International Symposium on Authenticity and Quality of Bee Products
 and
the 2nd World Symposium on Honeydew Honey

A. ORAL PRESENTATIONS

1. IMPROVEMENTS OF PHADEBAS^® FOR THE PHADEBAS^® HONEY DIASTASE TEST

Eric Jonsson

Image Life Science

Product Manager

ej@magle.se

The Phadebas^® method is one of two methods recommended by the International Honey Commission for determination of Diastase activity in honey. Phadebas^® Amylase test, used for the method, is manufactured and sold by Magle Life Sciences. Diastase activity given by this method is based on fixed equations determined through a study comparing the Schade method with one specific batch of Phadebas^® tablets[1]. However, since there may be differences in reactivity between Phadebas^® batches, using predetermined equations will lead to varying results. For other Phadebas^® applications this is not a problem as quantification is based on the batch specific standard curve supplied with each kit. Adding to this problem, the change of dye in the Phadebas reagent also caused a change in the assay sensitivity.

To meet the demand from honey analysing labs, Magle has developed a product that mimics the behaviour of the batch used in the original study. The product, which will be released later this year under the tradename Phadebas^® Honey Diastase Test (PHDT), has been developed in accordance to Magle’s quality management system (ISO 13485 and pharmaceutical GMP). The result is a product with very narrow specifications wherein all batches of PHDT will behave alike and adhere to the set of equations in the IHC guidelines. PHDT will be launched as an improved and cost effective alternative to the Schade method with; better precision, fewer reagents, fewer steps and less labour time. This presentation will discuss the quality assurance program as well as present the new product and the company.

2. BIPEA - ORGANISER OF PROFICIENCY TESTING SCHEMES FOR HONEY

Bruno Berken
Directeur / Director BIPEA
6/14 Avenue Louis Roche -
92230 Gennevilliers

FRANCE

Created in 1970, Bipea is an international non-profit organization of testing laboratories. Bipea regularly organizes 56 Proficiency Testing Schemes (PTS) in the following fields of interest:

-         Cereals, oilseeds and animal feed

-         Food industry and beverages

-         Environmental issues (quality of water) and contaminants

-         Cosmetics and fragrances.

Bipea's missions are:

-         To organize interlaboratory comparisons tests for the laboratories to check their performance.

-         To assist laboratories in the management, maintenance and improvement of their performances.

For a laboratory, the participation in a P.T. Scheme is a tool for self-checking and allows:

-         To control the accuracy of a result of the laboratory analysis

-         To quantify the bias of the laboratory.

Since 2001, Bipea organizes a Proficiency testing scheme on honey: five times per year, the laboratories receive homogeneous samples of honey. Thus the laboratories are given the opportunity to compare themselves, anonymously, for various physico-chemical parameters, but also for the microscopic examinations (floral and geographic origins), the sensorial analysis and the honey identification. With 30 international participants, located all around the world, we adapt our program to the different kinds of products that can be found in these different countries in order to stick to their needs.

3.PARTICIPATION RESULTS OF APICULTURE DIVISION LABORATORY IN PULAWY (POLAND) IN PROFICIENCY TEST FOR HONEY ORGANISED BY BIPEA

 Ewa Waś*, Helena Rybak-Chmielewska, Teresa Szczęsna, Katarzyna Kachaniuk

Research Institute of Pomology and Floriculture Apiculture Division, Department of Bee Products, Kazimierska 2 Str., 24-100 Puławy,

*e-mail: ewa.was@man.pulawy.pl

POLAND

 Bee Products Quality Testing Laboratory has participated in proficiency test organised by Bipea (Bureau InterProfessionnel d'Etude Analytique) since 2004. In 2008/2009 campagne the laboaratory participated in all five rounds and received satisfactory results (except sucrose content in September 2008) for the accreditated methods of honey quality determination: moisture (refractometric method), pH and free acidity (potentiometric method), electrical conductivity, 5-hydroxymethylfurfural (with HPLC and after Winkler), diastase activity (with Phadebas), sugar content (with HPLC) and pollen analysis.

