三棱乙酸乙酯部位HPLC指纹图谱及4种酚类成分含量测定

目的：建立醋炙三棱乙酸乙酯部位HPLC指纹图谱，并同时测定对羟基苯甲酸、香兰素、对香豆酸和阿魏酸4种酚类物质含量，为三棱药材的质量控制提供借鉴。方法：富集醋炙三棱乙酸乙酯部位，采用高效液相色谱（HPLC）法，应用Inert Sustain C₁₈色谱柱（4.6 mm×250 mm，5 μm），流动相甲醇-0.1%甲酸溶液进行梯度洗脱，检测波长254 nm，流速0.80 mL·min^-1，测定对羟基苯甲酸、香兰素、对香豆酸和阿魏酸4种酚类物质的含量。结果：建立了三棱乙酸乙酯部位的HPLC指纹图谱，检出共有峰16个，各批次药材的相似度均大于0.9；通过与对照品比对，确定了指纹图谱中5，8，9，10号峰分别为对羟基苯甲酸、香兰素、对香豆酸和阿魏酸，并对其进行定量分析，结果对羟基苯甲酸的线性范围为0.008~0.924 μg，平均回收率为98.5%，香兰素的线性范围为0.039~4.712 μg，平均回收率为101.1%，对香豆酸的线性范围为0.054~6.572 μg，平均回收率为100.3%，阿魏酸的线性范围为0.018~2.236 ng，平均回收率为101.0%。结论：HPLC指纹图谱方法与含量测定方法简便，结果准确，重复性好，可作为三棱的质量控制方法。

HPLC fingerprints of EtOAc extracts and determination of four phenolic compounds in Rhizoma Sparganii

OBJECTIVE To establish HPLC fingerprints of EtOAc extracts and simultaneously determine the contents of p-hydroxybenzoic acid, vanillin, p-coumaric acid and ferulic acid in Rhizoma Sparganii for quality control.METHODS Inert Sustain C₁₈ column (4.6 mm×250 mm, 5 μm) was used to elute with methanol-0.1% formic acid as mobile phase, at the detection wave length of 254 nm and flow rate at 0.80 mL·min^-1.RESULTS The chromatographic fingerprints of EtOAc extracts in Rhizoma Sparganii were established, and 16 peaks were detected. The similarity of each batch was more than 0.9. By comparing the reference substance, P-hydroxybenzoic acid, vanillin, p-coumaric acid and ferulic acid were confirmed. The linear range of p-hydroxybenzoic acid was 0.008-0.924 g and recovery was 98.5%, the linear range of vanillin was 0.039-4.712 g and recovery was 101.1%, the linear range of coumaric acid was 0.054-6.572 g and recovery was 100.3%, the linear range of ferulic acid was 0.018-2.236 g and recovery was 101.0%, respectively.CONCLUSION HPLC fingerprinting method and content determination method are simple, accurate and reproducible, and can be used as quality control methods for Rhizoma Sparganii.