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7 µm, 100 mm x 2.1 mm); mobile phase was 0.05% formic acid - acetonitrile (955 v/v); flow rate 0.1 mL/minute; gradient elution for 6 minutes; and detection at m/z 165.95 149 for O -methylguanine and m/z 136.9 110 for allopurinol. The present study has fulfilled the FDA validation parameter requirements. The method provides rapid, sensitive, and selective analysis of O -methylguanine using UPLC-MS/MS with a linear concentration range between 0.5-20 ng/mL. The present study has fulfilled the FDA validation parameter requirements. T