LC-MS/MS法检测人头发中利培酮及其代谢物9-羟利培酮含量的实验研究  

Determination of Risperidone and Its Metabolite of 9-hydroxyrisperidone in Hair by LC-MS/MS

作　　者：孙秀佳[1] 林治光[1] 任娟娟[1] 蒋平[1] 刘红梅[1] 李春波[1] 李华芳[1] 

机构地区：[1]上海交通大学医学院附属精神卫生中心生化药理研究室,上海200030

出　　处：《现代生物医学进展》2015年第12期2221-2224,2241,共5页Progress in Modern Biomedicine

基　　金：国家"重大新药创制"科技重大专项(2012ZX09303-003);上海市卫生系统优秀人才(XBR2011049);上海市卫生系统优秀学科带头人培养计划(XBR2011005);上海市精神卫生中心院级课题(2012-YJ-09)

摘　　要：目的:建立高效液相色谱-三重四级杆质谱联用(LC-MS/MS)法检测人头发中利培酮(RIP)及其代谢物9-羟利培酮(9-OH-RIP)含量的方法。方法:采用同位素内标氘4-利培酮(RIP-d4)及氘4-9-羟利培酮(9-OH-RIP-d4),流动相A为10 mmol/L醋酸胺溶液(甲酸调p H值为4.0),流动相B为乙腈,A/B=70/30,流速为0.3 m L/min,等度洗脱4.00 min。色谱柱为安捷伦Zorbax SB C18(2.1×50 mm,1.8μm),柱温30℃。准确称取20 mg丙酮清洗过晾干剪碎成粉末的头发样本,加1N氢氧化钠(Na OH)超声2 h,等量酸中和后,加200μL1N氢氧化钠溶液及5.0 m L的甲基叔丁基醚(MTBE)提取涡旋1分钟,3000×g离心5 min后取上清液在40度水浴下氮气吹干后用100μL流动相复溶,进样2μL经LC-MS/MS检测。MRM监测离子对:RIP:m/z 411.2→191.0,9-OH-RIP:m/z 427.2→207.1,RIP-d4:m/z 415.2→195.2,9-OH-RIP-d4:m/z 431.2→211.1。结果:利培酮及9-羟利培酮线性范围分别为0.5-25 ng/mg,0.0025-0.15 ng/mg,提取回收率均>70.0%,方法回收率均在85.0%-115.0%之间,线性r均>0.999,精密度和重现性RSD均<15%。结论:本研究建立了采用LC-MS/MS法检测人头发中利培酮及9-羟利培酮含量的方法,该法快速、简单、准确、重现性好。Objective： To establish an HPLC-MS/MS method for the determination of risperidone （RIP） and its metabolite 9-hydroxy-risperidone （9-OH-RIP） in the hair. Methods： The isptope-labeled internal standards （risperidone-d4 and 9-hydroxyrisperidone-d4） were used. The mobile phase A 10 mm ammonium buffer （tune pH to 4.0 by formic acid）; B： Acetonitrile; A/B-70/30, flow rate of 0.3 mL/min, the analytical column was Agilent Zorbax SB C18 （2.1 × 50 mm, 1.8/zm）, temperature was 30℃. 20 mg sample of hair was weighed accurately, which was washed by acetone and cuted into powder, ultrasonic for 2 h with added 1 mL of 1N NaOH, then neutralized by 1 mL 1N HCL, added 200 μL 1N NaOH and 5.0 mLof Methyl Tertiary Butyl Ether vortex for 1 min, 3000 g centrifugation for 5 min, supemate was dried under 40℃ with nitrogen, and redissolved by 100 I～L mobilephase, inject 2 ILL into LC-MS/MS. MRM： RIP： m/z 411.2→191.0, 9-OH-RIP： m/z 427.2--→207.1, RIP-d4：m/z 415.2→195.2, 9-OH-RIP-d4：rrdz431.2→211.1. Results： The results showed that the linear was 0.5-25 ng/mg for RIP and 0.0025-0.15 ng/mg for 9-OH RIP, both of the linearly dependent coefficient （r） were no less than 0.999, the recoverys of extraction 〉70.0 %, the recoverys of method was between 85.0 % and 115.0 %, and the RSDs of precision and accuracy were all no more than 15 %. Conclusion： A determination method of the concentration of RIP and 9-OH-RIP in hair was established, which was simple and rapid, and had superior sensitivity and selectivity.

关 键 词：利培酮 9-羟利培酮 LC-MS/MS 药物浓度 头发 

分 类 号：R969.1[医药卫生—药理学] R917[医药卫生—药学]