Ricciutelli Massimo, Marconi Shara, Boarelli Maria Chiara, Caprioli Giovanni, Sagratini Gianni, Ballini Roberto, Fiorini Dennis
HPLC-MS Laboratory, University of Camerino, V. S. Agostino 1, I-62032 Camerino, MC, Italy.
School of Science and Technology, Chemistry Division, University of Camerino, V. S. Agostino 1, I-62032 Camerino, MC, Italy.
J Chromatogr A. 2017 Jan 20;1481:53-63. doi: 10.1016/j.chroma.2016.12.020. Epub 2016 Dec 11.
In order to assess if an extra virgin olive oil (EVOO) can be acknowledged with the health claim related to olive oil polyphenols (Reg. EU n.432/2012), a new method to quantify these species in EVOO, by means of liquid-liquid extraction followed by HPLC-DAD/MS/MS of the hydroalcoholic extract, has been developed and validated. Different extraction procedures, different types of reverse-phase analytical columns (Synergi Polar, Spherisorb ODS2 and Kinetex) and eluents have been tested. The chromatographic column Synergi Polar (250×4.6mm, 4μm), never used before in this kind of application, provided the best results, with water and methanol/isopropanol (9/1) as eluents. The method allows the quantification of the phenolic alcohols tyrosol and hydroxytyrosol, the phenolic acids vanillic, p-coumaric and ferulic acids, secoiridoids derivatives, the lignans, pinoresinol and acetoxypinoresinol and the flavonoids luteolin and apigenin. The new method has been applied to 20 commercial EVOOs belonging to two different price range categories (3.78-5.80 euros/L and 9.5-25.80 euros/L) and 5 olive oils. The obtained results highlight that acetoxypinoresinol, ferulic acid, vanillic acid and the total non secoiridoid phenolic substances resulted to be significantly higher in HEVOOs than in LEVOOs (P=0.0026, 0.0217, 0.0092, 0.0003 respectively). For most of the samples analysed there is excellent agreement between the results obtained by applying the HPLC method adopted by the International Olive Council and the results obtained by applying the presented HPLC method. Results obtained by HPLC methods have been also compared with the ones obtained by the colorimetric Folin-Ciocalteu method.
为了评估特级初榨橄榄油（EVOO）是否可以获得与橄榄油多酚相关的健康声明（欧盟法规第432/2012号），已经开发并验证了一种通过液液萃取，然后对水醇提取物进行HPLC-DAD/MS/MS来定量测定EVOO中这些成分的新方法。测试了不同的萃取程序、不同类型的反相分析柱（Synergi Polar、Spherisorb ODS2和Kinetex）以及洗脱剂。此前从未在此类应用中使用过的色谱柱Synergi Polar（250×4.6mm，4μm）提供了最佳结果，以水和甲醇/异丙醇（9/1）作为洗脱剂。该方法能够定量测定酚醇酪醇和羟基酪醇、酚酸香草酸、对香豆酸和阿魏酸、裂环环烯醚萜衍生物、木脂素、松脂醇和乙酰氧基松脂醇以及黄酮类化合物木犀草素和芹菜素。该新方法已应用于20种属于两个不同价格范围类别的市售EVOO（3.78 - 5.80欧元/升和9.5 - 25.80欧元/升）以及5种橄榄油。所得结果表明，在高等级初榨橄榄油（HEVOOs）中，乙酰氧基松脂醇、阿魏酸、香草酸和总非裂环环烯醚萜酚类物质的含量显著高于低等级初榨橄榄油（LEVOOs）（P值分别为0.0026、0.0217、0.0092、0.0003）。对于大多数分析样品，应用国际橄榄理事会采用的HPLC方法获得的结果与应用本文提出的HPLC方法获得的结果之间具有极好的一致性。通过HPLC方法获得的结果也与通过比色福林 - 酚试剂法获得的结果进行了比较。
