Assessing the detectability of antioxidants in two-dimensional high-performance liquid chromatography.
This paper explores the analytical figures of merit of two-dimensional high-performance liquid chromatography for the separation of antioxidant standards. The cumulative two-dimensional high-performance liquid chromatography peak area was calculated for 11 antioxidants by two different methods--the areas reported by the control software and by fitting the data with a Gaussian model; these methods were evaluated for precision and sensitivity. Both methods demonstrated excellent precision in regards to retention time in the second dimension (%RSD below 1.16%) and cumulative second dimension peak area (%RSD below 3.73% from the instrument software and 5.87% for the Gaussian method). Combining areas reported by the high-performance liquid chromatographic control software displayed superior limits of detection, in the order of 1 × 10(-6) M, almost an order of magnitude lower than the Gaussian method for some analytes. The introduction of the countergradient eliminated the strong solvent mismatch between dimensions, leading to a much improved peak shape and better detection limits for quantification.
Centre for Chemistry and Biotechnology, School of Life and Environmental Science, Deakin University, Geelong, Australia.
Danielle N
DN
Bassanese
Centre for Chemistry and Biotechnology, School of Life and Environmental Science, Deakin University, Geelong, Australia.
Xavier A
XA
Conlan
Centre for Chemistry and Biotechnology, School of Life and Environmental Science, Deakin University, Geelong, Australia.
Neil W
NW
Barnett
Centre for Chemistry and Biotechnology, School of Life and Environmental Science, Deakin University, Geelong, Australia.
Paul G
PG
Stevenson
Antioxidants; Countergradient; Limit of detection; Solvent strength mismatch; Two-dimensional high-performance liquid chromatography