Planta Med 2019; 85(18): 1574
DOI: 10.1055/s-0039-3400411
Main Congress Poster
Poster Session 2
© Georg Thieme Verlag KG Stuttgart · New York

Simultaneous determination of malondialdehyde, glutathione, and glutathione disulfide with UPLC-Q-TOF/PDA in human lung cancer cells (A549)

WT Jeong
1   Department of Industrial Plant Science & Technology, Chungbuk National University,, Cheongju, Chungbuk 28644, Republic of Korea
,
HK Cho
1   Department of Industrial Plant Science & Technology, Chungbuk National University,, Cheongju, Chungbuk 28644, Republic of Korea
,
HR Lee
1   Department of Industrial Plant Science & Technology, Chungbuk National University,, Cheongju, Chungbuk 28644, Republic of Korea
,
HB Lim
1   Department of Industrial Plant Science & Technology, Chungbuk National University,, Cheongju, Chungbuk 28644, Republic of Korea
› Author Affiliations
Further Information

Publication History

Publication Date:
20 December 2019 (online)

 

To evaluate the antioxidative effects of medicinal plants in vitro, intracellular glutathione (GSH), oxidized glutathione (GSSG), and malondialdehyde (MDA) levels after treatment with an oxidizing agent and a medicinal plant extract are determined. Quantification is generally carried out with bio-assay kits and high-performance liquid chromatography (HPLC). As the compounds are measured individually, the procedure becomes time-consuming and expensive, which pose a disadvantage. Here, our aim was to establish a method to simultaneously determine GSH, GSSG, and MDA levels in cultured cells with ultra-performance liquid chromatography quadrupole time-of-flight/photodiode diode array (UPLC-Q-TOF/PDA).

All cellular analytes were purified using self-manufactured solid-phase extraction (SPE) cartridges, and SPE reproducibility was noted to be less than 7.26 %. Instrumental analysis methods verified by ICH guidelines demonstrated excellent linearity and sensitive detection limits. tert-Butyl hydroperoxide treatment was applied to induce oxidative stress because an increase in MDA and GSSG levels and decrease in GSH levels were assumed to be due to oxidative stress. GSH and GSSG exhibited significant changes with respect to the oxidative stress levels; however, MDA levels remained unchanged.

We simultaneously analyzed in cellular MDA, GSH, and GSSG levels in A549 cells. This study is the first attempt of this fast, time-saving, and economical method. In future, to evaluate medicinal plant antioxidant effect(s), it would be used in vitro.

 

  • References

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