The study of DNA adduct 8-hydroxy-2′deoxyguanosine (8-OHdG) formation of butylated hydroxyanisole (BHA) and its metabolite ter-butyl hydroquinone (TBHQ) through in vitro reaction with Calf Thymus DNA and 2′deoxyguanosine

Budiawan, S. S. Purwaningsih, D. I. Cahaya

Research output: Contribution to journalConference articlepeer-review

2 Citations (Scopus)

Abstract

Butylated Hydroxyanisole (BHA) and its metabolite Tert-Butyl Hydroquinone (TBHQ) are synthetic antioxidants, commonly used as food and beverage preservatives. Although WHO declared their safety, the use of these preservatives are still controversial because some studies showed that BHA induced proliferative effects in animal testing and TBHQ is considered as carcinogenic and causes DNA cleavage. This study is aimed to analyze the interaction between Calf Thymus DNA with BHA and TBHQ which are mediated with Copper (II) Chloride. The result of the study in spectrophotometric showed there was bathochromic shift as much as 2-3 nm in DNA treated with TBHQ. The next analysis used HPLC method in stationary phase of ODS, mobile phase of 10mM Natrium Hydrogen Phosphate Buffer and Methanol (85 : 15) for DNA adduct formation, 8-Hydroxy-2-Deoxyguanosine (8-OHDG) as biomarker of risk cancer. The resultof the study showed the formation of DNA adduct 8-OHDG in the interaction between DNA and 20-500 ppm of TBHQ. The 8-OHdG formation was greatly increased by the higher concentration of TBHQ. The relative amount of 8 OHDG which formed was reached 946/105 deoxyguanosine in DNA bases. Confirmation test by LCMS/MS was characterized with the detection of mother ion peak (m/z 284); fragment ion peaks at m/z 167.9, and 139.9; at retention time 3.52 min. Meanwhile the interaction between DNA and 50-250 ppm BHA did not induce 8-OHDG.

Original languageEnglish
Article number012010
JournalIOP Conference Series: Materials Science and Engineering
Volume188
Issue number1
DOIs
Publication statusPublished - 2 May 2017
EventInternational Symposium on Current Progress in Functional Materials 2016, ISCPFM 2016 - Bali, Indonesia
Duration: 26 Jul 201627 Jul 2016

Keywords

  • 8- hydroxy-2-deoxyguanosine (8-OHDG)
  • Butylated hydroxyanisole (BHA)
  • DNA adduct
  • HPLC
  • LCMS/MS
  • spectrophotometric
  • tert-butyl hydroquinone (TBHQ)

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