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This study investigated approaches for the profiling of coffee using two multidimensional approaches: (1) a multi-detection process and (2) a multi-separation process employing HPLC. The first approach compared multidetection techniques of conventional High Performance Liquid Chromatography (HPLC) hyphenated with a detector (DPPH•, UV-Vis and MS), and multiplexed mode via HPLC with an Active Flow Technology (AFT) column in Parallel Segmented Flow (PSF) format with DPPH• detection, UV-Vis and MS running simultaneously. Multiplexed HPLCPSF enabled the determination of key chemical entities by reducing the data complexity of the sample whilst obtaining a greater degree of molecule-specific information within a fraction of the time it takes using conventional multi-detection processes. DPPH•, UV-Vis and MS (TIC) were multiplexed for the analysis of espresso coffee and decaffeinated espresso coffee. Up to 20 DPPH• peaks were detected for each sample, and with direct retention time peak matching, 70% of DPPH• peaks gave a UV-Vis response for the espresso coffee and 95% for the decaffeinated espresso coffee. The second approach involved the use of a two-dimensional (2D) HPLC system to expand the separation space and separation power for the analysis of coffee, focusing on the resolution and detection of coeluting and overlapping peaks, which was beyond the limits of conventional HPLC in resolving complex samples. The 2DHPLC analysis resulted with the detection of 176 peaks and a closer observation showed the presence of an additional 17 peaks in a cut section where in 1D mode only one peak was observed.
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