基于多波长融合HPLC指纹图谱结合多成分定量的桑叶质量控制研究

Study on the quality control of mulberry leaves based on multi-wavelengths fusion HPLC fingerprint combined with multi-components quantification

目的建立多波长融合高效液相指纹图谱结合多成分定量测定桑叶的分析方法,为桑叶的质量评价和质量控制提供参考。方法桑叶水提取物制成干浸膏粉后,用体积分数70%甲醇超声提取30 min,色谱柱为ZORBAX SB-C_(18)(250 mm×4.6 mm,5μm),流动相为体积分数0.1%甲酸甲醇溶液-体积分数0.02%甲酸水溶液,梯度洗脱,流速为0.8 mL·min^(-1),进样体积为10μL,柱温为35℃,指纹图谱检测波长为254、285、300、312和344 nm,含量测定检测波长为254 nm。结果运用多波长融合技术对5波长指纹图谱进行拟合,建立桑叶融合指纹图谱,确立52个共有峰,指认6个特征峰,并采用系统指纹定量法对桑叶进行定性及整体定量分析;同时结合6个特征组分的准确含量测定,对桑叶品质进行综合评价。结果显示,10批不同产地的桑叶质量差异较大。结论所建立的方法全面的反映不同地域来源的桑叶品质,可为桑叶药材的质量控制提供更检测依据。

Objective Multi-wavelengths fusion high performance liquid chromatography(HPLC)fingerprint combined with multi-components quantification method was established to assess the quality of mulberry leaves.Methods The dry water extract powder of mulberry leaf was ultrasonically extracted for 30 min using 70%methanol solution,which was then analyzed by HPLC in a gradient elution mode.The chromatographic separation was performed on a ZORBAX C_(18) column(250 mm×4.6 mm,5μm)at the column temperature of 35℃.The mobile phase was composed of methanol with 0.1%formic acid and water with 0.02%formic acid at a flow rate of 0.8 mL·min^(-1).The injection volume was 10μL.The detection wavelengths for fingerprint were respectively set at 254,285,300,312 and 344;while quantitative wavelength was set at 254 nm.Results The multi-wavelengths fusion fingerprint was developed with five fingerprints followed by 52 common peaks,and 6 characteristic peaks were identified.Subsequently,the quality of mulberry leaves from 10 different geographical sources was comprehensively evaluated by systematic quantified fingerprint method from the aspects of qualification and integrated quantification.Moreover,the simultaneous determination of 6 characteristic components was also performed.The results showed that the quality of 10 batches of mulberry leaves from different geographical sources was significantly different.Conclusions The established method in our study is comprehensive for the quality evaluation of mulberry leaves from different regions,and can provide a more effective detection strategy for the quality control of mulberry leaf.