作者：夏伯姗^1,2, 王振磊², 李岱庆³, 王曼^1,2, 王鸿芡², 张梅¹, 秦永平², 南峰², 向瑾²

作者单位：1. 成都中医药大学药学院 中药材标准化教育部重点实验室 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 四川 成都 611137;
2. 四川大学华西医院国家药物临床试验机构临床药理研究室, 四川 成都 610041;
3. 扬子江药业集团四川海蓉药业有限公司, 四川 成都 610041

关键词：药动学;UPLC-MS/MS;氯沙坦;氯沙坦羧酸

摘要：

建立UPLC-MS/MS法测定人血浆中氯沙坦及代谢物氯沙坦羧酸的浓度，以用于氯沙坦的药代动力学研究。采用SepaxGP-C18（2.1 mm50 mm，1.8 m）色谱柱，以0.15%甲酸-0.04%氨水-水（超声20 min后，加入0.01%氨水）为水相，0.15%甲酸-0.04%氨水-98%乙腈为有机相。梯度洗脱，流量为0.3 mL/min。以d4-氯沙坦、d4-氯沙坦羧酸作内标，血浆样本用乙腈一步沉淀蛋白，采用多反应离子监测，正离子模式扫描，电喷雾离子源。结果表明：氯沙坦在2.0~2 000 ng/mL范围内，氯沙坦羧酸在2.4~2 400 ng/mL范围内线性良好；氯沙坦及内标的保留时间为1.16 min左右，氯沙坦羧酸及内标的保留时间为1.26 min左右；氯沙坦、氯沙坦羧酸提取回收率分别为109.9%、97.55%；精密度、基质效应经内标校准后均合格；稳定性各项数据均符合相关要求。该法快速、灵敏、专属性强、重现性好，适用于氯沙坦及其代谢物的血药浓度测定。

An UPLC-MS/MS method for simultaneous determination of losartan and metabolites in human plasma

XIA Boshan^1,2, WANG Zhenlei², LI Daiqing³, WANG Man^1,2, WANG Hongqian², ZHANG Mei¹, QIN Yongping², NAN Feng², XIANG Jin²

1. The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine, Key Laboratory of Systematic Research, Development and Utilization of Chinese Medicine Resources in Sichuan Province, Key Laboratory Breeding Base of Co-founded by Sichuan Province and MOST, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China;
2. GCP Center/Institute of Drug Clinical Trials, West China Hospital, Sichuan University, Chengdu 610041, China;
3. Yangtze River Pharmaceutical Group-Sichuan Hairong Pharmaceutical Co., Ltd., Chengdu 610041, China

Abstract: To develop an accurate and sensitive UPLC-MS/MS method for simultaneous determination of losartan and its active metabolites losartan carboxylic acid in human plasma to study pharmacokinetic of losartan in human. Using SepaxGP-C18(2.1 mm×50 mm, 1.8 μm) column with 0.15% formic acid-0.04% ammonia-water (ultrasonic for 20 min, add 0.01% ammonia water) as the aqueous phase, 0.15% formic acid-0.04% ammonia-98% acetonitrile as the organic phase, the flow rate was 0.3 mL/min, gradient elution. D4-losartan and d4-losartan carboxylic acid were used as the internal standard. Human plasma samples were deproteinized with acetonitrile. An electron spray ionization (ESI) was applied and operated in the positive multiple reaction monitoring (MRM) mode. In the plasma samples, the calibration curve for losartan and losartan carboxylic acid was in the range of 2.0-2 000 ng/mL and 2.4-2 400 ng/mL, respectively. The retention time of losartan and its internal standard were 1.16 min while the retention time of losartan carboxylic acid and its internal standard were 1.26 min. Pretreatment recovery was 109.9%, 97.55%. Accuracy, precision and matrixes effect after internal standard calibration are comply with relevant requirements. The method is simple, rapid, sensitive and accurate for the determination of losartan and metabolites in human plasma and to be suitable for the pharmacokinetic study.

