HPLC波长切换联合梯度洗脱法同时测定荆防败毒丸中10个成分的含量

Simultaneous determination of ten components in Jingfang Baidu pills by HPLC method with wavelength switching and gradient elution

目的:HPLC波长切换联合梯度洗脱法同时测定荆防败毒丸中的5-O-咖啡酰莽草酸、落新妇苷、花旗松素、黄杞苷、白藜芦醇、洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、瑟丹酸内酯和藁本内酯含量。方法:采用Shiseido C_(18)色谱柱(4.6 mm×250 mm,5μm),以甲醇-乙腈(3∶1)(A)与0.1%冰醋酸水溶液(B)为流动相,梯度洗脱,流速1.2mL·min^(-1),柱温30℃,5-O-咖啡酰莽草酸、落新妇苷、花旗松素、黄杞苷和白藜芦醇的检测波长为291 nm,洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、瑟丹酸内酯和藁本内酯的检测波长为280 nm。结果:5-O-咖啡酰莽草酸、落新妇苷、花旗松素、黄杞苷、白藜芦醇、洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、瑟丹酸内酯和藁本内酯10个成分的线性范围分别为9.13~182.60μg·mL^(-1)(r=0.999 4)、20.75~415.00μg·mL^(-1)(r=0.999 9)、3.54~70.80μg·mL^(-1)(r=0.999 5)、4.78~95.60μg·mL^(-1)(r=0.999 8)、2.42~48.40μg·mL^(-1)(r=0.999 3)、3.38~67.60μg·mL^(-1)(r=0.999 9)、4.94~98.80μg·mL^(-1)(r=0.999 6)、6.36~127.20μg·mL^(-1)(r=0.999 8)、3.07~61.40μg·mL^(-1)(r=0.999 7)、9.66~193.20μg·mL^(-1)(r=0.999 5);平均加样回收率(n=6)分别为97.3%(RSD=1.1%)、99.2%(RSD=1.3%)、96.8%(RSD=0.44%)、98.8%(RSD=0.86%)、96.9%(RSD=1.1%)、98.3%(RSD=1.6%)、97.1%(RSD=1.2%)、100.0%(RSD=0.60%)、97.9%(RSD=1.3%)、99.2%(RSD=1.1%)。3批样品中上述10个成分的含量范围分别为0.542~0.579、1.519~1.663、0.105~0.117、0.314~0.331、0.072~0.081、0.101~0.113、0.164~0.183、0.406~0.437、0.163~0.182、0.585~0.607 mg·g^(-1)。结论:本文建立的HPLC波长切换联合梯度洗脱法可为荆防败毒丸质量标准的提高提供依据。

Objective：To develop an HPLC method with wavelength switching and gradient elution for simultaneous determination of 5-O-caffeoylshikimic acid,astilbin,taxifolin,engeletin,resveratrol,senkyunolide H,senkyunolide I,senkyunolide A,sedanolide and ligustilide in Jingfang Baidu pills.Methods：The Shiseido C_（18） column（4.6 mm×250 mm,5 μm）was adopted,the mobile phase was methanol-acetonitrile（3∶1）（A）and 0.1% acetic acid glacial（B）with gradient elution at a flow rate of 1.2mL·min（-1）,the column temperature was set at 30 ℃.5-O-caffeoylshikimic acid,astilbin,taxifolin,engeletin and resveratrol were detected at 291 nm;senkyunolide H,senkyunolide I,senkyunolide A,sedanolide and ligustilide were detected at 280 nm.Results：The linear ranges of 5-O-caffeoylshikimic acid,astilbin,taxifolin,engeletin,resveratrol,senkyunolide H,senkyunolide I,senkyunolide A,sedanolide and ligustilide were 9.13（-1）82.60 μg·mL（-1）（r=0.999 4）,20.75-415.00 μg·mL（-1）（r=0.999 9）,3.54-70.80 μg·mL（-1）（r=0.999 5）,4.78-95.60 μg·mL（-1）（r=0.999 8）,2.42-48.40 μg·mL（-1）（r=0.999 3）,3.38-67.60 μg·mL（-1）（r=0.999 9）,4.94-98.80 μg·mL（-1）（r=0.999 6）,6.36（-1）27.20 μg·mL（-1）（r=0.999 8）,3.07-61.40 μg·mL（-1）（r=0.999 7）,9.66（-1）93.20 μg·mL（-1）（r=0.999 5）,respectively.The average recoveries（n=6）were 97.3%（RSD=1.1%）,99.2%（RSD=1.3%）,96.8%（RSD=0.44%）,98.8%（RSD=0.86%）,96.9%（RSD=1.1%）,98.3%（RSD=1.6%）,97.1%（RSD=1.2%）,100.0%（RSD=0.60%）,97.9%（RSD=1.3%）,99.2%（RSD=1.1%）,respectively.The contents of ten components in 3 batches of samples were 0.542-0.579,1.519（-1）.663,0.105-0.117,0.314-0.331,0.072-0.081,0.101-0.113,0.164-0.183,0.406-0.437,0.163-0.182,0.585-0.607 mg·g（-1）,respectively.Conclusion：HPLC method with wavelength switching and gradient elution can be used for the quality control of Jingfang Baidu pills.