OBJECTIVE: To establish a method for the contents determination and cluster analysis of free and combined anthraquinone of aloe emodin, rhein, emodin, rhubarb phenol and physcion in commercially available Yiqing granule from different manufacturers. METHODS: HPLC was performed on the column of Eclipse plus C18 with mobile phase of methanol-0.1% Phosphoricacid solution (gradient elution) at a flow rate of 0.8 ml/min, the detection wavelength was 254 nm, the column temperature was 25 ℃, and the injection volume was 20 μl. RESULTS: The linear range was 0.001 6-0.128 0 μg for aloe emodin(r=0.999 9), 0.003 4-0.273 6 μg for rhein(r=0.999 9), 0.003 7-0.299 2 μg for emodin (r=0.999 9), 0.006 7-0.536 0 μg for rhubarb phenol (r=0.999 9) and 0.001 7-0.134 4 μg (r=0.999 9) for physcion; RSDs of precision, stability and reproducibility tests were lower than 3%; recoveries were 97.77%-101.47%(RSD=1.30%,n=6), 98.09%-100.26%(RSD=0.76%,n=6), 96.42%-100.38%(RSD=1.58%,n=6), 96.63%-102.50% (RSD=2.17%,n=6) and 97.10%-103.70%(RSD=2.47%,n=6), respectively; the content range of total anthraquinone was 0.042-0.218 mg/g, 0.029-0.448 mg/g, 0.022-0.167 mg/g, 0.032-0.284 mg/g and 0.006-0.060 mg/g, combined anthraquinone was 0.010-0.111 mg/g, 0.013-0.092 mg/g, 0.011-0.097 mg/g, 0.030-0.246 mg/g and 0.001-0.034 mg/g, and free anthraquinone was 0.022 3-0.143 mg/g, 0.015-0.356 mg/g, 0.008-0.071 mg/g, 0.006-0.075 mg/g and 0.003-0.032 mg/g. Cluster analysis showed there were 4 batches belonged to Category 1, 2 batches belonged to Category 2 and 4 batches belonged to Category 3 for the total anthraquinone in 5 components; 4 bathes belonged to Category 1, 2 batches belonged to Category 2 and 4 batches belonged to Category 3 for the combined anthraquinone; and 6 bathes belonged to Category 1 and 4 batches belonged to Category 2 for the free anthraquinone. CONCLUSIONS: The method is simple and accurate, and can provide reference for improving the quality and diarrhea efficacy control of Yiqing granule; there were great differences in Rheum palmatum and combined anthraquinone from different manufacturers.
关键词
一清颗粒大黄游离蒽醌结合蒽醌高效液相色谱法聚类分析
Keywords
Yiqing granuleRhei Radix Et RhizomaFree anthraquinoneCombined anthraquinoneHPLCCluster analysis