Advantages of the IROA Protocol

The IROA technology directly addresses the majority of metabolomic bottlenecks:

  • Variability is removed. In a typical IROA experiment, the same cell pool is used to create both control and experimental samples. This removes the biological variability.
  • Removes sample-to-sample variance, fewer samples are required. The Control and Experimental samples are run as a single composite sample, reducing the number of analytical samples in half and removing all analytical variance.
  • Overcomes suppression. Identical molecules are labeled with either C12 or C13 media and during MS analysis experience the same ionization efficiency and suppression.
  • Artifacts and noise are removed resulting in greater clarity and accuracy. Molecules from either biological system (Experimental or Control) have a unique isotopic signature; therefore all unsigned molecules can be classified as artifacts or noise. This means all artifactual signals can be removed from the dataset (usually the largest portion of the dataset), generally not possible in current metabolomics datasets where the high number of "observations" increases the false discovery rate.
  • Compound identification of unknowns. The IROA technology enables compound identification. Because samples are isotopically-labeled, a molecular formula for each biochemical compound can be calculated using accurate mass and the number of compounds and the compound readily identified. Generally, these compounds would remain unnamed.
  • Quantitative accuracy. The IROA technology uses a central point of reference (biochemical) embedded in every sample thereby enabling measurements of samples with quantitative exactness.
  • Amenable to a completely automated solution (via software). The IROA ClusterFinder software is capable of interpreting these datasets, and will allow for a very dramatic reduction in data size as it easily sorts through the dataset removing irrelevant data, identifying and quantitating molecules of interest.
  • Very broadly applicable. The IROA experimental system can be used to test many stressing agents including chemical, genetic, environmental or any element or combination of elements that induces physiological alteration.
  • Saves time, effort and related costs. Smaller, simpler experiments are now possible since effectively there is an internal standard for every compound measured so the number of replicates required for each experiment is considerably reduced. IROA "makes metabolic profiling easy".