Characterization of the pigment fraction in sweet bell peppers (Capsicum annuum L.) harvested at green and overripe yellow and red stages by offline multidimensional convergence chromatography/liquid chromatography–mass spectrometry

Abstract

Offline multidimensional supercritical fluid chromatography combined with reversed-phase liquid chromatography was employed for the carotenoid and chlorophyll characterization in different sweet bell peppers (Capsicum annuum L.) for the first time. The first dimension consisted of an Acquity HSS C18 SB (100 × 3 mm id, 1.8 μm particles) column operated with a supercritical mobile phase in an ultra-performance convergence chromatography system, whereas the second dimension was performed in reversed-phase mode with a C30 (250 × 4.6 mm id, 3.0 μm particles) stationary phase combined with photodiode array and mass spectrometry detection. This approach allowed the determination of 115 different compounds belonging to chlorophylls, free xanthophylls, free carotenes, xanthophyll monoesters, and xanthophyll diesters, and proved to be a significant improvement in the pigments determination compared to the conventional one-dimensional liquid chromatography approach so far applied to the carotenoid analysis in the studied species. Moreover, the present study also aimed to investigate and to compare the carotenoid stability and composition in overripe yellow and red bell peppers collected directly from the plant, thus also evaluating whether biochemical changes are linked to carotenoid degradation in the nonclimacteric investigated fruits, for the first time.

The genusCapsicum: a phytochemical review of bioactive secondary metabolites

“…As ripening goes on, there is two possible scenarios in the yellow fruits, the accumulation of lutein and maintenance of b-carotene or the lowering of the levels of both previously cited carotenoids concomitantly with the increase in violaxanthin and a-carotene. 139,143,144,150,151 Furthermore, violaxanthin and lutein are described as the main carotenoids in ripe yellow-fruited species. 152 In species in which violaxanthin is the major carotenoid in ripe fruit, it comprises approximately 30-50% of the total content, 139 while lutein can comprises 41-67%.…”

“…The major compounds from capsaicinoids, carotenoids and avonoids classes described during this review were used as variables to perform a Principal Component Analysis (PCA) of samples analyzed by other researchers. 64,78,127,150,156,158,160,162,176,[201][202][203][204][205] Before the PCA analysis, data were normalized to reduce the inuence of different analytical methods applied by each researcher (see ESI material †).…”

Supercritical Fluid Chromatography in Natural Product Analysis – An Update

“…Since then, a large number of papers was published, which emphasizes the important role of SFC as an alternative analysis technique in this sector [36]. There are different forms of carotenoids: free carotenoids and more stable forms, esterified with fatty acids [39]. To release all esters and to simplify the analysis, most of the investigations were performed after a saponification step [40].…”

“…Bonaccorsi et al [39] identified more than 100 different compounds belonging to chlorophylls, free carotenes, free xanthophylls, and xanthophyll mono-and diesters in sweet bell peppers through the offline coupling of SFC and LC. The first dimension was performed on a SFC system using an Acquity UPC² HSS C 18 SB column and ethanol as modifier, while the second dimension consisted of a RP-HPLC combined with DAD and MS detection and was performed on a C30 stationary phase.…”

A new strategy of studying protein–protein interactions: Integrated strong anion exchange/reversed‐phase chromatography/immunoprecipitation coupled with mass spectrometry for large‐scale identification of proteins interact with immunoglobulin G in HeLa cells

“…Multidimensional chromatography provides enhanced resolution and separation power and an extended measured dynamic concentration range to analyze complex proteomics samples [13]. Moreover, this advantageous method offers a convenient online proteomics approach to reduce the complexity of the proteomic samples encountered during fractionation.…”