Method for determining concentration of olanzapine in blood plasma through high-performance liquid chromatography-tandem mass spectrometry

Abstract

The invention discloses a method for determining concentration of olanzapine in blood plasma through high-performance liquid chromatography-tandem mass spectrometry. The method comprises the followingsteps: preparing standard curve working fluid with a variety of concentrations by dissolving olanzapine with methanol, and preparing internal standard working fluid by dissolving stable isotope internal standard substances with the methanol; separately adding 0 microliter to 5 microliters of the standard curve working fluid with the variety of concentrations into blank plasma and complementing to100 microliters; swirling to prepare standard curve blood plasma samples with the variety of concentrations; adding the internal standard working fluid into the standard curve blood plasma samples, carrying out centrifugation, taking supernatant liquid and carrying out LC-MS / MS quantitative analysis, and drawing the standard curve; precisely sucking 100 microliters of to-be-tested blood plasma, adding the internal standard working fluid into the to-be-tested blood plasma, carrying out centrifugation, taking supernatant liquid and carrying out LC-MS / MS quantitative analysis, and reading the concentration of the olanzapine on the standard curve. The method disclosed by the invention has the characteristics of simpleness and convenience in sample pre-processing, short analysis time, small volumes of required samples, high sensitivity, high recycling ratio, small matrix effect and the like.

Description

technical field [0001] The present invention relates to methods for detecting drug concentrations. More specifically, the present invention relates to a method for determining the concentration of olanzapine in plasma by high performance liquid chromatography tandem mass spectrometry. Background technique [0002] Olanzapine (Olanzapine) is an atypical antipsychotic drug developed and marketed by Eli Lilly Pharmaceutical Company of the United States in 1996. The chemical name is 2-methyl-4-(4-methyl-1-piperazinyl)-10H-thieno[2,3-b][1,5]benzodiazepine The molecular formula is C17H20N4S, and the molecular weight is 456.32. Olanzapine is a second-generation antipsychotic drug that acts on multiple receptor systems, such as 5-HT receptors and histamine receptors. Olanzapine is mainly metabolized by hepatic cytochrome P450 enzymes (such as CYP4501A2) and glutamic acid transferase. [0003] For the quantitative determination of small molecular compounds, high-performance liqu...

AI Technical Summary

Problems solved by technology

At present, the LC-MS / MS method reported in the literature to determine the concentration of olanzapine in biological matrices is mainly used in the field of therapeutic drug monitoring (TDM), and has a wide linear range (such as 5-500nM, 0.25-50ng / mL, 1-300ng / mL), and cannot meet the requirements of clinical trials for biological sample detection limit, quality control and sample processing efficiency

In addition, the published methods also have problems such as large sample volume (0.2-0.5mL / sample), long analysis time per sample (6-11min), high lower limit of quantitation (such as 1-5ng / mL), sample pretreatment time-consuming liquid-liquid extraction and solid-phase extraction

Although there are reports in the literature that some LC-MS / MS methods use protein precipitation, however, the process of these protein precipitation still requires operations such as drying and reconstitution, which significantly reduces the efficiency of biological sample pretreatment, and has great impact on clinical pharmacokinetics / biology. High-throughput processing of large numbers of plasma / serum samples in equivalence studies presents challenges

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