基于UPLC-DAD指纹图谱结合化学计量学及6种成分含量测定的桑叶质量研究

Study on the quality of mulberry leaves based on UPLC-DAD fingerprint combined with chemometrics and determination of 6 index components

关键词:  桑叶  UPLC-DAD  指纹图谱  相似度评价  化学计量学  含量测定

Key words:  mulberry leaf  UPLC-DAD  fingerprint atlas  similarity evaluation  chemometrics  determination of content

摘要:

[目的] 建立超高效液相色谱法（UPLC）-DAD指纹图谱方法，结合化学计量学与多指标含量测定综合评价桑叶质量。[方法] 采用Agilent EC-C₁₈（2.1 mm×100 mm，1.7 μm），柱温35℃；检测波长分别为290 nm（检测没食子酸甲酯、隐绿原酸）和360 nm（检测芦丁、异槲皮苷、山奈酚-3-O-芸香糖苷、紫云英苷）；流动相为乙腈-0.2%磷酸水溶液，梯度洗脱，流速0.2 mL/min，进样体积2 μL。建立22批桑叶药材的UPLC-DAD指纹图谱，采用聚类分析和主成分分析等化学计量学方法，以没食子酸甲酯、隐绿原酸、芦丁、异槲皮苷、山奈酚-3-O-芸香糖苷、紫云英苷对桑叶进行质量评价。[结果] 22批桑叶UPLC-DAD指纹图谱指认了16个共有峰，不同批次的桑叶药材与对照指纹图谱相似度差异较大；通过对照品指认6个共有峰，6种指标性成分分离度良好，各成分均具有较宽的线性范围和良好的线性关系（r2>0.999），有良好的精密度、稳定性、重复性和加样回收率。[结论] 采用UPLC-DAD技术所建立指纹图谱联合6种指标成分含量测定方法操作便捷、结果准确、重复性好，为桑叶的质量控制提供科学依据。

Abstract:

[Objective] A UPLC-DAD fingerprint method was established to comprehensively evaluate the quality of mulberry leaf combined with chemometrics and multi-index content determination.[Methods] The chromatographic column was Agilent EC-C₁₈ column (2.1 mm×100 mm, 1.7 μm), and the column temperature was set at 35℃. The detection wavelength were set at 290 nm (for methyl gallate and cryptochlorogenic acid) and 360 nm (for rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin). The mobile phase was composed of acetonitrile-0.2% phosphate acid solution in gradient elution manner at a flow rate of 0.2 mL/min and Sample volume 2 μL. Establish UPLC-DAD fingerprints of 22 batches of mulberry leaf medicinal materials, using cluster analysis and principal component analysis and other chemometric methods, as well as methyl gallate, cryptochlorogenic acid, rutin, isoquercitrin, kaempferol-3-O-rutinoside and astragalus glycosides were used to evaluate the quality of mulberry leaves.[Results] Twenty-two batches of mulberry leaves UPLC-DAD fingerprints confirmed 16 common peaks, the similarities of mulberry leaf medicinal materials from different batches and the control fingerprints are quite different. And 6 shared peaks were identified by the reference substance, the results showed that the separation of the 6 index components was good, and each component had a wide linear range and a good linear relationship (r2>0.999), with good precision, stability, repeatability and sample recovery.[Conclusion] Using UPLC-DAD technology to establish a fingerprint spectrum combined with 6 index component content determination method is convenient, accurate and reproducible, which could be used for quality evaluation of mulberry leaves.