Research articles
ScienceAsia 52 (2026): 1-10 |doi:
10.2306/scienceasia1513-1874.2026.043
Development of a paper-based microfluidic biosensor for the
colorimetric detection of malate dehydrogenase in maize
seeds
Jakkaphan Kumsaba,b,*, Yodying Yingchutrakula, Nattapon Simanona, Chutikarn Butkinareea,
Sithichoke Tangphatsornruanga, Sudkate Chaiyoc,d
ABSTRACT: Seed deterioration is a major constraint on germination efficiency and overall crop productivity, while
conventional quality assessment methods are often laborious and time-consuming. In this study, a microfluidic paper
based analytical device (?PAD) was developed for rapid and cost-effective evaluation of maize seed quality via malate
dehydrogenase (MDH) activity. The colorimetric assay is based on NAD+ (nicotinamide adenine dinucleotide) reduc
tion and subsequent redox reaction with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)/PMS
(phenazine methosulfate), producing a purple signal proportional to enzyme activity. Optimal conditions included
a 3 mm flow aperture, 10 mg/ml MTT, 20 min incubation, and 30 ?l sample volume. The biosensor exhibited a
concentration-dependent response and a clear decline in signal with seed aging, correlating well with germination rate.
Statistical analysis confirmed significant differences (p < 0.015). The proposed ?PAD demonstrated high analytical
sensitivity for detecting MDH in low-concentration seed extracts, achieving a limit of detection (LOD) and limit of
quantification (LOQ) of 0.23 mg/ml and 0.70 mg/ml, respectively. This ?PAD provides a portable and reliable platform
for real-time seed vigor assessment, offering a practical alternative to conventional laboratory methods.
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| a |
National Center for Genetic Engineering and Biotechnology, NSTDA, Pathum Thani 12120 Thailand |
| b |
Department of Biotechnology, Faculty of Science and Technology, Thammasat University (Rangsit Center),
Pathum Thani 12120 Thailand |
| c |
The institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok 10330 Thailand |
| d |
Center of Excellence for Food and Water Risk Analysis (FAWRA), Department of Veterinary Public Health,
Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330 Thailand |
* Corresponding author, E-mail: jkkumsab@tu.ac.th
Received 5 Apr 2026, Accepted 2 May 2026
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