HPLC波长切换法同时测定清清颗粒中9个成分的含量

目的: 建立HPLC波长切换法同时测定清清颗粒中9个活性成分(白芍苷、芍药苷、甘草苷、甘草酸、小檗碱、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素)的含量. 方法: 采用Promosil C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱(0~10 min,18%A→23%A;10~20 min,23%A→30%A;20~35 min,30%A→48%A;35~40 min,48%A→18%A),流速1.0 mL·min^-1,柱温30 ℃,变换波长检测(230 nm,白芍苷、芍药苷;237 nm,甘草苷、甘草酸;345 nm,小檗碱;280 nm,黄芩苷、汉黄芩苷、黄芩素、汉黄芩素). 结果: 白芍苷、芍药苷、甘草苷、甘草酸、小檗碱、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素9个成分的线性范围分别为0.007~0.14 μg(r=0.999 6)、0.011~0.22 μg(r=0.999 9)、0.011 2~0.224 μg(r=0.999 6)、0.030 6~0.612 μg(r=0.999 9)、0.002 4~0.048 μg(r=0.999 8)、0.107 2~2.144 μg(r=0.999 9)、0.017 9~0.358 μg(r=0.999 9)、0.005 12~0.102 4 μg(r=0.999 9)、0.002 7~0.054 μg(r=0.999 6);平均加样回收率(n=6)为95.7%~101.0%.3批样品中上述9个成分的含量分别为 0.289~0.312、0.515~0.539、1.031~1.047、2.494~2.518、1.960~2.020、9.970~10.02、2.190~2.300、0.559~0.570、0.609~0.612 mg·g^-1. 结论: 本文建立的方法符合方法学验证要求,可用于清清颗粒的9个指标性成分的同时测定.

Objective: To develop an HPLC method for simultaneous determination of 9 components(albiflorin,paeoniflorin,liquiritin,glycyrrhizic acid,berberine,baicalin,wogonoside,baicalein,wogonin)in Qingqing granules. Methods: The Promosil C₁₈ (250 mm×4.6 mm,5 μm)column was adopted;the mobile phase was acetonitrile(A)-0.1% phosphoric acid solution(B)with gradient elution(0-10 min,18%A→23%A;10-20 min,23%A→30%A;20-35 min,30%A→48%A;35-40 min,48%A→18%A)at a flow rate of 1.0 mL·min^-1,the column temperature was set at 30 ℃,and the detection wavelengths were 230 nm(albiflorin,paeoniflorin),237 nm(liquiritin,glycyrrhizic acid),345 nm(berberine),280 nm(baicalin,wogonoside,baicalein,wogonin). Results: The linear ranges of albiflorin,paeoniflorin,liquiritin,glycyrrhizic acid,berberine,baicalin,wogonoside,baicalein,wogonin fell within the ranges of 0.007-0.14 μg(r=0.999 6),0.011-0.22 μg(r=0.999 9),0.011 2-0.224 μg(r=0.999 6),0.030 6-0.612 μg(r=0.999 9),0.002 4-0.048 μg(r=0.999 8),0.107 2-2.144 μg(r=0.999 9),0.017 9-0.358 μg(r=0.999 9),0.005 12-0.102 4 μg(r=0.999 9),0.002 7-0.054 μg(r=0.999 6),respectively.The average recoveries(n=6)were 95.7%-101.0%.The contents of albiflorin,paeoniflorin,liquiritin,glycyrrhizic acid,berberine,baicalin,wogonoside,baicalein,wogonin in three batches of samples were 0.289-0.312 mg·g^-1,0.515-0.539 mg·g^-1,1.031-1.047 mg·g^-1,2.494-2.518 mg·g^-1,1.960-2.020 mg·g^-1,9.970-10.02 mg·g^-1,2.190-2.300 mg·g^-1,0.559-0.570 mg·g^-1,0.609-0.612 mg·g^-1,respectively. Conclusion: This method is consistent with the method validation requirements,and can simultaneously determine the above 9 index components of Qingqing granules.