摘要
目的建立HPLC-MS/MS测定癫痫患儿血浆中吡仑帕奈(perampanel,PER)浓度的方法。方法血浆样品经有机溶剂沉淀蛋白后,以PER-D5为内标,采用反相色谱柱,以1 mmoL乙酸铵-水(A)和乙腈(B)作为流动相进行梯度洗脱,流速为0.45 mL·min-1,柱温为50℃,进样量为5μL。采用电喷雾离子源正离子模式,多离子反应监测模式扫描。考察该方法的专属性、定量下限(lower limit of quantitation,LLOQ)、标准曲线、残留效应、稀释效应、准确度精密度、基质效应和稳定性。结果PER在1~1500 ng·mL^(-1)线性关系良好(r=0.9988),LLOQ为1ng·mL^(-1)。批内和批间精密度RSD≤5.80%,准确度为92.07%~106.07%,稳定性良好,不受正常血浆和溶血血浆(溶血程度≤5%)基质的影响,方法学验证均符合2020年版《中国药典》规定。结论该方法快速、简便、稳定、灵敏度高、专属性强,可应用于PER在癫痫儿童中的治疗药物监测和药动学-药效学研究。
OBJECTIVE To establish a method for determination of perampanel(PER)in plasma of epileptic children by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).METHODS The plasma samples were pre-treated by protein precipitation with organic solvent.Perampanel-deuterium-5(PER-D5)was used as isotope internal standards(SIL-IS).Chromatographic column was Hypersil GOLDTM(2.1 mm×50 mm,3μm);flow phase was 1 mmoL ammonium acetate-water(A)and acetonitrile(B),and gradient elution was conducted at a flow rate of 0.45 mL·min-1.Electric spray ion(ESI)source was used with positive ion monitoring mode,and multireaction monitoring(MRM)mode was adopted for quantitative analysis.The specificity,lower limit of quantitation(LLOQ),standard curve,residual effect,dilution effect,precision,matrix effects and stability were investigated.RESULTS The linear range of levetiracetam was 1-1500 ng·mL^(-1)(r=0.9989),and the LLOQ was 1ng·mL^(-1).The intra-day and inter-day precision RSDs were equal or less than 5.80%,the accuracy was 92.07%-106.07%,with good stability.This method was not affected by common matrix and hemolysis matrix(hemolysis degree below 5%).The methodology validation was in line with the provisions of Chinese Pharmacopoeia(2020 edition).CONCLUSION This study establishes an HPLC-MS/MS method for determination of the blood concentration of PER in children,which is rapid,simple,stable,economical,with high specificity.It can be applied to the therapeutic drug monitoring of PER in children with epilepsy.
作者
孟翔飞
马英华
赵宜乐
武佳璇
东蕾
MENG Xiang-fei;MA Ying-hua;ZHAO Yi-le;WU Jia-xuan;DONG Lei(College of Pharmacy,Hebei Medical University,Shijiazhuang 050017,China;Department of Pharmacy,Children′s Hospital of Hebei Province,Shijiazhuang 050031,China)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2023年第11期1020-1025,共6页
Chinese Pharmaceutical Journal
基金
2021年度河北省卫生健康委医学科学研究课题计划项目资助(20211469)
2022年度河北省省级科技计划项目资助(202230714010408)。
