Clinical Chemistry
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Clinical Chemistry 23: 1581-1584, 1977;
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Clinical Chemistry, Vol 23, 1581-1584, Copyright © 1977 by American Association for Clinical Chemistry

Lactate dehydrogenase inhibitors in NADH preparations

SA Margolis, BF Howell and R Schaffer

The presence of a new lactate dehydrogenase inhibitor on the trailing edge of the NADH peak from chromatography on diethylaminoethyl-celluose [Loshon et al., Clin. Chem., this issue] was verified. It was resolved from the NADH by high-performance liquid chromatography on muBondapak C18. When the new inhibitor was present in a reaction mixture to the extent that, of the initial 260-nm absorbance, about 5% was contributed by the inhibitor, the rate of NADH oxidation by lactate dehydrogenase decreased by 65%. The inhibitor absorbs at 260 and 340 nm, and is different from the Strandjord-Clayson inhibitor [J. Lab. Clin. Med. 67, 144 (1966)] by both types of chromatography. Because this new inhibitor has ultraviolet properties similar to those of NADH and chromatographs with the NADH on DEAE-cellulose, the high-performance liquid chromatographic method must be used to ensure its absence in preparations of NADH used for clinical assay.


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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 © 1977 by the American Association for Clinical Chemistry.