Table 1. Results estimation obtained by Bee Products Quality Testing Laboratory in interlaboratory comparisons in 2008/2009 campagne.

*  z-score = (X_lab – X_tot)/S, (x_lab – laboratory result; X_tot – mean from all quantified results; S – standard deviation with all quantified results.  Interpretation criteria of z-score: z │≤ 2│– satisfactory result; 2<│z│≤ 3 – doubtful result; │z│ > 3 – unsatisfactory result. 

Participation in interlaboratory comparisons is one of the few way used in Bee Products Quality Testing Laboratory for quality control and allows maintaining a high level of proficiency of the accreditated methods there.

4. A REVIEW OF THE BIOLOGICAL AND FUNCTIONAL PROPERTIES OF HONEYDEW HONEYS IN COMPARISON TO BLOSSOM HONEYS

Stefan Bogdanov, Bee Product Science,

SWITZERLAND INFO@BEE-HEXAGON.NET 

Honeydew honeys and blossom honeys differ in regards to different physico-chemical properties. In comparison to blossom honeys honeydew honeys have generally a darker colour, higher electrical conductivity, higher pH, lower amount of fructose and glucose linked to a higher amount of oligosaccharides, and higher concentration of phenolics.

In one study the glycemic index of honeydew honey was found to be higher that that of different blossom honeys (Berg et al. 2008).

The prebiotic effect of honeydew is expected to be higher than that of blossom honeys due to its higher content of oligosaccharides, thought to be the principle prebiotic components (Sanz et al. 2005).

The antioxidant effect of honey depends mainly on the phenolic conent of honey. Dark honeys, and also honeydew honeys, have a higher content of phenolics and their antioxidant activity is higher than that of light blossom honeys. Oak honey, the darkest honeydew honey seems to have the highest antioxidant activity of all honeydew honeys (Bobis et al. 2008).

The antibacterial activity depends on the botanical origin of honey. Honeydew honeys have been reported to have generally higher antibacterial activity than light blossom honeys (Molan, 1991). This difference seems to be true both for the peroxide and non-peroxide antibacterial system.

On the other hand, the health enhancing properties of honeydew and blossom honeys have not been the object of comparative clinical studies.

5. DIFFERENTIATION OF HONEYDEW HONEY AND BLOSSOM HONEY ACCORDING TO THEIR SUGAR PROFILE

 Bentabol Manzanares, A. ^*1;  Hernández García, Z. ¹, Rodríguez Galdón, B.²; Rodríguez Rodríguez E.²; and Díaz Romero, C.²

¹ The House of the Honey, Cabildo de Tenerife, 38630 El Sauzal, Santa Cruz de Tenerife, Spàin

² Food Science and Nutrition Area. Department of Analytical Chemistry, Food Science and Nutrition. University of La Laguna. Avda. Astrofísico Sánchez s/n, 38201, La Laguna, Santa Cruz de Tenerife,

 SPAIN

In the present study we determined HPLC- differential refractive index (DRI) detector) the concentrations of sugars in multifloral and honeydew honeys produced in Tenerife in order to establish differences between them. In both cases fructose, glucose, sucrose, maltose, trehalose, turanose, isomaltose and melezitose/erlose were identified and quantified. The peaks corresponding to the monosaccharides glucose and fructose, were well defined and these monosaccharides were clearly the predominant sugars, which indicates that all the samples analyzed are genuine honeys. All honey samples had a content of sucrose lower than 5 g/100g and the value of glucose plus fructose was higher than 60 g/100g, which are the limit values for honeys allowed by European Community Directive. Similar chromatograms but with peaks of different heights for individual sugars were obtained for both types of honeys considered. The suspected honeydew honeys had significantly higher mean values of thehalose, turanose and isomaltose, and lower of total sugars, glucose, sucrose and erlose + melecitose, than the blossom honeys. No significant differences were found in fructose and maltose contents. Both types of honeys presented significant differences between all the carbohydrate ratios such as fructose/glucose, maltose/isomaltose, maltose/turanose and sucrose/turanose. Therefore, the profile of sugars can contribute in the differentiation between honeydew and blossom honeys.

