RP-HPLC法研究刺五加注射液中刺五加苷E、刺五加苷B在大鼠体内的药代动力学和组织分布特性

RP-HPLC determination of pharmacokinetics of eleutherococcuss eleutheroside E and eleutheroside B in rats plasma and tissues

目的:建立大鼠血浆中刺五加苷 E 和刺五加苷 B 的 RP-HPLC 分析方法,并对这2种成分在大鼠体内过程特性进行分析研究。方法:用乙腈沉淀生物样品中的蛋白质,同时又将生物样品中的刺五加苷 B 和苷 E 提取出来,然后用固相萃取法分离提取物,用60%甲醇将刺五加苷 B 和苷 E 从固相萃取小柱洗脱下来,用高效液相色谱法提取出来测定,色谱柱为 KromasilODS(4.6mm×250mm,5pan),柱温25℃,水-乙腈梯度流动相(0→15min,90:10→80:20;15→25min,80:20→50:50),流速0.8mL·min^(-1),一次进样,分别在220nm和206nm 波长下检测刺五加苷 E 和苷 B。结果:Wister 大鼠一次股静脉给药后血药浓度时间曲线呈三室模型,刺五加苷 E 和苷 B 的消除半衰期 t_(1/2)分别为4.66和2.49h。在主要组织中的分布特点是:刺五加苷 E 在血液、肾脏、心脏、肝脏和脾脏中都有分布,刺五加苷 B 在血液、肾脏、心脏、肝脏中都有分布,在脾脏中没有分布,刺五加苷 E 和苷 B 主要由肝、肾代谢、排泄。结论:本法可用于刺五加苷 E 和刺五加苷 B 体内过程的研究。样品直接用固相萃取小柱处理,可消除内源性成分干扰。

Objective： To establish an HPLC method for the analysis of pharmacokinetics and tissues distribution of eleutheroside E （ ELU E） and eleutheroside B （ ELU B） in rat after having taken eleutherococcuss injection. Methods：Protein from the biological sample was deposited using acetonitrile. ELU E and ELU B were extracted from the biological samples using acetonitrile, separated by solid - phase extraction, and eluted from the cartridge using 60% methanol. The analysis was performed on a Kromasil ODS column（4. 6 mm ×250 mm,5 μm）at 25℃ ,using water - acetonitrile as the gradient mobile phase and 0. 8 mL · min^ -1 flow rate. The gradient mobile phase consisted of water- acetonitrile（beginning from 90：10 programmed to 80：20 with 15 minutes,then programmed to 50：50 with 10 minutes） at a flow rate of 0. 8 mL · min^ -1. The detection wavelength was 220 nm and 206 nm for ELU E and ELU B. Results： Blood drug level - time cuvers of ELU E and ELU B in Wister rats following administration of an eleutherococcus injection into femoral vein were shown to fit a three - compartment model. The half - life （ t1/2 ） was 4. 66 h for ELU E and 2. 49 h for ELU B. After a single injected of eleutherococcuss injection, the concentration of ELU E and ELU B was followed as in order Cliver〉 Ckidney 〉 Cspleen 〉 Cheart and Ckidney 〉 Cliver 〉 Cheart. Concentrations of ELU E and ELU B were higher in the liver and kidney, this fact shows that both ELU E and ELU B are metabolized and excreted primarily from the liver and kidney. Conclusion：We believe the method described in the present paper can be used for the pharmacokinetic studies of ELU E and ELU B in rats.