make a 5% matrix solution.

[0097] 4) The trimethylamine oxide calibrator reserve liquid, the oxidized trimethylamine quality control liquid, and the deuterium oxide trimethylamine internal standard reserve liquid are diluted with the matrix aqueous solution to obtain a series of trimethylamine oxide calibration working liquid with a concen...

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Abstract

According to the kit for determining trimethylamine oxide and the preparation method and application of the kit, the current situation that a kit prepared through a solution technology is poor in stability can be effectively solved, and freeze-drying and long-term stability of an internal standard substance can be achieved. The kit comprises a series of trimethylamine oxide calibrators and a deuterated trimethylamine oxide internal standard substance, the series of trimethylamine oxide calibrators are freeze-dried substances of a series of trimethylamine oxide calibrator working solutions with concentration gradient, and the series of trimethylamine oxide calibrator working solutions are trimethylamine oxide calibrator working solutions with concentration gradient formed by diluting a trimethylamine oxide standard solution with known concentration with a matrix solution; the deuterated trimethylamine oxide internal standard substance is a lyophilized substance of a deuterated trimethylamine oxide internal standard substance working solution with a predetermined concentration, wherein the lyophilized substance is formed by diluting a deuterated trimethylamine oxide standard substance solution with a known concentration by using a matrix solution; a matrix in the matrix solution is serum albumin.

Description

Technical field [0001] The present invention relates to the field of detection technology for endogenous biomarkers, in particular to a kit for determining trimethylamine oxide and preparation methods and applications thereof. Background [0002] As an endogenous biomarker, trimethylamine oxide (TMAO), like other biomarkers, needs to first address matrix interference to ensure quantitative accuracy. Therefore, when choosing the right substrate, developers often take two main approaches: [0003] 1) Deduct the zero concentration blank blood matrix, that is, select the mixed blood as the matrix, subtract the different concentrations of calibrators and control products from the zero concentration mixed blood matrix, remove the TMAO interference in the mixed blood matrix itself, and then perform curve fitting and sample content determination. Although this practice ensures the "original taste" of the matrix, it ignores the fact that there are significant differences in TMAO content ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/72G01N30/04
CPCG01N30/02G01N30/06G01N30/34G01N30/72G01N2030/045
Inventor 侯利平王婷朱永瑞阳艳李超鹏李晓晖欧阳冬生
Owner 长沙都正医学检验有限责任公司
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