6. FIRST ESTIMATIONS FOR THE CHESNUT HONEYDEW TYPIFICATION IN SPAIN

 León-Ruiz, V.*; González-Porto, A.V.*; Rodrigo-Medina, S*; Escuredo O.**, Chouza M.** & Seijo C.**

*Centro Agrario de Marchamalo (Centro Apícola Regional) de la Junta de Comunidades de Castilla-La Mancha. 19180 Marchamalo (Guadalajara)

** Dpt. Vegetal Biology and Soil Sciences. Faculty of Sciences. University of Vigo. As Lagoas. 32004 Ourense.

SPAIN

Castanea sativa Miller presents a great interest for honey production in Northern and some regions of Central Spain. The bees obtain from it an important quantity of pollen, nectar and frequently honeydew. 

 Chestnut honeys present some sensorial characteristics very appreciated by a good number of consumers, as its dark amber colour sometimes with reddish tone; its strong, durable and woody olfative perception and its sweet-salted flavor with certain bitter aftertaste. Astringency is other common mouth perception. Some of these aspects are common to honeydew honey. The same occurs with some physico-chemical characteristics: high electrical conductivity, color, pH, etc.

 The aim of this work is to analyze honeys with similar sensorial attributes and differentiate their source: nectar or honeydew. We study spanish honeys from different geographical origin, where Chesnut honey is typical, like Galicia, Villuercas-Ibores (Extremadura) and Montes de Toledo (Castilla-La Mancha).

 Taking into account physico-chemical parameters as: electrical conductivity, HMF, diastase, pH, free acidity, moisture, and other like sugars or palynological analysis.   

Main differences between floral honey and honeydew were observed in electrical conductivity and sugars contents. 

This work has been financed by Junta de Comunidades de Castilla-La Mancha ( proyect PAI09-0018-9267 and predoctoral scholarship).

7. COMPARISON OF VOLATILE AND SEMI-VOLATILE COMPOUNDS ISOLATED FROM SIX TYPES OF GREEK MONOFLORAL HONEYS

 Tananaki Ch., Thrasyvoulou A.
 Laboratory of Apiculture-Sericulture,

School of Agriculture, Aristotle University of Thessaloniki, ,

etananaki@agro.auth.gr
GREECE

Volatile compounds contribute to consumer's preferences and also can be used as a useful tool for the determination of honey’s botanic origin. In this work  the volatile profiles of six types of greek monofloral honeys were determined using a Purge & Trap – Gas Chromatography – Mass Spectrometer system. Totally 166 compounds were detected in all samples, 81 of them in pine,  95 in oak,  83 in fir, 72 in knotgrass, 85 in erica and 71 in thorn honeys. Twenty eight substances were detected in every sample independently of their origin, while most important were octane, nonane, benzaldehyde, phenylacetadehyde, nonanal and decanal. Totally 86 substances were isolated in all samples at least at one type of honey and these compounds used to form characteristic combinations for each kind of honey and for the statistical analysis as well. Some substances isolated only from all samples with the same origin, like 2,3-dihydro-1,8-cineol and α-calacorene from oak honey, methyl-benzyl-methanol from fir honey, 2-hydroxy-isophorone and α-ionone from erica honey. The characteristic compounds and combinations can be used together with the physicochemical microscopic and organoleptic characteristics in order to determine the botanical origin of the studied Greek honeys.

