论文部分内容阅读
目的:建立广泛适用、灵敏快速的人血浆中二甲双胍高效液相色谱测定方法,并应用于不同来源制剂的生物利用度研究。方法:血浆样品中加入适量内标,以乙腈沉淀蛋白后用HPLC进行分析。采用Waters XTERRAMS C18色谱柱(5μm,4.6mm×250 mm),柱温为40℃,优化的流动相组成为乙腈-0.03 mol·L-1磷酸氢二钾-0.02 mol·L-1SDS溶液(33∶29∶38),流速1.0 mL·min-1,紫外检测器测定波长为234 nm;以两制剂双周期交叉试验设计方法,测定20名自愿受试者口服不同来源的2种二甲双胍片后的药动学参数并进行生物等效性评价。结果:优化方法的线性范围为49.85~2493 ng·mL-1,最低定量限为49.85 ng·mL-1,准确度在91.9%~94.5%之间,日内、日间精密度(RSD)≤8.5%,蛋白沉淀回收率86.4%~93.2%之间,稳定性考察结果良好;实验中,口服给药二甲双胍片受试制剂与参比制剂后,受试者体内药物的Cmax分别为(1234.58±189.11)、(1152.57±238.29)ng·mL-1;Tmax分别为(2.40±0.81)、(2.73±0.85)h;t1/2分别为(4.27±0.86)、(3.92±0.42)h;AUC0-t分别为(8589.85±1649.85)、(7749.71±1303.61)ng·h·mL-1;AUC0-∞分别为(9234.77±1646.80)、(8390.94±1392.35)ng·h·mL-1。以AUC0-t计算,受试制剂盐酸二甲双胍片的相对生物利用度为(112.08±18.38)%;以AUC0-∞计算,受试制剂盐酸二甲双胍片的相对生物利用度为(111.15±16.37)%。结论:优化的方法适用性广,快速灵敏,专属性强,可用于盐酸二甲双胍制剂的临床药动学研究及其制剂的生物等效性评价。不同来源的2种二甲双胍片具有生物等效性。
OBJECTIVE: To establish a widely applicable, sensitive and rapid method for the determination of metformin in human plasma and to study its bioavailability from different sources. Methods: The plasma samples were spiked with appropriate amount of internal standard and the proteins were precipitated with acetonitrile and analyzed by HPLC. The mobile phase consisted of acetonitrile-0.03 mol·L-1 dipotassium hydrogen phosphate-0.02 mol·L-1 SDS solution with a Waters XTERRA® MS C18 column (5 μm, 4.6 mm × 250 mm) (33:29:38) at a flow rate of 1.0 mL · min-1. The wavelength of UV detector was 234 nm. Two volunteers were randomized to receive two metformin tablets from different sources by a two-cycle crossover design method After the pharmacokinetic parameters and bioequivalence assessment. Results: The linear range of the optimized method was 49.85 ~ 2493 ng · mL-1, the lowest limit of quantification was 49.85 ng · mL-1, the accuracy was between 91.9% and 94.5%, the intra- and inter-day precision was less than or equal to 8.5 %, And the recoveries of protein precipitation ranged from 86.4% to 93.2%. The results of stability study were good. In the experiment, after oral administration of metformin tablets and reference preparations, the Cmax of the drugs in the subjects were (1234.58 ± 189.11 ) And (1152.57 ± 238.29) ng · mL-1, respectively. The Tmax values were (2.40 ± 0.81) and (2.73 ± 0.85) h respectively, and were 4.27 ± 0.86 and 3.92 ± 0.42 h (8589.85 ± 1649.85) and (7749.71 ± 1303.61) ng · h · mL-1, respectively. The AUC0-∞ were (9234.77 ± 1646.80) and (8390.94 ± 1392.35) ng · h · mL-1, respectively. The relative bioavailability of metformin hydrochloride tablets was (112.08 ± 18.38)% with AUC0-t. The relative bioavailability of metformin hydrochloride tablets was (111.15 ± 16.37)% with AUC0-∞. CONCLUSION: The optimized method has wide applicability, quick and sensitive, strong specificity and can be used in the clinical pharmacokinetic study of metformin hydrochloride preparation and the bioequivalence evaluation of its preparation. Two metformin tablets from different sources have bioequivalence.