Morazzoni Paolo, Vanzani Paola, Santinello Sandro, Gucciardi Antonina, Zennaro Lucio, Miotto Giovanni, Ursini Fulvio
Distillerie Bonollo Umberto S.p.A., Nutraceutical Division, Mestrino, 35035 Padova, Italy.
Department of Molecular Medicine, University of Padova, 35129 Padova, Italy.
Antioxidants (Basel). 2021 Mar 9;10(3):418. doi: 10.3390/antiox10030418.
A "green" solvent-free industrial process (patent pending) is here described for a grape seed extract (GSE) preparation (Ecovitis™) obtained from selected seeds of Veneto region wineries, in the northeast of Italy, by water and selective tangential flow filtration at different porosity. Since a comprehensive, non-ambiguous characterization of GSE is still a difficult task, we resorted to using an integrated combination of gel permeation chromatography (GPC) and electrospray ionization high resolution mass spectrometry (ESI-HRMS). By calibration of retention time and spectroscopic quantification of catechin as chromophore, we succeeded in quantifying GPC polymers up to traces at = 30. The MS analysis carried out by the ESI-HRMS method by direct-infusion allows the detection of more than 70 species, at different polymerization and galloylation, up to = 13. This sensitivity took advantage of the nanoscale shotgun approach, although paying the limit of missed separation of stereoisomers. GPC and MS approaches were remarkably well cross-validated by overlapping results. This simple integrated analytical approach has been used for quality control of the production of Ecovitis™. The emerging feature of Ecovitis™ vs. a popular benchmark in the market, produced by a different technology, is the much lower content of species at low and the corresponding increase of species at high .
本文描述了一种“绿色”无溶剂工业生产工艺(专利正在申请中),用于制备葡萄籽提取物(GSE,即Ecovitis™),该提取物来自意大利东北部威尼托地区酒庄挑选的葡萄籽,通过水和不同孔隙率的选择性切向流过滤获得。由于对GSE进行全面、明确的表征仍然是一项艰巨的任务,我们采用了凝胶渗透色谱法(GPC)和电喷雾电离高分辨率质谱法(ESI-HRMS)相结合的综合方法。通过校准保留时间并以儿茶素作为发色团进行光谱定量,我们成功地对聚合度高达30的痕量GPC聚合物进行了定量。通过直接进样的ESI-HRMS方法进行的质谱分析能够检测到70多种不同聚合度和没食子酰化程度的物质,聚合度最高可达13。这种灵敏度得益于纳米级鸟枪法,尽管存在立体异构体分离不完全的局限性。GPC和MS方法的结果相互重叠,得到了非常好的交叉验证。这种简单的综合分析方法已用于Ecovitis™生产的质量控制。与市场上采用不同技术生产的一种流行基准产品相比,Ecovitis™的一个新特点是低聚合度物质的含量低得多,而高聚合度物质的含量相应增加。