8. COMPOSITION AND PROPERTIES OF TWO UNIQUE HONEYS FROM THE BOREAL CONIFEROUS FOREST ZONE: FIREWEED AND RASPBERRY HONEY

Anneli Salonen, Riitta Julkunen-Tiitto

Natural Product Research Laboratories, Department of Biology, University of Eastern Finland, Joensuu,

FINLAND

 The aim of this work was to characterize the composition and properties of two unique and aromatic Finnish unifloral honeys: fireweed and raspberry honey. Honey from fireweed (Epilobium angustifolium L.) and from raspberry (Rubus idaeus L.) is collected especially in Eastern, Central and Northern Finland. Due to silviculture clear cutting practices these plants have become more common in Finnish forests. Organoleptic, physico-chemical and melissopalynological analyses were conducted according to the method descriptions of the International Honey Committee. These unifloral honeys are mild in flavour and odour, have low electrical conductivity and are rich in fructose. Until now the only way to distinguish between these unifloral honeys have been sensorial analyses.  In this study the content of phenolic compounds in fireweed and raspberry honeys was examined and considered.

9. INTERPRETATION OF INDIAN UNIFLORAL HONEY QUALITY DATA BY PATTERN RECOGNITION TECHNIQUES

Vikas Nanda and Bahadur Singh

Department of Food Engineering and Technology,

INDIA

 Sant Longowal Institute of Engineering and Technology, Longowal-148106, India

In this study, it was attempted to determine the quality variables capable of promoting a characterization of the honey from different plant sources viz. Trifolium alexandrinum (berseem clover), Brassica sp. (mustard), Helianthus annuus (sun flower), and Eucalyptus lanceolatus. Among the sources of honey significant difference were found in moisture content, free acidity, pH, fructose glucose ratio, ash content, diastase activity, hydroxymethylfurfural content and mineral content (Potassium, sodium, calcium, zinc and copper). Pattern recognition techniques such as principal component analysis (PCA) and linear discriminate analysis were performed to classify honeys according to their type on the basis of physicochemical parameters and mineral content. The variables with high discrimination were potassium, sodium and total acidity.

10. SLOVENE HONEY - PROTECTED GEOGRAPHICAL INDICATION

 Andreja Kandolf, Adriana Pereyra Gonzales, Nataša Lilek

Slovenian Beekeepeers Association

andreja.kandolf@czs.si

SLOVENIAN

Around 500 honey samples representing various floral and honeydew honeys were collected over four consecutive harvest seasons from various areas of Slovenia. The following physico-chemical properties were tested: electrical conductivity, acidity, diastaze number, water content, insoluble solids, HMF, fructose, glucose and sucrose and also residues of acaricides. It has been shown that most honeys lie within the limits set by legislation and most of those lie far below the limit. According to these results, the Slovenian Bekeepeers Association decided to get a Protected geographical indication (PGI) for honey produced in Slovenia and to respect all required parameters (honey quality, pollen spectrum, GMP, etc.).  

11. UNI-FLORAL HONEY ANALYSIS COMMERCIALIZED IN CATALONIA (SPAIN)

Garcés, S.¹; González-Porto, A.V.²  & J. Cambra¹

¹Dept. Biologia Vegetal. Fac. Biologia. Univ. Barcelona. Av. Diagonal, 645, 08028 Barcelona.

² Centro de Experimentación Agraria de Marchamalo Extramuros, s/n 19180 Marchamalo (Guadalajara).

lunaestrellada84@gmail.com

SPAIN.

 The characterization of 24 samples of Catalonian commercial brands of uni-floral honeys was carried out on the basis of melissopalynological analysis, physico-chemical properties (colour, water content, HMF, diastase, pH, electrical conductivity and poly-phenols), enzyme analysis (diastase and peroxide),  amino-acid test (proline) and residues from antibiotic treatments (tetracyclines). The botanical origins of the examined honeys were Castanea sativa Mill, Citrus spp, Erica spp, Eucalyptus spp, Lavandula spp, Medicago sativa, Quercus spp (honeydew), Robinia pseudoacacia L, Rosmarinus officinalis, Thymus vulgaris, Tilia spp and multifloral honey. Pattern recognition methods such as principal component analysis (PCA) were performed to classify honeys according to their type on the basis of physico-chemical parameters. These combinations of methods could be a promising approach to prove authenticity of honey.

