There is no standard cleanliness evaluation procedure for reusable medical devices, only recommended guidelines. Our aim was to develop an efficient and reproducible method to recover proteins in greater amounts than conventional procedures. In addition to the solvents indicated in the guidelines, sample buffers (SB1 and SB2) containing the detergent sodium dodecyl sulfate (SDS) were tested. Protein recovery was compared using pseudo-blood-contaminated samples heat-treated under dry and wet conditions. Protein recovery from dry-heated samples did not change significantly regardless of extraction conditions, whereas the recovery rate from wet-heated samples using Milli-Q water or 1% SDS decreased proportionately to the increase in temperature. SB1 and SB2 extraction showed excellent protein recovery up to 95°C. In quantifying the protein using bicinchoninic acid (BCA), the interference of tris (2-carboxyethyl) phosphine in SB2, but not dithiothreitol in SB1, was quenched by pretreatment with iodoacetamide (IAM). After cleaning samples wet-heated at 95°C for 10 min with a washer-disinfector, the protein recovery rate per sample was 8.67 ± 3.0 µg using SB2 vs. 6.37 ± 2.5 µg using 1% SDS. These results indicate that residual proteins binding strongly to the surface of medical devices by thermal coagulation can be almost completely recovered and quantified by combining SB2 extraction, pretreatment with IAM, and BCA quantification.
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