Development of an enzymatic assay system of d-lactate using d-lactate dehydrogenase and a UV-LED fluorescent spectrometer

Chien Ming Chen, Shih Ming Chen, Po Jen Chien, Han Yin Yu

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In this study, we aimed to develop a new enzymatic assay system of d-lactate with good precision, accuracy, and sensitivity for the determination of d-lactate concentrations in rat serum. d-Lactate dehydrogenase (d-LDH) was utilized to catalyze d-lactate and NAD+ to pyruvate and NADH, respectively. The generated NADH was excited by using a 340-nm UV-light-emitting diode (LED), and the fluorescence at 491nm was detected to determine the concentration of d-lactate in rat serum. The optics, consisting of the sample cuvette, were set on three-dimensional stages to receive the most intensive fluorescence signal into the spectrometer. The optimal conditions of the d-LDH reaction were pH 8.5 and 25°C for 90min. The results showed that the new d-lactate assay system had good linearity (R2=0.9964) in the calibration range from 5 to 150μM. Intra-day and inter-day accuracies were in the range of 103.96-109.09% and 102.84-104.59%, respectively, and the intra-day and inter-day precision was 4.28-6.82% and 4.04-12.40%, respectively. Finally, serum d-lactate concentrations determined by the proposed enzymatic assay system were compared with those obtained by a conventional HPLC method. The newly developed d-lactate assay system could detect 10-15 samples in 90min, whereas the HPLC method could detect only one sample over the same time period.

Original languageEnglish
Pages (from-to)150-155
Number of pages6
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume116
DOIs
Publication statusPublished - Mar 2 2015

Fingerprint

Enzyme Assays
Ultraviolet Rays
L-Lactate Dehydrogenase
Ultraviolet radiation
Spectrometers
Lactic Acid
Assays
Diodes
NAD
Rats
Fluorescence
Serum
High Pressure Liquid Chromatography
Pyruvic Acid
Calibration
Optics

Keywords

  • D-Lactate
  • D-Lactate dehydrogenase
  • Enzymatic assay system
  • NADH
  • UV-light-emitting diode

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry

Cite this

Development of an enzymatic assay system of d-lactate using d-lactate dehydrogenase and a UV-LED fluorescent spectrometer. / Chen, Chien Ming; Chen, Shih Ming; Chien, Po Jen; Yu, Han Yin.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 116, 02.03.2015, p. 150-155.

Research output: Contribution to journalArticle

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