IROA TruQuant/IQQ is a high-quality quantitation system that utilizes two (2) complex Internal Standards to make accurate biological measurements on several hundred biochemicals simultaneously in small quantities of biological samples and measure day-to-day (QA/QC) platform reproducibility.
 
(1) A spiked in Internal Standard (IROA-IS) is used to:  a) quantify 100's of biochemicals in a sample;  b) suppression-correct each compound and c) normalize sample to sample variances. (2) A Long-Term Reference Standard (IROA-LTRS) is analyzed every 10 samples to create a daily RT library of all compounds to be found in the IROA-IS for reproducible ID and to measure day-to-day instrument performance (QA/QC). 
 
The system is completely automated using IROA ClusterFinder software.
 
IROA TruQuant IQQ Workflow 3 STEP PROCESS:
(1) LC-MS analysis of experimental samples spiked 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 and dictionary identifies/quantitates compounds in experimental samples.

IROA LONG-TERM REFERENCE STANDARD  IROA-LTRS (paired 1:1 mixture of labeled U-5% and - 95% U-13C metabolites): ID and QC - used 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.

IROA INTERNAL STANDARD – IROA-IS (labeled 95% U-13C metabolites): - spiked into experimental samples and used as a yardstick in which to QUANTIFY all biochemicals in the sample relative to their counterparts in the IROA-IS. Even if analyzed on a different chromatographic system, results can be equated using dictionary because of IROA-IS. Experimental compounds may have different RTs but will exhibit the same mass and physical characteristics.

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

  • Cost effective simultaneous metabolite identification and quantitation
  • Spiked-in standard containing 100’s of identified compounds allows accurate compound formula ID for MS alone; complete ID with addition of SWATH, or IM, even at low concentrations
  • Unique IROA patterns discriminate peaks of biological origin from artefactual peaks allowing the removal of false data
  • All IROA-based fragments will have the IROA ratio pattern of their parent peaks and can be identified as such using the “peak correlation” ClusterFinder module
  • Alignment of all peaks in the chromatogram using IROA-IS generated Retention Time (RT) ladder
  • Retention Index within the sample allows “unknowns” to be tracked with accuracy and precision
  • Suppression-corrected measurements for significantly better quantitation
  • Normalizes (suppression-corrected) experimental samples to overcome day-to-day, instrument-to-instrument variances
  • ClusterFinder software solution builds libraries, IDs/quantitates compounds and normalizes data
  • Ensures high level QC for accurate and reproducible results

   

For background reading:  WHITEPAPER - Challenges in Metabolomics Analysis (and a solution)

TAKE THE IROA TruQuant IQQ CHALLENGE:  Generate a Calibration curve using the IROA IQQ Workflow and get a free kit!