iroa patterns  (IROA Technologies LLC)

Journal: Nature Communications

Article Title: Ion suppression correction and normalization for non-targeted metabolomics

doi: 10.1038/s41467-025-56646-8

Figure Lengend Snippet: In this protocol, the experimental plasma samples ( a ) are prepared and dried and example spectra are shown ( a′ ). They are then reconstituted with a solvent containing the IROA-IS ( b ) and its example spectral pattern ( b′ ) to yield the analytical samples ( c ) and its spectral pattern ( c′ ). The analytical samples are randomized and injected within a sequence that starts and ends with injections of the IROA-LTRS ( d ) & and its spectral pattern ( d′ ), which is also injected approximately every 10 injections. In IROA MS/MS fragmentation, the IROA peaks retain their patterns ( d2′ ) because wide windows are used. Based on the presence of the IROA-IS, each sample can be suppression-corrected and normalized despite significant differences in sample input (original sample aliquot volume prior to dry down) ( e ). Source data are provided as a Source Data file. Data are shown as mean ± s.d. ( e ).

Article Snippet: IROA-LTRS (Fig. , red/yellow samples) is a 1:1 mixture of chemically equivalent IROA-IS standards at 95% 13 C and 5% 13 C. The combination produces the IROA-LTRS isotopic pattern illustrated in Fig. Metabolite 12 C and 13 C isotopologs co-elute, and the signature IROA peak pattern distinguishes real metabolites from artifacts, which lack the IROA pattern.

Techniques: Solvent, Injection, Sequencing, Tandem Mass Spectroscopy

Journal: Nature Communications

Article Title: Ion suppression correction and normalization for non-targeted metabolomics

doi: 10.1038/s41467-025-56646-8

Figure Lengend Snippet: a The IROA ion suppression correction workflow. b Number of MSTUS peaks detected across analytical conditions. c–k Raw MSTUS-12C (blue lines) and suppression-corrected MSTUS-12C (red lines) values are shown for: c HILIC positive mode, uncleaned source; d HILIC positive mode, clean source; e HILIC negative mode, uncleaned source; f HILIC negative mode, clean source; g RPLC positive mode, uncleaned source; h RPLC positive mode, clean source; ( i ) RPLC negative mode, uncleaned source; j RPLC negative mode, clean source; and k IC negative mode, cleaned source. l Ratio of raw MSTUS-12C to suppression-corrected MSTUS-12C peak intensity across chromatographic methods and experimental conditions. m Raw and suppression-corrected phenylalanine values in RPLC positive ionization mode with cleaned source. n Raw and suppression-corrected pyroglutamylglycine values in IC negative ionization mode. o Identified chemical composition in entire RPLC clean dataset, as an example. p Kohonen Self Organizing Maps (SOM) show suppression patterns in the RPLC Clean raw dataset for all 539 compounds. q Density map shows compounds associated with each of the patterns discovered in ( o ). r Raw MSTUS-12C (blue lines) and suppression-corrected MSTUS-12C (red lines) values are shown for RPLC positive mode and RPLC negative mode ( s ) for urine. t Ratio of raw MSTUS-12C to suppression-corrected 12 C peak intensity for urine matrix in positive and negative ion modes. u Plasma and urine MSTUS-12C signals for 4 common metabolites before and after suppression correction. 12C SC = 12C suppression corrected MSTUS; 13C raw = 13C raw MSTUS. Colors represent percent peak intensity as indicated by the color bar. Source data are provided as a Source Data file. Data are shown as mean ± s.d. ( c – n , r , s ).

Article Snippet: IROA-LTRS (Fig. , red/yellow samples) is a 1:1 mixture of chemically equivalent IROA-IS standards at 95% 13 C and 5% 13 C. The combination produces the IROA-LTRS isotopic pattern illustrated in Fig. Metabolite 12 C and 13 C isotopologs co-elute, and the signature IROA peak pattern distinguishes real metabolites from artifacts, which lack the IROA pattern.

