A method for detecting 5-aminolevulinic acid by high performance liquid chromatography tandem mass spectrometry

Abstract

The invention discloses a method for detecting 5-aminolevulinic acid by using a high performance liquid phase tandem mass spectrometer. It uses a substitute matrix that does not contain 5-aminolevulinic acid, and adds different concentrations of 5-aminolevulinic acid solutions to prepare a standard curve; adds different concentrations of 5-aminolevulinic acid solutions to a real blank matrix to prepare a quality control sample. After adding a stable isotope-labeled internal standard to the standard curve and quality control samples, the samples were deproteinized and derivatized, and then measured with a high-performance liquid chromatography tandem mass spectrometer to establish a relationship between the mass spectral response and the concentration of 5-aminolevulinic acid. Linear relationship, so as to deduce the concentration of 5-aminolevulinic acid in the sample to be tested with unknown concentration. The invention solves the problems of difficult accurate quantification of endogenous 5-aminolevulinic acid in the biological matrix, many interference components and low sensitivity, and has the advantages of simple operation, high sensitivity, high flux and low cost, and can meet the requirements of large-scale biological Concentration determination of 5‑aminolevulinic acid in matrix samples.

Description

technical field [0001] The invention belongs to the technical field of detection, and in particular relates to a method for detecting 5-aminolevulinic acid by using high performance liquid phase tandem mass spectrometry. Background technique [0002] Photodynamic therapy (PDT) of photosensitizers is a novel non-invasive treatment method for tumors and inflammations, which relies on irradiation of light sources with specific wavelengths to activate photosensitizers in tumors and inflammatory tissues to produce biologically toxic singlet oxygen, etc. Active oxygen species, and then oxidize and damage tumors, kill bacteria, fungi, and eliminate inflammation. 5-Aminolevulinic acid (5-ALA) is one of the known photosensitizers at this stage and has been widely reported in the literature. [0003] Photodynamic therapy based on 5-aminolevulinic acid has less trauma in the treatment of skin diseases, and has become the first-line treatment for skin precancerous lesions, severe acne ...

AI Technical Summary

Problems solved by technology

[0008] In order to overcome the analysis and detection research: 5-aminolevulinic acid is highly polar and has weak retention on conventional reversed-phase chromatographic columns; the concentration of 5-aminolevulinic acid in biological matrices is low, usually at the concentration level of nanograms per milliliter; 5 -Aminolevulinic acid is an endogenous compound, and there are high concentrations of endogenous or exogenous interferences in the body that are similar to the chemical structure of 5-aminolevulinic acid. The technical problem to be solved by the present invention is to provide a A method that can accurately detect trace amounts of 5-aminolevulinic acid in biological matrices. This method uses high-performance liquid chromatography tandem mass spectrometry to detect 5-aminolevulinic acid, which solves the problem that endogenous 5-aminolevulinic acid in biological matrices is difficult to accurately detect. Quantitative, many interfering components, and low sensitivity. It has the advantages of high sensitivity, high throughput, easy operation, good stability, and can save a lot of economic and time costs. It can meet the requirements of 5-aminolevulinic acid in large quantities of biological matrix samples. The concentration determination can be widely used in the analysis and detection of biological samples in drug development

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