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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 UPLCBEH 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 a method for the determination of rosuvastatin by ultra performance liquid chromatography-tandem mass spectrometry and investigate its uptake on Caco-2 cells. Methods: ACQUITY UPLCBEH C8 column (50 mm × 2.1 mm, 1.7 μm) was used as the mobile phase. The mobile phase consisted of 0.1% formic acid in water (55:45) and the flow rate was 0.2 mL · min -1. MS conditions: positive ion detection, MRM mode. The effects of time, temperature, drug concentration, pH value and various inhibitors on uptake of rosuvastatin on Caco-2 cells were investigated. Results: Rosuvastatin showed a good linear relationship (r2> 0.99) in the range of 0.1-500 nmol·L -1. The uptake of rosuvastatin by Caco-2 cells was linear within 10 min, 10 min after the absorption gradually saturated, the drug intake time was set at 10 min; pH value of rosuvastatin uptake significantly increased under acidic conditions, increased intake, pH was 6.0; concentration and temperature-dependent results Showed that rosuvastatin was absorbed by two modes of active transport and passive diffusion; verapamil, a P-glycoprotein inhibitor, had little effect on the uptake of rosuvastatin; the drug-resistant protein 2 (MRP2) inhibitor MK -571 and breast cancer drug resistance protein (BCRP) inhibitor Ecljeta increased the uptake of rosuvastatin (P <0.05). OATP2B1 inhibitor estrone-3-sulfate reduced Caco -2 cells to rosuvastatin uptake (P <0.05). Conclusions: The method is sensitive, rapid, simple and specific. Rosuvastatin is absorbed by both active and passive forms in Caco-2 cells and partially translocated into the cells through OATP2B1. MRP2 and BCRP inhibit rosuvastatin Absorption efflux, and P-gp may not affect its absorption.