Frequency division multiplex HPLC-MS for simultaneous analyses

Multiplex high-performance liquid chromatograph-mass spectrometry (HPLC-MS), in which multiple HPLCs and one MS are hyphenated, is an approach for high throughput analysis in HPLC-MS. A general multiplex HPLC-MS method employs a column-switching technology, and only one HPLC is connected to one MS at a time. In the present study, we propose a novel multiplex HPLC-MS system for simultaneous HPLC-MS analyses. In this study, multiple HPLCs are hyphenated with one MS without a column-switching mechanism, and a mixed-chromatogram is observed by the MS. Here, we employ a frequency division multiplexing (FDM) technique used in communication engineering to extract any chromatogram from the mixed-chromatogram. When a modulator (chopper or ion-gate type) is set between each ion source and the MS, each modulator blocks each sample stream with an individual frequency. In theory, each chromatogram can be extracted from the mixed-chromatogram via a signal processing based on a Fourier transform (FT), frequency-based signal extraction, and reversed FT. In the actual experiment, two HPLCs are hyphenated with one MS (2HPLC-1MS). The use of chopper type modulators leads to the extraction and restoration of each chromatogram from the mixed-chromatogram. However, each restored-chromatogram involves signal interference. On the other hand, the ion-gate modulation system successfully resulted in restored-chromatograms without interference. The potential of the novel multiplex HPLC-MS system based on FDM is confirmed with respect to the simultaneous and continuous analyses of plural samples.