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Quantitative analysis method and kit for detecting various intestinal microbial metabolites

A technology for gut microbes and detection kits, applied in the field of detection, can solve problems such as indicators that cannot be checked at one time

Pending Publication Date: 2021-07-20
上海脉示生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, although the current detection method also uses liquid chromatography-mass spectrometry, it can only detect one or a small number of indicators, and cannot jointly detect all indicators at once.

Method used

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  • Quantitative analysis method and kit for detecting various intestinal microbial metabolites
  • Quantitative analysis method and kit for detecting various intestinal microbial metabolites
  • Quantitative analysis method and kit for detecting various intestinal microbial metabolites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0149] 1. Main instruments and equipment:

[0150] Shimadzu LC-MS 8050; LabSolutions Insight software.

[0151] 2. Reagents and materials:

[0152] Acetyl-L-Carnitine, Betaine Aldehyde, Betaine, Choline Chloride, Crotonobetaine Hydrochloride, Gamma-Butyl Betaine, L-Carnitine, Trimethylamine Hydrochloride, Trimethyllysine, and Trimethylamine Oxide Standard Products, Acetyl L-carnitine-D3, Betaine-D11, Choline Chloride-D9, L-Carnitine-D9, Trimethylamine-D10, Trimethylamine Oxide-D9 Internal Standard, Methanol, Acetonitrile, Ethanol, Formic Acid, Ammonia , sodium carbonate, and carbon-adsorbed human plasma are all commercially available reagents.

[0153] 3. Liquid chromatography and mass spectrometry analysis conditions:

[0154] Chromatographic conditions:

[0155] Column: ACQUITY UPLC BEH HILIC ( 1.7μM, 2.1×100mm);

[0156] Guard pre-column: ACQUITY UPLC BEH HILIC VanGuard pre-column( 1.7μM, 2.1×5mm);

[0157] Mobile phase A: the aqueous solution of 0.1% (v / v) formic...

Embodiment 2

[0179] Example 2 By selecting different concentrations of sodium carbonate solution in the enhancer component B, the influence of the enhancer composed of tert-butyl bromoacetate and sodium carbonate solution on the pretreatment of quality control I, quality control II and samples was verified again. The results of Table 1, Table 2, and Table 3 show that the different concentrations of sodium carbonate solution in the enhancer have an impact on the detection results of acetyl-L-carnitine, trimethylamine oxide, and trimethylamine in quality control I, quality control II, and samples, and have an impact on the detection results of betaine Aldehyde, betaine, choline chloride, croton betaine, γ-butyl betaine, L-carnitine, and trimethyllysine were not affected.

[0180] The different concentrations of the sodium carbonate aqueous solution in the embodiment 2 of table 1 affect the detection of quality control I (unit: uM)

[0181]

[0182] The different concentrations of the sodi...

Embodiment 3

[0189]Example 3 By selecting different concentrations of acetic acid in the extraction solvent acetonitrile acetic acid, the effect of the extraction solvent composed of acetonitrile and acetic acid on quality control I, quality control II and plasma sample pretreatment was verified again. As shown in Table 4, Table 5, and Table 6 below, the different concentrations of acetic acid in the extraction solvent have an impact on the detection results of crotonbetaine, trimethylamine oxide, and trimethylamine in quality control I, quality control II, and plasma samples. Aldehyde, betaine, choline chloride, croton betaine, γ-butyl betaine, L-carnitine, and trimethyllysine had no effect.

[0190] The different concentrations of acetic acid in acetonitrile acetic acid in the embodiment 3 of table 4 affect the detection of quality control I (unit: uM)

[0191]

[0192] The different concentrations of acetic acid in the acetonitrile acetic acid in the embodiment 3 of table 5 affect th...

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Abstract

The invention provides a quantitative analysis method and a kit for one-time detection of multiple intestinal microbial metabolites for the first time. The kit is composed of a reference substance, a quality control substance, an isotope internal standard extracting solution, a reinforcing agent, an extracting solvent, a mobile phase additive A, a mobile phase additive B, a 96-hole reaction plate, a 96-hole filter plate and a specification. The kit can be used for simultaneously detecting acetyl L-carnitine, betaine aldehyde, betaine, choline chloride, crotonbetaine hydrochloride, gamma-butyrobetaine, L-carnitine, trimethylamine hydrochloride, trimethyl lysine and trimethylamine oxide, the consumed time is short, and the detection efficiency is improved.

Description

technical field [0001] The invention belongs to the detection field, and in particular relates to a quantitative analysis method and a kit for detecting various intestinal microbial metabolites. Background technique [0002] Cardiovascular disease is currently the number one cause of death among urban and rural residents in China, higher than tumors and other diseases. The newly released "China Cardiovascular Disease Report 2018" shows that the prevalence and mortality of cardiovascular disease in China are still on the rise. It is estimated that there are 290 million patients with cardiovascular disease nationwide, including 13 million strokes and 11 million coronary heart diseases. 5 million for pulmonary heart disease, 450,000 for heart failure, 2.5 million for rheumatic heart disease, and 270 million for high blood pressure. 30% of the population in China have ≥3 cardiovascular risk factors, that is, nearly 400 million people have ≥3 cardiovascular risk factors at the s...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/89G01N30/72G01N30/04G01N30/06G01N30/34G01N30/88
CPCG01N30/02G01N30/89G01N30/72G01N30/04G01N30/06G01N30/34G01N30/88G01N2030/045G01N2030/067G01N2030/8822
Inventor 詹红铃木亨霍志远孔志霞蔡洪玉
Owner 上海脉示生物技术有限公司
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