GC-MS ANALYSIS OF TURKISH HONEYDEW HONEYS PRODUCTION OF MARCHALINA HELLENICA ON PINUS BRUTIA ASLI ÖZKÖK1, KADRİYE SORKUN1, BEKİR SALİH2 [email protected] [email protected] [email protected] 1Hacettepe University Faculty of Sciences, Department of Biology, 06800, Beytepe-Ankara-Turkey 2Hacettepe University Faculty of Sciences, Department of Chemistry, 06800, Beytepe-Ankara-Turkey Abstract MATERIAL AND METHODS 78 honey samples were collected from ten areas of Muğla city in western Turkey. These Collection of Honey Samples points were Merkez, Milas, Ortaca, Köyceğiz, Marmaris, Fethiye, Yatağan, Bodrum, Ula 78 honey samples were collected from ten areas of the Muğla city in western and Datça where honeydew beekeeping is practiced extensively, from September to Turkey, Merkez, Milas, Ortaca, Köyceğiz, Marmaris, Fethiye, Yatağan, Bodrum, November in the years of 2005 and 2006. All honey samples were examined under the Ula and Datça where honeydew beekeeping is practiced extensively, from light microscope. The Number of Honeydew Element (NHE) and the Number of Total Pollen (NTP) in these samples were also investigated. The quality of the samples was September to November in the years of between 2004 and 2006. In this study, determined on the basis of the NHE/NTP ratio. Each honey sample having the NHE/NTP suitable apiaries were chosen from villages (Figure 7-9). It was important that the ratio higher than 4.5 was accepted as a high quality honeydew honey. Following villages are separated with enough distance and there are enough vegetation microscopic studies, 50 honey samples were determined as honeydew honey samples and differences, which also represent Muğla statistically. these samples were prepared for GC-MS analysis to screening the chemical composions. As a result of organic chemical substances quantified by GC-MS of 50 honeydew honeys, it was found that the main organic compound groups were mainly aliphatic and aromatic aldehydes, aromatic alcohols, hydrocarbons with linear chains, acid esters with linear chains, aromatic hydrocarbons, flavanones, aromatic acids, aromatic acid esters, aromatic amines and other minor chemical substances. Figure 11. The GC-MS Chromatography of one of the pine honey samples INTRODUCTION The origin of honeydew honey which includes pine honey as well is honeydew. DISCUSSION Honeydew is the rectal secretion with high sugar content of insects feeding on At this study generally aromatic aldehydes and aromatic alcohols have been identified at all honey samples. Radovic et al. (2001) also determined aromatic phloem juice (Zander and Koch, 1994). Insects provide for their food needs from dense sugar solution in phloem juice and after sufficient intake excrete the rest. Figure 7. Muğla-Datça-Sındı village Figure 8. Muğla-Milas-Pinar village aldehydes and aromatic alcohols at honey samples. Allissandrakis et al. (2007) Microscopic fungus and algae emerge on sticky sugary excreta which may then also found aromatic alcohols at honey samples. pass to honey (D’Albore, 1997). Bees carry this dense sugary substance to their Hydrocarbons with linear chains were found at hives and transform it into honey. Honey produced by bees this way is called 1,3,4,6,9,10,12,14,15,16,17,18,19,20,21,22,24,27,28,30,31,32,35,36,37,38,39,40,4 honeydew honey (Zander and Koch, 1994). 1,42,43,46 and 49 honey samples. Bentivenga et al. (2004) also determined The origin of pine honey produced in Turkey Pinus brutia Ten. and it is produced hydrocarbons with linear chains at honey and beeswax. by Marchalina hellenica (syn. Monophlebus hellenicus) which lives on it (Figure Aromatic hydrocarbons shows environmental pollution. These kind of components 1 and Figure 2). The habitat of this insect living on Pinus brutia Ten. consists especially indicate environmental pollution at water. (Bentivenga et al., 2004). It only of Greece and Turkey (Santas, 1979). In Turkey Marchalina hellenica has been determined aromatic hydrocarbons at 1,4,5,13,14,18,25,27,38,40,42,44 habitat covers Southern Marmara, Aegean and Western Mediterranean Regions honey samples. (Gürkan, 1989). Antioxidant activity is the ability and potential of honey in reducing oxidative Pine honey is produced in the world only in Turkey (90% of total amount) and in Figure 9. Muğla-Yatağan-Bencik village Figure 10. Muğla-Fethiye-Ören village reactions within the food systems and human health. Notably, these oxidative Greece (10% of total amount). In Turkey, a large part of honey produced by reactions can cause deleterious reactions in food products (e.g. lipid oxidation in Marchalina hellenica from Pinus brutia and marketed commonly as “pine Microscopic Analysis of Honey Samples meat, and enzymic browning in fruits and vegetables) and adverse health effects, honey” comes from Muğla region (Şahin, 2000) (Figure 3-5). For microscopic analysis of the honey samples which were collected from 33 such as chronic diseases and cancers (Gheldof and Engeseth, 2002). The villages in the Muğla city, methods by Moar (1985) and Sorkun (2008) were antioxidants that naturally occur in honey contribute to antioxidant capacity followed. Honey preparations were examined with a Nikon Eclipse E400 (Nicholls and Miraglio, 2003). It has been found flavanones at honey sample microscope. As a result of microscopic analysis of the honey samples, 50 honey numbers 4,6,8,9,13,17,20,21,22,24,26,27,28,29,30,33,34,35,39,43,45,46,47,48,50. samples were selected because of the fact that their NHE to NTP ratios were Sample 35 has the highest flavanone amount with %3,31. Shimoda et al. (1996), bigger than 4,5. These honeydew honeys were found to be appropriate for GC- also determined flavanones at honey samples. MS analysis study. Lidocaine, which is used local anesthetic component, was found at all honey samples. Yan and Yu-Hong, (2007) also was determined this component at honey GC-MS Analysis of Honey Samples samples. Chemical analysis of honey samples have been made by GC-MS according to This study showed that the 50 samples of Muğla Turkish Pine Honey contained a Barcarola et al, (1998); Radovic et al.,(2001); Soria et al.,(2003); Cuevas-Glory et lot of different components. Some of them good quality but some of them bad. So al.,(2007) methods. According to this method 10 gr honey was weighed to a glass it is need to make different experiments and studies. tube and 10 ml methanol was added. This mixture have been kept waiting until it ACKNOWLEDGEMENTS Figure 1. Pinus brutia and Marchalina Figure 2. Mature Marchalina hellenica was dry. After drying the mixture 10 ml ethanol was added and closed by lid. We are indebted to the TÜBİTAK for financial support, and also to MUĞLA ARI YETİŞTİRİCİLERİ hellenica in Muğla (Erlinghagen, 2001) This mixture was waited 24 hours. After 24 hours this mixture was depleted BİRLİĞİ and local people who helped us during our fieldwork. another tube and waited ethanol dry. 0,5 ml ethanol was added and mixed with REFERENCES pipet. 2 µl mixture was injected to GC-MS. A GC 6890N from Hewlett-Packard Alissandrakis, E., Tarantalis, P. A., Harizanis, P. C. 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