Hollow-fiber flow field-flow fractionation for mass spectrometry: from proteins to whole bacteria

Mass spectrometry (MS) provides analyte identification over a wide molar-mass range. However, particularly in the case of complex matrices, this ability is often enhanced by the use of pre-MS separation steps. A separation, prototype technique for the “gentle” fractionation of large/ultralarge analytes, from proteins to whole cells, is here described to reduce complexity and maintain native characteristics of the sample before MS analysis. It is based on flow field-flow fractionation, and it employs a micro-volume fractionation channel made of a ca. 20 cm hollow-fiber membrane of sub-millimeter section. The key advantages of this technique lie in the low volume and low-cost of the channel, which makes it suitable to a disposable usage. Fractionation performance and instrumental simplicity make it an interesting methodology for in-batch or on-line pre-MS treatment of such samples.