title: A fast and sensitive method for the separation of carotenoids using ultra-high performance supercritical fluid chromatography-mass spectrometry.
abstract: In this study, a rapid and sensitive ultra-high performance supercritical fluid chromatography-mass spectrometry (UHPSFC-MS) method has been developed and partially validated for the separation of carotenoids within less than 6 min. Six columns of orthogonal selectivity were examined, and the best separation was obtained by using a 1-aminoanthracene (1-AA) column. The length of polyene chain as well as the number of hydroxyl groups in the structure of the studied carotenoids determines their differences in the physiochemical properties and thus the separation that is achieved on this column. All of the investigated carotenoids were baseline separated with resolution values greater than 1.5. The effects of gradient program, back pressure, and column temperature were studied with respect to chromatographic properties such as retention and selectivity. Electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) were compared in both positive and negative mode, using both direct infusion and hyphenated with UHPSFC. The ESI in positive mode provided the highest response. The coefficient of determination (R (2)) for all calibration curves were greater than 0.998. Limit of detection (LOD) was in the range of 2.6 and 25.2 ng/mL for α-carotene and astaxanthin, respectively, whereas limit of quantification (LOQ) was in the range of 7.8 and 58.0 ng/mL for α-carotene and astaxanthin, respectively. Repeatability and intermediate precision of the developed UHPSFC-MS method were determined and found to be RSD < 3 % and RSD < 6 %, respectively. The method was applied in order to determine carotenoids in supercritical fluid extracts of microalgae and rosehip. Graphical Abstract Ultra-high performance supercritical fluid chromatography-a rapid separation method for the analysis of carotenoids in rosehip and microalgae samples.
authors.affiliation: Centre for Analysis and Synthesis, Department of Chemistry, Lund University, P.O. Box 124, 22100, Lund, Sweden.
authors.forename: Firas
authors.initials: F
authors.lastname: Jumaah
authors.affiliation: Centre for Analysis and Synthesis, Department of Chemistry, Lund University, P.O. Box 124, 22100, Lund, Sweden.
authors.forename: Merichel
authors.initials: M
authors.lastname: Plaza
authors.affiliation: Centre for Analysis and Synthesis, Department of Chemistry, Lund University, P.O. Box 124, 22100, Lund, Sweden.
authors.forename: Victor
authors.initials: V
authors.lastname: Abrahamsson
authors.affiliation: Centre for Analysis and Synthesis, Department of Chemistry, Lund University, P.O. Box 124, 22100, Lund, Sweden.
authors.forename: Charlotta
authors.initials: C
authors.lastname: Turner
authors.affiliation: Centre for Analysis and Synthesis, Department of Chemistry, Lund University, P.O. Box 124, 22100, Lund, Sweden. margareta.sandahl@chem.lu.se.
authors.forename: Margareta
authors.initials: M
authors.lastname: Sandahl
mesh_terms: D002338:Carotenoids / Q000032:analysis* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D025924:Chromatography, Supercritical Fluid / Q000379:methods*; D057230:Limit of Detection; D058086:Microalgae / Q000737:chemistry; D027826:Rosa / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D024341:Xanthophylls / Q000032:analysis / Q000302:isolation & purification
chemical_list: D024341:Xanthophylls; D002338:Carotenoids; C041635:alpha-carotene; C005948:astaxanthine
keywords: Carotene; Mass spectrometry; Method development; Orthogonal column screening; Supercritical fluid; Xanthophyll