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Published on: February 24, 2018
Lactate dehydrogenase inhibitors in NADH preparations
Clinical Chemistry
|September 1, 1977
Summary
A newly discovered lactate dehydrogenase inhibitor interferes with NADH assays. This compound, which absorbs UV light similarly to NADH, requires high-performance liquid chromatography for accurate clinical measurements.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Lactate dehydrogenase (LDH) assays are crucial in clinical diagnostics.
- NADH is a key component in these assays, and its purity is essential for accurate results.
Purpose of the Study:
- To identify and characterize a novel inhibitor affecting NADH in LDH assays.
- To establish a reliable method for detecting and quantifying this inhibitor.
Main Methods:
- Chromatography on diethylaminoethyl-celluose (DEAE-cellulose).
- High-performance liquid chromatography (HPLC) on muBondapak C18.
- UV spectrophotometry at 260 nm and 340 nm.
Main Results:
- A new LDH inhibitor was identified co-eluting with NADH on DEAE-cellulose.
- HPLC successfully resolved the inhibitor from NADH.
- The inhibitor caused a 65% decrease in NADH oxidation rate at low concentrations.
- The inhibitor exhibited distinct chromatographic behavior compared to the Strandjord-Clayson inhibitor.
Conclusions:
- The new inhibitor possesses UV absorbance properties similar to NADH, complicating its detection.
- HPLC is essential for ensuring the purity of NADH preparations for clinical LDH assays.
- Accurate clinical assays necessitate the removal or accurate accounting of this novel inhibitor.
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