BACKGROUND: Wheat bran contains high levels of phenolic acids having important roles in the prevention and treatment of diabetes, heart condition and many cancer diseases. Ancient wheats contain high levels of these valuable ingredients. Therefore, an ultrasound-assisted extraction (UAE) technique was optimized using response surface methodology for the extraction of major phenolic acids (ferulic, salicylic, 2-hydroxycinnamic, p-coumaric, t-ferulic, caffeic and vanillic acids) from bran layers of some important ancient wheat species. Optimum combinations were determined for each phenolic acid. RESULTS: Various experimental conditions, extraction time (5.86-34.14 min) and temperature variation (21.71-78.28°C) were examined to optimize the UAE. Solvent type (ethanol, methanol and acetone) and solid-to-liquid ratio (1/90-1/30) were also evaluated. Among the solvent types, the highest extraction yield was obtained in ethanol for all analyzed phenolic acids. Extraction temperature of 44.57°C and extraction time of 23.00 min were found to be an optimal combination for multiple response prediction. To evaluate the usefulness of the method, 18 ancient wheat (einkorn and emmer wheat) bran samples were analyzed. Various levels of individual phenolic acids were detected in the samples. CONCLUSIONS: Analytical conditions giving the highest amount of each phenolic acid were determined in the study. The method can be a good choice because of its simplicity, rapid processing time and low solvent requirements. UAE could be considered in the food, nutrition and pharmaceutical industries for the extraction of some valuable phenolic acids.

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Bulgur, as one of the earliest processed foods, has been intensely consumed in the Middle East, Central Asia, and Turkey cuisines for a long time (Bayram, 2000), and its consumption has also become widespread in the USA and European countries in recent years (Stone et al., 2020). Tetraploid wheat (Triticum turgidum ssp. durum) is used for macaroni or pasta production in worldwide and it is also used for bulgur production especially in Turkey. Leading country in the world, Turkey produced approximately 1.5 million tons of bulgur and 283.026 tons of these amounts were exported in 2020 (TUIK, 2021).Bulgur, one of the important traditional products in Turkish culture, is actually a functional food because of its nutritional and bioactive components in relation to dietary fiber, vitamins B, micronutrients (Donmez et al., 2004;Hayta, 2002;Saka et al., 2020) and low glycemic index (Jenkins et al., 1986), and has long shelf life since it is resistant to insect and microorganism damages (Bayram, 2000;Savas & Basman, 2016). More than 250 food stuffs such as bulgur balls, kısır, desserts, and pilaf are produced by using bulgur for carbohydrate sources in daily diet of Turkish society (Bayram, 2000;

3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazoliumbromide (MTT) assay is a widely used assessment method for the determination of anticancerogenic effects of active compounds including plant secondary metabolites. Recently, some important plant active ingredients have been widely investigated for anticancerogenic properties on melanoma cancer lines which are the most lethal type of skin cancer. Although some methods including DNA assay, 3H-thymidine incorporation and flow cytometry have been recommended for counting cells in the culture, MTT is one of the most frequent method and therefore, MTT assay needs to be optimized for melanoma cell lines. In this study, the MTT analytical procedure for determination of cell viability of human melanoma cell line (SK-Mel-30) was divided into nine steps and various parameters in each step (reagent amount, incubation time, centrifugation, solvent type, waiting time before spectrophotometric analysis and spectrophotometric parameters) were optimized. Optimum amount of MTT reagent and incubation time after MTT addition were determined as 10 µL and 4 h for 96 well plate, respectively. Various solvents were evaluated for solubility effectiveness of the formed crystals and DMSO was found to be the best solvent to dissolve the crystals. Waiting time before spectrophotometric reading and Uv-vis spectrums were also evaluated. At the end of the study a flowchart, presented the best analytical conditions, was constructed. Obtained findings can be used for the determination of anticancerogenic properties of plant ingredients.