Within-day reproducibility of an HPLC-MS-based method for metabonomic analysis: application to human urine.
Self-evidently, research in areas supporting "systems biology" such as genomics, proteomics, and metabonomics are critically dependent on the generation of sound analytical data. Metabolic phenotyping using LC-MS-based methods is currently at a relatively early stage of development, and approaches to ensure data quality are still developing. As part of studies on the application of LC-MS in metabonomics, the within-day reproducibility of LC-MS, with both positive and negative electrospray ionization (ESI), has been investigated using a standard "quality control" (QC) sample. The results showed that the first few injections on the system were not representative, and should be discarded, and that reproducibility was critically dependent on signal intensity. On the basis of these findings, an analytical protocol for the metabonomic analysis of human urine has been developed with proposed acceptance criteria based on a step-by-step assessment of the data. Short-term sample stability for human urine was also assessed. Samples were stable for at least 20 h at 4 degrees C in the autosampler while queuing for analysis. Samples stored at either -20 or -80 degrees C for up to 1 month were indistinguishable on subsequent LC-MS analysis. Overall, by careful monitoring of the QC data, it is possible to demonstrate that the "within-day" reproducibility of LC-MS is sufficient to ensure data quality in global metabolic profiling applications.
Gika
Helen G
HG
AstraZeneca, Department of Drug Metabolism and Pharmacokinetics, Mereside, Alderley Park, Macclesfield, Cheshire SK10 4TG, United Kingdom.
Theodoridis
Georgios A
GA
Wingate
Julia E
JE
Wilson
Ian D
ID
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D020543:Proteome / Q000378:metabolism*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014556:Urine / Q000737:chemistry*
D020543:Proteome