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ctrum of a fragmentation related to Figure 9B. The solution ion spectrum of 7 is also reported 7 is also10C) for comparison: a number of fragmentations similar to those present in the IL-6 Inhibitor Accession Product (Figure reported (Figure 10c) for comparison: several fragmentations comparable to these present in the product ion spectrum of are alleged observed. are indeed observed. ion spectrum of its alleged metabolite its indeed metaboliteAntioxidants 2022, 10, x FOR PEER Review Antioxidants 2022, 11,14 of 21 13 ofFigure ten. (a) Superimposed mass chromatograms ofof the m/z 288.0 precursor ion, obtained from (A) Superimposed mass chromatograms the m/z 288.0 precursor ion, obtained from the rat liver microsomal fraction at t =t 0 (dotted line) and t t==22h (continuous line) incubation with all the rat liver microsomal fraction at = 0 (dotted line) and h (continuous line) incubation with compound 7. (B) Solution ion spectrum of in the selected m/z 288.0 precursor, collected at min, from 7. (b) Item ion spectrum the selected m/z 288.0 precursor, collected at 3.78 3.78 min, compound in the latter analysis. (c) Product ion spectrum of the selected m/z 333.26, a precursor on the latter evaluation. (C) Solution ion spectrum on the chosen m/z 333.26, a precursor of compound 7. compound 7.The identical experiment was also executed on a rat liver microsomal fraction incubated together with the exact same experiment was also executed on a rat liver microsomal fraction incubated compound six. The m/z 274.0 precursor ion was isolated on Q1, representing the with compound 6. The m/z compound 6 metabolite obtained by a single de-nitration molecular ion of a hypothetic274.0 precursor ion was isolated on Q1, representing the molecular ion of a in this case, no CysLT2 Antagonist manufacturer signal 6 metabolite within the chromatogram. in the side chain:hypothetic compound was observedobtained by a single de-nitration fromFurther and much more this case, no signal waswere carried out chromatogram. two most the side chain: in sensitive experiments observed inside the by deciding on the Further and more sensitiveSRM transitions, namely m/z 274228 (lossthe NO2 ) and probable precursor roduct ion experiments had been carried out by deciding on of two most probable precursor roduct side chain), likewise fragmentation is observed in compound m/z 274167 (losses from theion SRM transitions, namely m/z 274228 (loss of NO2) and m/z 274167 (losses from the side chain), the two fragmentation obtained from 6. Figure 11A reports the comparison between likewise SRM transitionsis observed in compound microsomal fraction, just before the incubation together with the two SRM transitions the rat liver six. Figure 11a reports the comparison betweencompound six and after two obtained from the From this experiment of enhanced sensitivity, a high chromatographic hours, respectively.rat liver microsomal fraction, just before the incubation with compound six and is evident in the retention time of this min only within the profile sensitivity, a two peakafter two hours, respectively. From two.61 experiment of enhancedcollected following high chromatographic The is evident at the retention time of rat min only inside the profile hours’ incubation. peaksame experiment was executed on the2.61liver microsomal fraction collected right after two hours’ incubation. The same experiment was executed on the rat liver incubated with compound 7, to confirm that m/z 274.0 corresponds to a metabolite of microsomal fraction incubated with compound 7, to 11B). compound six, not made from compound 7 (Figureconfirm that m/z 274.0 corresponds t

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