LC-MS/MS法同时测定人乳汁中贝他斯汀和羟氯喹的浓度
Simultaneous determination of bepotastine and hydroxychloroquine concentrations in human breast milk by LC-MS/MS
白梦如 1申潜 2马志媛 1王刚1
作者信息
- 1. 杭州市第一人民医院药学部,杭州 310006
- 2. 浙江省食品药品检验研究院国家药品监督管理局仿制药评价关键技术重点实验室,杭州 310052
- 折叠
摘要
目的 建立同时测定人乳汁中贝他斯汀和羟氯喹浓度的液相色谱-质谱联用(LC-MS/MS)方法并应用于临床.方法 向50 μL乳汁样品中加入200 μL含内标(100 ng/mL氯喹)的甲醇,涡旋沉淀蛋白后离心取上清液进样分析.色谱柱为Waters ACQUITY UPLC HSS T3柱,流动相为0.1%甲酸-10 mmol/L乙酸铵溶液(A相)和甲醇(B相)(梯度洗脱),流速为0.35 mL/min,进样量为2 μL,分析时间为4 min.采用电喷雾离子源,正离子多反应监测模式扫描,定量分析离子对分别为m/z 388.9→201.9(贝他斯汀)、m/z 336.3→247.1(羟氯喹)和m/z 320.2→247.2(氯喹).对所建LC-MS/MS法进行方法学考察,并用于检测1例哺乳期患者乳汁中的药物浓度.结果 贝他斯汀在2～200 ng/mL(r=0.999)、羟氯喹在50～1 000 ng/mL(r=0.998)范围内线性关系良好.贝他斯汀和羟氯喹质控样本的批内、批间精密度均小于15%,准确度、提取回收率、基质效应和稳定性均符合生物样品定量分析方法验证要求.哺乳期患者乳汁中的药物浓度检测结果显示,该患者服药2 h和14 h后乳汁中贝他斯汀的浓度分别为34.95、5.72 ng/mL,羟氯喹分别为211.92、104.18 ng/mL,推算出相对婴儿剂量分别为1.83%、0.56%.结论 该方法操作简便、快速、灵敏度高,适用于人乳汁中贝他斯汀和羟氯喹浓度的同时测定,可为临床哺乳期的安全用药提供参考.
Abstract
OBJECTIVE To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the simultaneous determination of bepotastine and hydroxychloroquine concentrations in human breast milk and apply it in clinical practice.METHODS The milk samples(50 μL)were precipitated with 200 μL methanol containing the internal standard(100 ng/mL chloroquine),and the supernatant was taken for analysis after vortexing and centrifugation.The separation was performed on a Waters ACQUITY UPLC HSS T3 column with mobile phase consisted of 0.1%formic acid-10 mmol/L ammonium acetate solution(phase A)and methanol(phase B)at gradient elution of 0.35 mL/min.The injection volume was 2 μL,and the analysis time was 4 min.The detection of the analytes was performed by electrospray ionization in positive mode by multiple reaction monitoring with the transition of m/z 388.9→201.9(bepotastine),m/z 336.3→247.1(hydroxychloroquine),and m/z 320.2→247.2(chloroquine).The established LC-MS/MS method was researched in methodology and used to determine the drug concentrations in the breast milk of 1 case of lactating patient.RESULTS The linear range of bepotastine was 2-200 ng/mL(r=0.999),and hydroxychloroquine was 50-1 000 ng/mL(r=0.998).The intra-assay and inter-assay precisions were both≤15%,and the accuracy,extraction recovery,matrix effect,and stability all met the acceptance criteria for bioanalytical method validation.The concentration result of bepotastine and hydroxychloroquine in the breast milk of the lactating patient showed,after 2 h and 14 h,the concentrations of bepotastine in the breast milk of the patient were 34.95 ng/mL and 5.72 ng/mL;those of hydroxychloroquine were 211.92 ng/mL and 104.18 ng/mL,respectively.The relative infant doses were 1.83%and 0.56%,respectively.CONCLUSIONS The established method is simple,rapid,and sensitive.It is suitable for simultaneous determination of bepotastine and hydroxychloroquine concentrations in human milk and can provide reference for safe drug use during lactation.
关键词
贝他斯汀/羟氯喹/乳汁/哺乳期/药物浓度/液相色谱-质谱联用技术Key words
bepotastine/hydroxychloroquine/breast milk/lactation/drug concentration/LC-MS/MS引用本文复制引用
基金项目
国家自然科学基金(82003838)
浙江省自然科学基金(LY22H310006)
浙江省卫生健康科技计划(2021RC021)
杭州市医药卫生科技项目(A20210336)
出版年
2024
国家科技期刊平台
NSTL
万方数据