Ion mobility spectrometry combined with ultra performance liquid chromatography/mass spectrometry for metabolic phenotyping of urine: Effects of column length, gradient duration and ion mobility spectrometry on metabolite detection

The need for rapid and efficient high throughput metabolic phenotyping (metabotyping) in metabolomic/metabonomic studies often requires compromises to be made between analytical speed and metabolome coverage. Here the effect of column length (150, 75 and 30 mm) and gradient duration (15, 7.5 and 3 min respectively) on the number of features detected when untargeted metabolic profiling of human urine using reversed-phase gradient ultra performance chromatography with, and without, ion mobility spectrometry, has been examined. As would be expected, reducing column length from 150 to 30 mm, and gradient duration, from 15 to 3 min, resulted in a reduction in peak capacity from 311 to 63 and a similar reduction in the number of features detected from over ca. 16,000 to ca. 6500. Under the same chromatographic conditions employing UPLC/IMS/MS to provide an additional orthogonal separation resulted in an increase in the number of MS features detected to nearly 20,000 and ca. 7500 for the 150 mm and the 30 mm columns respectively. Based on this limited study the potential of LC/IMS/MS as a tool for improving throughput and increasing metabolome coverage clearly merits further in depth study. [Display omitted] •Ion mobility spectrometry (IMS) significantly increased the number of analytes detected during the LC-MS of urine.•Nearly ca. 20,000 features were seen for urine using LC-IMS-MS in a 15 min analysis compared to ca. 16,000 by LC-MS alone.•In a 3 min analysis using a 30 mm column nearly 7600 features were detected with combined IMS and MS.•For high throughput analysis a 75 mm column and a 3 min analysis was a good compromise between speed and features detected.•The use of IMS also improved the quality of the mass spectra obtained.