Departament de Química, Universitat de Lleida, 25198 Lleida, Spain.
J Chromatogr A. 2012 Feb 10;1224:1-10. doi: 10.1016/j.chroma.2011.12.025. Epub 2011 Dec 13.
High-performance liquid chromatography (HPLC) has become the method of choice for carotenoid analysis. Although a number of normal-phase columns have been reported, reverse-phase columns are the most widely used stationary phases for the analysis of these molecules. C18 and C30 stationary phases have provided good resolution for the separation of geometrical isomers and carotenoids with similar polarity. More recently ultra high-performance liquid chromatography (UHPLC) has been used. UHPLC has a number of distinct advantages over conventional HPLC. These include: faster analyses (due to shorter retention times), narrower peaks (giving increased signal-to-noise ratio) and higher sensitivity. High strength silica (HSS) T3 and C18 and ethylene bridged hybrid (BEH) C18 stationary phases, with sub-2 μm particles have been used successfully for UHPLC analysis and separation of carotenoids. A number of spectroscopic and mass spectrometric techniques have also been used for carotenoid qualitative and quantitative analysis. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF-MS), atmospheric-pressure solids-analysis probe (ASAP) and Raman spectroscopy are used to profile rapidly and qualitative carotenoids present in different crude extracts. Such detection methods can be used directly for the analysis of samples without the need for sample preparation or chromatographic separation. Consequently, they allow for a fast screen for the detection of multiple analytes. Quantitative carotenoid analysis can be carried out using absorbance or mass detectors. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is efficient for carotenoid identification through the use of transitions for the detection of analytes through precursor and daughter ions. This approach is suitable for the identification of carotenoids with the same molecular mass but different fragmentation patterns. Here we review critically the latest improvements for carotenoid resolution and detection and we discuss a number of analytical techniques for qualitative and quantitative analysis of carotenoids.
高效液相色谱（HPLC）已成为类胡萝卜素分析的首选方法。尽管已经报道了许多正相柱，但反相柱是分析这些分子最广泛使用的固定相。C18 和 C30 固定相为分离几何异构体和极性相似的类胡萝卜素提供了良好的分辨率。最近，超高效液相色谱（UHPLC）已被用于分析。UHPLC 相对于传统 HPLC 具有许多明显的优势。这些优势包括：更快的分析（由于保留时间较短）、更窄的峰（增加信噪比）和更高的灵敏度。高强度硅胶（HSS）T3 和 C18 以及乙烯桥联混合（BEH）C18 固定相，使用亚 2μm 颗粒已成功用于 UHPLC 分析和类胡萝卜素的分离。许多光谱和质谱技术也被用于类胡萝卜素的定性和定量分析。基质辅助激光解吸电离飞行时间质谱（MALDI/TOF-MS）、常压固体分析探头（ASAP）和拉曼光谱用于快速定性和定性分析不同粗提物中存在的类胡萝卜素。这些检测方法可以直接用于分析样品，而无需样品制备或色谱分离。因此，它们允许快速筛选检测多种分析物。可以使用吸光度或质量检测器进行定量类胡萝卜素分析。液相色谱-串联质谱（LC-MS/MS）通过使用用于检测分析物的前体离子和子离子的跃迁，可有效地用于类胡萝卜素的鉴定。这种方法适用于鉴定具有相同分子量但不同碎片模式的类胡萝卜素。在这里，我们批判性地回顾了类胡萝卜素分辨率和检测的最新进展，并讨论了几种定性和定量分析类胡萝卜素的分析技术。