12. NATURAL HONEY: A HEALTHY SOURCE OF DIETARY FRUCTOSE.

Ajıbola Abdulwahid & Erlwanger Kennedy

School of Physiology, Faculty of Health Sciences

University of the Witwatersrand, Johannesburg,

SOUTH AFRİCA

PURPOSE OF REVIEW: Consumption of fructose as an industrial sweetener is rising globally. Excess dietary fructose can induce a complex metabolic syndrome which includes insulin resistance, elevated triglycerides, abdominal obesity, high blood pressure, inflammation, oxidative stress, fatty liver, renal injury and cardiovascular disease. These pathological changes are absent with high intake of honey, despite its high sugar content, which is mainly fructose.

FINDINGS: Honey is a ‘health food’ releasing fructose slowly into the blood stream for energy to facilitate body functions. The slow release leads to a fairly constant plasma sugar level and low glycaemic index. Antioxidants, sugars, hydrogen peroxide, minerals, vitamins and phytonutrients found in honey provide nutrients, curb pathology, improve haematological profiles and blood clinical chemistry parameters as well as enhance the immune status in both human and animals models.  Control of blood pressure has also been recorded in honey-fed hypertensive patients.

SUMMARY: The synergy of nutraceuticals in honey curbs the potential metabolic dysfunction that could be induced by high fructose intake. Thus preventing pre-diabetes, excessive lipogenesis, oxidative damage, and reducing the risk of developing cardiovascular diseases. In addition, honey enhances carbohydrate metabolism and maintains energy homeostasis. Further studies to identify the mechanisms involved in this metabolic and energy homeostasis are underway. Natural honey is a healthy dietary substitute for refined carbohydrates.

13. VOLATILE AND SEMIVOLATILE COMPONENTS OF Mesembryanthemum crystallinum HONEYS FROM GRAN CANARIA (SPAIN)

 Jiménez-Pulido, A¹; Sancho, MT¹, Sanjuán, E.²; Millán, R.², and Fernández-Muiño MA¹*

¹University of Burgos (Spain). Department of Biotechnology and Food Science. Faculty of Sciences. Plaza de Misael Bañuelos s/n. 09001 Burgos. Spain.

²University of Las Palmas de Gran Canaria. Campus of Arucas. Faculty of Veterinary. 35416 Arucas. Las Palmas (Canarias).

*Presenting author. E-mail: mafernan@ubu.es

SPAIN.

 As a preliminary study for the characterization of Mesembryanthemum crystallinum honeys from Gran Canaria (Spain), we have analyzed the profile of volatile and semivolatile compounds of six samples. Aroma-compounds isolation was performed by solvent (ethyl acetate) extraction and the concentrated unmethylated extracts were analyzed by GC-MS. In the samples analyzed 71 compounds were found, the most important of which are the following: Among aliphatic compounds: levo and meso-butane-2,3-diol isopropanol and 1,3-dihydroxy-2-propanone. Among monoterpenes, both isomers E and Z of 2,6-dimethyl-2,7-octadien-1,6-diol were found. Among benzene derivatives: 3,4,5-trimethoxy-benzoic-methyl-ester was found as a major component in most samples. Other benzene derivatives found as important components were 4-vinyl-2-methoxy-phenol and phenylacetic acid. Many Norisoprenoids were also found being the most representative vomifoliol. Among compounds related with heating or degradation processes HMF, 2-hidroxy-2-ciclopenten-1-one and 2,3-dihydro-3,5-dihydroxy-6 methyl-4-H pyranone were found. Finally, caffeine was found in all samples with area percentages ranging from 0,53% to 7,72%.

