液相色谱串联质谱法测定人血浆中磺胺多辛及乙胺嘧啶浓度

Concentration determination of sulfadoxine and pyrimethamine in human plasma by LC-MS/MS method

目的建立测定血浆中磺胺多辛和乙胺嘧啶的液相色谱串联质谱(LC-MS/MS)方法。方法血浆样品中加入内标氘3-磺胺多辛和氘3-乙胺嘧啶,直接沉淀法处理样品。色谱柱为XBridgeTM Phenyl分析柱(150mm×2.1mm,5μm),流动相为含0.1%甲酸的水溶液-含0.1%甲酸的乙腈溶液(50∶50,V/V),流速为0.3mL·min-1。正离子多离子反应监测(MRM)扫描分析,离子通道分别为m/z311.2→156.2(磺胺多辛),m/z249.2→177.2(乙胺嘧啶),m/z314.2→156.2(氘3-磺胺多辛),m/z252.2→177.2(氘3-乙胺嘧啶)。结果磺胺多辛的线性范围为0.5～250mg·L-1,定量下限为0.5mg·L-1;乙胺嘧啶的线性范围为0.005～2.5mg·L-1,定量下限为0.005mg·L-1,提取回收率在88.8%～100.2%,批内、批间RSD均小于11%。结论本方法操作简便,特异性强,灵敏度高,可用于磺胺多辛和乙胺嘧啶的药动学研究。

AIM To establish a LC-MS/MS method for determination of sulfadoxine and pyrimethamine in human plasma. METHODS After addition of sulfadoxine-D3 （internal standard, IS） and pyrimethamine-D3 （IS） in combination, the serum analyte was processed by direct protein precipitation. The chromatographic separation was performed on XBridgeTM Phenyl （150 mm × 2.1 mm, 5μm） column with a mobile phase of 0.1% formic acid solution - 0.1% formic acid acetonitrile （50 ： 50, V/V） at a flow rate of 0.3 mL·min-1. The protonated ions of analytes were detected in positive ionization through multiple reaction monitoring mode （MRM） . The mass transition pairs of m/z 311.2 →156.2, m/z 314.2 → 156.2 and m/z 249.2 -→ 177.2, m/z 252.2 → 177.2 were used to detect sulfadoxine, sulfadoxine＇-D3 and pyrimethamine, pyrimethamine-D3, respectively. RESULTS The linear concentration ranges of the calibration curves and low limit of quantitation were 0.5 - 250 mg ＂L-I and 0.5 mg·L-1 for sulfadoxine, 0.005 - 2.5 mg ·L-1 and 0.005 mg·L-1 for pyrimethamine, respectively. The extraction recovery rates were ranged from 88.8% to 100%; within-batch RSD and between-batch RSD were less than 11%. CONCLUSION The pretreatment with LC-MS/MS method is simple and quick, with high specificity and sensitivety, suitable for the research of sulfadoxine and pyrimcthamine pharmacokinetics.