Abstract
The conventional approach to the measurement of peak capacity of reserved-phase columns under gradient elution assumes that all peaks have constant peak width, but this assumption can lead to inaccurate measurement of peak capacity. An integration approach employing a series of alkylphenones as model compounds was employed to more accurately measure the peak capacity for small molecule compounds under gradient elution. The base peak width of alkylphenones was plotted against retention time and a peak width function over retention time was established by polynomial regression. The peak capacity was then calculated by integrating the inverse of the peak width function over a gradient window. Compared to the conventional method, the integration method is not based on the assumption of equal peak width, thus providing a more accurate measurement of the peak capacity of reserved-phase columns, especially shorter ones packed with sub-2 μm particles under gradient elution.
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References
Snyder LR, Dolan JW (1998) In: Brown PR, Grushka E (eds) Advances in chromatography, Marcel Dekker, New York, 38:115–187
Okafo GN, Roberts JK (2003) In: Lee DC, Webb M (eds) Pharmaceutical analysis, Blackwell, Oxford, 31–48
Giddings JC (1967) Anal Chem 39:1027–1028 doi:10.1021/ac60252a025
Grushka E (1970) Anal Chem 42:1142–1147 doi:10.1021/ac60293a001
David JM, Giddings JC (1983) Anal Chem 55:418–424 doi:10.1021/ac00254a003
Giddings JC (1991) Unified Separation Science. Wiley, New York, 105–106
Dolan JW, Snyder LR, Djordjevic NM, Hill DW, Waeghe TJJ (1999) Chromatogr A 857:1–20 Medline doi:10 1016/S0021-9673(99)00765-7
Lan K, Jorgenson JW (1999) Anal Chem 71:709–714 Medline doi:10.1021/ac980702v
Minakuchi H, Ishizuka N, Nakanishi K, Soga N, Tanaka NJ (1998) Chromatogr A 828:83–90 doi:10.1016/S0021-9673(98)00605-0
Cheng Y, Lu Z, Neue U (2001) Rapid Commun Mass Spectrom 15:141–151 Medline doi:10.1002/1097-0231(20010130)15:2 ≤ 141::AID-RCM201 ≥ 3.0.CO;2-I
McCalley DVJ (2004) Chromatogr A 1038:77–84 Medline doi:10.1016/j.chroma.2004.03.038
Gilar M, Daly AE, Kele M, Neue UD, Gebler JCJ (2004) Chromatogr A 1061:183–192 Medline doi:10.1016/j.chroma.2004.10.092
Wren SACJ (2005) Pharm Biomed Anal 38:337–343 Medline doi:10.1016/j.jpba.2004.12.028
Shen Y, Zhang R, Moore R, Kim J, Metz TO, Hixson KK, Zhao R, Livesay EA, Udseth HR, Smith RD (2005) Anal Chem 77:3090–3100 Medline doi:10.1021/ac0483062
Wang X, Barber WE, Carr PWJ (2006) Chromatogr A 1107:139–151 Medline doi:10.1016/j.chroma.2005.12.050
Liu H, Finch JW, Lavallee MJ, Collamati RA, Benevides CC, Geber JCJ (2007) Chromatogr A 1147:30–36 Medline doi:10.1016/j.chroma.2007.02.016
Medina JC, Wu N, Lee ML (2001) Anal Chem 73:1301–1306 Medline doi:10.1021/ac001031f
Shen Y, Lee ML (1998) Anal Chem 70:3853–3856 doi:10.1021/ac9802426
Neue UD (2005) J Chromatogr A 1079:153–161 Medline doi:10.1016/j.chroma.2005.03.008
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The authors would like to thank ChemPharm Management at Johnson & Johnson Pharmaceutical Research and Development for providing financial support to Srinivasan Sriram through a co-op program.
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Guo, Y., Srinivasan, S., Gaiki, S. et al. Measuring Peak Capacity of Reversed-Phase Columns for Small Molecule Compounds Under Gradient Elution. Chroma 68, 19–25 (2008). https://doi.org/10.1365/s10337-008-0636-0
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DOI: https://doi.org/10.1365/s10337-008-0636-0