Amasya, Akdağ'dan toplanan Galanthus fosteri Baker bitkisi üzerinde farmakognozik çalışmalar
Küçük Resim Yok
Tarih
2015
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, Türkiye'de yabani olarak yetişen bir Galanthus türü olan Galanthus fosteri Baker bitkisi çeşitli açılardan ele alınarak ayrıntılı olarak incelenmiştir. İlk çalışmamızda, Herba ve Bulbus Galanthi drogları üzerinde anatomik ve mikroskobik araştırmalar yapılmıştır. Drog örneklerinin kalite kontrol araştırmaları (kurutmada kayıp, total kül, sülfat külü ve hidroklorik asitte çözünmeyen kül) Avrupa Farmakopesi'ne göre gerçekleştirilmiştir ve sonuçlar % değerler olarak rapor edilmiştir. Buna ilaveten drogların total alkaloit miktarları titrimetrik bir yöntemle tayin edilmiştir. Galanthus genusunun önemli alkaloitleri arasında yer alan galantamin ve likorin, G. fosteri bitkisinden hazırlanan dört farklı drog örneğinde, yüksek basınçlı sıvı kromatografisi (HPLC) yöntemiyle kalitatif ve kantitatif olarak araştırılmıştır. Bunlara ek olarak, G. fosteri'den hazırlanan dört farklı drog örneğinin içermiş olduğu yirmi üç farklı alkaloit kapiler gaz kromatografisi-kütle spektrometresi (CGC-MS) yöntemiyle tespit edilmiştir. G. fosteri bitkisinin alkaloit içeriği GC-MS yöntemiyle ilk kez bu çalışmada rapor edilmiştir. Biyolojik aktivite rehberliğinde izolasyon ve yapı aydınlatma çalışmalarının yapıldığı araştırmalarda, oda sıcaklığında kurutulmuş ve toz edilmiş total bitkisel materyalden hazırlanan total alkaloit ekstresinin silika jel sütunda fraksiyonlandırılmasıyla elde edilen ana fraksiyonların in vitro Ellman metoduna dayalı mikroplaka deneyinden yararlanılarak asetilkolinesteraz ve butirilkolinesteraz inhibitör aktiviteleri tespit edilmiştir. Antikolinesteraz aktivite gösteren fraksiyonlar, preparatif sütun kromatografisi, preparatif İ.T.K. ve diğer saflaştırma yöntemlerinden faydalanılarak izolasyona tabi tutulmuşlardır. Elde edilen bileşiklerin yapıları spektroskopik yöntemler (UV, IR, CD, 1D ve 2D NMR, ESIMS, HR-ESIMS) kullanılarak aydınlatılmıştır. Bu çalışmaların sonucunda Galanthus fosteri Baker'dan, Amaryllidaceae alkaloitlerinin likorin alt grubundan (-)-galantin, (-)- inkartin, (-)-oksoinkartin ve (-)-9-O-metilpsödolikorin, hemantamin alt grubundan (+)-vittatin, (-)-3,11-O-diasetil-9-O-demetilmaritidin ve (+)-11-O-asetil-9-O-demetilmaritidin, galantamin alt grubundan (-)-galantamin, bunlara ilaveten hordenin ve O,N-dimetilnorbelladin adlı bileşikler izole edilmiş ve yapıları aydınlatılmıştır. Bunlardan (-)-oksoinkartin, (-)-3,11-O-diasetil-9-O-demetilmaritidin ve (+)-11-O-asetil-9-O-demetilmaritidin, ilk kez bu çalışmada izole edilmiştir. (-)-9-O-metilpsödolikorin ve O,N-dimetilnorbelladin genus bazında yeni bileşikler olup, (-)-galantin, (-)- inkartin ve (+)-vittatin ise tür seviyesinde yenidir. Ayrıca izole edilmiş olan her bir bileşiğin in vitro Ellman metoduna dayalı mikroplaka deneyi kullanılarak asetilkolinesteraz ve bütirilkolinesteraz inhibitör aktiviteleri çalışılmıştır. Bunlardan (-)-9-O-metilpsödolikorin, (-)-3,11-O-diasetil-9-O-demetilmaritidin, (+)-11-O-asetil-9-O-demetilmaritidin, hordenin ve O,N-dimetilnorbelladin anti-asetilkolinesteraz aktivite gösterirken, (-)-galantin, (-)- inkartin, (-)-9-O-metilpsödolikorin, (+)-vittatin, (+)-11-O-asetil-9-O-demetilmaritidin, hordenin ve O,N-dimetilnorbelladin ise anti-butirilkolinesteraz aktivite göstermiştir. Aktif olan tüm bileşiklerin IC50 değerleri hesaplanmıştır. Anahtar Kelimeler: Galanthus fosteri Baker; Kalite kontrol çalışmaları; Antikolinesteraz aktivite rehberliğinde izolasyon
