பயோஅனாலிசிஸ் & பயோமெடிசின் ஜர்னல்

Sodium Deoxycholate Micelles Activated Separation of Coexisting Fivenucleobases by High-performance Thin-layer Chromatography

Abstract

Ali Mohammad, Sameen Laeeq and Abdul Moheman

A novel high-performance thin-layer chromatographic method has been developed for the resolution of fi ve-coexisting nucleobases (adenine, guanine, cytosine, thymine, and uracil). The nucleobases were separated on aluminum-backed cellulose 60 F 254 plates with the aid of 5.0% aqueous sodium deoxycholate (NaDC)-acetonitrile (AcN), 1:3 (v/v) as mobile phase. All the nucleobases were viewed on HPTLC plates under 254nm UV light. The order of R F value given in parentheses was guanine (0.12) < adenine (0.44) < cytosine (0.50) < uracil (0.72) < thymine (0.84). The effect of pH (acidity or basicity) of the mobile phase on the retention of individual nucleobases was examined. Furthermore, the effect of interference of mono- (Li + , Na + ), and bivalent (Mg 2+ , Ba 2+ , Co 2+ , Ni 2+ , Cu 2+ , Zn 2+ , Cd 2+ , Pb 2+ ) cations; mono- (Br - , CH 3 COO - , NO 3 - , IO 4 - ) , and bivalent (CO 3 2- , SO 4 2- MoO 4 2- ) anions, and complexing ligands (urea, and EDTA) on the retention behavior of nucleobases were also assessed. The chromatography of nucleobases was also performed on silica 60 F 254 , RP-18 F 254 , and kieselgel 60 F 254 HPTLC plates. These TLC plates failed to separate the coexisting purines and pyrimidines. The detection limit of all nucleobases on cellulose 60 F 254 layers was 5.4 × 10 -2 μ g spot -1 . The proposed method is rapid, easy, and reliable. It can be applied for routine analysis of DNA, and RNA nucleobases.