Keywords: pharmacokinetics; UPLC-MS/MS; losartan; losartan carboxylic acid

2018, 44(7): 61-67,82  收稿日期: 2018-03-09;收到修改稿日期: 2018-04-20

基金项目: 

作者简介: 夏伯姗(1994-),女,四川成都市人,硕士研究生,专业方向为中药学。

参考文献

[1] CHEUNG T T, CHEUNG B M. Managing blood pressure control in Asian patients:safety and efficacy of losartan[J]. Clinical Interventions in Aging, 2014(9):443-450.
[2] LO M W, MICHAEL R. Pharmacokinetics of losartan, an angiotensin Ⅱ receptor antagonist, and its active metabolite EXP3174 in humans[J]. Clinical Pharmacology & Therapeutics, 1995, 58(6):641-649.
[3] WANG Y H, PAN P P, DAI D P, et al. Effect of 36 CYP2C9 variants found in the Chinese population on losartan metabolism in vitro[J]. Xenobiotica; the fate of foreign compounds in biological systems, 2014, 44(3):270.
[4] YASAR U, FORSLUNDBERGENGREN C, TYBRING G, et al. Pharmacokinetics of losartan and its metabolite E-3174 in relation to the CYP2C9 genotype[J]. Clinical Pharmacology & Therapeutics, 2002, 71(1):89-98.
[5] 田自有, 陈赛贞, 徐利君, 等. RP-HPLC法测定人血浆中氯沙坦钾、厄贝沙坦和格列齐特浓度[J]. 中国临床药学杂志, 2013, 22(4):230-233.
[6] 韦阳, 黄慧芳, 邵庆翔, 等. 国产氯沙坦钾片的生物等效性研究[J]. 中国临床药理学杂志, 2000, 16(2):106-110.
[7] 金杰. 健康志愿者氯沙坦钾片的生物等效性研究[J]. 中国医药指南, 2013(2):3-5.
[8] 杨平, 王艳娇, 李琳, 等. 氯沙坦钾胶囊与片在健康人体的生物等效性[J]. 中国临床药理学杂志, 2006, 22(6):436-439.
[9] 陈露露, 谭志荣, 阳国平, 等. 氯沙坦钾片的生物等效性及CYP2C9*3基因多态性对氯沙坦药代动力学的影响[J]. 中国临床药理学与治疗学, 2015, 20(2):175-181.
[10] 苑菲, 许剑安, 汪红, 等. LC-MS/MS法测定氯沙坦及其代谢物的血浆浓度及氯沙坦钾片生物等效性评价[J]. 中国临床药学杂志, 2013(1):13-18.
[11] 商丹丹, 邸欣, 王鑫, 等. 液相色谱-串联质谱法同时测定人血浆中氯沙坦及其活性代谢物氯沙坦酸的浓度[J]. 中国新药与临床杂志, 2013, 32(12):975-980.
[12] 王嫣然, 王彩平, 谢荟茹, 等. HPLC-MS/MS同时测定人体血浆中氯沙坦钾与其代谢物氯沙坦羧酸(E-3174)的浓度及其药动学应用[J]. 中国药学杂志, 2014, 49(12):1056-1061.
[13] 高荧, 乔亚楠, 陆宇婷, 等. 液相色谱-串联质谱法同时测定人血浆中氯沙坦、5-羧酸氯沙坦和氢氯噻嗪的浓度及药动学[J]. 中国新药与临床杂志, 2014, 33(12):884-889.
[14] SHAH H J, KUNDLIK M L, PATEL N K, et al. Rapid determination of losartan and losartan acid in human plasma by multiplexed LC-MS/MS[J]. Journal of Separation Science, 2009, 32(20):3388-3394.
[15] 黄明, 张全英, 宗顺麟. LC-MS/MS快速测定人血浆中氯沙坦浓度及其生物等效性研究[J]. 实用药物与临床, 2017, 20(4):443-446
[16] 马萍, 李鹏飞, 童卫杭, 等. 氯沙坦钾片在健康人体的生物等效性[J]. 中国临床药理学杂志, 2012, 28(3):179-182.
[17] 方翼, 关鑫, 顾景凯, 等. 国产氯沙坦钾片在中国健康志愿者的药动学及生物等效性研究[J]. 中国药学杂志, 2010, 45(20):1572-1577.
[18] PRASAJA B, SASONGKO L, HARAHAP Y, et al. Simultaneous quantification of losartan and active metabolite in human plasma by liquid chromatography-tandem mass spectrometry using irbesartan as internal standard[J]. Journal of Pharmaceutical & Biomedical Analysis, 2009, 49(3):862.
[19] 李三妮, 张古英, 韩华. UPLC-MS/MS法同时测定人血浆中氯沙坦、氨氯地平和辛伐他汀的浓度[J]. 药学与临床研究, 2015(6):539-542.
[20] 邹晓华, 王双虎, 胡国新, 等. UPLC-MS/MS快速测定大鼠血浆中的氯沙坦及其代谢产物[J]. 中国现代应用药学, 2014, 31(6):727-731.