IROA metabolomics workflow for improved accuracy, identification and quantitation

A method of quality assurance and/or quality control on the operational constancy of a mass spectral apparatus and associated ion mobility channel and chromatographic apparatus, when present, that comprises the steps of:

• a) carrying out one or more mass spectral analyses of experimental samples;
• b) carrying out a mass spectral analysis on multiple isotopic ratio outlier analysis (IROA) standard samples each of which is an aliquoted composition of biologically-produced metabolite compounds, each of said metabolite compounds having a molecular weight of about 2000 AMU or less, and each of said metabolite compounds being present as paired first and second isotopically-labeled compounds that are equally present at two predetermined balances for a non-hydrogen/deuterium atom present in said compounds, said first isotopically-labeled compounds containing about 2 to about 10% of a first isotope and about 90 to about 98% of a second isotope of said non-hydrogen/deuterium atom, and said second isotopically-labeled compounds containing about 90 to about 98% of said first isotope and about 2 to about 10% of said second isotope of the same non-hydrogen/deuterium atom, said first and second isotopes being stable to radioactive decay and other than hydrogen and deuterium, wherein mass spectral analysis of each IROA standard sample provides symmetrical patterns of peaks of said paired first and second isotopically-labeled compounds present that reflect i) the identity and number of peak pairs found, ii) their chromatographic retention time, iii) the amplitude of each peak pair and the sum of all peak pairs, iv) the resolution and sensitivity of the mass spectral analysis and v) allows the elimination of artifacts and noise during the course of carrying out analyses of both the standard and experimental samples,
• c) storing the values determined from the standard sample analyses to determine trends in instrumentation, and
• d) evaluating the mass spectral analyses of step b) to determine the existence of differing results, and if found, using those differing results to correct for variances in measured aspects in the results of the experimental samples.