Techniques: Hydrophilic Interaction Liquid Chromatography

Journal: Nature Communications

Article Title: Ion suppression correction and normalization for non-targeted metabolomics

doi: 10.1038/s41467-025-56646-8

Figure Lengend Snippet: a Optimization of cancer cell count for the IROA-IS ion suppression correction workflow. Raw MSTUS-12C (blue lines), suppression-corrected MSTUS-12C (red lines), and DUAL-MSTUS normalized (green lines) values are shown for: b RPLC positive mode, cleaned source; and c RPLC negative mode, clean source. d Optimization of IROA-IS during extraction and during reconstitution for ion suppression correction workflow. e Identified chemical composition in OVCAR-8 cell using IROA-IS in extraction solvent for entire RPLC clean dataset by both positive and negative mode. f Identified chemical composition in OVCAR-8 cell using IROA-IS as reconstitute solvent for entire RPLC clean dataset by both positive and negative mode. g Identified chemical composition in OVCAR-4 cell using IROA-IS in extraction solvent for entire RPLC clean dataset by both positive and negative mode. h Identified chemical composition in OVCAR-4 cell using IROA-IS as reconstitute solvent for entire RPLC clean dataset by both positive and negative mode. i Percent coefficient of variation (%CV) for raw, suppression-corrected, and normalized data from OVCAR-8 cell using IROA-IS in extraction solvent for entire RPLC clean dataset by both positive and negative mode. j Percent coefficient of variation (%CV) for raw, suppression-corrected, and normalized data from OVCAR-8 cell using IROA-IS as reconstitute solvent for entire RPLC clean dataset by both positive and negative mode. k Percent coefficient of variation (%CV) for raw, suppression-corrected, and normalized data from OVCAR-4 cell using IROA-IS in extraction solvent for entire RPLC clean dataset by both positive and negative mode. l Percent coefficient of variation (%CV) for raw, suppression-corrected, and normalized data from OVCAR-4 cell using IROA-IS as reconstitute solvent for entire RPLC clean dataset by both positive and negative mode. Percent coefficient of variation (%CV) for non-IROA-driven raw, and IROA-driven raw, suppression-corrected, and normalized data from OVCAR-4 ( m ) and OVCAR-8 ( n ) cell lines, respectively, for entire RPLC clean dataset by both positive and negative mode. Colors represent percent peak intensity as indicated by the color bar. Source data are provided as a Source Data file. Data are shown as mean ± s.d. ( b , c ).

Article Snippet: IROA-LTRS (Fig. , red/yellow samples) is a 1:1 mixture of chemically equivalent IROA-IS standards at 95% 13 C and 5% 13 C. The combination produces the IROA-LTRS isotopic pattern illustrated in Fig. Metabolite 12 C and 13 C isotopologs co-elute, and the signature IROA peak pattern distinguishes real metabolites from artifacts, which lack the IROA pattern.

Techniques: Cell Counting, Extraction, Solvent

Journal: Nature Communications

Article Title: Ion suppression correction and normalization for non-targeted metabolomics

doi: 10.1038/s41467-025-56646-8

Figure Lengend Snippet: a IROA-IS Workflows for demonstration of ovarian cancer cell metabolism with or without ASNase treatment. Heatmap demonstrate most altered metabolite networks includes amino acid and peptide metabolism using IROA-IS and no IROA-IS in OVCAR-8 ( b , c ) and OVCAR-4 cell ( d , e ), respectively. f Prospective mechanism of peptide elevation in ASNase resistant ovarian cancer cell (OVCAR-4). Colors represent percent peak intensity as indicated by the color bar.

Article Snippet: IROA-LTRS (Fig. , red/yellow samples) is a 1:1 mixture of chemically equivalent IROA-IS standards at 95% 13 C and 5% 13 C. The combination produces the IROA-LTRS isotopic pattern illustrated in Fig. Metabolite 12 C and 13 C isotopologs co-elute, and the signature IROA peak pattern distinguishes real metabolites from artifacts, which lack the IROA pattern.

Techniques: