Discovery-based analysis and quantification for comprehensive three-dimensional gas chromatography flame ionization detection data

Abstract Basic principles are introduced for implementing discovery-based analysis with automated quantification of data obtained using comprehensive three-dimensional gas chromatography with flame ionization detection (GC3-FID). The GC3-FID instrument employs dynamic pressure gradient modulation, providing full modulation (100% duty cycle) with a fast modulation period (PM) of 100 ms. Specifically, tile-based Fisher-ratio analysis, previously developed for comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC×GC-TOFMS), is adapted and applied for GC3-FID where the third chromatographic dimension (3D) is treated as the “spectral” dimension. To evaluate the instrumental platform and software implementation, ten “non-native” compounds were spiked into a ninety-component base mixture to create two classes with a concentration ratio of two for the spiked analyte compounds. The Fisher ratio software identified 95 locations of potential interest (i.e., hits), with all ten spiked analytes discovered within the top fourteen hits. All 95 hits were quantified by a novel signal ratio (S-ratio) algorithm portion of the F-ratio software, which determines the time-dependent S-ratio of the 3D chromatograms from one class to another, thus providing relative quantification. The average S-ratio for spiked analytes was 1.94 ± 0.14 mean absolute error (close to the nominal concentration ratio of two), and 1.06 ± 0.16 mean absolute error for unspiked (i.e., matrix) components. The appearance of the S-ratio as a function of 3D retention time in the GC3 dataset, referred to as an S-ratiogram, provides indication of peak purity for each hit. The unique shape of the S-ratiogram for hit 1, α-pinene, suggested likely 3D overlap. Parallel factor analysis (PARAFAC) decomposition of the hit location confirmed that overlap was occurring and successfully decomposed α-pinene from a highly overlapped (3Rs = 0.1) matrix interferent.