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T.C. Harran Üniversitesi Fen Bilimleri Enstitüsü T.C. HARRAN ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ YÜKSEK LİSANS TEZİ HELLEBORUS VESICARIUS (RANUNCULACEAE) BİTKİSİNİN İÇERDİĞİ UÇUCU YAĞLARIN TAYİNİ VE FARKLI EKSTRELERİNİN ANTİMİKROBİYEL AKTİVİTELERİ Tarık KARABULUT KİMYA ANABİLİM DALI ŞANLIURFA 2014 ..........................................................danışmanlığında,....................................’ın hazırladığı “...........................................” konulu bu çalışma ........./......../ ........ tarihinde aşağıdaki jüri tarafından oy birliği / oy çokluğu ile Harran Üniversitesi Fen Bilimleri Enstitüsü ............................................................................................ Anabilim Dalı’nda YÜKSEK LİSANS / DOKTORA TEZİ olarak kabul edilmiştir. İmza Danışman : Unvan Adı SOYADI ………………………………. Üye : Unvan Adı SOYADI ………………………………. Üye : Unvan Adı SOYADI ………………………………. Bu Tezin ........................ Anabilim Dalında Yapıldığını ve Enstitümüz Kurallarına Göre Düzenlendiğini Onaylarım Prof. Dr. ..................................... Enstitü Müdürü Bu Çalışma HÜBAK Tarafından Desteklenmiştir. Proje No: 12122 Not: Bu tezde kullanılan özgün ve başka kaynaktan yapılan bildirişlerin, çizelge, şekil ve fotoğrafların kaynak gösterilmeden kullanımı, 5846 sayılı Fikir ve Sanat Eserleri Kanunundaki hükümlere tabidir. ÖZET Yüksek Lisans Tezi HELLEBORUS VESICARIUS (RANUNCULACEAE) BİTKİSİNİN İÇERDİĞİ UÇUCU YAĞLARIN TAYİNİ VE FARKLI EKSTRELERİNİN ANTİMİKROBİYEL AKTİVİTELERİ Tarık KARABULUT Harran Üniversitesi Fen Bilimleri Enstitüsü Kimya Anabilim Dalı Danışman : Yrd. Doç. Dr. Zafer UYAR Yıl:2014, Sayfa:56 Bu çalışmada Helleborus vesicarius (ranunculaceae) bitkisi kimyasal ve mikrobiyolojik olmak üzere iki alanda incelendi. Türü belirlendikten sonra Helleborus vesicarius (ranunculaceae) bitkisi güneş ışığından uzak kurutuldu ve sonra öğütüldü. Daha sonra bitki bileşenleri metanol ile ekstrakte edilerek ham metanol ekstresi hazırlandı. Daha sonra bir dizi seçici katı-sıvı ve sıvı-sıvı ekstraksiyon kullanılarak ham metanol ekstresi polaritesine göre sırasıyla heksan, diklorometan, bütanol ve su fraksiyonlarına ayrıldı. Bu ekstrelerden uçucu ve esansiyel yağ asitlerinin zengin olduğu Heksan ekstresi metillenerek GC-MS analizi yapıldı ve elde edilen sonuçlar faydalı yağ asitleri açısından irdelendi. Ayrıca tüm ekstrelerin (metanol, heksan, diklorometan, bütanol ve su) Salmonella thyphimurium CCM 5445, Escherichia coli ATCC 25922, Escherichia coli 0157H7 gram-negatif bakterilerine ve Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis ATCC 12228, Enterococcus faecalis ATCC 29212, Enterococcus faecium DSM 13590 gram-pozitif bakterilerine karşı antibakteriyel aktiviteleri ve Candida albicans ATCC 10239 mayasına karşı antifungal aktiviteleri agar-disk difüzyon ve minimum inhibisyon konsantrasyonu tayin yöntemleriyle referans ilaçlarla karşılaştırılarak ve incelendi. ANAHTAR KELİMELER: Helleborus vesicarius (Ranunculaceae), Uçucu yağlar, Antimikrobiyel aktivite, Doğal Bileşikler, Biyolojik Aktivite. ii ABSTRACT MSc Thesis Tarık KARABULUT Harran University Graduate School of Natural and Applied Sciences Department of Chemistry Supervisor: Assistant Professor Zafer UYAR Year: 2014, Page: 56 In this study, Helleborus vesicarius (ranunculaceae) plant was evaluated in two areas including chemical and microbiological. After identified taxanomically, Helleborus vesicarius (ranunculaceae) plant was dried away from sunlight and then finely ground. Then the crude methanol extract was prepared by extraction of the plant components with methanol. Then crude methanol extract was separated into hexane, dichloromethane, butanol, and water fractions respectively based on increasing polarity using a series of selective solid-liquid and liquid-liquid extractions. Out of all extracts, Hexane extract, which is rich in essential fatty acids and volatile fatty acids, was methylated and its GC-MS analysis was performed and the results obtained were evaluated in terms of beneficial fatty acids. Also antibacterial activity against Salmonella thyphimurium CCM 5445, Escherichia coli ATCC 25922, Escherichia coli 0157H7 gram-negative bacteria and Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis ATCC 12228, Enterococcus faecalis ATCC 29212, Enterococcus faecium DSM 13590 gram-pozitive bacteria and antifungal activity against Candida albicans ATCC 10239 yeast of all extracts (methanol, hexane, dichloromethane, butanol and water) were examined by agar- disk diffusion assay and minimum inhibitory concentration assay compared with the reference drugs. KEY WORDS: Helleborus vesicarius (Ranunculaceae), Essential oils, Antimicrobial activity, Natural Compounds, Biological Activity. iii TEŞEKKÜR Danışmanım Yrd. Doç. Dr. Zafer UYAR’ a sonsuz sabrı, araştırmaya çalışmaya