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IROA TruQuant IQQ (CAT NO: WORKFLOW $200) 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.
MATERIALS:
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.
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 software and dictionary identifies/quantitates compounds in experimental samples.
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