High performance liquid chromatography detection method of free methoxy group noradrenaline and methoxy group in human plasma

Abstract

The invention discloses a high performance liquid chromatography detection method of free methoxy group noradrenaline and methoxy group in human plasma. The high performance liquid chromatography detection method of the free methoxy group noradrenaline and the methoxy group in the human plasma comprises the following steps that: (1) sample pre-treatment is conducted, interior label working solution and spiritus mindereri are successively added into a plasma sample, vortex is mixed, and centrifugation is conducted; (2) weak positive ion exchange solid phase extraction 96 pore plate (SPE 96 poreplate) is activated; (3) solid phase extraction treatment is conducted, after the step (1) is conducted, the sample is loaded into the treated SPE 96 pore plate, a formic acid acetonitrile solution is eluted, and eluent is dried by nitrogen; and (4) redissolution is conducted, analysis and detection are conducted by the high performance liquid chromatography detection method. The high performanceliquid chromatography detection method of the free methoxy group noradrenaline and the methoxy group in the human plasma has the advantages that accuracy is high, reproducibility is high, sensitivityis high, specificity is high, the efficiency is high, flux is high, the cost is low, and the operation is easy, convenient and fast.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a high-accuracy, high-reproducibility, high-sensitivity, high-specificity, high-efficiency, high-throughput, low-cost, simple and fast operation method for removing free methoxy in human plasma. High performance liquid chromatography tandem mass spectrometry detection method for norepinephrine and methoxyepinephrine. Background technique [0002] Catecholamines are amine compounds with catechol (ie catechol) structure, including dopamine, norepinephrine, epinephrine and their derivatives. Norepinephrine and epinephrine are both hormones secreted by the adrenal medulla and neuromediators of noradrenergic fibers in the sympathetic and central nervous systems. Noradrenaline is widely distributed in the central nervous system, and its content is relatively large, while that of epinephrine is relatively small. Dopamine is mainly concentrated in the extrapyramidal syst...

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Problems solved by technology

[0004] 1. The reagents of the radioenzyme method are expensive and cannot meet the needs of clinical mass detection;

[0005] 2. The pretreatment of capillary electrophoresis includes operations such as dialysis and centrifugation, and capillary modification is required to prevent catecholamines from interacting with the capillary wall. Therefore, this method is cumbersome and time-consuming, and the detection is difficult and expensive;

[0006] 3. The fluorescence of catecholamines is weak, and the content in biological samples is extremely low, so the fluorescence method needs additional derivatization treatment to strengthen the fluorescence signal, which is tedious and time-consuming;

[0007] 4. Liquid chromatography requires a relatively complicated pretreatment process to separate impurities in biological samples, which is time-consuming and laborious, and has low sensitivity, long analysis time, low detection throughput, serious matrix interference, poor specificity, and low degree of automation

[0009] 1. The "Method for detecting catecholamines and metabolites by liquid chromatography tandem mass spectrometry" published on the website of the State Intellectual Property Office, which includes solid phase extraction and mass spectrometry positive ion mode scanning can simultaneously detect six kinds of catecholamines. It has the following defects: (1) It does not stipulate the use of internal standard of stable isotope markers with consistent structure

Compared with the MRM mode, the positive ion scan mode is susceptible to interference from the complex matrix of biological samples, and the specificity and specificity of the method are relatively poor

[0010] 2. "High-throughput liquid chromatography tandem mass spectrometry detection method and method for detection of four catecholamine metabolites" disclosed on the website of the State Intellectual Property Office, the method includes S1 sample preparation, S2 derivatization reaction, S3 sample pretreatment, S4 Liquid chromatography separation and S5 tandem mass spectrometry detection and other steps, this method has the following defects: (1) The pretreatment process is complicated, including protein precipitation, vacuum freeze-drying treatment and derivatization reactions, etc., which is cumbersome and time-consuming, and the cost of instruments and reagents is relatively high High, not suitable for clinical large-scale sample analysis; (2) For the detection of NMN, no isotope-labeled internal standard with consistent structure is used, which may affect the accuracy and precision of actual clinical sample detection; (3) The instrument analysis time of samples Longer, the analysis of a single sample takes 15 minutes, the efficiency is low, and it is not suitable for the detection of clinical large-scale samples

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