Analysis of Deoxyribonucleotide in Plasmid Deoxyribonucleic Acid Using High Performance Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometry

摘要: An ion pair reverse phase high performance liquid chromatography (RP-HPLC) coupled with inductively plasma mass spectrometry (ICP-MS) method has been developed for the determination of deoxyribonucleotide in plasmid DNA. Optimum separation conditions four deoxyribonucleotides Including 2'-deoxycytidine-5'-monophosphate (dCMP), 2'-deoxythymidine-5'-monophosphate 2'-deoxyguanosine-5'-monophosphate (dGMP) and 2'-deoxyadenosine-5'-monophosphate (dAMP) are obtained using a mobile containing 10 mmol/L ammonium acetate (pH 4.8) 2.5% methanol. The hexapole collision/reaction cell technique (CCT) by adding O-2 is employed detection (PO+)-P-31-O-16 to avoid potential interferences polyatomic isobaric at m/z 31 directly detecting P-31(+). By such HPLC-TCP-MS hyphenation technique, limits 0.211 mu mol/L dCMP, 0.204 dTMP, 0.173 dGMP 0.225 dAMP, respectively. This hyphenated HPLC-ICP-MS successfully applied analysis DNA, which contained 152.9 +/- 2.4 228.2 4.0 125.3 3.0 222.9 3.3 dAMP.