Generation of ultrahigh peak capacity LC separations via elevated temperatures and high linear mobile-phase velocities.
The use of a combination of ultraperformance liquid chromatography at approximately 11,000 psi on sub 2-microm particles combined with reversed-phase gradient chromatography at a temperature of 90 degrees C is described as applied to the analysis of endogenous and drug metabolites in human and animal urine. By using elevated temperatures, back pressures can be reduced while maintaining high flow rates and chromatographic efficiency, with peaks 1-3 s wide at the base. Application to urine samples provided a peak capacity of approximately 700 for a 10-min analysis and greater than approximately 1000 in 1 h. Despite the narrow nature of the peaks, good quality mass spectra were also obtained, allowing the identification of typical drug and endogenous metabolites. These ultra-high-resolution chromatograms should be ideal for the analysis of complex samples in, for example, metabolite identification, impurity identification, and metabonomic/metabolomic studies. Applications in natural product analysis and proteomics can also be envisaged.
Plumb
Robert S
RS
Biological Chemistry, Division of Biomedical Sciences, Sir Alexander Fleming Building, Imperial College London, SW7 2AZ, London, UK. r.plumb@ic.ac.uk
Rainville
Paul
P
Smith
Brian W
BW
Johnson
Kelly A
KA
Castro-Perez
Jose
J
Wilson
Ian D
ID
Nicholson
Jeremy K
JK
D000082:Acetaminophen / Q000737:chemistry / Q000652:urine; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D020719:Glucuronides / Q000737:chemistry / Q000652:urine; D006801:Humans; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature; D014556:Urine / Q000737:chemistry
D020719:Glucuronides; D000082:Acetaminophen