14. EUCALYPTUS OCCIDENTALIS ENDL. UNIFLORAL HONEY: PHYSICOCHEMICAL, MELISSOPALYNOLOGICAL AND ORGANOLEPTIC CHARACTERISTICS

 Piana Maria Lucia *, Belligoli Paola**, Sesta Giulio**, Lusco Lorenzo**, Persano Oddo Livia**

*Piana Ricerca e Consulenza – Castel San Pietro Terme (BO), lucia.piana@pianaricerca.it** C.R.A. - Istituto Sperimentale per la Zoologia Agraria, Sezione di Apicoltura (presently abolished) – Roma,

ITALY

In Italy Eucalyptus unifloral honey is mainly produced on E. camaldulensis Dehnh.; its analytical profile is well known and currently used for authenticity control of the unifloral denomination. In the last years we observed some honey batches, marketed as Italian Eucalyptus honey, that do not fulfil the required outline, mainly for the melissopalynological characteristics. The productive information reveals that these products originate from another Eucalyptus species, blooming in late summer - early autumn, Eucalyptus occidentalis Endl. . A study was then carried out on about 50 samples of this kind of honey, purchased directly from the producers, in order to describe its physicochemical, melissopalynological and organoleptic characteristics. The results showed that physicochemical and organoleptic characteristics are quite similar to those of E. camaldulensis unifloral honey, except for sucrose content, slightly higher in E. occidentalis honey. On the contrary, melissopalynological features, with an absolute pollen number (PG/10 g) between 20.000 and 50.000 and a relative frequency mainly between 40 and 80%, are specific of this species, also characterized by bigger pollen grains. Mixed honeys from both Eucalyptus species, with intermediate features, are quite frequent.

15. LEAD AND CADMIUM IN HONEY DEW AND BLOSSOM  HONEYS  ORIGINATED FROM DIFFERENT REGIONS OF GREECE IN THE  YEAR  2009

 S. Samargi ¹ and  E. Gouta²

1.      Residue  analysis  lab , Intitute  of  Food  Hygiene  of  Athens, Center  of  Athenian  Veterinary  Institutions , Ministry  of  Rural  Development  and  Food.

2.      Department  of  Hygiene  of  bee-hive  product  control , Institute  of  Food  Hugiene  of  Athens , Center  of  Athenian  Veterinary  Institutions, Ministry  of  Rural  Development  and  Food.

GREECE

 Lead (Pb)  and  Cadmium (Cd)  are  considered  the  principal  toxic  heavy  metals  of  environmental  contaminants.The  proposed  to  the  E.U  MRL (Maximum Residue Limit) (Byrne,2000) , is  0.1mg/kg  for  Cd  and 1mg/kg  for         Pb in food products.

 In  the  present  study  22  honey  samples (14  honeydew  ,  5  blossom  honeys  and  3  blossom  and  Honeydew) , originated  from  diferent  parts  of  Greece , were  examined.  These  samples  come  from  the  annual  national  residue  program  for  the  honey , of  the  year  2009..

The  method  used  was  from  our  lab  of  Residue  Analysis  in  food  of  animal  origin  and  the  samples  were  determined  by  atomic  absorption  spectrometry.

The  LOQ  for  Pb  was  0.0016  mg/kg  and  for  the  Cd  0.00055  mg/kg.    .

For  Pb  in  the  honeydew  honeys  the  measurements  ranged  from  0.0021  mg/kg  to  0.055  mg/kg.  and   all  values  were  found  below  the  proposed  limit  of  1mg/kg..MRL (Maximum Residue  Limit), whereas  for  Cd  the  measurements  ranged  from  below  the  LOQ  (Limit  of  Quantitation)  of  the  method  to  0.0014  mg/kg ,  with   all  the  values  below  the  proposed  limit  of  the  MRL  0.1 mg/kg.

For  blosson  honeys  ,  for  Pb  the  measurements  ranged  from  0.0032   mg/kg  to  0.018    mg/kg  and  all  values  were  found  below  the  proposed  limit  of  1mg/kg , whereas  for  Cd  the  measurements  ranged  from   below  the  LOQ  of  the  method  , with  all  the  values  below  the  proposed  MRL  limit  of  0.1  mg/kg.

