Lin Hai-Shu, Ho Paul C
Department of Pharmacy, National University of Singapore, Singapore 119260, Singapore.
J Pharm Biomed Anal. 2009 Feb 20;49(2):387-92. doi: 10.1016/j.jpba.2008.10.042. Epub 2008 Nov 5.
A rapid HPLC-UV method had been developed and validated to quantify 3,5,4'-trimethoxy-trans-stilbene (TMS), a naturally occurring and pharmacologically active analog of resveratrol in rat plasma. The samples were mixed with three volumes of acetonitrile to precipitate protein. Chromatographic separation was achieved on a RP-HPLC column (Agilent ZORBAX Eclipse Plus C18: 250 mm x 4.6mm i.d., 5microm), which was protected by a guard column (Agilent ZORBAX Eclipse Plus C18: 12.5 mm x 4.6mm i.d., 5microm) through isocratic delivery of a mobile phase of acetonitrile: water (75:25, v/v) at a flow rate of 1.2ml/min. The assay was executed at 30 degrees C and the UV absorbance at 320nm was monitored. The retention time of TMS and trans-stilbene (internal standard) was 6.5 and 8.3min, respectively. The calibration curve was linear within the range of 15-1000ng/ml (R(2)>0.998) and 15ng/ml was the lower LOQ. The intra- and inter-day precisions were good and the RSD was all lower than 7.3%. The mean absolute recovery of TMS in plasma ranged from 99.2 to 104.1%. This HPLC method had been successfully applied to study the pharmacokinetics of TMS, which was fully dissolved with hydroxypropyl-beta-cyclodextrin (HP-beta-CyD). In comparison with resveratrol, TMS had greater plasma exposure, longer elimination half-life and lower clearance. As TMS had superior pharmacokinetic characteristics, its potential as a preventive or therapeutic agent in resveratrol-effective conditions or diseases should be considered.
已开发并验证了一种快速高效液相色谱-紫外法，用于定量大鼠血浆中白藜芦醇的天然存在且具有药理活性的类似物3,5,4'-三甲氧基反式芪（TMS）。将样品与三倍体积的乙腈混合以沉淀蛋白质。在反相高效液相色谱柱（安捷伦ZORBAX Eclipse Plus C18：250 mm×4.6mm内径，5微米）上实现色谱分离，该柱由保护柱（安捷伦ZORBAX Eclipse Plus C18：12.5 mm×4.6mm内径，5微米）保护，通过等度输送乙腈：水（75:25，v/v）的流动相，流速为1.2ml/分钟。该测定在30℃下进行，并监测320nm处的紫外吸光度。TMS和反式芪（内标）的保留时间分别为6.5和8.3分钟。校准曲线在15 - 1000ng/ml范围内呈线性（R²>0.998），15ng/ml为较低的定量下限。日内和日间精密度良好，相对标准偏差均低于7.3%。TMS在血浆中的平均绝对回收率为99.2%至104.1%。该高效液相色谱法已成功应用于研究TMS的药代动力学，TMS与羟丙基-β-环糊精（HP-β-CyD）完全溶解。与白藜芦醇相比，TMS具有更高的血浆暴露量、更长的消除半衰期和更低的清除率。由于TMS具有优异的药代动力学特性，应考虑其在白藜芦醇有效治疗的病症或疾病中作为预防或治疗剂的潜力。
