肖榕+林艳+雷思敏+章莹+黄杰+夏伯候+李春+廖端芳+吴萍+林丽美

[摘要] 采用超高效液相色谱（UPLC），建立不同产地生/制何首乌二苯乙烯苷（TSG）的含量测定方法和指纹图谱。大孔树脂柱处理各样品，UPLC分析，Waters ACQUITY UPLC@BEH C₁₈色谱柱（2.1 mm×50 mm，1.7 μm），乙腈-0.5%甲酸溶液梯度洗脱，流速0.3 mL·min^-1，波长254 nm，柱温（25±5） ℃。结果发现大孔树脂分离后药材二苯乙烯苷含量均在25.0%以上；成功建立不同产地生/制何首乌药材UPLC指纹图谱；标定了17个色谱峰，聚类分析不能完全将生/制首乌分开，而主成分分析可明显地将其分为两类；二苯乙烯苷是两类何首乌自变量投影重要性指标（VIP）差异最大的组分。前处理方法可获得高含量的二苯乙烯苷，测定方法简单，灵敏，可靠，可用于生/制何首乌的快速鉴别，可作为何首乌药材整体质量控制的方法之一。

[关键词] 何首乌； 二苯乙烯苷； UPLC指纹图谱； 聚类分析； 主成分分析

[Abstract] To establish a content determination method for 2，3，5，4′-tetrahydroxy stilbene-2-O-β-D-glucoside （TSG） of the crude/processed root of Polygonum multiflorum from different habitats in China and set up the fingerprint by using UPLC. Various samples were pretreated by macro-porous resin. Then UPLC analysis was performed on Waters ACQUITY UPLC@BEH C₁₈ chromatographic column （2.1 mm×50 mm，1.7 μm） at （25±5） ℃. A binary gradient elution system was composed of acetonitrile （phase A） and 0.5% acetic acid solution （phase B）. Detection was performed at the wavelength of 254 nm， and the mobile flow rate was set at 0.3 mL·min^-1. Results showed that the yield of extraction of the 2，3，5，4′-tetrahydroxy stilbene-2-O-β-D-glucoside from root of P. multiflorum was all over 25.0% after macro-porous resin separation； an exclusive UPLC fingerprint method of the crude/processed root of P. multiflorum from different habitats was successfully set up and 17 chromatographic peaks were calibrated. Cluster analysis can not entirely distinguish the crude one from the processed one， while principal component analysis absolutely can. 2，3，5，4′-tetrahydroxy stilbene-2-O-β-D-glucoside is the composition that has largest differences in variable importance in projection （VIP） between crude and processed root of P. multiflorum. The separating method can gain high-purity 2，3，5，4′-tetrahydroxy stilbene-2-O-β-D-glucoside， and the determination method is simple， sensitive， reliable and can be used in fast identifying the crude/processed root of P. multiflorum or as a method for overall quality control of root of P. multiflorum.

[Key words] Polygonum multiflorum； 2，3，5，4′-tetrahydroxy stilbene-2-O-β-D-glucoside； UPLC fingerprint； cluster analysis； principal components analysis

何首烏为蓼科植物何首乌Polygonum multiflorum Thunb.的干燥块根[1]，具有保护神经、抗衰老、降血脂、抗动脉粥样硬化和促进生长等多方面的药理作用[2]，主要活性成分包含蒽醌类、二苯乙烯苷类、黄酮类、磷脂类等[3]。其中2，3，5，4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（简称二苯乙烯苷，TSG）是何首乌最具代表性的特征性成分，有降低胆固醇，增强免疫，抗骨质疏松，调节脂质，抗动脉粥样硬化等功效[4]，现已被2015年版《中国药典》采纳作为何首乌质量控制的指标性成分[5]。传统上，何首乌有生首乌和制首乌之分。生首乌是取原药材，除去杂质，洗净，润透，切块，干燥后所得；制首乌是生首乌经不同炮制方法而成[2]。……