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1 2012

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New microfluidic device for lactate dehydrogenase (LDH) activity analysis

Jędrych E., Mazur M., Grabowska-Jadach I., Brzózka Z.

Challenges of Modern Technology

2012

Vol. 3, no. 4

3--8

In this paper, we present cytotoxicity analysis (determination of lactate dehydrogenase — LDH activity performed in a designed and fabricated microfl uidic system. This method allowed for analysis of a supernatant collected from A549 (human lung cancer) and HT-29 (human colon cancer epithelial) cells, which were incubated for 24 h with selected compounds. LDH is an intracellular enzyme present in tissues, which is released into the supernatant caused by membrane damage or cell lyses. In our tests, LDH-Cytotoxicity Assay Kit (BioVision) was used. The toxic eff ect of drugs was measured in the developed microsystem made of PDMS (poly(dimethylsiloxane)). Analytical reaction took place in the special designed microchannel geometry. Then, the LDH activity was measured at 490 nm using spectrophotometer. In subsequent experiments, appropriate conditions for measurements using a microfl uidic system were chosen. It was found that the selected reagent is sensitive to temperature changes and light exposure. Reaction time in the microsystem was determined by changes of fl ow rates of reagents. Afterwards, for the various reaction time, the toxic eff ect of 5-fl uorouracil, celecoxib and 1,4-dioxane was performed. The obtained results were compared with the results carried out in 96-well plates. Based on these results, it was noted that the enzymatic reaction time in the microsystem is shorter than in 96-well plate. Moreover, the advantage of using microsystem is also the small amount of reagents.