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Tez Yazım Klavuz Ve Şablonu T.C. PAMUKKALE ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ BİYOLOJİ ANABİLİM DALI FARKLI GEOFİT CİNSLERİNDEN SEÇİLMİŞ TAKSON EKSTRAKTLARININ BİYOLOJİK AKTİVİTELERİ ÜZERİNE ARAŞTIRMALAR DOKTORA TEZİ MURAT TURAN DENİZLİ, TEMMUZ 2020 T.C. PAMUKKALE ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ BİYOLOJİ ANABİLİM DALI FARKLI GEOFİT CİNSLERİNDEN SEÇİLMİŞ TAKSON EKSTRAKTLARININ BİYOLOJİK AKTİVİTELERİ ÜZERİNE ARAŞTIRMALAR DOKTORA TEZİ MURAT TURAN DENİZLİ, TEMMUZ 2020 Bu tez çalışması Pamukkale Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından 2019FEBE001 nolu proje ile desteklenmiştir. Bu tezin tasarımı, hazırlanması, yürütülmesi, araştırmalarının yapılması ve bulgularının analizlerinde bilimsel etiğe ve akademik kurallara özenle riayet edildiğini; bu çalışmanın doğrudan birincil ürünü olmayan bulguların, verilerin ve materyallerin bilimsel etiğe uygun olarak kaynak gösterildiğini ve alıntı yapılan çalışmalara atfedildiğine beyan ederim. Murat TURAN ÖZET FARKLI GEOFİT CİNSLERİNDEN SEÇİLMİŞ TAKSON EKSTRAKTLARININ BİYOLOJİK AKTİVİTELERİ ÜZERİNE ARAŞTIRMALAR DOKTORA TEZİ MURAT TURAN PAMUKKALE ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ BİYOLOJİ ANABİLİM DALI (TEZ DANIŞMANI: PROF. DR. RAMAZAN MAMMADOV) DENİZLİ, TEMMUZ 2020 Bu çalışmada, Arum rupicola var. virescens (Stapf) P.C. Boyce, Cyclamen cilicium Boiss. & Heldr, Gagea bohemica (Zauschn.) Schult. & Schult.f., Narcissus tazetta L. subsp. tazetta, Paeonia kesrouanensis J. Thiébaut türlerinin yer üstü ve yer altı kısımlarının antioksidan aktiviteleri, total fenolik, flavonoid ve tanen madde miktar tayinleri, UPLC-ESI-MS/MS ile fenolik bileşenlerin analizi, asetilkolinesteraz ve tirozinaz enzimleri inhibisyonu, bu türlerin Artemia salina L. üzerindeki toksik etkileri, Musca domestica L. (ev sineği) ve Culex pipiens L. (sivrisinek) üzerindeki larvasidal etkileri incelenmiştir. Su, metanol ve aseton çözücüleri kullanarak elde edilen ekstraktların antioksidan aktivitelerini belirlemek için DPPH, ABTS, β-karoten-linoleik asit yöntemi, fosfomolibdenyum yöntemi, metal şelatlama, FRAP, CUPRAC, ve miktar tayinleri için toplam fenolik, flavonoid ve tanen madde miktarları deneyleri yapılmıştır. İçeriğindeki fenolik bileşenlerin belirlenmesi için UPLC-ESI-MS/MS yöntemi kullanılmıştır. Sulu ekstraktların A. salina üzerinde toksik etki deneyi, M. domestica ve Cx. pipiens üzerinde larvasidal etki deneyleri yapılmıştır. Antioksidan aktivite deneylerinde en yüksek aktivite P. kesrouanensis yer üstü kısmında % 68.52 ± 0.12 (0.06 ± 0.07 mg/mL, IC50) gözlenirken, en düşük aktivite G. bohemica türünün yer altı kısmında % 6.80 ± 0.16 (6.60 ± 0.002 mg/mL, IC50) gözlenmiştir. Toplam fenolik ve flavonoid madde miktarı tayinlerinde en fazla P. kesrouanensis yer üstü kısmında sırasıyla 11.62 ± 0.11 mg GAE/g ekstrakt, 15.90 ± 0.13 mg QE/g ekstrakt değeri ile gözlenmiştir. M. domestica ve Cx. pipiens üzerinde yapılan larvasidal etki deneyinde P. kesrouanensis yer üstü kısmı en güçlü etkiyi göstermiştir. A. salina’ya karşı toksik etki deneyinde P. kesrouanensis daha çok toksik olduğu (% 44.44 ± 2.78) tespit edilmiştir. ANAHTAR KELİMELER: Antioksidan, Asetilkolinesteraz enzimi, Tirozinaz enzimi, UPLC-ESI-MS/MS, Musca domestica, Culex pipiens i ABSTRACT INVESTIGATIONS ON THE BIOLOGICAL ACTIVITIES OF SELECTED TAXON EXTRACTS FROM DIFFERENT GEOPHYTE GENERA PH.D THESIS MURAT TURAN PAMUKKALE UNIVERSITY INSTITUTE OF SCIENCE BIOLOGY (SUPERVISOR: PROF. DR. RAMAZAN MAMMADOV) DENİZLİ, JULY 2020 In this study, antioxidant activities, total phenolic, flavonoid and tannin content determination, analysis of phenolic components with UPLC-ESI-MS/MS, inhibition of acetylcholinesterase and tyrosinase enzymes, toxic effects against Artemia salina L., larvicidal effects against Musca domestica L. (house fly) and Culex pipiens L. (mosquito) were investigated from fresh and underground parts of Arum rupicola var. virescens (Stapf) P.C. Boyce, Cyclamen cilicium Boiss. & Heldr, Gagea bohemica (Zauschn.) Schult. & Schult.f., Narcissus tazetta L. subsp. tazetta, Paeonia kesrouanensis J. Thiébaut taxons. DPPH, ABTS, β- carotene-linoleic acid method, phosphomolybdenum method, metal chelation, FRAP, CUPRAC and total total phenolic, flavonoid and tannin content tests was performed to determine antioxidant activities and determination of contents of extracts obtained using water, methanol and acetone solvents. The UPLC-ESI- MS/MS method was used to determine the phenolic compounds. Toxic effects of aqueous extracts on A. salina, larvicidal effects on M. domestica and Cx. pipiens were performed. In antioxidant activity assays, the highest activity was observed in fresh part of P. kesrouanensis % 68.52 ± 0.12 (0.06 ± 0.07 mg/mL, IC50), while the lowest activity was observed in underground part of G. bohemica % 6.80 ± 0.16 (6.60 ± 0.002 mg/mL, IC50). In the determination of total phenolic and flavonoid, fresh part of P. kesrouanensis was the most abundant with 11.62 ± 0.11 mg GAE/g extract, 15.90 ± 0.13 mg QE/g extract, separately. Fresh part of P. kesrouanensis showed the strongest effect in the larvicidal effect test against M. domestica and Cx. pipiens. In the toxic effect assay against A. salina, P. kesrouanensis was found to be more toxic (% 44.44 ± 2.78). KEYWORDS: Antioxidant, Acetylcholinesterase enzyme, Tyrosinase enzyme, UPLC-ESI-MS/MS, Musca domestica, Culex pipiens ii İÇİNDEKİLER Sayfa ÖZET ................................................................................................................... i ABSTRACT ....................................................................................................... ii İÇİNDEKİLER ................................................................................................ iii ŞEKİL LİSTESİ .............................................................................................. vii TABLO LİSTESİ ............................................................................................. ix SEMBOL LİSTESİ .......................................................................................... xi ÖNSÖZ ............................................................................................................. xii 1. GİRİŞ ............................................................................................................. 1 1.1 Tez Kapsamında Çalışılan Bitkiler Hakkında Botanik Bilgiler ........... 3 1.1.1 Araceae Familyası .......................................................................... 4 1.1.1.1 Arum Cinsi ................................................................................ 4 1.1.1.1.1 Arum rupicola var. virescens Türü ve Taksonomik Sınıflandırılması ................................................................................... 5 1.1.2 Primulaceae Familyası ................................................................... 6 1.1.2.1 Cyclamen Cinsi ......................................................................... 6 1.1.2.1.1 Cyclamen cilicium Türü ve Taksonomik Sınıflandırılması 7 1.1.3 Liliaceae Familyası ........................................................................ 8 1.1.3.1 Gagea Cinsi ............................................................................... 8 1.1.3.1.1 Gagea bohemica Türü ve Taksonomik Sınıflandırılması ... 9 1.1.4 Amaryllidaceae Familyası ........................................................... 10 1.1.4.1 Narcissus Cinsi ....................................................................... 10 1.1.4.1.1 Narcissus tazetta subp. tazetta Türü ve Taksonomik Sınıflandırılması ................................................................................. 11 1.1.5 Paeoniaceae Familyası ................................................................. 12 1.1.5.1 Paeonia Cinsi .......................................................................... 12 1.1.5.1.1 Paeonia kesrouanensis Türü ve Taksonomik Sınıflandırılması ................................................................................. 13 1.2 Serbest Radikaller ............................................................................... 14 1.2.1 Serbest Radikal Bileşiklerinin Kaynakları ................................... 15 1.2.1.1 Endojen Kaynaklar .................................................................. 16 1.2.1.1.1 Mitokondriyal Elektron Transportu .................................. 16 1.2.1.1.2 Otooksidasyon .................................................................. 16 1.2.1.1.3 Enzimatik Oksidasyon ...................................................... 16 1.2.1.1.4 Geçiş Metalleri .................................................................. 16 1.2.1.1.5 Fagositoz ve Solunum Sırasındaki Parçalanmalar ............ 17 1.2.1.2 Eksojen Kaynaklar .................................................................. 17 1.2.2 Serbest Oksijen Türleri ................................................................ 17 ∙- 1.2.2.1 Süperoksit (O2 ) Radikali ........................................................ 18 1.2.2.2 Hidrojen Peroksit (H2O2) ........................................................ 18 1.2.2.3 Hidroksil (OH∙) radikali .......................................................... 19 1.2.2.4 Hipokloröz Asit (HOCl) .......................................................... 19 1 1.2.2.5 Singlet oksijen ( O2) ................................................................ 19 iii 1.2.3 Serbest Radikallerin Etkileri ........................................................ 19 1.2.3.1 Serbest Radikallerin Lipitlere Etkileri .................................... 20 1.2.3.2 Serbest Radikallerin Proteinlere
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