HPLC波长切换联合梯度洗脱法同时测定参苏宣肺丸中的6种成分

Determination of Six Components in Shensu Xuanfei Wan by HPLC with Wavelength Switching Combined Gradient Elution Method

目的建立HPLC波长切换联合梯度洗脱法同时测定参苏宣肺丸中咖啡酸、迷迭香酸、3＇-羟基葛根素、葛根素、3＇-甲氧基葛根素、大豆苷的含量。方法采用ZOBAX SB-C18色谱柱（4.6 mm×250 mm,5μm）;流动相A为甲醇-乙腈（1∶1）,流动相B为0.1%冰醋酸溶液,进行梯度洗脱;流速为1.0 m L·min-1;咖啡酸和迷迭香酸的检测波长为320 nm,3＇-羟基葛根素、葛根素、3＇-甲氧基葛根素和大豆苷的检测波长为250 nm;进样量为20μL。结果咖啡酸、迷迭香酸、3＇-羟基葛根素、葛根素、3＇-甲氧基葛根素、大豆苷的浓度与峰面积分别在2.32-46.40μg·m L-1（r=0.999 8）、3.06-61.20μg·m L-1（r=0.999 5）、4.45-89.00μg·m L-1（r=0.999 9）、14.48-289.60μg·m L-1（r=0.999 9）、4.86-97.20μg·m L-1（r=0.999 7）、3.69-73.80μg·m L-1（r=0.999 6）内具有较好的线性关系,平均加样回收率和相应的RSD分别为99.0%（1.6%）,97.5%（0.6%）,99.3%（0.9%）,98.6%（1.3%）,97.6%（1.3%）,97.0%（0.9%）。结论方法操作准确、简便,可用于参苏宣肺丸的质量控制。

OBJECTIVE To develop an HPLC with wavelength switching combined gradient elution method for simultaneous determination of caffeic acid, rosmarinic acid, 3＇-hydroxy puerarin, puerarin, 3＇-methoxypuerarin and daidzin in Shensu Xuanfei Wan. METHODS ZOBAX SB-C18 column（4.6 mm×250 mm,5 μm） was applied with methanol-acetonitrile（1∶1）（A） and 0.1% glacial acetic acid solution（B） as mobile phase in gradient mode. The flow rate was 1.0 m L·min-1. Caffeic acid and rosmarinic acid were detected at 320 nm, 3＇-hydroxy puerarin, puerarin, 3＇-methoxypuerarin and daidzin were detected at 250 nm. Injection volume was 20 μL. RESULTS The concentration and peak area of caffeic acid, rosmarinic acid, 3＇-hydroxy puerarin, puerarin, 3＇-methoxypuerarin and daidzin were at 2.32-46.40 μg·m L-1（r=0.999 8）, 3.06-61.20 μg·m L-1（r=0.999 5）, 4.45-89.00 μg·m L-1（r=0.999 9）, 14.48-289.60 μg·m L-1（r=0.999 9）, 4.86-97.20 μg·m L-1（r=0.999 7）, 3.69-73.80 μg·m L-1（r=0.999 6）, respectively, and showed a good linear relationship. The average recoveries and the corresponding RSD were 99.0%（1.6%）, 97.5%（0.6%）, 99.3%（0.9%）, 98.6%（1.3%）, 97.6%（1.3%）, 97.0%（0.9%）, respectively. CONCLUSION The method is accurate, quick, simple. It could be reliable for the quality control of Shensu Xuanfei Wan.