A Novel Analysis Method for Lactate Dehydrogenase Activity in Serum Samples Based on Fluorescence Capillary Analysis

Abstract

Based on fluorescence capillary analysis technology, a method for quantitating lactate dehydrogenase (LDH) activity in a micro-volume sample was developed. Sample and reagent consumptions were merely 2 and 16 μL per time, respectively. The optimized test conditions were as follows. The reaction reagent consisted of 0.10 M phosphate buffer (pH 6.5), 0.30 mM NADH and 1.20 mM pyruvate. NADH standard was prepared with a phosphate buffer of pH 8.0, and its linear response was controlled in 0.05–0.30 mM. LDH standards containing 2.0 mM PEG could exhibit long-term stability. Under the optimized conditions, a linear response for LDH from 50 to 1200 U L^–1 and a detection limit of 31 U L^–1 were obtained with good precision (RSD: 2.1–2.2%, n = 10) and better recovery of 96–105%. The method’s characteristics was high sensitivity, low consumptions, simple operations, good precision and reliability, lending itself to the miniaturization of fluorophotometer which transformed into a bedside instrument in the hospital.