Comparison of methanol and perchloric acid extraction procedures for analysis of nucleotides by isotachophoresis

Abstract

During the past decade, isotachophoresis has become increasingly popular for the determination of nucleotide spectra owing to its high sensitivity and reproducibility. At present, this is one of the more important applications of the technique in view of the number of publications in the field. In a review article in 1980 [l] , Holloway and Luestorff cited 57 references on the subject. Gower and Woledge [2] and Woledge and Reilly [ 31 reported the determination of nucleotides in frog muscles by isotachophoresis. Oerlemans et al. [4] introduced a screening method of inborn errors based on nucleotide profiling in human muscles. Aomine and co-workers [5, 61 studied cardiac and skeletal muscles in different species. Perez et al. [7] investigated the ATP/ADP ratio in different rat tissues. Eriksson [S] and Eriksson and Straath [9] analysed nucleotides in rat liver and Talbot [lo, 111 measured nucleotides in red blood cells. We have previously reported on the analysis of first-cleavage-stage embryos of Nassarius reticulatus (trefoils) [12, 131. An important step in nucleotide analysis of biological tissues is the extraction procedure. Extraction procedures must fulfil certain criteria: enzymatic activity should be suppressed quickly and efficiently, and the components of interest should be extracted with high and reproducible recovery. Numerous extraction procedures are described in the literature, all of which