In  the 3   blossom  and  honeydew  honeys  the  measurements  ranged  from  0.0093  up  to  0.070  mg/kg  for  Pb  and  for  Cd  the  measurements  were  found  below  the  LOQ  of  the  method. All  values  were  found  below  the  proposed  MRLs.

All  values  were  found  very  low  and  it  is  noticed  the  relatively  low  concentration  of  honeys    of  Pb  which  is  probably  due  to  the  increased  use  of  car-engine  catalysts.

16. HONEY BEES OF WESTERN TURKEY AND PINE HONEY

 İrfan Kandemir and Ayça Ozkan

Department of Biology, Faculty of Science, Ankara University, 06100, Ankara-TURKEY 

Turkey holds more than 20% of all known honey bee species in its geography. Among these Apis mellifera anatoliaca predominantly distributed all over the geography. Apis mellifera caucasica has its own geography naturally in north east of Turkey along the border to Georgia. Apis mellifera meda is distributed in the south east of Turkey on the border to Iraq and Iran. Apis mellifera syriaca is found on the southern Turkey bordering to Syria. Honey bees of Thrace are different than other bees diagnostic to carnica group.

 The bees of western Turkey according to Ruttner (1988) is found different than the rest of Turkey and associated with the pine honey. Here we studied the new methodology in order to distinguish these bees with using reference samples. We found that the bees from this region is statistically different (P<0.05) than the subspecies distributed from the other parts of Turkey based on geometric morphometric methods of both types landmark based and shape analysis of wing cells. Here we report the honey bee differences, beekeeping in western Anatolian and the economics of the region on Turkish beekeeping sector.

17. PINE AND FIR TREE HONEYS: A REVIEW OF THE TWO PILLARS OF BEEKEEPING IN GREECE

 Spyros Skareas

GRECCE

 Pine and Fir honey provide more than 70% of Greece’s annual honey production. We will present important facts about these two major honey sources, focusing on the plants themselves, their insect pests that provide these valuable honeydews, as well as the beekeeping techniques involved to exploit them. Special interest will be given to Vanilla-Fir Honey from Mt Mainalo of Peloponnesus, the only Greek honey certified as to its origin (PTO – Protected Designation of Origin)

18. SUPPORTED COLONY MANAGEMENT SYSTEM FOR INCREASİNG HONEY PRODUCTİON.

Prof. Dr. Muhsin Doğaroğlu

TURKEY

Colony managements applied by beekeepers have effective roles in maintaining the increase in colony size and productivity. Some specific measurements used with this criteria have selective importance in honeydew honey production at the and of the season. The characteristics of honeydew honey production is completely different to floral honey production. Incorrect applications may cause decreases in crop production and lead to winter loses. The best colony management technics and colony support management systems are discussed under this topic. The Colony support management system is based on setting pair colonies which produce honey from each colony in every pair. Of this colony pair, one is called the supporter and the other is called the producer. Observations showed producer colonies able to produce honey more than any other usual colonies in all varying conditions and the system is able to increase total honey production of the apiary.

19. CHROMATOGRAPHY ANALYSYS OF CARDOHYDRATES USED IN QUALITY CONTROL OF HONEYS

 Helena Rybak-Chmielewska*, Teresa Szczęsna, Ewa Waś, Katarzyna Kachaniuk

*e-mail: helena.chmielewska@man.pulawy.pl

Research Institute of Pomology and Floriculture Apiculture Division, Department of Bee Products, Kazimierska 2 Str., 24-100 Puławy,

POLAND

 Honey is a natural sweet substance composed mainly of easily digestible simple sugars and of some small percentage of di- and trisaccharides. The qualitative and quantitative composition of sugars is peculiar to the honey. Chromatographic spectrum of honey carbohydrates is guarantee of the product authenticity. The aim of the study was comparison of sugar composition of honey and several syrups (bee feeds) before and after they were processed by bees. Another objective was to find some significant differences between the bee-processed syrups and honey. According to the received results the differences were determined for fructose content, fructose-to-glucose ratio, and within disaccharides, for sucrose and maltose content. Fructose content below 32%,  maltose content over 5% and fructose-to-glucose ratio  (F/G) below 1 can be used as a distinguisher of honey adulteration with winter stores processed from starch syrups. For identification of inverts processed by bees from sucrose syrups, erlose content (over 2%), sucrose content (over 3%) and sucrose-to-maltose ratio (approaching 1 or higher) can be used.

