Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Material Science, Shaanxi Normal University, Xi'an 710062, China.
Talanta. 2010 Sep 15;82(4):1212-7. doi: 10.1016/j.talanta.2010.06.038. Epub 2010 Jul 3.
Response surface methodology (RSM) was applied to the optimization of on-line solid-phase extraction (SPE) parameters, and an automated system of on-line SPE coupled with high-performance liquid chromatography (HPLC) with fluorescence detection was developed for the determination of puerarin and daidzein in human serum. The human serum sample of 50 microL was injected into a conditioned C18 SPE cartridge, and the matrix was washed out with acetonitrile-KH(2)PO(4)-triethylamine buffer (0.01 M, pH 7.4) (3:97, v/v) for 3 min at a flow rate of 0.25 mL/min. Then the target analytes were eluted and transferred to the analytical column. A chromatographic gradient elution was programmed with the mobile phase consisting of acetonitrile and KH(2)PO(4)-triethylamine buffer, and the analytes were determined with a fluorescence detector at excitation wavelength of 350 nm and emission wavelength of 472 nm, respectively. The proposed method presented good linear relations (0.85-170 microg/mL for puerarin and 0.2-40 microg/mL for daidzein), satisfactory precision (RSD<8%), and accredited recovery (92.5-107.8%).
响应面法（RSM）被应用于在线固相萃取（SPE）参数的优化，并且建立了一种自动化的在线 SPE 与高效液相色谱（HPLC）与荧光检测相结合的系统，用于测定人血清中的葛根素和大豆苷元。50 μL 的人血清样品注入到预处理的 C18 SPE 柱中，用乙腈-KH（2）PO（4）-三乙胺缓冲液（0.01 M，pH 7.4）（3:97，v/v）以 0.25 mL/min 的流速冲洗 3 分钟以洗脱基质。然后将目标分析物洗脱并转移到分析柱上。采用乙腈和 KH（2）PO（4）-三乙胺缓冲液作为流动相进行色谱梯度洗脱，并用荧光检测器分别在激发波长为 350nm 和发射波长为 472nm 处测定分析物。所提出的方法呈现出良好的线性关系（葛根素为 0.85-170μg/mL，大豆苷元为 0.2-40μg/mL），令人满意的精密度（RSD<8%）和准确的回收率（92.5-107.8%）。
