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目的 建立兔血浆中DL111 IT的高效液相色谱荧光检测法。方法 血浆样品和内标格列本脲经氯仿提取后 ,选用DiamonsilODS C18柱 (15 0mm× 4 . 6mm ,5 μm) ,乙腈 - 0 . 0 2 5mol·L-1磷酸氢二铵缓冲溶液 (磷酸调pH 5 .0 ) (6 0 .4 0 ,V/V)为流动相 ,流速 1 0mL·min-1,荧光检测 (λex=2 5 0nm ,λem=332nm)。结果 DL111 IT在浓度 1 .0 0～ 2 0. 0 0ng·mL-1范围内呈良好线性 ,r=0. 9996 ,n =5。高 (2 0 . 0 0ng·mL-1)、中 (10 . 0 0ng·mL-1)、低 (1 .0 0ng·mL-1) 3个质控样本平均提取回收率分别为85 . 3%± 1 .3% ,84 .9%± 2 . 7% ,85 . 8%± 1 .8% ,方法回收率分别为 99 .5 %± 0 . 4 % ,10 2 . 1%± 1. 8% ,10 .1 3%± 2 .4 % ;日内(n =5 )和日间 (n =5 )RSD分别低于 3 .0 %和 7. 0 %。血浆样品检测限为 0 3ng·mL-1(S/N =3) ,定量限为 1ng·mL-1(S/N =10 ,RSD <7 .0 % )。结论 本法准确、灵敏、操作简便 ,为DL111 IT药代动力学研究提供了方法学基础。
OBJECTIVE To establish a high performance liquid chromatography (HPLC) fluorescence detection method for DL111 IT in rabbit plasma. Methods After the plasma samples and the glyburide internal standard were extracted by chloroform, Diamonsil ODS C18 column (15 0 mm × 4.6 mm, 5 μm), acetonitrile-0.0225 mol·L -1 diammonium phosphate buffer (phosphoric acid The pH was adjusted to 6.0 (V / V). The flow rate was 10 mL · min-1 and the fluorescence was detected (λex = 250 nm, λem = 332 nm). Results DL111 IT showed good linearity in the range of 1 0 0 ~ 2 0 0 ng · mL-1, r = 0. 9996, n = 5. The average recoveries of the three control samples with high (2.0 0 ng · mL -1), medium (10.0 ng · mL -1) and low (1.0 ng · mL -1) were 85.3 % ± 1 .3%, 84 .9% ± 2. 7% and 85. 8% ± 1 .8%, respectively. The recoveries were 99.5% ± 0.4% and 102.1% ± 1, respectively. (N = 5) and daytime (n = 5) RSDs were lower than 3.0% and 7.0%, respectively. The detection limit of plasma samples was 0 3 ng · mL-1 (S / N = 3). The limit of quantitation was 1 ng · mL-1 (S / N = 10, RSD <7.0%). Conclusion The method is accurate, sensitive and easy to operate, which provides a methodological foundation for the study of DL111 IT pharmacokinetics.