Identification of a photooxygenation product of chlortetracycline in hog lagoons using LC/ESI-ion trap-MS and LC/ESI-time-of-flight-MS

In the present work, a photoproduct of chlortetracycline (CTC) was identified for the first time in hog lagoon samples from confined animal feeding operations. Screening of several samples by LC/ESI-MS indicated the presence of a potential photooxygenation product of CTC with a nominal mass that was 32 Da higher than the parent drug. Generation of this assumed photoproduct (designated M510) was achieved in a 24-h irradiation experiment of an oxygenated alkaline medium containing 50 mg L-1 CTC. Accurate mass measurements with an ESI-TOF-MS of the protonated isomerization product of CTC (iCTC) and the postulated photooxygenation product, bearing two oxygen atoms more than iCTC, were m/z 479.1229 and 511.1109, respectively. These corresponded to errors of -2.8 ppm for iCTC and +1.0 ppm for M510 relative to the theoretical masses. The generation of second- and third-stage mass spectra in an ESI-ion trap-MS showed similar characteristic fragmentation patterns for iCTC and the photoproduct M510, leading to the conclusion that the M510 structure consisted of an iCTC-like skeleton bearing two additional hydroxy groups. The site and configuration of one hydroxylation was confidently assigned, while the position of the other hydroxy group was tentatively assigned. Comparison of the(+)-ESI-MS3 spectrum and the retention time of M510 in the sample from the irradiation experiment with those from the hog lagoon samples yielded excellent agreement, suggesting that the compounds were identical. Quantitative analysis of seven hog lagoon samples showed iCTC concentrations of 1910-15 800 mug L-1, while the levels of M510 were found to be between 46 and 303 mug L-1.