The purpose of the present study was to develop a simple, rapid and sensitive high-performance thin-layer chromatography (HPTLC) method for the simultaneous estimation of 5-fluorouracil (5FU) and berberine (BER) in rabbit plasma. BER and 5-FU are currently under extensive studies for their synergistic effects in cancer chemotherapy. The HPTLC method was developed using TLC aluminum plates precoated with silica gel 60 F₂₅₄ as the stationary phase. A mixture of toluene:methanol:ethyl acetate:formic acid (6:2:6:1, v/v) was used as the solvent system for development and detection was carried out at 266 nm. The method was designed to obtain the most optimum retardation factors (Rf) with best resolution. The Rf values of BER and 5-FU were 0.28 ± 0.02 and 0.57 ± 0.02, respectively. The plasma extraction was optimized using ethyl acetate. The method was validated in compliance with the USFDA guideline for bioanalytical method validation. The method was found to be linear in the range of 20 ng/spot to 300 ng/spot with R² of 0.9964 for 5-FU and 0.9943 for BER. The lower limit for quantification (LLOQ) of BER and 5-FU in rabbit plasma were obtained as 29 ng/spot and 26 ng/spot, respectively. The stability of BER and 5-FU in plasma were confirmed during freeze-thaw cycles (−20 °C), on bench for 5 h and long term for 21 days. The proposed method was validated statistically and was precise, accurate and sensitive for the estimation of BER and 5-FU in rabbit plasma. This method can be extended for therapeutic drug monitoring and pharmacokinetic studies in rabbits.

中文翻译：

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一种简便，快速，灵敏的高效薄层色谱方法，可同时估算兔血浆中的小ber碱和5-氟尿嘧啶

本研究的目的是开发一种简单，快速，灵敏的高性能薄层色谱法（HPTLC），用于同时估算兔血浆中的5-氟尿嘧啶（5FU）和小ber碱（BER）。BER和5-FU在癌症化疗中的协同作用目前正在广泛研究中。HPTLC方法是使用预涂硅胶60 F _254的TLC铝板开发的作为固定相。甲苯：甲醇：乙酸乙酯：甲酸的混合物（6：2：6：1，v / v）用作显影的溶剂系统，并在266 nm处进行检测。该方法旨在获得具有最佳分辨率的最佳延迟因子（Rf）。BER和5-FU的Rf值分别为0.28±0.02和0.57±0.02。使用乙酸乙酯优化血浆提取。该方法已按照美国食品药品管理局（USFDA）的生物分析方法验证指南进行了验证。发现该方法在R ²为20 ng / spot到300 ng / spot范围内是线性的5-FU为0.9964，BER为0.9943。兔子血浆中BER和5-FU的定量下限（LLOQ）分别为29 ng / spot和26 ng / spot。在冻融循环（-20°C），卧推5 h和长期21天的过程中，确认了血浆中BER和5-FU的稳定性。该方法经统计学验证，准确，准确，灵敏，可用于测定兔血浆中的BER和5-FU。该方法可扩展用于兔子的治疗药物监测和药代动力学研究。