Clinical Chemistry
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Clinical Chemistry 22: 1322-1329, 1976;
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Clinical Chemistry, Vol 22, 1322-1329, Copyright © 1976 by American Association for Clinical Chemistry

Purification and analysis of the purity of NADH

SA Margolis, BF Howell and R Schaffer

We developed an analytical reverse-phase high-performance liquid chromatographic procedure for rapid assessment of the purity of NADH. The method completely separates adenosine monophosphate and adenosine diphosphoribose from NADH. By use of this analytical technique we found that preparative chromatography on DEAE-cellulose gives NADH that is free from adenine nucleotides as well as other impurities that commonly are present in NADH. The absorbance ratio at 260 and 340 nm of the purified NADH in 1.8 mmol/liter ammonium carbonate is 2.261 +/- 0.002 (+/- 1 SD).


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A. Kaplan, E. Weiss, S. Byrne, N. El-Torkey, and S. Margolis
Purified reduced nicotinamide adenine dinucleotide: responses to lactate dehydrogenase isozymes from three cell sources
Science, May 1, 1981; 212(4494): 553 - 555.
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Copyright © 1976 by the American Association for Clinical Chemistry.