Method for detecting metanephrines through liquid chromatography-tandem mass spectrometry

Abstract

The invention discloses a method for detecting metanephrines through liquid chromatography-tandem mass spectrometry. The method comprises the following steps that 1 sampling preprocessing is conducted, wherein a mixed internal standard solution is added into a to-be-detected sample for protein precipitation, then centrifugation is conducted to acquire a liquid supernatant, and the liquid supernatant is blow-dried with nitrogen gas and then subjected to double solving by using a double solvent to obtain a double solution; 2, the double solution obtained in the step 1 is purified; and 3, the metanephrines in a purified product obtained in the step 2 are determined by means of liquid chromatography-tandem mass spectrometry. The method is simple in pre-processing, high in sensitivity and specificity and good in repeatability, part of artificial operations are replaced with on-line automatic pre-processing, the detection efficiency is improved, the error rate is decreased, the requirement on detection personnel is lowered, the cost is reduced, and the method is suitable for being applied and popularized in a clinical chromatography-mass spectrometry laboratory with the low automation degree.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a method for detecting methoxyadrenaline substances by liquid chromatography-tandem mass spectrometry. Background technique [0002] Pheochromocytoma and paraganglioma are tumors characterized by massive secretion of catecholamines. Methoxyepinephrine substances in plasma include methoxyepinephrine, methoxynorepinephrine and 3-methoxytyramine. from chromaffin cells. Plasma concentrations of methoxyepinephrines were associated with elevated long-term catecholamine levels and were less affected by changes in short-term catecholamine secretion. The secretion of catecholamines in the blood is intermittent, affected by the sampling time point, etc., intermittently entering the blood may easily lead to false positive or false negative results. The half-life of plasma methoxyadrenaline substances is longer than that of catecholamine hormones, and they exist in t...

AI Technical Summary

Problems solved by technology

Among them, there are many interferences in the detection of biological samples by electrochemical methods, and the quantitative analysis of 3-methoxytyramine is also impossible. Some laboratories use derivatization pretreatment methods. The derivatization process is time-consuming and highly toxic. Extraction technology is the most commonly used sample pretreatment method reported in most literatures and in clinical mass spectrometry laboratories, but there are still some problems in this technology: 1. Off-line manual solid phase extraction technology (including traditional solid phase extraction and simplified solid phase extraction) Phase extraction technology) has many steps and complex operations (activation, equilibration, sample loading, rinsing, and elution), which have high requirements for laboratory technicians in all aspects, and the training period is long, which is prone to human-induced error results; 2 , The offline automatic solid-phase extraction instrument has achieved the liberation of manpower to a certain extent, but it is generally suitable for the processing of larger volume samples, and it is not suitable for micro-volume samples, and after the sample processing, it still needs to be concentrated and redissolved, which is more cumbersome ;3. Off-line SPE column activation, equilibration, sample loading, elution and elution volume are relatively large. To meet the requirements of reversed-phase chromatographic columns and low-sensitivity mass spectrometers, nitrogen blowing concentration and redissolution are often required in the pretreatment process 4. At present, most of the online solid-phase extraction technology is used, and ultrafiltration tubes are used to remove protein in sample pretreatment, and the cost of ultrafiltration tubes is relatively high. In the actual application process, organic Reagent protein precipitation is still the daily preferred method in clinical laboratories; 5. On-line solid-phase extraction technology is used for elution with a high proportion of organic solvents, and the eluate directly enters the chromatographic column for separation and analysis. If a reversed-phase column is used, there is no retention, and HILIC is used When using a chromatographic column for analysis, methoxyepinephrine and 3-methoxytyramine need to be chromatographically separated due to the same product ion to reduce co-elution interference. Failure to separate the two compounds may easily cause biased results for 3-methoxytyramine High, but the two substances have not been separated in the literature

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