利用高壓液相層析儀同時分析兔子血漿中的Caffeic Acid、Ferulic Acid 及Isoferulic Acid   全文下載

Simultaneous Determination of Caffeic Acid, Ferulic Acid and Isoferulic Acid in Rabbit Plasma by High Performance Liquid Chromatography

使用逆相的柱狀分析管同時分析兔子血液中的caffeic acid、ferulicacid 及isoferulic acid 的濃度。其方法是用ODS 的柱狀分析管（150 mm × 4.6 mm I.D.，5 μm）當作固定相，而以acetonitrile/glacial acetic acid/water（15:0.5:85，v/v，pH 調整至4.5）之溶液為移動相，在流速1 mL/min，偵測紫外光波長為321 nm 的條件下分析。先取100 μL 的血漿為同時偵測caffeic acid、ferulic acid 與isoferulic acid 濃度之樣品，而以10% trifluoroacetic acid （v/v）當作去除蛋白的步驟，可偵測至最低濃度為0.1 μg/mL。Caffeic acid、ferulic acid 及isoferulic acid 在0.1-100 μg/mL 的範圍內有很好的線性關係。以此方法處理樣品時，caffeic acid、ferulic acid 及isoferulic acid 三種成分之平均回收率分別為96.2%，98.0% 及98.3%。而此三種成分在室溫下或4° C的溫度下，不論是經過三次的冷凍及回融的步驟或製備後24 hr 等安定性測試，皆有很好的安定性。因此，這個分析方法可成功的運用在兔子經靜脈注射caffeic acid 後，有關caffeic acid、ferulic acid 與isoferulic acid 之藥物動力學研究。

A simple and sensitive high performance liquid chromatographic method with reverse-phase column for the simultaneous quantification of caffeic acid, ferulic acid, and isoferulic acid in rabbit plasma was developed. An ODS column (150 mm × 4.6 mm I.D., 5 μm) was used as the stationary phase and the mobile phase consisted of acetonitrile/glacial acetic acid/water (15:0.5:85, v/v, pH adjusted to 4.5). Flow-rate was 1.0 mL/min and UV absorbance was set at 321 nm. One hundred microliter of plasma was used to simultaneously measure the concentrations of caffeic acid, ferulic acid, and isoferulic acid. After a direct clean-up procedure with 10% of trifluoroacetic acid, the lower limits of quantification were 0.1 μg/mL and the standard curves were found to be linear over the concentration ranges of 0.1-100 μg/mL for caffeic acid, ferulic acid, and isoferulic acid. The average recoveries for caffeic acid, ferulic acid, and isoferulic acid were 96.2%, 98.0%, and 98.3%, respectively. The results for all compounds also showed good stability after three freeze/thaw cycles and storage at ambient temperature and 4°C for 24 hr. The assay method was successfully applied to the pharmacokinetic study of caffeic acid, ferulic acid, and isoferulic acid after an intravenous administration of caffeic acid to rabbits.