用高效液相色谱-同位素稀释质谱法测定川崎病患儿血浆中阿司匹林和水杨酸的药物浓度

Determination of aspirin and salicylic acid of children with Kawasaki disease by HPLC-IDMS

目的 建立高效液相色谱-同位素稀释质谱(HPLC-IDMS)法测定川崎病患儿血浆中阿司匹林(ASP)和水杨酸(SA)浓度的方法。方法 血浆样品经有机溶剂沉淀蛋白后,分别以ASP-D4和SA-D4为内标,采用Phenomenex Kinetex XB C18色谱柱(2.6 mm×50.0 mm, 3.0μm),以0.1%甲酸-水(A)和乙腈(B)作为流动相进行梯度洗脱,流速为0.5 mL·min^(-1),进样量为6μL。质谱检测方式为负离子模式,多反应监测扫描。结果 ASP在1.02~4 000 ng·mL^(-1)线性关系良好(r2=0.995 4),定量下限(LLOQ)为1.02 ng·mL^(-1)。SA在1.28~5 000 ng·mL^(-1)线性关系良好(r2=0.998 5),LLOQ为1.28 ng·mL^(-1)。准确度、精密度、基质效应和稳定性均符合《中国药典》(2020年版)规定。结论 该方法操作简便快速稳定,灵敏度专属性高,可用于ASP和SA在川崎病患儿中的治疗药物监测和群体药代动力学-药效学研究。

Objective To establish a method for determination of aspirin(ASP)and salicylic acid(SA)in plasma of children with kawasaki disease by high performance liquid chromatography-isotope dilution mass spectrometry(HPLC-IDMS).Methods The samples were pretreated by precipitation protein method with organic solvent.ASP-D4and SA-D4 were used as isotope internal standards.Chromatographic column:Phenomenex Kinetex XB C18(2.6 mm×50.0 mm,3.0μm);flow phase:0.1%formic acid-water(A)and acetonitrile(B),gradient elution;flow speed:0.5 mL·min^(-1);injection volume:6μL.The detection method of mass spectrometry was negative ion mode,multireaction monitoring mode for quantitative analysis.Results The linear range of aspirin was 1.02-4000 ng·mL^(-1)(r^(2)=0.9954),and lower limit of quantification(LLOQ)was 1.02 ng·mL^(-1).The linear range of salicylic acid was 1.28-5000 ng·mL^(-1)(r^(2)=0.9985),and LLOQ was 1.28 ng·mL^(-1).Accuracy,precision,matrix effect and stability were in line with the provisions of Chinese Pharmacopoeia(2020edition).Conclusion This study determined the blood concentration of aspirin and salicylic acid in children with HPLC-IDMS,which is rapid,simple,stable,economical,and high specificity.It can be applied to therapeutic drug monitoring of aspirin and salicylic acid in clinical children with kawasaki disease.