Investigating biomarkers of exposure to jet aircraft oil fumes using mass spectrometry

Most commercial passenger jet aircraft use very hot compressed engine air, after cooling as a source for ventilation and cabin pressurization onboard. This design means that engine oil and/or hydraulic fluid can contaminate the ventilation supply air during otherwise normal flights, exposing onboard crewmembers and passengers to the fumes. The oils and hydraulic fluids contain a complex mixture of triaryl phosphates (TAPs) and decomposition products. Although the health and flight safety consequences of inhaling these fumes have been widely documented, measures of onboard inhalation exposure have been lacking. An approach is presented for documenting exposure to engine oil fumes by using high-resolution mass spectrometry (MS) to monitor and quantify post-translational modifications of subjects butyrylcholinesterase (BChE) that are consistent with exposure to the engine oil TAPs. We hypothesized that plasma from exposed individuals would show modifications or adducts of these OPs on the active site serine (Ser198) of BChE. Plasma BChE from 64 exposed subjects was purified to near homogeneity and concentrated using antibodies immobilized on paramagnetic beads. The purified BChE was eluted at low pH, digested with trypsin, and analyzed by liquid chromatography (LC)-MS. In subjects reporting onboard oil fume exposures, the most consistent adduct modifying the Ser198-containing tryptic peptide had a mass value of +154.0031 Da. The normalized peak area (NPA) of the +154Da modification was determined by comparing the relative MS1 intensities of the +154Da-modified Ser198 containing peptide to the total observable peptides containing the active site, including missed cleavages. Notably, adducts from in vitro exposures of bioactivated TAPs to purified BChE conducted in this study (i.e., 80Da, 156Da, 170Da, and 186Da) as well as adducts reported in other earlier in vitro studies (i.e., 65Da, 80Da, 91Da, 107Da, 16Da, 17Da, and 180Da) were not detected in exposed subjects. Of 67 subjects in this study, the average NPA of +154Da-Ser198 resulted from fume event exposures that pre-dated 2012 (N=54; 0.46-17.8, X ̅ =4.0) was 7.4X higher than control subjects. These data are uncorrected for the time lag between the reported exposure and the blood draw. Samples from the remaining 13 subjects with exposures from 2016-2024 showed only the 154Da modification at background levels (0.24-1.13; X ̅=0.53), as confirmed in control plasma samples from individuals who had not flown in at least three months. The observed reduction in the 154Da adduct over time in exposed individuals is likely a function of the change in the formulation of the OP blends added to engine oils during the course of the study. Further investigation into other protein biomarkers and adducts correlated with exposure to the current oil additives and hydraulic fluid fumes on aircraft is warranted. The most satisfactory solution would be to eliminate the exposure hazard by implementing bleed-free systems or, at a minimum, to develop less toxic oil formulations, suitable bleed air filters, and modified designs.