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"Tersier-butil hidrokuinon (TBHQ) merupakan antioksidan sintetis yang sering digunakan dalam kehidupan sehari-hari sebagai pengawet dalam berbagai produk dan dinyatakan non toksik. Namun penelitian-penelitian terbaru menunjukkan TBHQ dianggap karsinogenik karena dapat menyebabkan kerusakan DNA, dan saat ini masih menjadi kontroversi. Penelitian ini bertujuan mengetahui mekanisme pembentukan DNA Adduct 8-OHdG akibat paparan senyawa TBHQ dengan pendekatan reaksi fenton. Penelitian dilakukan secara in vitro dan in vivo. Studi in vivo dilakukan dengan memberikan paparan TBHQ, CuCl dan CuCl dicampur TBHQ pada tikus. Studi in vitro dilakukan dengan mereaksikan 2-deoksiguanin dan TBHQ dengan penambahan CuCl dan H2O2 dengan variasi waktu inkubasi dan variasi pH. Studi in vitro dilanjutkan dengan menganalisis terbentuknya DNA Adduct 8-OHdG menggunakan LC-MS/MS. Setiap perbedaan variasi sangat mempengaruhi jumlah konsentrasi 8-OHdG yang terbentuk. Pada pH 7,4 inkubasi 12 jam menghasilkan 8-OHdG lebih banyak. Studi in vivo dilanjutkan uji Elisa-kit setelah 28 hari pemaparan kadar 8-OHdG rata-rata yang terdeteksi pada kelompok paparan TBHQ, kelompok paparan CuCl, dan kelompok paparan TBHQ + CuCl yang terdeteksi berturut-turut sebesar 0,3840; 0,8705; 1,2205 ppb. Pada in vitro dan in vivo, semakin lama waktu paparan semakin banyak pembentukan 8-OHdG yang dihasilkan. Pencampuran TBHQ, CuCl dan H2O2 pada in vitro, dengan pencampuran TBHQ dan CuCl pada in vivo keduanya menunjukkan sinergisitas menghasilkan 8-OHdG lebih banyak.

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Tert-butyl hydroquinone (TBHQ) is a synthetic antioxidant that is often used in daily life as a preservative in various products and is declared non-toxic. But recent studies show that TBHQ is considered carcinogenic because it can cause DNA damage, and currently controversy. This study aims to determine the mechanism for the formation of 8-OHdG DNA Adduct due to exposure to TBHQ compounds with the approach of the fenton reaction. The study was conducted in vitro and in vivo. In vivo studies were carried out by giving exposure to TBHQ, CuCl and CuCl mixed with TBHQ in mice. In vitro studies were carried out by reacting 2-deoksiguanin and TBHQ with the addition of CuCl and H2O2 with variations in incubation time and pH variation. In vitro studies were continued by analyzing the formation of the 8-OHdG DNA Adduct using LC-MS / MS. Any difference in variation greatly affects the amount of 8-OHdG concentration formed. At pH 7.4 incubation 12 hours produces more 8-OHdG. The in vivo study continued with the Elisa-kit test after 28 days of exposure to the average 8-OHdG levels detected in the exposure group TBHQ, the group exposed to CuCl, and detected groups of TBHQ + CuCl at 0.3840; 0.8705; 1,2205 ppb. In in vitro and in vivo, the longer the exposure time the more formation of 8-OHdG is produced. Mixing TBHQ, CuCl and H2O2 in vitro, by mixing TBHQ and CuCl in vivo both showed synergy to produce 8-OHdG more."

"Penelitian ini dilakukan untuk menganalisis pembentukan DNA Adduct 8-OHdG akibat kerusakan oksidatif DNA yang disebabkan oleh paparan akrilamida (1 mg/kg BB) dan Cu (II) (10 mg/kg BB). Studi in vivo dilakukan dengan menggunakan kelompok tikus (Rattus norvegicus) galur Sprague Dawley yang diberi paparan selama 28 hari dan dilakukan pengambilan sampel urin setiap 7 hari. Studi in vitro dilakukan dengan mereaksikan 2-„deoksiguanosin pH 7,4 dengan akrilamida, Cu (II), H2O2 melalui reaksi Fenton-like pada suhu 37 °C. Analisis 8-OHdG dilakukan dengan instrumentasi LC-MS/MS ionisasi positif, fasa terbalik, dengan gradien elusi campuran ammonium asetat 20mM dan asetonitril. Hasil studi in vivo menunjukkan bahwa paparan akrilamida, Cu, dan gabungan akrilamida + Cu (II) mengakibatkan adanya kerusakan DNA yang dapat menimbulkan risiko kanker. Kelompok paparan gabungan akrilamida + Cu (II) menunjukkan kadar yang paling tinggi, hal ini menunjukkan adanya kesinergisan antara akrilamida dan Cu (II) pada pembentukan kadar 8-OHdG. Pengujian kadar 8-OHdG secara berkala menunjukkan kadar 8-OHdG yang semakin meningkat seiring dengan lamanya waktu paparan. Hasil studi in vitro menunjukkan bahwa akrilamida tidak menginduksi pembentukan 8-OHdG secara langsung, melainkan perlu adanya proses metabolisme terlebih dahulu.

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This study was conducted to analyze the formation of 8-OHdG DNA Adduct due to oxidative DNA damage caused by exposure to acrylamide (1 mg / kg BB) and Cu (II) (10 mg / kg BW). In vivo studies were carried out using a group of Sprague Dawley rats (Rattus norvegicus) that were exposed for 28 days of exposure and urine samples were taken every 7 days. In vitro studies were carried out by reacting 2-oksdeoxiguanosine pH 7.4 with acrylamide, Cu (II), H2O2 through Fenton-like reaction at 37 ° C. The 8-OHdG analysis was performed with positive ionization LC-MS / MS instrumentation, reversed phase system, with a mixture of elution gradient of ammonium acetate 20mM and acetonitrile. The results of in vivo studies showed that acrylamide, Cu, and acrylamide combined Cu (II) exposure caused DNA damage that could cause cancer risk. The exposure group of acrylamide combined Cu (II) combined showed the highest levels, this indicates a synergy between acrylamide and Cu (II) in the formation of 8-OHdG levels. Periodic analysis of 8-OHdG levels shows that 8-OHdG levels are increasing along with the length of time of exposure. In vitro testing shows that acrylamide does not directly induce the formation of 8-OHdG, but rather requires a metabolic process first."