LC-MS/MS法测定瑞舒伐他汀在Caco-2细胞上的摄取特性  

Determination of uptake of rosuvastatin in Caco-2 cells by LC-MS/MS

作　　者：许青青[1,2] 黄晨蓉 王子腾[1,2] 张诗超[1,2] 缪丽燕[1,2] 

机构地区：[1]苏州大学药学院,苏州215006 [2]苏州大学附属第一医院药学部,苏州215006

出　　处：《中国新药杂志》2016年第13期1550-1554,共5页Chinese Journal of New Drugs

摘　　要：目的:建立瑞舒伐他汀的超高效液相色谱-串联质谱检测方法,研究其在Caco-2细胞上的摄取特性。方法:采用ACQUITY UPLCBEH C8柱(50 mm×2.1 mm,1.7μm),流动相为0.1%甲酸水溶液-乙腈(55∶45),流速为0.2 mL·min^(-1)。质谱条件:采用正离子检测,MRM模式。考察时间、温度、药物浓度、pH值及各类抑制剂对瑞舒伐他汀在Caco-2细胞上摄取的影响。结果:瑞舒伐他汀在0.1~500 nmol·L^(-1)的范围内呈良好的线性关系(r2>0.99);Caco-2细胞对瑞舒伐他汀的摄取在10 min内呈线性,10 min后吸收量逐渐趋于饱和,药物的摄取时间定为10 min;pH值对瑞舒伐他汀的摄取有显著影响,酸性条件下摄取量增加,pH定为6.0;浓度和温度依赖性结果显示,瑞舒伐他汀依靠主动转运和被动扩散2种方式吸收;P-糖蛋白抑制剂维拉帕米对瑞舒伐他汀的吸收基本没有影响;耐药相关性蛋白2(MRP2)抑制剂MK-571和乳腺癌耐药蛋白(BCRP)抑制剂依克立达的存在可增加瑞舒伐他汀的吸收(P<0.05);有机阴离子转运多肽OATP2B1抑制剂雌酮-3-硫酸钠可减少Caco-2细胞对瑞舒伐他汀的吸收(P<0.05)。结论:该法灵敏、快速、简单、专属性强,瑞舒伐他汀在Caco-2细胞中以主动和被动2种形式吸收,部分通过OATP2B1转运进细胞内,MRP2和BCRP对瑞舒伐他汀的吸收有外排作用,而P-gp可能不影响其吸收。Objective： To establish an LC-MS /MS method for the determination of rosuvastatin and study its uptake in Caco-2 cells. Methods： The samples were seperated on an ACQUITY UPLCBEH C8column（ 50 mm ×2. 1 mm,1. 7 μm） using mobile phase consisting of water（ 0. 1% formic acid） and acetonitrile（ 55∶ 45） at a flow rate of 0. 2 m L·min-（-1）. The protonated ions of analytes were detected by positive ionization with multiple reaction monitoring mode（ MRM）. The uptake of rosuvastatin in Caco-2 cells at various time,concentration,temperature,pH values,and various inhibitors was measured. Results： The calibration curve of rosuvastatin was linear over the range of 0. 1 - 500 nmol·L-（-1） in cells（ r2 0. 99）; The uptake of rosuvastatin in Caco-2 cells showed a linear trend within 10 min and saturated gradually after 10 min,therefore the uptake time was fixed at 10 min; pH values significantly affected the uptake of rosuvastatin and was fixed at 6. 0; the concentration- and temperature-dependent uptake behaviour indicated that rosuvastatin relied mainly on active transport and passive diffusion; verapamil almost had no effect on the uptake of rosuvastatin,while MK-571 and elacridar increased the uptake,and Estrone-3-sodium salt decreased the uptake significantly（ P 0. 05）. Conclusion： This validated LC-MS / MS method is sensitive,rapid,simple,and specific. The uptake of rosuvastatin in Caco-2 cells includes active and passive manners.OATP2B1 maybe plays an role in the absorption of rosuvastatin,and MRP2 and BCRP act on the efflux of rosuvastatin,while P-gp does not.

关 键 词：瑞舒伐他汀 CACO-2细胞 摄取 液相色谱-质谱联用 

分 类 号：R972.6[医药卫生—药品]