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Simultaneously characterizing and quantifying chloramphenicol and its metabolite using LC/MS/MS

r that the peak one is CAP-glucuronide, and peak two is the parent CAP in Figure 1. Single mass spectrum of CAP shows [M-H]- pair m/z 321 and 323 (Figure 4). Product ion spectrum (Figure 5) shows two predominant ions m/z 121 and 152, which correspond to [C6H4NO2-H]- and [C6H4NO2+CH2O]-. Ions m/z 194 [m/z 321-C2H2NOCl2]-, 176 [m/z 194-(H2O)]-, 219 [m/z 321-(HCOCl2)-2H]-, 249 [m/z 321-HCl2]-, and 257/259 [m/z 321-(HCOCl)]- also present in the spectrum (Figure 5). The product ion spectrum shows the isotopic ion pair of m/z 257 and 259, which gives approximately equal abundance, indicating the loss of one chlorine atom (either 35Cl or 37Cl) from the 35Cl 37Cl parent ion. The single MS spectrum (Figure 2.) of CAP metabolite and its product ion spectrum (Figure 3.) stated that the metabolite is glucuronidation of CAP. Figure 4. Single MS spectrum of peak 2 at t = 4.31 minutes from Figure 1. Figure 5. Product ion spectrum of peak 2 (M/z 323) at t = 4.31 minutes from Figure 1. Figure 6. MRM quantitation chromatogram of the same analysis run shown in Figure 1. Quantitative Confirmation: The MRM mode quantitation of CAP has been obtained within the same run with the qualitative information collections. The results show in Table 1 with an example chromatogram (t=60 minutes) shows in Figure 6. It is a fast run (CAP elution time t = 4.31minutes). T1/2 was calculated to be 46 minutes and its plot shows in Figure 7.

Conclusions

As stated previously, the advantages for this method are fast run time (about 5 minutes for a entire run) and simultaneously doing qualitative and quantitative work together. The instrument is based on a quadrupole ion path and is capable of doing conventional multiple reaction monitoring (MRM) mode quantitation as a tandem mass spectrometer. On top of that it also has several high sensitivity ion trap MS scans using the final quadrupole as a linear ion trap. MS3 spectrum can b
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