Department of Pharmaceutical Sciences, University of Modena and Reggio Emilia, Via G. Campi 183, 41125 Modena, Italy.
J Pharm Biomed Anal. 2011 Jul 15;55(5):934-48. doi: 10.1016/j.jpba.2011.03.024. Epub 2011 Mar 25.
In this study, the composition of polyphenols (phenolic acids and flavonoids) in propolis extracts was investigated by HPLC-DAD and HPLC-ESI-MS/MS by comparing the performance of ion trap and triple quadrupole mass analyzers. The analyses were carried out on an Ascentis C(18) column (250mm×4.6mm I.D., 5μm), with a mobile phase composed by 0.1% formic acid in water and acetonitrile. Overall, the UV spectra, the MS and MS/MS data allowed the identification of 40 compounds. In the case of flavonoids, the triple quadrupole mass analyzer provided more collision energy if compared with the ion trap, originating product ions at best sensitivity. The HPLC method was validated in agreement with ICH guidelines: the correlation coefficients were >0.998; the limit of detection was in the range 1.6-4.6μg/ml; the recovery range was 96-105%; the intra- and inter-day %RSD values for retention times and peak areas were found to be <0.3 and 1.9%, respectively. The developed technique was applied to the analysis of hydroalcoholic extracts of propolis available on the Italian market. Although the chromatographic profile of the analyzed samples was similar, the quantitative analysis indicated that there is a great variability in the amount of the active compounds: the content of total phenolic acids ranged from 0.17 to 16.67mg/ml and the level of total flavonoids from 2.48 to 41.10mg/ml. The proposed method can be considered suitable for the phytochemical analysis of propolis extracts used in phytotherapy.
在这项研究中，通过比较离子阱和三重四极杆质谱仪的性能，用 HPLC-DAD 和 HPLC-ESI-MS/MS 研究了蜂胶提取物中多酚（酚酸和类黄酮）的组成。分析在 Ascentis C(18)柱（250mm×4.6mm ID，5μm）上进行，流动相由水中的 0.1%甲酸和乙腈组成。总的来说，紫外光谱、质谱和质谱/质谱数据允许鉴定 40 种化合物。在类黄酮的情况下，与离子阱相比，三重四极杆质谱仪提供了更多的碰撞能量，从而在最佳灵敏度下产生了产物离子。HPLC 方法按照 ICH 指南进行了验证：相关系数>0.998；检测限在 1.6-4.6μg/ml 范围内；回收率范围为 96-105%；保留时间和峰面积的日内和日间%RSD 值分别发现<0.3 和 1.9%。所开发的技术应用于分析意大利市场上可获得的蜂胶水醇提取物。尽管分析样品的色谱图相似，但定量分析表明活性化合物的含量存在很大差异：总酚酸含量范围为 0.17-16.67mg/ml，总类黄酮含量范围为 2.48-41.10mg/ml。该方法可被认为适合于在植物疗法中使用的蜂胶提取物的植物化学分析。
