Fig. 3.

The benefits of integrating protein extraction with protein digestion. The final protocol for Mitra microsamplers with dried blood includes denaturation and reduction for one hour at 60 °C and 1200 RPM and digestion with trypsin for six hours at 37 °C and 1000 RPM and (1) improves recovery, as compared to VAMS samplers that are removed after denaturation and reduction (orange bars, n = 4) and (2) eliminates the need for longer or more rigorous incubation (grey bars, include two hours instead of one hour denaturation and reduction, n = 3) or sonication (blue bars, include one hour sonication and two hours denaturation and reduction, n = 3). Error bars indicate the standard deviation. All relative response ratios are normalized to the relative response ratio of the final protocol, indicated by the dashed line at y = 1.0. The inlets below the legend show the final color of the Mitra tips after the different protocols.