Citation: ZHANG Xiao-Dan, XU Yi, ZHANG Tao, LÜ Jun-Jiang. Assessing Plant Antioxidants by Cellular Antioxidant Activity Assay Based on Microfluidic Cell Chip with Arrayed Microchannels[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(4): 604-609. doi: 10.11895/j.issn.0253-3820.150596 shu

Assessing Plant Antioxidants by Cellular Antioxidant Activity Assay Based on Microfluidic Cell Chip with Arrayed Microchannels

ZHANG Xiao-Dan ^1,2,3 ,
XU Yi ^1,2,3,, ,
ZHANG Tao ^1,2,3 ,
LÜ Jun-Jiang ¹

1.
¹ College of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400030, China;
2.
² Key Disciplines Lab of Novel Micro-nano Devices and System Technology, Chongqing 400030, China;
3.
³ International R & D center of Micro-nano Systems and New Materials Technology, Chongqing 400030, China

Corresponding author: XU Yi,
Received Date: 27 July 2015
Available Online: 4 October 2015
Fund Project: 本文系国家自然科学基金项目(No.21375156) (No.21375156)国家高技术研究发展计划(863)项目(No.2015AA021104) (863)项目(No.2015AA021104)中央高校基本科研业务费专项重点项目(No.106112015CDJZR225512) (No.106112015CDJZR225512)重庆市科技攻关项目(No.cstc2015jcsf80001)资助 (No.cstc2015jcsf80001)

An integrated microfluidic chip with arrayed micro channels that consisted of eight repeat arrayed 6×6 cell culture chamber was designed and fabricated. The analytical microsystem combined with designed microchip, measuring device and environmental control unit was established for cell culture and parallel cellular antioxidant activity(CAA) analysis of plant antioxidants. The microfluidic chip included a polydimethylsiloxane(PDMS) cover and glass substrate that consisted of two hundreds and eighty-eight round cell culture microchambers and forty-eight independent parallel array channels. Eight groups of different samples with six different concentrations could be investigated simultaneously with multimode reader in one test. HepG2 cells were successfully cultured on the microchip. Moreover, the viability percentage of the HepG2 cells exposed to these plant antioxidants solutions at different concentrations for 24 h was higher than 90%. With 2', 7'-Dichlorofluorescin diacetate(DCFH-DA) as a fluorescence probe, 2, 2'-azobis(2-amidinopropane) dihydrochloride(ABAP) as the initiator of intracellular reactive oxygen species(ROS), we tested the inhibitory effect of several plant antioxidants such as quercetin, rutin and kaempferol on free radicals. The CAA units were calculated by the data measured from cellular morphology and fluorescence intensity over time. It was shown that the CAA units of quercetin, rutin and kaempferol were 71.42±0.19, 74.31±0.36 and 69.92±0.09(x±s, n=3), while the calculated IC₅₀ were(7.20±0.06) μmol/L,(52.06±0.14) μmol/L and(32.55±0.03) μmol/L(x±s, n=3), respectively.
- Microfluidic cell chip with arrayed microchannels,
- Cellular antioxidant activity,
- High throughput,
- Screening
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