Determination of the Concentration of Pyrazinamide in Human Plasma by LC-MS/MS
Objective:To establish a method for determining the concentration of pyrazinamide in human plasma.Methods:Using pyrazinamide-d3 as internal standard,the plasma concentrations of pyrazinamide were determined by LC-MS/MS after protein precipitation.The liquid chromatographic separation process was performed on an AQUASIL C18 column by solvent gradient combined with velocity gradient elution.The mobile phases were consisted of water(mobile phase A)and acetonitrile-water(5∶95,V/V,mobile phase B),both contained 0.1%formic acid and 2 mmol·L-1 ammonium.The column temperature was set at 40℃,and an injection volume of 10 μL for each sample was sufficiently detected.The electrospray ion source and multi-reaction monitoring mode were used for positive iron scanning.The ion pair used for quantitative analysis of pyrazinamide and internal standard were m/z 124.10→81.10 and m/z 127.10→84.10,respectively.Results:The linear range of pyrazinamide were 40 ng·mL-1～20000 ng·mL-1(R2＞0.9992),and the lower limit of quantification was 40 ng·mL-1.The precision and accuracy within and between batches,extraction recovery rate,matrix effect and stability all met relevant requirements.Conclusion:The established method is simple,rapid and suitable for the clinical pharmacokinetic studies.
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