20. HPLC DETERMINATION OF MALTODEXSTRINS USED FOR DETECTION OF HONEY ADULTERATION BY STARCH SYRUPS

 Teresa Szczęsna*, Helena Rybak-Chmielewska, Ewa Waś, Katarzyna Kachaniuk, Dariusz Gerula.

Research Institute of Pomology and Floriculture Apiculture Division, Department of Bee Products, Kazimierska 2 Str., 24-100 Puławy,

*e-mail: teresa.szczesna@man.pulawy.pl

POLAND

For several years in Poland various types of starch syrup have been produced and utilized chiefly by food industry. Some of those syrups have roused the interest of beekeepers due to their relatively low price and convenient ready-to-use form. The aim of the study was to develop a quantitative method for maltodextrins assay occur in starch syrup with an idea to use that method for identification of honey adulteration with admixtures of those syrups. The experimental material was comprised of starch syrups and winter stores processed from those syrups. Moreover, honey samples with an admixture of starch syrups prepared in the laboratory were analyzed. Separation, identification and quantity analysis of maltodextrins (maltotetraose - Dp4, maltopentaose - Dp5, maltohexaose - Dp6 and maltoheptaose - Dp7) were done by Shimadzu HPLC with refractometric detector. Chromatographic conditions were performed on Phenomenex Luna 5μm NH₂ 100A 250 x 4.60 mm column at a temperature of 40°C with the mobile phase of 65 : 35 acetonitrile-water and 3 ml/min flow rate. The quantitative assays were made using the external standard method. The detection limit for all maltodextrins was calculated on 0.05 g/100g. The precision and repeatability of the method were satisfactory. The  correlation coefficient of linear dependence of malto-compounds concentration on the peak area received value over 0.999. Maltodextrins content in starch syrups and in the winter stores processed thereof was 3.3 and 2.3 g/100g, respectively. The elaborated method for maltodextrins assay can be used for estimation of honey adulteration with winter stores processed from starch syrup or with the syrup per se. Method allows detection of 10% addition of starch syrup in honey.

21. DEVELOPMENT OF A SCREENING METHOD FOR THE AUTHENTICATION OF THE BOTANICAL ORIGIN OF HONEY BY MID-INFRARED SPECTROSCOPY

Martina Janke, Kathrin Kemper, Laila Ettalibi, Werner von der Ohe

GERMANY

 Fourier transform mid-infrared spectroscopy (Tensor 27, Bruker) using an attenuated total reflectance cell (ATR-MIR) was applied to model a spectral-library basing on the spectral characteristics of 6 different unifloral honey types (acacia, rape, lime, heather honeydew, n = 239) previously classified with official methods (chemical, pollen and sensory analysis).

The IDENT-method (OPUS 6.5, Bruker) was developed after multivariate data analysis and selection of significant spectral ranges (wave number range between 4000 and 550 cm-1). The developed IDENT-method (spectral library) consists of one headlibrary and 4 sublibraries on 3 different levels. The IDENT-method was tested for 72 honey samples, so far. Most of the unifloral honeys revealed very high rates of correct classification (Acacia 100%, Heather 92%, Lime 100%, Rape 97%, Honeydew 71%). The results demonstrate that the model used was robust.

Before this new method can be used for routine honey control, it has to be tested with a sufficient number of authentic unifloral samples. The range of the method has to be amplified for more honey types, unifloral and polyfloral.

22. SOLID-PHASE EXTRACTION FOR THE DETERMINATION OF ORGANIC ACIDS IN HONEY

 S. Oelschlaegel,  R. