Method for detecting level of metabolite of alkyl resorcinol in plasma through LC-MS/MS

A technology for alkyl resorcinol and metabolites, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome sample pretreatment, long analysis time, etc., achieve good chromatographic separation effect, high sensitivity, and reduce impurities Cumulative effect

Pending Publication Date: 2021-11-23
ZHUHAI PEOPLES HOSPITAL GUANGDONG PROVINCE
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Problems solved by technology

[0005] The present invention provides a method for detecting the level of plasma alkylresorcinol metabolites by LC-MS / MS, which is intended to solve the problems of long analysis time and complicated sample pretreatment in existing detection methods

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  • Method for detecting level of metabolite of alkyl resorcinol in plasma through LC-MS/MS
  • Method for detecting level of metabolite of alkyl resorcinol in plasma through LC-MS/MS
  • Method for detecting level of metabolite of alkyl resorcinol in plasma through LC-MS/MS

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Embodiment 1

[0048] Determination of Concentration of Alkyl Resorcinol Metabolites in Plasma by LC-MS / MS

[0049] 1. Plasma sample pretreatment:

[0050] a. After the plasma sample is thawed at room temperature and vortexed, use a microsampler to accurately pipette 50 μL of the plasma sample into a 1.5mL EP tube, and add the same volume of enzymatic digestion buffer (0.2mol / mL ammonium acetate buffer solution, β-glucuronidase ≥200U / mL, pH=5.0), mix well and incubate overnight at 37°C in a constant temperature water bath.

[0051] b. Take 50.0 μL of the enzymolyzed plasma sample in a 1.5 mL EP tube, add 20 μL of internal standard working solution (syringic acid concentration: 100 ng / ml), and 200 μL of acetonitrile.

[0052] c. Vortex for 1 min, centrifuge for 10 min (14000 rpm, 4° C.), transfer the supernatant of the centrifuge tube and add 200 μL of acetonitrile again, repeat the above centrifugation process twice, and then combine the supernatants transferred twice.

[0053] d. The comb...

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Abstract

The invention discloses a method for detecting the level of a metabolite of alkyl resorcinol in plasma through LC-MS/MS. The metabolite is 3-(3, 5-dihydroxyphenyl)-1-propionic acid (DHPPA), and belongs to the technical field of biomarker detection. A sample is pretreated by adopting a protein precipitation method, a BETASILPhenyl chromatographic column (2.1 * 150mm, 3mu m) is adopted for chromatographic separation, and gradient elution is carried out by adopting a mobile phase 0.1% acetic acid aqueous solution and 0.1% acetic acid acetonitrile; mass spectrometry employs electrospray ionization (ESI) and multiple reaction monitoring mode (MRM) to determine a target analyte. Comprehensive methodological verification is performed from indexes such as specificity, standard curve, linear range and LLOQ, precision and accuracy, extraction recovery rate and matrix effect, stability test and the like, and results all meet standard requirements. The method provided by the invention has the characteristics of high sensitivity, high specificity and high flux, and can realize accurate and rapid quantitative analysis of plasma DHPPA; sample pretreatment is simple and convenient, and large-scale sample analysis requirements of crowd research can be met.

Description

technical field [0001] The invention belongs to the technical field of biomarker detection, and in particular relates to a method for detecting the level of plasma alkylresorcinol metabolites by LC-MS / MS. Background technique [0002] Alkylresorcinols are a group of phenolic lipids that are abundant in the bran of wheat and rye and low or absent in refined wheat and rye and other common grains. Alkyl resorcinol has heat stability in food processing, about 60% is absorbed by the human body, and its metabolic pathway in the human body is similar to that of tocopherol, and it mainly undergoes β-oxidation in the liver through the cytochrome P450 enzyme system, and the resulting metabolism The products 3-(3,5-dihydroxyphenyl)-1-propionic acid (DHPPA) and 3,5-dihydroxybenzoic acid (DHBA) could be detected in human plasma. Both interventional and observational studies found that the concentrations of alkylresorcinols and their metabolites in the body were significantly affected by...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/54G01N30/72
CPCG01N30/02G01N30/06G01N30/54G01N30/72
Inventor 孙涛平占美晓耿旭洋邓瑶
Owner ZHUHAI PEOPLES HOSPITAL GUANGDONG PROVINCE
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