A multifunctional microfluidic droplet-array chip for analysis by electrospray ionization mass spectrometry

This paper describes a multifunctional semi-closed droplet-array chip coupled with electrospray ionization mass spectrometry (ESI-MS) detection for multiple sample pretreatment and analysis. A novel interfacing method for coupling droplet system with ESI-MS was proposed using a sampling probe-two-dimensional (2D) droplet-array strategy. The 2D droplet-array system was composed of an 8 x 8 microwell array chip for droplet storage and a layer of oil covering the droplets served as a "virtual wall" to avoid droplet evaporation or cross-contamination. An L-shaped capillary was adopted as the interface of the droplet array and ESI-MS, using its inlet end as a sampling probe for droplets and its outlet with a tip size of similar to 20 mm as an electrospray emitter, without the need for any droplet extraction device. The droplet analysis was performed by moving the droplet-array chip to allow the capillary sampling probe to sequentially enter into the droplets through the oil and introduce the sample solution into the capillary emitter for MS detection. The MS analysis time for each droplet sample was 40 s with a sample consumption of ca. 13 nL. A good repeatability of 5.7% (RSD, n = 9) was obtained for 10(-6) M reserpine droplet analysis. The uses of the semi-closed 2D droplet array and off-line interfacing mode provide the system with the substantial flexibility and controllability in droplet indexing, multi-step manipulating, and on-demand sampling for MS analysis. We applied the present system in multi-step pretreatment and identification of small amounts of proteomic samples of myoglobin and cytochrome C, including in-droplet protein reduction, alkylation, digestion, and purification based on solid-phase extraction, matrix modification, sample droplet introduction under flow injection mode, and ESI-MS detection.