In this study, Galanthus fosteri Baker, which is a wild-growing Galanthus species of Turkey, has been thoroughly investigated by several approaches. As the first study, the anatomical and microscobical investigations on Herba and Bulbous Galanthi have been carried out. The quality control investigations (loss on drying, total ash, sulphated ash and hydrochloric acid-insoluble ash) of drug specimens have been determined according to European Pharmacopoeia and the results were reported as percentage values. Moreover, the total alkaloidal contents of the drugs were determined by a titrimetric method. Four different drug specimens prepared from G. fosteri, were qualitatively and quantitatively analyzed for the two principle alkaloids of Galanthus genus, galanthamine and lycorine, by using High Pressure Liquid Chromatography (HPLC). Moreover,identification of twenty three alkaloids in four different drug specimens of G. fosteri were determined by capillary gas chromatography-mass spectrometry (CGC-MS). The alkaloid content of G. fosteri was reported for the first time by GC-MS in this study. In the studies where biological activity guided isolation and structure elucidation investigations are conducted, total alkaloidal extract prepared from plant material dried at room temperature and powdered, was fractionated on a column of silica gel and the obtained main fractions were subjected to the acetylcholinesterase and butrylcholinesterase inhibitor activity tests by using a microplate assay, based on in vitro Ellman method. Fractions which showed anticholinesterase activity, have been purified by preparative column chromatography, preparative T.L.C., and other purification methods. The structures of the compounds thus obtained were elucidated by using spectral techniques (UV, IR, CD, 1D and 2D NMR, ESIMS, HR-ESIMS). As a result of these studies, the Amaryllidaceae alkaloids (-)-galanthine, (-)- incartine, (-)-oxoincartine and (-)-9-O-methylpseudolycorine from the lycorine subgroup, (+)-vittatine, (-)-3,11-O-diacetyl-9-O-demethylmaritidine and (+)-11-O-acetyl-9-O-demethylmaritidine from the haemanthamine subgroup, (-)-galanthamine from the galanthamine subgroup, moreover hordenine and O,N-dimethylnorbelladine were isolated from Galanthus fosteri Baker and their structures were elucidated. Of these compounds, (-)-oxoincartine (-)-3,11-O-diacetyl-9-O-demethylmaritidine and (+)-11-O-acetyl-9-O-demethylmaritidine have been isolated for the first time in this study. The occurrence of (-)-9-O-methylpseudolycorine and O,N-dimethylnorbelladine are new at the genus level, (-)-galanthine, (-)- incartine and (+)-vittatine are new for the species. Additionally, acetylcholinesterase and butrylcholinesterase inhibitories activity of each compound were performed by using a microplate assay, based on in vitro Ellman method. (-)-9-O-methylpseudolycorine, (-)-3,11-O-diacetyl-9-O-demethylmaritidine and (+)-11-O-acetyl-9-O-demethylmaritidine, hordenine and O,N-dimethylnorbelladine showed anti-acetylcholinesterase activity; (-)-galanthine, (-)- incartine, (-)-9-O-methylpseudolycorine, (+)-vittatine, (+)-11-O-acetyl-9-O-demethylmaritidine, (-)-galanthamine, hordenine and O,N-dimethylnorbelladine displayed anti-butrylcholinesterase activity. The IC50 values of all the active compounds were calculated. Keywords: Galanthus fosteri Baker; Quality control investigations; Anticholinesterase activity guided isolation.
