Use of 2-mercaptopyridine for the determination of alkylating agents in complex matrices: application to dimethyl sulfate |
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Authors: | Hoogerheide J G Scott R A |
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Institution: | Pfizer Global Research and Development, Pfizer Inc., 7000 Portage Road, Kalamazoo, MI 49001, USA |
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Abstract: | A rapid and sensitive method for the determination of alkylating agents in complex reaction mixtures was developed and characterized. Analyses are based on the alkylation of 2-mercaptopyridine by the analyte; the derivative is separated by RP-HPLC and measured by fluorescence detection. When applied to the determination of dimethyl sulfate, the method is linear over four orders of magnitude: 0.01-10 μg mL−1. By using recrystallized 2-mercaptopyridine, quantitation limits of 10 ng mL−1 can be achieved. Precision of the assay is 2% R.S.D. in the 1-10 μg mL−1 range and about 15% R.S.D. at 10 ng mL−1. Studies on the pH dependence of the derivatization reaction were key to minimizing interference from the dimethyl sulfate degradation product, monomethyl sulfate, in quenched reaction samples. |
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Keywords: | Dimethyl sulfate 2-Mercaptopyridine Alkylating agent Reversed-phase liquid chromatography Fluorescence |
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