IROA WORKFLOW (CAT NO: WORKFLOW) - why should I care?

The use of stable isotopes (as internal standards) is not a new thing.  However using a 95% U-13C internal standard brings a lot to the table. Especially if the internal standard is a complex standard such as metabolite yeast extract which contains 100s of primary metabolism metabolites.  Labeled peaks can be easily found using software algorithms and their unlabeled counterparts identified and quantitated.  What especially important and unique to this Workflow that by using 95% U-13C internal standards the software can correct for ion suppression and normalize the samples.  And the metabolites in these internal standards have been characterized by LC-MS and incorporated into the ClusterFinder software. Workflow and Software all for $200 (for 90 samples).

In short, the Workflow is a high-quality quantitation system that utilizes two (2) complex Internal Standards to make accurate measurements of several hundred metabolites simultaneously in experimental samples and measure day-to-day (QA/QC) platform reproducibility.

Materials

  1. IROA LONG-TERM REFERENCE STANDARD  IROA-LTRS (paired 1:1 mixture of labeled U-5% and – 95% U-13C metabolites): ID and QC – analyze separately to build a triply redundant dictionary (library) of RT, m/z and physical characteristics including fragmentation data and to account for fluctuations in mass and RT.
  2. IROA INTERNAL STANDARD – IROA-IS (labeled 95% U-13C metabolites): – spike into experimental samples and use as a yardstick in which to QUANTIFY all biochemicals in the sample relative to their counterparts in the IROA-LTRS. Even if analyzed on a different chromatographic system, results can be equated using dictionary because of the IROA-IS. Experimental compounds may have different RTs but will exhibit the same mass and physical characteristics.
  3. CLUSTERFINDER SOFTWARE

IROA TruQuant IQQ Workflow 3 step process

  1. LC-MS analysis of experimental samples with IROA-IS.  Inject IROA-LTRS periodically i.e. 10 sample intervals, as QC Standard to account for fluctuations in mass and retention time drift.
  2. Generate “dictionary” of RT, m/z, formula and physical characteristics from the analysis of IROA-LTRS using ClusterFinder software.
  3. ClusterFinder software and dictionary identifies/quantitates, suppression corrects and normalizes compounds in experimental samples.

Summary of Benefits of the IROA TruQuant (IQQ) Measurement System

  1. Daily instrument QA/QC.
  2. Economical – $200 for 90 samples
  3. No false data; removal of artifactual peaks.
  4. Accurate compound formula ID for MS alone; complete ID with addition of SWATH, or IM.
  5. All source ionization losses corrected for significantly better quantitation.
  6. Sample normalization to a universal standard for complete comparability.
  7. Internal standard metabolites have been characterized by several LC-MS methods and incorporated into ClusterFinder software.
  8. ClusterFinder software solution builds libraries, IDs/quantitates compounds, corrects for ion-suppression and normalizes data.
  9. Mass-spec vendor agnostic; specific extensions for inclusion of Sciex SWATH, and Agilent IM.

IROA TECHNOLOGIES provides aneconomical yeast extract workflow solution (only $2/sample) for metabolic profiling.

The QC Workflow kit contains dilute-and-shoot uniquely labeled yeast extract for use as an internal standard in quantitative MS-based quantitative profiling studies. Yeast is an ideal internal standard as it contains 100s of key primary metabolites including sugars, coenzymes, nucleotides, organic acids,  amino acids, peptides and polyamines.

Uses

  • Targeted or untargeted, LC-MS-based analysis
  • Ion suppression correction and sample normalization
  • Accurate metabolite ID and quantitation
  • Numerous sample matrices
  • Method and instrument QC
  • Biomarker discovery

Advantages of the unique 95% U-13C label

  • Automated through the use of ClusterFinder software
  • Labeled peaks easily found using software algorithms and their unlabeled counterparts in any sample matrix are identified and quantitated
  • 100s of metabolite peaks in the yeast extract have been well characterized by LC-MS-MS and incorporated into ClusterFinder
  • Improves metabolite identification
  • Through the use of the 95% U-13C internal standards ClusterFinder can correct for ion suppression and normalize samples
  • Built-in measurement of day-to-day (QA/QC) platform reproducibility

 

Here at IROA Technologies, our metabolite yeast extract QC Workflow Kit simplifies the analysis for our clients. We providevery cost-efficient internal metabolite standards as well as metabolite libraries to achieve reproducible results and make metabolomics easier.

We utilize the most advanced technology and internal measurement systems in an effort to reduce noise, variability, and ion suppression to deliver accurate and reproducible datasets.

Reading materials

User Manual

POSTER – Correction of Ion Suppression and Normalization for Improved Quantitative Rigor and Reproducibility using IROA

WHITEPAPER – Integration of Standards for Ion Suppression Correction and QC in an Untargeted Metabolomics Workflow