Increased Sensitivity in the Detection of Adenine and Pyridine Nucleotides by Exposure to Ultraviolet Light at Low Temperature

Journal/Book: (Reprinted from Nature Vol. 206 No. 4988 pp. 1044-1045 June 5 1965). 1965;

Abstract: Department of Biochemistry School of Medicine State University of New York in Buffalo New York. IN 1957 Szent-Györgyi described a procedure for the enhancement of fluorescence of compounds spotted an paper by cooling with solid carbon dioxide or liquid nitrogen1. Gordon and South have applied this procedure to the action of aromatic acids and other aromatic ringcontaining compounds2. We have found that by observing absorption fluorescence and delayed fluorescence on exposure to long and short wave-length ultra-violet light NAD+ and NADH can be distinguished from each other and from ADP and ATP. Heat-resistant thin-layer chromatographic plates coated with 250-µ layera of silica gel (Merck-Darmstadt) or DEAE cellulose ("BIORAD") and Whatman No. 1 filter paper disks were spotted with 2 ml. each of adenosine diphosphate (ADP) adenosine triphosphate (ATP) niacin adenine dinucleotide (NAD+) and reduced niacin adenine dinucleotide (NADH) solutions. NADH showed a strong light blue fluorescence while the other compounds showed dark areas indicating absorption of the ultra-violet light when exposed to ultra-violet light at room temperature. On cooling with liquid nitrogen the results reported in Table 1 were observed. . . . .

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