Ultra high pressure in the second dimension of a comprehensive two-dimensional liquid chromatographic system for carotenoid separation in red chili peppers.
A comprehensive normal-phase × reversed-phase (NP-LC × RP-LC) liquid chromatography system was developed, and applied for analysis of the intact carotenoid composition of red chili peppers, with photodiode array and mass spectrometry detection. A micro-bore cyano column (250 mm × 1.0 mm, 5 μm d.p.) was chosen for the first dimension ((1)D) separation, interfaced to a second dimension ((2)D) C18 column (30 mm × 4.6 mm, 2.7 μm d.p.) packed with fused-core particles. Subsequently, two columns of the same stationary phase were coupled serially for second dimension separation, and operated under ultra high pressure LC conditions (UHPLC), within a cycle time of 1.50 or 1.00 min, and equal modulation times. Performances of the three different set-ups were evaluated, in terms of peak capacity values (n(c)), and afterwards corrected by taking into account both the under-sampling, and the orthogonality effects. After these adjustments, the peak capacity values were estimated as follows: n(c) 526, for the NP-LC × RP-LC system, n(c) 373, for the NP-LC × RP-UHPLC system with a 1.50 min modulation time, n(c) 639, for the NP-LC × RP-UHPLC system, with a 1.00 min modulation time. A total of 33 compounds were separated into 10 different chemical classes in the two-dimensional space, and identified by accurate IT-TOF (ion trap-time of flight) MS detection.
Chromaleont s.r.l. A spin-off of the University of Messina, c/o Dipartimento Farmaco-chimico, University of Messina, viale Annunziata, 98168 Messina, Italy.
Francesco
F
Cacciola
Paola
P
Donato
Daniele
D
Giuffrida
Germana
G
Torre
Paola
P
Dugo
Luigi
L
Mondello
D002212:Capsicum / Q000737:chemistry*; D002338:Carotenoids / Q000737:chemistry / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods
D002338:Carotenoids