Effect of first-dimension undersampling on effective peak capacity in comprehensive two-dimensional separations

Anal Chem. 2008 Jan 15;80(2):461-73. doi: 10.1021/ac071504j. Epub 2007 Dec 13.

Abstract

The objective of this work is to establish a means of correcting the theoretical maximum peak capacity of comprehensive two-dimensional (2D) separations to account for the deleterious effect of undersampling first-dimension peaks. Simulations of comprehensive 2D separations of hundreds of randomly distributed sample constituents were carried out, and 2D statistical overlap theory was used to calculate an effective first-dimension peak width based on the number of observed peaks in the simulated separations. The distinguishing feature of this work is the determination of the effective first-dimension peak width using the number of observed peaks in the entire 2D separation as the defining metric of performance. We find that the ratio of the average effective first-dimension peak width after sampling to its width prior to sampling (defined as <beta>) is a simple function of the ratio of the first-dimension sampling time (t(s)) to the first-dimension peak standard deviation prior to sampling (1sigma): <beta> = square root1+0.21(t /(s)(1) sigma(2) This is valid for 2D separations of constituents having either randomly distributed or weakly correlated retention times, over the range of 0.2 </= t(s)/1 sigma < or = 16. The dependence of <beta> on t(s)/1 sigma from this expression is in qualitative agreement with previous work based on the effect of undersampling on the effective width of a single first-dimension peak, but predicts up to 35% more broadening of first-dimension peaks than is predicted by previous models. This simple expression and accurate estimation of the effect of undersampling first-dimension peaks should be very useful in making realistic corrections to theoretical 2D peak capacities, and in guiding the optimization of 2D separations.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Chromatography / methods*
  • Chromatography, Gas
  • Chromatography, Liquid
  • Models, Statistical
  • Monte Carlo Method
  • Normal Distribution