Preparation method and detection method of TMAO biological negative sample and biological standard substance

Abstract

The invention discloses a preparation method and a detection method of a TMAO biological negative sample and a biological standard substance. The method comprises the following steps: taking a biological sample, adding a biological enzyme to act on TMAO so as to reduce or eliminate the content of TMAO in the biological sample, and terminating the reaction between the biological enzyme and TMAO soas to obtain the biological negative sample. The method comprises the following steps: preparing a TMAO biological negative sample by using a biological enzyme, so that the endogenous TMAO backgroundinterference can be effectively eliminated, the reaction specificity is strong, the efficiency is high, the operation is convenient, and the economic benefit is high; by means of the scheme, endogenous TMAO background interference can be effectively eliminated, an enzyme reaction is ingeniously utilized, oxidative TMAO is reduced, metabolized and decomposed or groups are added to change structuralmodification, and therefore background interference caused by TMAO in a blank matrix is eliminated; and excessive bio-enzyme can be removed by subsequently inactivating or filtering the bio-enzyme, so that the bio-enzyme can be further applied to preparation of TMAO standard substances.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a preparation method and a detection method of a TMAO biological negative sample and a biological standard. Background technique [0002] Trimethylamine oxide (Trimetlyl amine oxide, TMAO) is a product produced by the metabolism of alkaloid components such as choline or carnitine through intestinal flora. Related studies have shown that TMAO is closely related to the occurrence and development of various cardiovascular diseases, such as atherosclerosis, heart failure and hypertension. Therefore, it is of great significance to detect TMAO. When establishing a TMAO detection method, it is necessary to use human plasma, urine, etc. as a blank matrix. However, almost all human plasma contains TMAO. Therefore, it is difficult to obtain a blank biological matrix during method validation. Blank biological matrices such as water, bovine serum albumin solution, phosphate buffer saline...

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Problems solved by technology

Some studies have shown that samples with low TMAO values ​​can be found. However, due to the limited number of such samples and the complicated search process, the practicability is not strong

Chinese invention patent application document CN109387410A discloses a TMAO-negative sample and its preparation method and application. It specifically discloses that TMAO contained in the sample can be removed by adding Reynolds salt to the biological sample. Although this method can eliminate the TMAO in the blank matrix For the purpose of TMAO background interference, there are obvious defects in the reaction of chemical complexing agents, such as: 1. The reaction process is relatively violent, and a large amount of precipitation will be produced after the reaction, and these precipitations will affect other components in the sample; 2. Due to biological samples The content of TMAO in the medium is relatively low, the added reducing agent is easy to be excessive, and the excessive reducing agent is difficult to completely remove or just react completely, resulting in the negative sample obtained cannot be prepared into TMAO biological standard by adding TMAO in the later stage; It is also highly toxic and easily enters the eyes or adheres to the skin, causing strong irritation; the sample after the reaction is extremely harmful to the water body, and even a small amount of the product cannot contact groundwater, waterways or sewage systems, so it is harmful to the experimental operators and the environment. There are great security risks

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