HPLC法测定大鼠血浆中DX-11的质量浓度

目的建立测定大鼠血浆中DX-11N(β榄香烯13基)色氨酸]质量浓度的HPLC方法,并应用该法研究DX-11在大鼠体内的药动学。方法色谱柱:Kromasil C18柱(250 mm×4.6 mm,5μm),流动相:甲醇体积分数为0.2%的甲酸水溶液(体积比为75∶25),流速:1.0 mL.min-1,检测波长:220 nm,己烯雌酚作为内标物。结果 DX-11线性:0.2～20.0 mg.L-1,回归方程:Y=1.869×10-1ρ+9.830×10-2(r=0.998 1),定量下限:0.2 mg.L-1,回收率:84.1%～88.9%,日内和日间精密度均不大于10.2%。18只大鼠分别灌胃给予低、中、高3个剂量DX 12后,血浆中DX-11的tmax分别为(1.4±0.2)、(1.8±0.7)、(1.3±0.3)h,ρmax分别为(2.3±0.3)、(3.5±0.6)、(6.4±3.0)mg.L-1,t1/2分别为(1.7±0.7)、(2.3±1.0)、(1.4±0.8)h,AUC(0-∞)分别为(5.4±1.8)、(10.6±1.8)、(21.0±13.1)mg.h.L-1。结论建立并验证的HPLC法适用于DX-11在大鼠体内的药动学研究。

Determination of DX-11 in rat plasma by HPLC

Objective To establish a high performance liquid chromatographic method for determining the concentration of DX-11 N-(β-elemene-13-yl) tryptophan] in rat plasma. Methods The chromatographic separation was performed on a Kromasil C₁₈ column (250 mm×4.6 mm, 5 μm), and a mobile phase consisted of methanol -0.2% formic acid (V∶V = 75∶25) at a flow rate of 1.0 mL·min^-1. The detection wavelength was set at 220 nm, and the diethylstilbestrol was used as the internal standard. Results A good linearity was obtained from 0.2 mg·L^-1 to 20.0 mg·L^-1 of DX-11 in rat plasma with a regressive equation of Y = 1.869×10^-1ρ+9.830×10^-2 (r = 0.998 1). The limit of detection was 0.2 mg·L^-1and the recoveries were ranging from 84.1% to 88.9%. The RSDs of intra- and inter-day were less than 10.2%. The pharmacokinetic parameters of DX-11 in rat plasma after i.g. administration of different doses of DX-12 to 18 healthy rats were as follows: t_max = (1.4 ± 0.2), (1.8 ± 0.7), and (1.3 ± 0.3) h, ρ_max = (2.3 ± 0.3), (3.5 ± 0.6), and (6.4 ± 3.0) mg·L^-1, t_1/2 = (1.7 ± 0.7), (2.3 ± 1.0), and (1.4 ± 0.8) h, AUC_(0-∞) = (5.4 ± 1.8), (10.6 ± 1.8), and (21.0 ± 13.1) mg·h·L^-1, respectively. Conclusion The established method would be applied for the pharmacokinetic study of DX-11 in rat plasma.