HPLC Determination of Four Flavonoids of Aurantii Fructus Immaturus in Niuhuang Qingwei Pills and Genetic Analysis of Medicinal Materials / 中国实验方剂学杂志

ABSTRACT

Objective: To establish an high performance liquid chromatography (HPLC) method for the simultaneous determination of four constituents in Niuhuang Qingwei pill (narirutin,naringin,hesperidin,and neohesperidin), and identify the source of Fructus Aurantii Immaturus. Method: The analysis was performed on a Waters CORTECS C18 column (4.6 mm×50 mm,2.7 μm), with acetonitrile-0.12% formic acid solution as the mobile phase for gradient elution. The flow rate was 0.5 mL·min-1, the detection wavelength was set at 283 nm, and the column temperature was 27℃. Result: 12 batches of Niuhuang Qingwei pills showed the different content of flavonoids as Citrus aurantium and C. sinensis. Narirutin,naringin,hesperidin and neohesperidin showed good linear relationships within the ranges of 5.47-2 735 ng (r=0.999 6),7.25-3 625 ng (r=0.999 5),8.41-4 205 ng (r=0.999 4) and 8.36-4 180 ng (r=0.999 5),and their average recoveries were 101.3% (n=6,RSD 2.9%),98.0% (n=6,RSD 1.8%),95.9% (n=6,RSD 0.8%) and 96.0% (n=6,RSD 1.1%), respectively. The contents of narirutin,naringin,hesperidin,neohesperidin and total flavonoids were 0.36-1.28,2.66-4.87,1.02-11.07,3.58-6.41,and 7.98-13.34 mg·g-1, respectively. Conclusion: The developed method was simple,accurate and reliable,which can be used to identify the source of Aurantii Immaturus Fructus and simultaneously determine the content of four flavonoids in Niuhuang Qingwei pills. It could provide basic research for quality control and composition comparison of 2 kinds of Niuhuang Qingwei pills, showing more comprehensive indicators and reference value for the quality standard improvement of Niuhuang Qingwei pill.