In this study, Galanthus fosteri Baker, which is a wild-growing Galanthus species of Turkey, has been thoroughly investigated by several approaches. As the first study, the anatomical and microscobical investigations on Herba and Bulbous Galanthi have been carried out. The quality control investigations (loss on drying, total ash, sulphated ash and hydrochloric acid-insoluble ash) of drug specimens have been determined according to European Pharmacopoeia and the results were reported as percentage values. Moreover, the total alkaloidal contents of the drugs were determined by a titrimetric method. Four different drug specimens prepared from G. fosteri, were qualitatively and quantitatively analyzed for the two principle alkaloids of Galanthus genus, galanthamine and lycorine, by using High Pressure Liquid Chromatography (HPLC). Moreover,identification of twenty three alkaloids in four different drug specimens of G. fosteri were determined by capillary gas chromatography-mass spectrometry (CGC-MS). The alkaloid content of G. fosteri was reported for the first time by GC-MS in this study. In the studies where biological activity guided isolation and structure elucidation investigations are conducted, total alkaloidal extract prepared from plant material dried at room temperature and powdered, was fractionated on a column of silica gel and the obtained main fractions were subjected to the acetylcholinesterase and butrylcholinesterase inhibitor activity tests by using a microplate assay, based on in vitro Ellman method. Fractions which showed anticholinesterase activity, have been purified by preparative column chromatography, preparative T.L.C., and other purification methods. The structures of the compounds thus obtained were elucidated by using spectral techniques (UV, IR, CD, 1D and 2D NMR, ESIMS, HR-ESIMS). As a result of these studies, the Amaryllidaceae alkaloids (-)-galanthine, (-)- incartine, (-)-oxoincartine and (-)-9-O-methylpseudolycorine from the lycorine subgroup, (+)-vittatine, (-)-3,11-O-diacetyl-9-O-demethylmaritidine and (+)-11-O-acetyl-9-O-demethylmaritidine from the haemanthamine subgroup, (-)-galanthamine from the galanthamine subgroup, moreover hordenine and O,N-dimethylnorbelladine were isolated from Galanthus fosteri Baker and their structures were elucidated. Of these compounds, (-)-oxoincartine (-)-3,11-O-diacetyl-9-O-demethylmaritidine and (+)-11-O-acetyl-9-O-demethylmaritidine have been isolated for the first time in this study. The occurrence of (-)-9-O-methylpseudolycorine and O,N-dimethylnorbelladine are new at the genus level, (-)-galanthine, (-)- incartine and (+)-vittatine are new for the species. Additionally, acetylcholinesterase and butrylcholinesterase inhibitories activity of each compound were performed by using a microplate assay, based on in vitro Ellman method. (-)-9-O-methylpseudolycorine, (-)-3,11-O-diacetyl-9-O-demethylmaritidine and (+)-11-O-acetyl-9-O-demethylmaritidine, hordenine and O,N-dimethylnorbelladine showed anti-acetylcholinesterase activity; (-)-galanthine, (-)- incartine, (-)-9-O-methylpseudolycorine, (+)-vittatine, (+)-11-O-acetyl-9-O-demethylmaritidine, (-)-galanthamine, hordenine and O,N-dimethylnorbelladine displayed anti-butrylcholinesterase activity. The IC50 values of all the active compounds were calculated. Keywords: Galanthus fosteri Baker; Quality control investigations; Anticholinesterase activity guided isolation.
Açıklama
Anahtar Kelimeler
Eczacılık ve Farmakoloji, Pharmacy and Pharmacology
