益母草HPLC指纹图谱及多成分含量测定方法研究

Fingerprint analysis and multi-component determination of Leonurus japonicus by high performance liquid chromatography

建立益母草的HPLC指纹图谱及多成分含量同时测定方法,为益母草质量综合评价和控制提供参考。益母草用70%乙醇水浴回流提取2 h,提取物采用HPLC法分析,色谱柱为Kromasil C18(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%甲酸,梯度洗脱,流速为1.0 mL·min^-1,柱温30℃,检测波长为280 nm。25批样品得到共有峰12个,确认了其中5个峰对应的化学成分。丁香酸、盐酸益母草碱、芦丁、金丝桃苷和异槲皮苷分别在进样量0.4261~85.22 ng(r=0.9999)、7.948~1590 ng(r=0.9993)、10.20~2040 ng(r=1.0000)、2.018~403.6 ng(r=0.9999)、8.704~1741 ng(r=0.9999)内呈良好的线性关系;平均回收率分别为99.0%、97.6%、97.4%、96.9%和98.5%,RSD分别为1.1%、1.8%、1.4%、1.5%和1.3%。该指纹图谱特征性明显,并可同时测定5个成分的含量。

An HPLC fingerprint and multi-component determination method of Leonurus japonicus was established for comprehensive evaluation and quality control of Leonurus japonicus.The sample was incubated in70%ethanol in a water bath for 2 h,and the extract was analyzed by HPLC using Kromasil C18 column(250 mm×4.6 mm,5μm).The mobile phase consisted of acetonitrile-0.1%formic acid with gradient elution.The flow rate was1.0 mL·min^-1.The temperature of column was 30℃.The detection wavelength was 280 nm.HPLC fingerprint of characteristic components of Leonurus japonicus was established.There were 12 common peaks among 25 batches of samples,and 5 of them were identified and determined.Syringic acid,leonurine hydrochloride,rutin,hyperoside or isoquercitrin showed a good linearity in the ranges of 0.4261-85.22 ng(r=0.9999),7.948-1590 ng(r=0.9993),10.20-2040 ng(r=1.0000),2.018-403.6 ng(r=0.9999),or 8.704-1741 ng(r=0.9999),respectively.The average recoveries were 99.0%,97.6%,97.4%,96.9%and 98.5%with RSD of 1.1%,1.8%,1.4%,1.5%and 1.3%,respectively.The HPLC characteristic fingerprint of Leonurus japonicus was specific,and this method can simultaneously determine the content of 5 components.