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Öğe Amanitin and phallotoxin concentration in Amanita phalloides var. alba mushroom(Pergamon-Elsevier Science Ltd, 2013) Kaya, Ertuğrul; Yılmaz, İsmail; Sinirlioğlu, Zeynep Aydın; Karahan, Selim; Bayram, RecepAlthough rarely seen, Amanita phalloides var. alba, a variety of A. phalloides type mushrooms, causes mushroom poisoning resulting in death. Since it is frequently confused with some edible mushrooms due to its white colored cap and macroscopic appearance, it becomes important in toxicological terms. Knowledge of the toxin amount contained in this mushroom type is invaluable in the treatment of cases involving poisoning. In this study, we examined the toxin levels of various parts of the A. phalloides var. alba mushroom growing Duzce region of Turkey. Toxin analyses were carried out for A. phalloides var. alba, which were collected from the forests Duzce region of Turkey in 2011, as a whole and also separately in its spore, pileus, gills, stipe and volva parts. The alpha amanitin, beta amanitin, gamma amanitin, phalloidin and phallacidine analyses of the mushrooms were carried out using the RP-HPLC method. A genetic analysis of the mushroom showed that it had similar genetic characteristics as A. phalloides and was a variety of it. The lowest toxins quantity was detected in spores, volva and stipe among all parts of the mushroom. The maximum amount of amatoxins was measured in the gills. The pileus also contained a high amount of amatoxins. Generally, amatoxins and phallotoxin concentrations were lower as compared to A. phalloides, but interestingly all toxins other than gamma toxin were higher in the spores of A. phalloides var. alba. The amount of toxin in all of its parts had sufficient concentrations to cause death. With this study, the amatoxin and phallotoxin concentrations in A. phalloides var. alba mushroom and in its parts have been revealed in detail for the first time. (C) 2013 Elsevier Ltd. All rights reserved.Öğe Amatoxin and phallotoxin concentration in Amanita phalloides spores and tissues(Sage Publications Inc, 2015) Kaya, Ertuğrul; Karahan, Selim; Bayram, Recep; Yaykaşlı, Kürşat Oğuz; Çolakoğlu, SerdarMost of the fatal cases of mushroom poisoning are caused by Amanita phalloides. The amount of toxin in mushroom varies according to climate and environmental conditions. The aim of this study is to measure -, -, and -amanitin with phalloidin and phallacidin toxin concentrations. Six pieces of A. phalloides mushrooms were gathered from a wooded area of Duzce, Turkey, on November 23, 2011. The mushrooms were broken into pieces as spores, mycelium, pileus, gills, stipe, and volva. -, -, and -Amanitin with phalloidin and phallacidin were analyzed using reversed-phase high-performance liquid chromatography. As a mobile phase, 50 mM ammonium acetate + acetonitrile (90 + 10, v/v) was used with a flow rate of 1 mL/min. C18 reverse phase column (150 x 4.6 mm; 5 mu m particle) was used. The least amount of -amanitin toxins was found at the mycelium. The other toxins found to be in the least amount turned out to be the ones at the spores. The maximum amounts of amatoxins and phallotoxin were found at gills and pileus, respectively. In this study, the amount of toxin in the spores of A. phalloides was published for the first time, and this study is pioneering to deal with the amount of toxin in mushrooms grown in Turkey.Öğe Düzce yöresinde yetişen Amanita Phalloides mantarındaki Alfa Amanitin düzeyinin Hplc yöntemiyle ölçümü(2012) Kaya, Ertuğrul; Karahan, Selim; Hancı, Mustafa; Yaykaşlı, Kürşat Oğuz; Sarıtaş, Ayhan; Bayram, RecepAmaç: Düzce ili sınırlarında 2010 yılında toplanan Amanita phalloides mantarındaki alfa amanitin toksin düzeyinin HPLC yöntemiyle ölçümü amaçlanmıştır. Yöntem: Bir mantar bütün olarak, diğeri ise parçalara ayrılarak ekstraksiyon yapılmıştır. Ölçümler HPLC cihazında 303 nm UV dalga boyu, 250x4,6 mm C18 5 µm partikül içeren kolon kullanılarak gerçekleştirilmiştir. Mobil faz olarak amonyum asetat + metanol + asetonitril (80+10+10, v/v/v) kullanılmış ve akış hızı 1 mL/dakikaya ayarlanmıştır. Sonuçlar 1 g kuru mantardaki toksin miktarı olarak verilmiştir. Bulgular: Bütün mantardaki alfa amanitin miktarı 4,806 mg (±0,033), şapkada 3,522 mg (±0,024), lamelde 5,318 mg (±0,056), halkada 0,903 mg (±0,004), sapta 2,577 mg (±0,037), kapçıkta 0,698 mg (±0,008) olarak ölçüldü. Sonuç: Düzce yöresinde yetişen Amanita phalloides mantarlarındaki alfa amanitin düzeyleri, başka bölgelerde yetişenlerden farklılık göstermektedir. Bulduğumuz sonuçlardan daha yüksek ve daha düşük seviyede toksin düzeyi ölçülmüş araştırmalar literatürde mevcuttur. Bu farklılığın etkenleri arasında iklim şartları yanında ekstraksiyon ve analiz yöntemlerindeki farklılıklar da rol oynayabilir.