alexa Sodium Deoxycholate Micelles Activated Separation of Co
ISSN: 1948-593X

Journal of Bioanalysis & Biomedicine
Open Access

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Research Article

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

Ali Mohammad*, Sameen Laeeq and Abdul Moheman

 

Analytical Research Laboratory, Department of Applied Chemistry, Faculty of Engineering & Technology, Aligarh Muslim University, Aligarh, India

Corresponding Author:
Ali Mohammad
Analytical Research Laboratory
Department of Applied Chemistry
Faculty of Engineering & Technology
Aligarh Muslim University
Aligarh-202 002, India
Fax: +91-571-2702758
E-mail: [email protected]

Citation: Mohammad A, Laeeq S, Moheman A (2010) Sodium Deoxycholate Micelles Activated Separation of Coexisting Five-nucleobases by Highperformance Thin-layer Chromatography. J Bioanal Biomed 2: 055-059. doi:10.4172/1948-593X.1000022

Copyright: © 2010 Mohammad A, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

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.

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