Method for quantitatively detecting catecholamine and metabolites thereof by liquid chromatography-mass spectrometry

Abstract

The invention relates to a method for quantitatively detecting catecholamine and metabolites thereof by liquid chromatography-mass spectrometry. The method mainly comprises the following steps: (1) pretreating a sample; (2) carrying out solid-phase extraction on the pretreated sample solution, and extracting catecholamine and metabolites thereof in the plasma; and (3) specifically detecting each analyte and the internal standard thereof by liquid chromatography-mass spectrometry, and comparing the ratio of the analyte to the internal standard signal with the ratio of a calibration curve to determine the concentration of the analyte. The method can quantitatively detect six catecholamines hormones in a human plasma sample, and the detection result can assist a clinician in diagnosing diseases related to abnormal catecholamines hormone content in an adult.

Description

technical field [0001] The invention relates to a method for detecting catecholamines and their metabolites, more specifically a method for quantitatively detecting catecholamines and their metabolites by liquid chromatography mass spectrometry. Background technique [0002] Pheochromocytoma (PCC) and paraganglioma (PGL) are tumors originating from the adrenal medulla or the extra-adrenal sympathetic nerve chain, respectively, and mainly synthesize and secrete large amounts of catecholamines (CA), such as norepinephrine (NE), epinephrine (E), and dopamine (DA), causing a series of clinical syndromes such as elevated blood pressure in patients, and causing serious complications such as heart, brain, and kidney. PCC accounts for 80%-85%, and PGL accounts for 15%-20%. The two are collectively called PPGL. [0003] PPGL is a rare endocrine disease, and the prevalence of PPGL in general hypertension clinics is 0.2%-0.6%. The main clinical manifestation of PPGL is hypertension a...

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Problems solved by technology

[0006] At present, the "High Performance Liquid Chromatography Tandem Mass Spectrometry Method for the Detection of Free Methoxynorepinephrine and Methoxyepinephrine in Human Plasma" published on the website of the Intellectual Property Office includes solid-phase extraction to process plasma samples and electrospray cation multi- The reaction monitoring mode (MRM) can detect two kinds of catecholamine metabolites at the same time. This method has the following defects: 1) the amount of plasma required for sample pretreatment is large; 3) the elution volume is too large and the pretreatment time is prolonged; 4) the machine uses Larger injection volume is required to meet the detection of linear low-concentration samples; 5) The flow rate of the on-board detection is too high, and the cost of reagents is high; 6) The formic acid-ammonium acetate buffer salt system is used, and the separation is biased, making it difficult to ensure reproducibility; 7) The linear range is narrow and cannot be fully applied to the quantitative detection of clinical high-concentration samples; 8) The sensitivity is low

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