Cen Meifeng, Liang Heng, Xiong Xiaohong, Zeng Jie, Cheng Xuange, Wang Sujun
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Center, 280 Wai Huan East Road, Guangzhou 510006, China.
J Chromatogr Sci. 2017 Nov 1;55(10):1037-1042. doi: 10.1093/chromsci/bmx072.
A simple, optimized and sensitive high-performance liquid chromatograph method with ultraviolet (UV) detection (HPLC/UV) was developed and validated for determination of isochlorogenic acid A in rat plasma. The analytes were successfully separated on a Shodex C18 column (5 μm particle size, 250 mm × 4.6 mm, i.d.), the mobile phase contained 0.1% phosphoric acid aqueous solution (solvent A) and methanol (solvent B) (50:50, v/v) at a flow rate of 1.0 mL/min. The wavelength for UV detection was set at 300 nm and the column temperature was maintained in 30°C. Calibration curve for isochlorogenic acid A was found to be good linear over the range of 0.04-40 μg/mL (r = 0.9998). The intra- and inter-day precisions (relative standard deviation) were within 7.63% and the assay accuracy (RE) ranged from -1.41 to 3.25%. The limit of detection and the lower limit of quantification were 0.012 and 0.04 μg/mL, respectively. The validated method was successfully applied to pharmacokinetic study of isochlorogenic acid A in rats for the first time. The pharmacokinetic parameters were evaluated after the rats were administered intravenously and intragastrically isochlorogenic acid A at the single dose of 18 mg/kg, respectively. The absolute bioavailability was calculated to be 22.6%.
建立了一种简单、优化且灵敏的带紫外（UV）检测的高效液相色谱法（HPLC/UV），并对其进行验证，用于测定大鼠血浆中的异绿原酸A。分析物在Shodex C18柱（粒径5μm，250mm×4.6mm，内径）上成功分离，流动相包含0.1%磷酸水溶液（溶剂A）和甲醇（溶剂B）（50:50，v/v），流速为1.0mL/min。UV检测波长设定为300nm，柱温保持在30℃。异绿原酸A的校准曲线在0.04 - 40μg/mL范围内呈良好线性（r = 0.9998）。日内和日间精密度（相对标准偏差）在7.63%以内，测定准确度（RE）范围为-1.41%至3.25%。检测限和定量下限分别为0.012和0.04μg/mL。该验证方法首次成功应用于大鼠异绿原酸A的药代动力学研究。分别以18mg/kg的单剂量静脉注射和灌胃给予大鼠异绿原酸A后，评估药代动力学参数。计算得出绝对生物利用度为22.6%。
