Method and kit for determining amount of coenzyme Q10 in biological sample

By optimizing the mobile phase and mass spectrometry parameters through fuzzy logic and combining it with liquid chromatography-tandem mass spectrometry, the problems of insufficient accuracy and sensitivity in LC-MS/MS detection of coenzyme Q10 were solved, achieving efficient and accurate CoQ10 detection, reducing costs and improving detection reliability.

CN120820643APending Publication Date: 2025-10-21SICHUAN TAIKANG HOSPITAL CO LTD
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Patent Information

Application Number
CN202510837437.9
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-06-20
Publication Date
2025-10-21

AI Technical Summary

Technical Problem

Existing LC-MS/MS detection methods have problems with insufficient accuracy and sensitivity when detecting coenzyme Q10 in biological samples. In particular, reduced CoQ10 is easily oxidized during sample storage and pretreatment, resulting in increased detection errors. In addition, commercial kits have insufficient standard curve stability under specific conditions.

Method used

Fuzzy logic was used to optimize the mobile phase combination and mass spectrometry parameters. Combined with liquid chromatography tandem mass spectrometry (LC-MS/MS), 1,4-benzoquinone solution and internal standard were used for extraction. Isopropanol was optimized as the extractant. An APCI ion source was used. Fuzzy reasoning was used to determine the optimal mobile phase ratio of methanol:isopropanol = 8:2. Mass spectrometry conditions were optimized to achieve a balance between signal intensity, retention time and background noise.

Benefits of technology

The accuracy and sensitivity of the test were significantly improved, with the lower limit of quantification reaching 0.1 ng/mL, and the intra-batch and inter-batch CVs controlled within 10%, meeting the relevant standard requirements, reducing experimental costs and resource consumption, and enhancing the reliability of CoQ10 detection.

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Abstract

Methods and kits for determining the amount of coenzyme Q10 in a biological sample are provided. Relates to the technical field of medical biology. The method comprises the following steps: taking a biological sample to be determined, adding a 1, 4-benzoquinone solution and an internal standard substance, adding an extracting agent, extracting, taking an extracting solution, filtering, and performing liquid chromatography-tandem mass spectrometry determination, wherein the chromatographic conditions are as follows: a Waters C18 chromatographic column is adopted, and a 100% B phase is used as a mobile phase for elution; the mobile phase comprises 0.1% by volume of a component 1 and 99.9% by volume of a component 2, the component 1 is formic acid, and the component 2 is composed of methanol and isopropanol in a ratio of (7-9): (3-1). According to the embodiment of the invention, parameters are optimized through fuzzy logic, the efficient, accurate, sensitive and low-cost method for measuring the amount of the coenzyme Q10 in the biological sample is established, the mobile phase selection of LC-MS / MS detection of the CoQ10 is optimized by virtue of the capability of processing uncertain data through the fuzzy logic, and the precision of the method is remarkably improved compared with that of similar researches.
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