Abstract
Carotenoids contained in plant extracts were analyzed using liquid chromatography/positive-and negative-ion atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS) with a narrow-bore C 30 reversed-phase high-performance liquid chromatographic (HPLC) column and a gradient solvent system containing methanol-methyl tert-butyl ether-ammonium acetate at a flow rate of 300 μl min -1 . In addition to mass spectrometric detection, photodiode-array UV/visible absorbance detection was used between the HPLC column and mass spectrometer for additional carotenoid characterization. Positive-ion APCI produced protonated molecules and molecular ions for both xanthophylls and, unexpectedly, hydrocarbon carotenes ; and during negative-ion APCI, M -. and [M - H] - ions were observed. In order to investigate the origin of the unexpected [M + H] + ions, positive-ion APCI of β-carotene was investigated using deuterochloroform as the only solvent. Because β-carotene formed primarily deuterated ions, [M + D] + , during APCI in deuterochloroform, the mobile phase was determined to be the source of hydrogen for protonation. The hydroxylated xanthophyll lutein fragmented during positive-ion APCI to eliminate water from the protonated molecule and form the base peak of m/z 551. Using collision-induced dissociation in the ion source, additional fragmentation pathways characteristic of tandem mass spectra of carotenoids were observed such as retro-Diels-Alder fragmentation, [M - 56] +. , for α-carotene and loss of toluene from the molecular ion, [M-92] +. for lutein, α-and β-carotene. The limits of detection for protonated molecules of α-carotene and lutein were approximately 3 and 13 pmol, respectively. In negative-ion APCI, the limits of detection were approximately 3 and 1 pmol for M -1 ions of α-carotene and lutein, respectively.
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