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High resolution carotenoid separations of serum and natural products using UHPLC
Author(s) -
Craft Neal E.
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.640.6
Subject(s) - resolution (logic) , chromatography , carotenoid , chemistry , high resolution , elution , high performance liquid chromatography , selectivity , analytical chemistry (journal) , computer science , organic chemistry , remote sensing , food science , artificial intelligence , geology , catalysis
Carotenoids have been measured using HPLC with pressures up to 6000 psi for nearly 35 years. Recently, UHPLC, offering pressures up to 18,000 psi, has become readily available. These high pressures permit the use of <2um particle packings to produce very high efficiencies and rapid separations. Alternatively, UHPLC can be used to produce much higher resolution separations of carotenoids. Currently few sub‐2um column packings are available. Objective To compare several commercial columns and mobile phase combinations to determine the separation efficiency and selectivity using UHPLC and establish high resolution separations. Results A good HPLC separation with a 3um column produces about 20,000 theoretical plates. Multiple UHPLC columns were used in series to achieve unique selectivity and nearly 50,000 plates. The data from carotenoid standards was used to predict elution of other carotenoids and choose conditions for high resolution separations. Selected conditions were used to separate carotenoids in serum and natural products. Conclusions UHPLC was successfully used in high‐resolution mode to achieve separations previously unattainable with conventional HPLC.