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Abstrak :
Akrilamida merupakan monomer hasil depolimerisasi poliakrilamida akibat adanya pengaruh suhu, cahaya dan pH. Akrilamida juga dapat ditemukan pada produk pangan dengan pengolahan suhu tinggi. Akrilamida dapat menginduksi terbentuknya Reactive Oxygen Species (ROS) seperti radikal hidroksil. Radikal hidroksil dapat berinteraksi dengan basa guanin membentuk DNA adduct 8-hidroksi 2'-deoksiguanin (8-OHdG). Penelitian ini bertujuan untuk mengetahui pembentukan 8-OHdG akibat paparan akrilamida yang berinteraksi dengan logam Cu (I). Penelitian ini dilakukan secara in vitro dan in vivo. Studi dilakukan secara in vitro melalui reaksi Fenton antara 2'- deoksiguanosin (2dG) dengan akrilamida dan Cu (I). Reaksi dilakukan pada suhu 37 °C, variasi pH 7,4 dan 8,4, serta variasi waktu inkubasi 4, 8, 12 dan 24 jam. Studi in vivo dilakukan pada tikus putih jantan (Rattus norvegicus) galur Sprague-Dawley yang diberikan paparan akrilamida serta kombinasi akrilamida dan Cu (I) dengan dosis yang berbeda selama 21 hari. Hasil studi in vitro dianalisis menggunakan LC-MS/MS QTOF dan plasma darah hasil studi in vivo dianalisis menggunakan ELISA. Inkubasi pada pH 7,4, suhu 37 °C dan waktu inkubasi 24 jam memberikan konsentrasi 8-OHdG tertinggi sebesar 19,09 ng/mL. Konsentrasi tertinggi 8-OHdG dalam plasma darah terdapat pada kelompok paparan akrilamida 25 mg/kg BB dan Cu (I) 10 mg/kg BB sebesar 1,31 ng/mL. Berdasarkan hasil studi in vitro dan in vivo yang telah dilakukan, hal ini menunjukkan terdapat sinergisitas antara akrilamida dan Cu (I) dalam pembentukan 8-OHdG.

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Acrylamide is a monomer as depolymerization results caused by the effect of temperature, light and pH. Acrylamide can be found on food products processed with high temperature. Acrylamide can induce the formation of reactive oxygen species (ROS), e.g. hydroxyl radical. Hydroxyl radical can interact with guanine to form DNA Adduct 8-hydroxy-2'-deoxyguanosine (8-OHdG). The aim of this study is to investigate the formation of 8-OHdG due to co-exposure of acrylamide and copper (I). The study was carried out in vitro through the Fenton reaction and in vivo. In vitro study was undergone through Fenton reaction between 2'-deoxyguanosine and acrylamide at temperature 37 °C, various of pH 7,4 and 8,4, and time of incubation 4, 8, 12 and 24 hours. While in vivo study was carried out by administering orally of acrylamide single dose and combination of acrylamide and copper (I) to male rats (Rattus norvegicus) strain Sprague-Dawley for 21 days exposure. The results of in vitro study were analyzed by LC-MS/MS QTOF and plasma resulted from in vivo study were analyzed by ELISA to determine the concentration of 8-OHdG. Incubation on pH 7,4, 37 °C and time of incubation 24 hours gave highest concentration of 8-OHdG. Meanwhile the highest concentration of 8-OHdG in plasma found in group with the exposure of acrylamide 25 mg/kg BW and copper (I) 10 mg/kg BW. According to the results from in vitro and in vivo studies, it was shown that both acrylamide and copper (I) gave synergistic effect towards the formation of 8-OHdG.

Abstrak :
Pada penelitian ini dilakukan studi pembentukan DNA adduct 8-Hidroksi-2';-Deoksiguanosin 8-OHdG sebagai biomarker kerusakan DNA dengan mereaksikan basa DNA 2';-deoksiguanosin dengan t-BHQ melalui Fenton Like Reaction Cr III dan H2O2 pada variasi suhu 37°C dan 60°C, pH 7,4 dan pH 8,4, dengan waktu inkubasi 7 jam dan 12 jam. Hasil adduct dianalisis menggunakan instrumen HPLC reversed phase dengan detektor UV pada panjang gelombang 254 nm. Pembentukan DNA Adduct 8-OHdG dari senyawa t-BHQ terdeteksi pada reaksi 2'dG dan t-BHQ pada suhu 60°C waktu inkubasi 12 jam, reaksi antara 2'dG, Cr III terdeteksi pada waktu inkubasi 7 jam, pH 8,4, 37°C, pada suhu 60°C pH 7,4 dan 8,4 serta waktu inkubasi 12 jam pada suhu 37°C dan 60°C. Reaksi antara 2'dG, t-BHQ, H2O2 hanya terdeteksi pada suhu 60°C waktu inkubasi 12 jam, dan reaksi antara 2'dG, Cr III, H2O2, dan 2'dG, t-BHQ, Cr III, serta 2'dG dengan reaksi Fenton terdeteksi baik pada waktu inkubasi 7 dan 12 jam. ...... ChemistryTitle In Vitro Study of DNA Adduct 8 Hydroxy 2' Deoxyguanosine 8 OHdG Formation with Tert Butyl Hydroquinone t BHQ Through Fenton Like Reaction In this research, DNA adduct formation of 8 hydroxy 2 39 Deoksiguanosin 8 OHdG as biomarkers of DNA damage which conducted by reacting the DNA bases 2'-deoxyguanosine base DNA with t BHQ through the Fenton Like Reaction Cr III and H2O2 with variation of temperature 37°C and 60°C, pH pH 7,4 and pH 8,4, and incubation time 7 hours, and 12 hours studied. The adduct results were analyzed using instruments reversed phase HPLC with UV detector at a wavelength of 254 nm. The formation of DNA Adduct 8 OHdG of t BHQ compound is detected in the reaction of 2'dG, t BHQ at the temperature of 60°C with incubation time at 12 hours, reaction of 2'dG, Cr III is detected when incubation time at 7 hours, pH 8,4 and temperature at 37°C, when the temperature at 60°C with pH 7,4 and 8,4 and when incubation time is at 12 hours with the temperature at 37°C and 60°C. Reaction of 2' dG, t BHQ, H2O2 only detected at the temperature of 60 C with incubation time at 12 hours, and the reaction of 2'dG, Cr III, H2O2, and 2'dG, t BHQ, Cr III, also 2'dG with Fenton reaction are detected either when incubation time at 7 and 12 hours.

Abstrak :
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. ......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.