Şeyma Özçirak Ergün


3 Prediction of Ionizing Radiation Doses in Irradiated red Pepper (Capsicum annuum) and Mint (Mentha piperita) by Gel Electrophoresis

Authors: Ergün Şakalar, Şeyma Özçirak Ergün, Emrah Yalazi̇, Nebahat Şahi̇n


Food irradiation is a usage of exposing food to ionising radiation (IR) such as gamma rays. IR has been used to decrease the number of harmful microorganisms in the food such as spices. Excessive usage of IR can cause damage to both food and people who consuming food. And also it causes to damages on food DNA. Generally, IR detection techniques were utilized in literature for spices are Electron Spin Resonance (ESR), Thermos Luminescence (TL). Storage creates negative effect on IR detection method then analyses of samples have been performed without storage in general. In the experimental part, red pepper (Capsicum annuum) and mint (Mentha piperita) as spices were exposed to 0, 0.272, 0.497, 1.06, 3.64, 8.82, and 17.42 kGy ionize radiation. ESR was applied to samples irradiated. DNA isolation from irradiated samples was performed using GIDAGEN Multi Fast DNA isolation kit. The DNA concentration was measured using a microplate reader spectrophotometer (Infinite® 200 PRO-Life Science–Tecan). The concentration of each DNA was adjusted to 50 ng/µL. Genomic DNA was imaged by UV transilluminator (Gel Doc XR System, Bio-Rad) for the estimation of genomic DNA bp-fragment size after IR. Thus, agarose gel profiles of irradiated spices were obtained to determine the change of band profiles. Besides, samples were examined at three different time periods (0, 3, 6 months storage) to show the feasibility of developed method. Results of gel electrophoresis showed especially degradation of DNA of irradiated samples. In conclusion, this study with gel electrophoresis can be used as a basis for the identification of the dose of irradiation by looking at degradation profiles at specific amounts of irradiation. Agarose gel results of irradiated samples were confirmed with ESR analysis. This method can be applied widely to not only food products but also all biological materials containing DNA to predict radiation-induced damage of DNA.

Keywords: Electrophoresis, Gel electrophoresis, Dna, ionizeradiation

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2 Triplex Detection of Pistacia vera, Arachis hypogaea and Pisum sativum in Processed Food Products Using Probe Based PCR

Authors: Ergün Şakalar, Şeyma Özçirak Ergün, Emrah Yalazi̇, Emine Altinkaya, Cengiz Ataşoğlu


In recent years, food allergies which cause serious health problems affect to public health around the world. Foodstuffs which contain allergens are either intentionally used as ingredients or are encased as contaminant in food products. The prevalence of clinical allergy to peanuts and nuts is estimated at about 0.4%-1.1% of the adult population, representing the allergy to pistachio the 7% of the cases of tree nut causing allergic reactions. In order to protect public health and enforce the legislation, methods for sensitive analysis of pistachio and peanut contents in food are required. Pea, pistachio and peanut are used together, to reduce the cost in food production such as baklava, snack foods.DNA technology-based methods in food analysis are well-established and well-roundedtools for species differentiation, allergen detection. Especially, the probe-based TaqMan real-time PCR assay can amplify target DNA with efficiency, specificity, and sensitivity.In this study, pistachio, peanut and pea were finely ground and three separate series of triplet mixtures containing 0.1, 1, 10, 100, 1000, 10,000 and 100,000 mg kg-1 of each sample were prepared for each series, to a final weight of 100 g. DNA from reference samples and industrial products was successfully extracted with the GIDAGEN® Multi-Fast DNA Isolation Kit. TaqMan probes were designed for triplex determination of ITS, Ara h 3 and pea lectin genes which are specific regions for identification pistachio, peanut and pea, respectively.The real-time PCR as quantitative detected pistachio, peanut and pea in these mixtures down to the lowest investigated level of 0.1, 0.1 and 1 mg kg-1, respectively. Also, the methods reported here are capable of detecting of as little as 0.001% level of peanut DNA, 0,000001% level of pistachio DNA and 0.000001% level of pea DNA. We accomplish that the quantitative triplex real-time PCR method developed in this study canbe applied to detect pistachio, peanut and peatraces for three allergens at once in commercial food products.

Keywords: Allergens, Dna, real-time PCR, TaqMan probe

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1 The Qualitative and Quantitative Detection of Pistachio in Processed Food Products Using Florescence Dye Based PCR

Authors: Ergün Şakalar, Şeyma Özçirak Ergün


Pistachio nuts, the fruits of the pistachio tree (Pistacia vera), are edible tree nuts highly valued for their organoleptic properties. Pistachio nuts used in snack foods, chocolates, baklava, meat products, ice-cream industries and other gourmet products as ingredients. Undeclared pistachios may be present in food products as a consequence of fraudulent substitution. Control of food samples is very important for safety and fraud. Mix of pistachio, peanut (Arachis hypogaea), pea (Pisum sativum L.) used instead of pistachio in food products, because pistachio is a considerably expensive nut. To solve this problem, a sensitive polymerase chain reaction PCR has been developed. A real-time PCR assay for the detection of pea, peanut and pistachio in baklava was designed by using EvaGreen fluorescence dye. Primers were selected from powerful regions for identification of pea, peanut and pistachio. DNA from reference samples and industrial products were successfully extracted with the GIDAGEN® Multi-Fast DNA Isolation Kit. Genomes were identified based on their specific melting peaks (Mp) which are 77°C, 85.5°C and 82.5°C for pea, peanut and pistachio, respectively. Homogenized mixtures of raw pistachio, pea and peanut were prepared with the ratio of 0.01%, 0.1%, 1%, 10%, 40% and 70% of pistachio. Quantitative detection limit of assay was 0.1% for pistachio. Also, real-time PCR technique used in this study allowed the qualitative detection of as little as 0.001% level of peanut DNA, 0,000001% level of pistachio DNA and 0.000001% level of pea DNA in the experimental admixtures. This assay represents a potentially valuable diagnostic method for detection of nut species adulterated with pistachio as well as for highly specific and relatively rapid detection of small amounts of pistachio in food samples.

Keywords: real-time PCR, pistachio, peanut, pea

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