Content Determination of 6 Active Constituents in Compound Huangqin Tablets by HPLC-QAMS / 中国药房

ABSTRACT

OBJECTIVE:To establish quantitative analysis of HPLC-quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of 6 active constituents in Compound huangqin tablets. METHODS:Using baicalin as internal content,HPLC method was used to calculate relative correction factor (RCF) of baicalin to polydatin,wogonoside, baicalein,andrographolide and dehydroandrographolide. The contents of above-mentioned 5 components in Compound huangqin tablets were calculated by using RCF(calculation value). The external standard method(ESM)was used to determine the contents of 6 constituents (measured value). The differences between calculation value and measured value were compared. The determination was performed on ZORBAX Eclipse XDB C18 column with mobile phase consisted of acetonitrile-0.05％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 306 nm for polydatin,225 nm for andrographolide,275 nm for baicalin,wogonoside and baicalein,254 nm for dehydroandrographolide. The column temperature was set at 25 ℃ and the injection volume was 10 μL. RESULTS:The linear range was 4.555-81.99 μg/mL for polydatin, 47.03-846.5 μg/mL for baicalin,12.73-229.1 μg/mL for wogonoside,12.88-231.8 μg/mL for baicalein,2.632-47.38 μg/mL for andrographolide,3.508-63.14 μg/mL for dehydroandrographolide (r=0.9992-0.9997),respectively. Limits of detection were 2.25,6.92,1.76,4.69,1.37,1.51 ng;limits of quantitation were 6.89,19.98,5.28,13.81,4.15,4.71 ng. Average recoveries were 98.2％,98.8％,97.9％,98.9％,98.8％,98.7％(RSD=0.91％-1.32％,n=9). RSDs of precision,reproducibility and stability tests (48 h) were lower than 2.0％ (n=6 or n=7). RCFs of baicalin to polydatin, wogonoside, baicalein, andrographolide and dehydroandrographolide were 0.529,0.506,1.004,0.831 and 1.087,respectively. There was no statistical significance between calculated values and measured ones(P>0.05). CONCLUSIONS:The method is simple,effective,accurate and cost saving. It can be used for simultaneous determination of 6 active constituents in Compound huangqin tablets.