teşviği, yaklaşımı ve bu çalışma dışında dahi hayata dair azim ve kararlılıkta bana ilham olması sebebiyle şükranlarımı sunarım. Bu çalışmada bana tavsiyeleri ile desteklerini esirgemeyen Prof. Dr. Mustafa DEĞİRMENCİ ve Doç. Dr. Zülfiye İLTER’ e, ayrıca gösterdiği anlayıştan dolayı 4. Sınıf Emniyet Müdürü Rasim UZUN’ a teşekkürü bir borç bilirim. Laboratuar çalışmalarında yardımlarını esirgemeyen ve dostlukları ile bu çalışmada yanımda olan Fatma TUMBUL, Ayşe YILMAZ ve Ülkü ARSLAN’ a tüm yaptıkları için teşekkür ederim. Nesrin KARABULUT’ a canı gönülden desteği, ilgisi ve tükenmeyen sabrından dolayı her daim teşekkürü bir borç bilirim. iv İÇİNDEKİLER Sayfa No ÖZET ..................................................................................................................................................... ii ABSTRACT .......................................................................................................................................... iii TEŞEKKÜR .......................................................................................................................................... iii İÇİNDEKİLER ...................................................................................................................................... v SİMGELER DİZİNİ ........................................................................ Hata! Yer işareti tanımlanmamış. ŞEKİLLER DİZİNİ .............................................................................................................................. vii ÇİZELGELER DİZİNİ ........................................................................................................................ vii 1.GİRİŞ .................................................................................................................................................. 1 1.1. Botanik Kısım ......................................................................................................................... 1 1.2. Kimyasal Kısım ...................................................................................................................... 7 1.3. Biyolojik Kısım .................................................................................................................... 10 1.3.1. Antimikrobiyel aktivite çalışmaları ..................................................................... 10 2. ÖNCEKİ ÇALIŞMALAR ................................................................................................................ 16 3. MATERYAL ve YÖNTEM ............................................................................................................. 18 3.1. Genel .................................................................................................................................... 18 3.2.Bitki Materyali ...................................................................................................................... 18 3.3.Ekstrelerin Hazırlanması ....................................................................................................... 19 3.4.Heksan Ekdtrelerinin GC-MS Analizi ................................................................................... 20 3.5.Mikrobiyoloj,Analiz: Antimikrobiyel Aktivite...................................................................... 21 3.5.1.Disk difüzyon yöntemi ......................................................................................... 22 3.5.2.Minimum inhibitor konsantrasyonu (MİK) ......................................................... 22 4.ARAŞTIRMA BULGULARI veTARTIŞMA .................................................................................. 24 4.1.Heksan Ekstresinin GC-MS Analizi ...................................................................................... 24 4.2.Mikrobiyolojik Analiz: Antimikrobiyel Aktivite .................................................................. 41 4.2.1.Disk difüzyon testi analizi .................................................................................... 42 5.SONUÇLAR ve ÖNERİLER ............................................................................................................ 45 KAYNAKLAR .................................................................................................................................... 46 ÖZGEÇMİŞ ......................................................................................................................................... 55 v SİMGELER DİZİNİ BuOH Bütanol CH2Cl2 Diklorometan DMSO Dimetil sülfoksit KOH Potasyum hidroksit MeOH Metanol vi ŞEKİLLER DİZİNİ Sayfa No Şekil 1.1. Helleborus vesicarius (Ranunculaceae) bitkisinin Türkiye’de yetiştiği bölgeler. ................... 6 Şekil 1.3a. Bakterilerin Sınıflandırılması .............................................................................................
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