Bachmann Stefan, Huck Christian W, Bakry Rania, Bonn Günther K
Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innsbruck, Austria.
Electrophoresis. 2007 Mar;28(5):799-805. doi: 10.1002/elps.200600228.
A CE method employing capacitively coupled contactless conductivity (C(4)D) compared to indirect UV-detection was developed for the analysis of phytochemically relevant flavonoids, such as 6-hydroxyflavone, biochanin A, hesperetin and naringenin. To ensure fast separation at highest selectivity, sensitivity and peak symmetry, the pH value and the concentration of the running BGE had to be optimized regarding both co- and counter-EOF mode. Optimum conditions were found to be 1.0 and 5.0 mM chromate BGE (pH 9.50) in the counter- and co-EOF mode, respectively. Validation of the established CE-C(4)D method pointed out to be approximately seven times more sensitive compared to indirect UV-detection applying the same conditions. The lower LOD defined at an S/N of 3:1 was found between 0.12 and 0.21 microg/mL for the analytes of interest using C(4)D and between 0.77 and 1.20 microg/mL using indirect UV-detection. Compared to an earlier published CE method employing direct UV-detection, C(4)D was found to be approximately two times more sensitive. Due to the lower baseline noise, C(4)D showed an excellent regression coefficient >0.99 compared to 0.93 when using indirect UV detection calibrating within a concentration range between 1 and 10 microg/mL. The influence of the sugar moiety on the conductivity of a flavonoid was studied upon the analysis of the aglycon hesperetin and the rutinosid hesperidin. The sugar moiety in hesperedin shows a higher conductivity compared to hesperetin. Finally, the optimized established CE-C(4)D method was applied to the determination and quantification of naringenin in Sinupret.
开发了一种采用电容耦合非接触式电导检测(C(4)D)的毛细管电泳(CE)方法,用于分析与植物化学相关的黄酮类化合物,如6-羟基黄酮、鹰嘴豆芽素A、橙皮素和柚皮素。为确保在最高选择性、灵敏度和峰对称性下实现快速分离,必须针对共电渗流和反电渗流模式优化运行缓冲液的pH值和浓度。发现最佳条件分别是在反电渗流模式下为1.0 mM铬酸盐缓冲液(pH 9.50),在共电渗流模式下为5.0 mM铬酸盐缓冲液(pH 9.50)。已建立的CE-C(4)D方法的验证表明,在相同条件下,与间接紫外检测相比,其灵敏度大约高7倍。对于感兴趣的分析物,使用C(4)D时,在信噪比为3:1时定义的较低检测限在0.12至0.21 μg/mL之间,使用间接紫外检测时在0.77至1.20 μg/mL之间。与早期发表的采用直接紫外检测的CE方法相比,C(4)D的灵敏度大约高2倍。由于基线噪声较低,在1至10 μg/mL的浓度范围内进行校准时,C(4)D显示出优异的回归系数>0.99,而使用间接紫外检测时为0.93。在分析苷元橙皮素和芸香苷橙皮苷时,研究了糖基对黄酮类化合物电导率的影响。橙皮苷中的糖基显示出比橙皮素更高的电导率。最后,将优化建立的CE-C(4)D方法应用于仙璐贝中柚皮素的测定和定量分析。