Mantarlarda nükleik asit bileşiklerinin sıvı kromatografisi yöntemi ile tayini
Küçük Resim Yok
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Mantarlardaki dikkat çekici nükleosit rezervuarı nedeniyle, son zamanlarda bunların dağılımının nitel ve nicel olarak belirlenmesi önem kazanmıştır. Bu çalışmada Tekirdağ yöresinden toplanan yenilebilir mantar türlerindeki bazı nükleosit ve nükleobazların HPLC sistemi ile analiz edilmesi amaçlanmıştır. Materyal ve Metot: Analizi yapılan mantar türleri sırasıyla Trüf (Tuber borchii), Trüf (Tuber aestivum), Borazan (Craterellus fallax), Kuzu Göbeği (Morchella esculenta), Porçini (Boletus edulis), Sığır Dili (Hydnum repandum), Sarı Kız (Cantharellus cibarius) ve Kanlıca (Lactarius Salmonicolor) mantarlarıdır. Mantarlar liyofilize edildikten sonra -20°C'de muhafaza edilmişlerdir. Mantarda bulunan nükleotidler ters faz sıvı kromatografisi yöntemi ile analiz edilmiştir. Bileşiklerin ayrılması gradient elüsyonla SWEA C18 kolonu (4.6 mm x 250 mm, 5µm) ile gerçekleştirilmiştir. A çözücüsü olarak pH'sı 3.7 olan MeOH: 50 mM sodyum fosfat tamponu (1:99, h/h) karışımı; B çözücüsü olarak pH'sı 6.2 olan MeOH: 50 mM sodyum fosfat tamponu (20:80) kullanılmıştır. İç standart 3-metil ksantindir. Yöntem valide edilmiştir. Liyofilize edilen mantar örneklerini ekstrakte etmek için ses dalgaları destekli sıvı ekstraksiyonu ve basınçlı sıvı ekstraksiyonu olmak üzere iki farklı ekstraksiyon yöntemi ile kullanılmıştır. Ekstraksiyon çözücüsü olarak deiyonize su kullanılmıştır. Sıcaklık, zaman gibi parametreler optimize edilmiştir. Bulgular: 14 nükleotit 25 dakika sürede 254 nm dalgaboyunda eş zamanlı olarak ayrılmıştır. Ses dalgaları ekstraksiyon yönteminin daha etkin olduğu saptanmıştır. Bunun sebebi basınçlı sıvı ekstraksiyonu sisteminin kapalı bir sistem olması ve matriks çözücü etkileşiminin yetersiz kalması olabilir. Çalışılan yenilebilir mantar türleri arasında en yüksek nükleotit miktarına sahip olan tür ün Kuzu Göbeği olduğu bulunmuştur. Sonuç: Çalışılan tüm mantarlarda en fazla bulunan bileşiklerin adenosin, guanosin ve üridin olduğu tespit edilmiştir. Anahtar kelimeler: HPLC-DAD, nükleobaz, nükleosit, yenilebilir mantarlar, ses dalgaları destekli sıvı ekstraksiyonu
Aim: The remarkable reservoir of nucleotides in mushroom has led to a recent interest in the qualitative and quantitative determination of their distribution. In the present research, it was intended to analyse some nucleosides and nucleobases in edible mushroom species collected from Tekirdağ region by HPLC system. Material and Method: The analyzed mushroom species are Truffle (Tuber borchii), Truffle (Tuber aestivum), Black Trumpet (Craterellus fallax), Lamb Belly (Morchella esculenta), Porcini (Boletus edulis), Sığır Dili (Hydnum repandum), Chanterelle (Cantharellus cibarius) and Kanlıca (Lactarius salmonicolor) mushrooms, respectively. After the mushrooms were lyophilized, they were stored at -20°C. The nucleotides in the mushrooms were analyzed by the reverse phase liquid chromatography method. Separation of compounds was accomplished by gradient elution on a SWEA C18 column (4.6 mm x 250 mm, 5µm). A mixture of MeOH: 50 mM sodium phosphate buffer (1:99, v/v) at pH 3.7 as solvent A; MeOH: 50 mM sodium phosphate buffer (20:80) at pH 6.2 was used as solvent B. The internal standard is 3-methyl xanthine. The method has been validated. Two different extraction methods were used to extract the lyophilized mushroom samples, namely, sonication-assisted liquid extraction and pressurized liquid extraction. Deionized water was used as the extraction solvent. Parameters such as temperature and time have been optimized. Results: The 14 nucleotides were simultaneously separated at a wavelength of 254 nm in 25 minutes. Sonication-assisted liquid extraction method has been found to be more effective. This is because the pressurized liquid extraction system is a closed system and matrix-solvent interaction is insufficient. Among the edible mushroom species studied, Lamb's Belly was found to have the highest nucleotide content. Conclusion: Adenosine, guanosine and uridine were found to be the most abundant compounds in all mushrooms studied. Keywords: HPLC-DAD, nucleobase, nucleoside, edible mushrooms, sound wave assisted liquid extraction
Aim: The remarkable reservoir of nucleotides in mushroom has led to a recent interest in the qualitative and quantitative determination of their distribution. In the present research, it was intended to analyse some nucleosides and nucleobases in edible mushroom species collected from Tekirdağ region by HPLC system. Material and Method: The analyzed mushroom species are Truffle (Tuber borchii), Truffle (Tuber aestivum), Black Trumpet (Craterellus fallax), Lamb Belly (Morchella esculenta), Porcini (Boletus edulis), Sığır Dili (Hydnum repandum), Chanterelle (Cantharellus cibarius) and Kanlıca (Lactarius salmonicolor) mushrooms, respectively. After the mushrooms were lyophilized, they were stored at -20°C. The nucleotides in the mushrooms were analyzed by the reverse phase liquid chromatography method. Separation of compounds was accomplished by gradient elution on a SWEA C18 column (4.6 mm x 250 mm, 5µm). A mixture of MeOH: 50 mM sodium phosphate buffer (1:99, v/v) at pH 3.7 as solvent A; MeOH: 50 mM sodium phosphate buffer (20:80) at pH 6.2 was used as solvent B. The internal standard is 3-methyl xanthine. The method has been validated. Two different extraction methods were used to extract the lyophilized mushroom samples, namely, sonication-assisted liquid extraction and pressurized liquid extraction. Deionized water was used as the extraction solvent. Parameters such as temperature and time have been optimized. Results: The 14 nucleotides were simultaneously separated at a wavelength of 254 nm in 25 minutes. Sonication-assisted liquid extraction method has been found to be more effective. This is because the pressurized liquid extraction system is a closed system and matrix-solvent interaction is insufficient. Among the edible mushroom species studied, Lamb's Belly was found to have the highest nucleotide content. Conclusion: Adenosine, guanosine and uridine were found to be the most abundant compounds in all mushrooms studied. Keywords: HPLC-DAD, nucleobase, nucleoside, edible mushrooms, sound wave assisted liquid extraction
Açıklama
Anahtar Kelimeler
Eczacılık ve Farmakoloji, Pharmacy and Pharmacology
