Magiera Sylwia, Baranowska Irena, Lautenszleger Anna
Department of Inorganic, Analytical Chemistry and Electrochemistry, Silesian University of Technology, 7M. Strzody Str., 44-100 Gliwice, Poland.
Department of Inorganic, Analytical Chemistry and Electrochemistry, Silesian University of Technology, 7M. Strzody Str., 44-100 Gliwice, Poland.
J Pharm Biomed Anal. 2015 Jan;102:468-75. doi: 10.1016/j.jpba.2014.10.004. Epub 2014 Oct 14.
A simple and rapid ultra-high performance liquid chromatographic (UHPLC) method coupled with an ultraviolet detector (UV) has been developed and validated for the separation and determination of 14 major flavonoids ((±)-catechin, (-)-epicatechin, glycitin, (-)-epicatechin gallate, rutin, quercitrin, hesperidine, neohesperidine, daidzein, glycitein, quercetin, genistein, hesperetin, and biochanin A) in herbal dietary supplements. The flavonoids have been separated on a Chromolith Fast Gradient Monolithic RP-18e column utilizing a mobile phase composed of 0.05% trifluoroacetic acid in water and acetonitrile in gradient elution mode. Under these conditions, flavonoids were separated in a 5 min run. The selectivity of the developed UHPLC-UV method was confirmed by comparison with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis. The validation parameters such as linearity, sensitivity, precision, and accuracy were found to be highly satisfactory. The optimized method was applied to determination of flavonoids in different dietary supplements. Additionally, the developed HPLC-UV method combined with 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay was used in the evaluation of antioxidant activity of the selected flavonoids.
已开发并验证了一种简单快速的超高效液相色谱（UHPLC）方法，该方法与紫外检测器（UV）联用，用于分离和测定草本膳食补充剂中的14种主要黄酮类化合物（（±）-儿茶素、（-）-表儿茶素、甘草素、（-）-表儿茶素没食子酸酯、芦丁、槲皮苷、橙皮苷、新橙皮苷、大豆苷元、甘草苷元、槲皮素、染料木黄酮、橙皮素和鹰嘴豆芽素A）。这些黄酮类化合物在Chromolith Fast Gradient Monolithic RP-18e柱上进行分离，流动相由0.05%三氟乙酸水溶液和乙腈组成，采用梯度洗脱模式。在这些条件下，5分钟内即可分离出黄酮类化合物。通过与超高效液相色谱-串联质谱（UHPLC-MS/MS）分析比较，证实了所开发的UHPLC-UV方法的选择性。发现线性、灵敏度、精密度和准确度等验证参数非常令人满意。该优化方法用于测定不同膳食补充剂中的黄酮类化合物。此外，所开发的HPLC-UV方法结合2,2-二苯基-1-苦基肼自由基（DPPH）测定法用于评估所选黄酮类化合物的抗氧化活性。
