RP-HPLC同时测定大鼠血浆中4种丹参酮的浓度及药动学研究

Simultaneous Determination of Four Tanshinones in Rat Plasma by High Performance Liquid Chromatography and Its Application to Pharmacokinetic Studies

目的建立了同时测定大鼠血浆中4种丹参酮浓度的反相高效液相色谱方法,并将其应用于丹参醇提物中4种丹参酮的药动学研究。方法采用VP-ODS C18柱(4.6mm×250mm,5μm),以甲醇-0.05mol.L-1醋酸铵缓冲溶液(醋酸调pH至2.5)(70:30)为流动相,流速1.0mL.min-1,检测波长263nm,柱温40℃,以丙酸睾丸素为内标,测定血浆4种丹参酮的浓度。结果隐丹参酮、丹参酮Ⅰ、次甲基丹参酮和丹参酮ⅡA的线性范围分别为:0.01～12.75mg.L-1;0.01～16.25mg.L-1;0.02～13.75mg.L-1;0.02～13.25mg.L-1,其回收率均大于88.7%。药动学参数表明,隐丹参酮和丹参酮ⅡA符合二室开放模型,次甲基丹参酮和丹参酮Ⅰ符合一室开放模型。结论该方法简便、可靠、准确,可用于丹参提取物中丹参酮类的血药浓度分析和药动学研究。

OBJECTIVE To develop a reversed phase high performance liquid chromatographic method for simultaneous determination of four tanshinones in rat plasma in the pharmacokinetic study. METHODS The assay was conducted oil a VP-ODS C18 column （4. 6 mm× 250 ram,5 μm）, and the mobile phase consisted of methanol and 0. 05 mol·L^-1 ammonium acetate buffers volume ratio of 70：30 （pH 2. 5 ）. The flow rate was 1.0 mL·min^-1. The detection wavelength was set at 263 nm. Testosterone propionate was used as internal standard. RESULTS The response was linear in the concentration range of 0. 01 - 12. 75 mg·L^-1 for cryptotan- shinone, 0. 01- 16. 25 mg·L^-1 for tanshinone Ⅰ 0. 02 - 13.75 mg·L^-1 for methltanshinone and 0. 02-13.25 mg·L^-1 for tanshinone Ⅱ A, respectively. The recovery of assay was above 88.7%. The pharmacokinetie paramenters showed that cryptotanshinone and tanshinone Ⅱ after oral administration were fitted with a two-compartment open model while the methltanshinone and tanshinone Ⅰ were fitted with one -compartment open model. CONCLUSION determine the four kinds of tanshinones in rat plasma.