HPLC-Q-TOF-MS技术在预知子配方颗粒指纹图谱建立和化学成分归属中的应用

Establishment of Fingerprint and Identification of the Components in Akebiae Fructus Formula Granule by HPLC-Q-TOF-MS

关键词:  预知子配方颗粒  HPLC-Q-TOF-MS  指纹图谱  4,5-O-二咖啡酰基奎宁酸

Key words:  Akebiae Fructus formula granule  HPLC-Q-TOF-MS  fingerprint  4,5-O-dicaffeoylquinic acid

摘要:

目的 建立预知子配方颗粒的HPLC指纹图谱，并使用Q-TOF-MS技术鉴定共有峰成分。方法 采用Agilent Poroshell 120 EC-C₁₈色谱柱（4.6 mm×50 mm，2.7 mm），以乙腈-0.05%甲酸水溶液为流动相进行梯度洗脱，检测波长210 nm，柱温30℃，流速0.5 mL·min^-1。采用相似度分析软件对指纹图谱进行分析。以标准物质和文献数据为参照，使用Q-TOF-MS负离子检测模式对特征峰进行质谱分析并指认。结果 建立了预知子配方颗粒的HPLC指纹图谱，以4，5-O-二咖啡酰基奎宁酸峰为参照峰，标定了14个共有峰，样品间相似度范围为0.943~0.994，样品与对照指纹图谱相似度范围为0.976~0.988。使用Q-TOF-MS对共有峰进行了指认，确定了8个峰成分，分别为新绿原酸、绿原酸、木通苯乙醇苷B、3，4-O-二咖啡酰基奎宁酸、3，5-O-二咖啡酰基奎宁酸、4，5-O-二咖啡酰基奎宁酸、川续断皂苷Ⅵ、α-常春藤皂苷。结论 该方法灵敏、准确、可靠，可用于预知子配方颗粒的质量评价。

Abstract:

OBJECTIVE To establish the HPLC fingerprint of Akebiae Fructus formula granule and identify common peak components by Q-TOF-MS technique. METHODS Acetonitrile-0.05% formic acid solution was used as a mobile phase to gradient elute Agilent Poroshell 120 EC-C₁₈ chromatographic column(4.6 mm×50 mm, 2.7 μm). The detection wavelength was 210 nm, the column temperature was 30℃, and the flow rate was 0.5 mL·min^-1. At the same time, similarity analysis software was used to analyze the fingerprint. In addition, the characteristic peaks were analyzed and identified by mass spectrometry using Q-TOF-MS anion detection mode with reference substances and literature data. RESULTS The HPLC fingerprint of Akebiae Fructus formula granule has been established. Based on the 4,5-O-Dicaffeoylquinic acid peak as the standard peak, 14 common peaks were identified. Specifically, the similarity range between samples and the similarity range between samples and reference fingerprint were 0.943-0.994 and 0.976-0.988, respectively. Furthermore, high resolution Q-TOF-MS was used to identify common peaks. It was indicated that a total of 8 peak components were identified, including neochlorogenic acid, chlorogenic acid, calceolarioside B, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, Asperosaponin VI and α-Hederin. CONCLUSION This method is sensitive, accurate and reliable, and can be used to predict the quality evaluation of Akebiae Fructus formula granule.