基于HPLC波长切换法同时测定温经养血合剂中8个主要成分含量

Simultaneous determination of eight main components in Wenjing Yangxue Heji by the HPLC wavelength switching method

目的建立HPLC波长切换法同时测定温经养血合剂(WYH)中8个成分含量的方法。方法采用Agilent Eclipse XDB-C18色谱柱(250 mm×4.6 mm,5μm);以乙腈-0.1%磷酸溶液为流动相,梯度洗脱;检测波长分别为215 nm(柠檬苦素、吴茱萸碱、吴茱萸次碱)、230 nm(氧化芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷)和290 nm(桂皮醛)。结果所测8个成分依次的质量浓度分别在3.62~90.50μg/ml(r=0.9999)、1.66~41.50μg/ml(r=0.9998)、2.34~58.50μg/ml(r=0.9991)、1.98~49.50μg/ml(r=0.9994)、5.51~137.75μg/ml(r=0.9997)、8.69~217.25μg/ml(r=0.9995)、2.05~51.25μg/ml(r=0.9999)和3.86~96.50μg/ml(r=0.9996)范围内线性关系良好;平均回收率分别为98.52%(RSD=0.61%)、97.27%(RSD=0.83%)、98.18%(RSD=0.65%)、97.67%(RSD=0.48%)、99.16%(RSD=0.90%)、99.96%(RSD=0.36%)、96.93%(RSD=0.99%)和98.82%(RSD=1.00%)。结论本文建立的HPLC波长切换法同时测定WYH中的8个成分,方法简便,可为其质量控制提供参考。

Objective To develop an HPLC wavelength switching method for the simultaneous determination of eight main components in Wenjing Yangxue Heji（WYH）. Methods The Agilent Eclipse XDB-C18（250 mm×4.6 mm,5 μm）column was used.The mobile phase consisted of acetonitrile and 0.1% phosphoric acid aqueous solution by gradient elution.The detection wavelengths were 215 nm（limonin,evodiamine and rutaecarpine）,230 nm（oxypaeoniflorin,albiflorin,paeoniflorin and benzoylpaeoniflorin）and290 nm（cinnamaldehyde）. Results The present HPLC wavelength switching method had a good linearity within the concentration ranges of 3.62-90.50 mg/L（r=0.9999）for limonin,1.66-41.50 μg/ml（r=0.9998）for elvodiamine,2.34-58.50 μg/ml（r=0.9991）for rutaecarpine,1.98-49.50 μg/ml（r=0.9994）for oxypaeoniflorin,5.51-137.75 μg/ml（r=0.9997）for albiflorin,8.69-217.25 μg/ml（r=0.9995）for albiflorin,2.05-51.25 μg/ml（r=0.9999）for paeoniflorin,and 3.86-96.50 μg/ml（r=0.9996）for benzoylpaeoniflorin. The average recoveries in the order of the compounds were 98.52%（RSD=0.61%）,97.27%（RSD=0.83%）,98.18%（RSD=0.65%）,97.67%（RSD=0.48%）,99.16%（RSD=0.90%）,99.96%（RSD=0.36%）,96.93%（RSD=0.99%）,and 98.82%（RSD=1.00%）. Conclusion An HPLC wavelength switching method has been successfully established for the simultaneous determination of 8 components in WYH,which is simple,practical and thus could be used for the quality control of WYH.