Öğe The measurement of alpha amanitin levels using hplc method in Amanita phalloides from Düzce province(Duzce University Medical School, 2012) Kaya, Ertu?rul; Karahan, Selim; Hanci, Mustafa; Yaykaşli, Kürşat O?uz; Saritaş, Ayhan; Bayram, Recep; Yilmaz, IsmailAim: The aim of this study is to measure the level of alpha amanitin toxin using HPLC method from Amanita phalloides mushroom collected in the province of Düzce in 2010. Method: One of the mushrooms as a whole body. The other one was extracted after seperated into parts. The measurement was done by HPLC using 303 nm UV wavelength and 250x4,6 mm C18 5 ?m particle included column. Ammonium acetate+methanol+acetonitrile (80+10+10, v/v/v) was used as a mobile phase, and the flow rate was set 1 ml/min. The results were given as a toxin quantity in 1 g dry mushroom. Results: The amount of alpha amanitin was measured as 4,806 mg (±0,033) in the whole body, 3,522 mg (±0,024) in the cap, 5,318 mg (±0,056) in the lamellar, 0,903 mg (±0,004) in the ring, 2,577 mg (±0,037) in the stipe, 0,698 mg (±0,008) in the volva. Conclusion: The level of alpha amanitin in Amanita phalloides from Duzce Province is differ from different countries. Higher and lower levels of toxin than our data obtained investigations are present in the literature. The reason of this differences might be several factors like extraction methods, analysis methods and environmental conditions. © 2012 Düzce Medical Journal.Öğe Production of high purity beta amanitin(Duzce University Medical School, 2012) Kaya, Ertu?rul; Yaykaşli, Kürşat O?uz; Karahan, Selim; Bayram, Recep; Saritaş, Ayhan; Yaykaşli, EmineObjective: Beta amanitin has been used in experiment and has been purified only around 90% purity using existing methods. In this study, it has been aimed to describe the method in order to produce high-purity beta amanitin using preparative HPLC. Methods: Amanita phalloides mushrooms hve been collected, extracted and purified 2 times using preparative HPLC. Validation of the toxin has been performed by comparison of retention time at HPLC and ultraviolet spectrum. Results: Beta amanitin was obtained with 91% (±2.36) purity after first purification process. Beta amanitin was obtained with 99,2% (±0.38) purity after second purification process. It seemed that purified toxin and standard were given maximum absorbance at 303 nm and minumum absorbans at 263 nm, and the structure of the spectrums for both was similar. Conclusion: Beta amanitin with >%99 purity can be produced by this method. © 2012 Düzce Medical Journal.Öğe Purification of High Purity Alpha Amanitin Using Preparative HPLC Method(Duzce Univ, 2012) Kaya, Ertugrul; Karahan, Selim; Yaykasli, Kursat Oguz; Bayram, Recep; Saritas, AyhanObjective: Alpha-amanitin is purified only around 90% purity using existing methods. In this study, it was aimed to describe the method in order to obtain high-purity alpha-amanitin using preparative HPLC. Methods: Amanita phalloides mushroom was collected, extracted and purified 2 times using preparative HPLC. Validation of the toxin was performed by comparison of retention time and ultraviolet spectrum at HPLC. Results: Alpha-amanitin was obtained with 93% (+/- 1.24) purity after first purification process. Alpha-amanitin was obtained with 99,8% (+/- 0.26) purity after second purification process. It was seemed that purified toxin and standard were given maximum absorbance at 303 nm and minimum absorbance at 263 nm, and the structure of the spectrums for both was similar. Conclusion: Alpha-amanitin with >99% purity can be obtained by this method at low cost.Öğe Yüksek saflıkta beta amanitin üretimi(2012) Kaya, Ertuğrul; Yaykaşlı, Kürşat Oğuz; Karahan, Selim; Bayram, Recep; Sarıtaş, Ayhan; Yaykaşlı, EmineAmaç: Beta amanitin nadiren bilimsel araştırmalarda kullanılmakta ve kullanımı gün geçtikçe artmaktadır. Piyasada bu ürün %90 saflıkta ticari olarak satılmaktadır. Bu araştırmada yüksek saflıkta beta amanitin üretme yöntemi tanımlanmıştır. Yöntem: Saflaştırma işlemi Amanita phalloides mantarlarından ekstraksiyonla yapılmıştır. Öncelikle bu mantarlar toplanmış, ekstrakte edilmiş, 2 defa preparatif HPLC ile saflaştırma işlemi uygulanmıştır. Toksinin karşılaştırması, analitik HPLC sisteminde tutulma zamanı ve ultraviyole spektrumu karşılaştırılması yöntemleriyle yapılmıştır. Bulgular: İlk saflaştırma sonucunda elde edilen beta amanitin saflık oranı %91(±2,36) olarak ölçülmüştür. İkinci saflaştırma sonucunda elde edilen beta amanitin saflık oranı %99,2(±0,38) olarak ölçülmüştür. Saflaştırma sonucu elde ettiğimiz toksin ile beta amanitin standardın UV spektrumlarında her ikisinde de 303 nm’de maksimum, 263 nm’de minimum absorbans verdiği ve spektrum yapısının aynı olduğu görülmüştür. Sonuç: Tanımladığımız bu yöntemle, >%99 saflıkta beta amanitinin üretilmesi mümkündür.
