高效液相色谱-串联质谱法测定人腹腔积液中伏立康唑浓度及临床应用  

Determination of voriconazole in human peritoneal fluid by high-performance liquid chromatography-tandem mass spectrometry and its clinical application

作　　者：林晓彬 梁桃 胡晓光[2] 陈攀[1] 蔡常洁[2] 陈孝[1] 闫佳佳[1] LIN Xiao-bin;LIANG Tao;HU Xiao-guang;CHEN Pan;CAI Chang-jie;CHEN Xiao;YAN Jiajia(Department of Pharmacy,First Affiliated Hospital of Sun Yat-sen University,Guangdong Guangzhou 510080,China;Second Department of Critical Care Medicine,First Affiliated Hospital of Sun Yat-sen University,Guangdong Guangzhou 510080,China)

机构地区：[1]中山大学附属第一医院药学部,广东广州510080 [2]中山大学附属第一医院重症二科,广东广州510080

出　　处：《中国医院药学杂志》2022年第21期2244-2248,共5页Chinese Journal of Hospital Pharmacy

基　　金：广东省医院药学研究基金(澳美基金)(编号:2022A06)。

摘　　要：目的:建立高效液相色谱-串联质谱(HPLC-MS/MS)法测定人腹腔积液中伏立康唑浓度,并应用于临床样本检测。方法:腹腔积液样品经纯乙腈沉淀蛋白后,通过Hypersil GOLD C_(18)色谱柱分离;流动相为0.1%甲酸水-纯乙腈(60:40,V/V);流速0.4 mL·min^(-1);柱温30℃。质谱采用电喷雾电离、正离子多反应监测模式扫描定量:m/z 350.0→281.0(伏立康唑)和m/z353.1→284.1(伏立康唑-d3,内标)。结果:腹腔积液样品中伏立康唑浓度在0.05~10.00μg·mL^(-1)范围内线性良好(R2=0.9999)。日内和日间精密度均小于2%,提取回收率为(100.35±4.37)%~(107.68±3.97)%,内标归一化基质效应因子为(98.36±1.69)%~(100.57±1.10)%且RSD小于2%。此外,各项稳定性考察结果均合格。利用该方法测得4例重度肝硬化患者静脉用伏立康唑首剂给药(约滴注1 h)的腹腔积液0~12 h的浓度-时间曲线下面积(AUC0-12)和血浆AUC0-12的比值平均为0.54(范围0.49~0.61),提示肝硬化患者中伏立康唑具有较好的腹膜腔渗透性。结论:该方法简单、快速、灵敏、准确,适用于人腹腔积液中伏立康唑的检测和临床药动学研究。OBJECTIVE To develop a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for the determination of voriconazole in human peritoneal fluid,and apply it to the detection of clinical samples.METHODS The peritoneal fluid sample was pretreated with precipitating by pure acetonitrile.Chromatographic separation was performed on a Hypersil GOLD C_(18)column with mobile phase consisting of A(aqueous phase:0.1%formic acid)and B(organic phase:pure acetonitrile)(A:B=60:40,V/V),the flow rate was 0.4 mL·min^(-1),the column temperature was maintained at 30℃.The multiple reaction monitoring transitions with an electrospray ionization source in the positive mode with m/z350.0→281.0for voriconazole and 353.1→284.1for D3-voriconazole(internal standard)were conducted on a triple-quadrupole tandem mass spectrometer to detect voriconazole.RESULTS The method exhibited good linearity(R^(2)=0.9999)over the voriconazole concentration range of 0.05-10.00μg·mL^(-1)in peritoneal fluid.The intra-and inter-day precisions were within2%.The extraction recovery was(100.35±4.37)%-(107.68±3.97)%.The internal standard normalized matrix effect factor ranged from(98.36±1.69)%to(100.57±1.10)%and the RSD was within 2%.The method showed acceptable results in stability tests.Using this method,the mean ratio of AUC0-12in peritoneal fluid to AUC0-12in plasma of four patients with severe liver cirrhosis after the first dose of intravenous voriconazole(approximately 1-h infusion)was 0.54(ranged from0.49 to 0.61),suggesting good penetration of voriconazole into the peritoneal cavity in patients with liver cirrhosis.CONCLUSION The method is simple,fast,sensitive and accurate,which can be used for the determination and clinical pharmacokinetic study of voriconazole in human peritoneal fluid.

关 键 词：伏立康唑 高效液相色谱-串联质谱 腹腔积液 药动学 

分 类 号：R969[医药卫生—药理学]