HPLC法同时测定狗脊中5种成分含量

Simultaneous determination of five components in Cibotii Rhizoma by HPLC

目的建立HPLC定量方法,同时测定狗脊饮片中曲酸、5-羟甲基糠醛(5-HMF)、原儿茶酸、麦芽酚及原儿茶醛5种成分含量。方法应用HPLC技术,采用岛津Shim-pak C18(4.6 mm×250 mm,5μm),流动相以甲醇为有机相、0.1%甲酸-甲酸铵溶液为水相,流速1.0 mL/min,检测波长280 nm(曲酸、原儿茶酸、麦芽酚、原儿茶醛)及260 nm (5-HM F),柱温35℃。结果曲酸、5-HM F、原儿茶酸、麦芽酚及原儿茶醛浓度分别在0.53~21.17、9.64~482.00、2.52~100.88、0.52~21.09、0.52~20.87μg/mL范围内与峰面积线性关系良好(r^2=0.999 9,n=6);定量限/检测限(LOQ/LOD)分别为0.03/0.01、0.20/0.05、0.35/0.07、0.05/0.02、0.05/0.01μg/mL;平均回收率为85.2%~102.8%。结论采用C18柱为固定相、双波长检测、梯度洗脱HPLC方法测定狗脊中5种成分含量,方法灵敏度高,准确性好,可用于狗脊饮品及其制品的质量控制。

Objective To establish a validated HPLC method for quantitative analysis of kojic acid,5-HMF,protocatechuic acid,maltol and protocatechuic aldehyde in Cibotii Rhizoma.Methods The samples were determined on a Shimatsu Shim-pak C18 column(4.6 mm×250 mm,5μm)by gradient elution with methanol as organic phase,0.1%formic acid-ammonium formate aqueous solution as aqueous phase,at a flowrate of 1.0 mL/min.The detection wavelength was 280 nm for kojic acid,protocatechuic acid,maltol and protocatechuic aldehyde,while 260 nm for 5-HMF.The column temperature was controlled at 35℃.Results Kojic acid,5-HMF,protocatechuic acid,maltol and protocatechuic aldehyde exhibited good linearity(r^2=0.999 9,n=6)among the ranges of 0.53-21.17,9.64-482.00,2.52-100.88,0.52-21.09,0.52-20.87μg/mL.The limit of quantitation(LOQ)/limit of detection(LOD)were 0.03/0.01,0.20/0.05,0.35/0.07,0.05/0.02,0.05/0.01μg/mL,respectively.The average recoveries were 85.2%-102.8%.Conclusion C18 column was employed as stationary phase,dual wavelength detection and gradient elution HPLC method were used to determine the contents of five components in Cibotii Rhizoma.The method performs with high sensitivity and good accuracy,and can be used for quality control of Cibotii Rhizoma and its products.