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Development of a paper-based microfluidic biosensor for the colorimetric detection of malate dehydrogenase in maize seeds
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DOI：10.2306/scienceasia1513-1874.2026.043.png)
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Abstract
En 中文
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 (mu 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) reduction 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 mu 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 mu 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 mu PAD provides a portable and reliable platform for real-time seed vigor assessment, offering a practical alternative to conventional laboratory methods.
Keywords:
seed quality
microfluidic paper-based analytic device (mu PAD)
malate dehydrogenase
seed deterioration
colorimetric detection
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