Berbegal Carmen, Pardo Isabel, Ferrer Sergi
ENOLAB, ERI-ISIC BioTecMed/IViSoCa, Universitat de València, Burjassot, E46100, Valencia, Spain.
J Sci Food Agric. 2016 Mar 30;96(5):1556-61. doi: 10.1002/jsfa.7249. Epub 2015 Jun 30.
HPLC column technology has been improved, providing better resolution of closely eluting compounds, better analyte sensitivity, and shorter analysis times. The core-shell technology columns offer a faster analysis through the use of shorter columns without compromising resolution. The aim of this work was to improve the methods for determination of biogenic amines (BAs) in wine using the new HPLC PFP core-shell column technology.
Two different elution programs were designed to quantify BAs with the core-shell PFP column. Program I flow rate was 2 mL min(-1). The total elution time was 10 min. In elution program II, the flow rate was 0.8 mL min(-1) and the total elution time was 25 min. The two elution programs used with the core-shell PFP HPLC column showed differences related mainly to the histamine peak. The chromatograms showed that when a temporary isocratic elution was added in the gradient (program II), the histamine peak was eluted later, causing its isolation, and therefore its quantification was easier.
Compared to the previous C18 HPLC column for the BAs determination in wine, the main advantage of the presented technique is the reduction of the run times and solvent volumes, and has a better sensitivity and selectivity as peaks are higher and sharper.
高效液相色谱(HPLC)柱技术已经得到改进,能够更好地分离保留时间相近的化合物,提高分析物的灵敏度,并缩短分析时间。核壳技术柱通过使用更短的柱实现更快的分析,同时不影响分离度。本研究的目的是利用新型HPLC五氟苯基(PFP)核壳柱技术改进葡萄酒中生物胺(BAs)的测定方法。
设计了两种不同的洗脱程序用于使用核壳PFP柱对生物胺进行定量。程序I的流速为2 mL min⁻¹。总洗脱时间为10分钟。在洗脱程序II中,流速为0.8 mL min⁻¹,总洗脱时间为25分钟。与核壳PFP HPLC柱一起使用的这两种洗脱程序主要在组胺峰方面表现出差异。色谱图显示,当在梯度洗脱(程序II)中添加一段等度洗脱时,组胺峰洗脱时间更晚,从而实现了分离,因此其定量更容易。
与之前用于葡萄酒中生物胺测定的C18 HPLC柱相比,本技术的主要优点是运行时间和溶剂量减少,并且具有更好的灵敏度和选择性,因为峰更高且更尖锐。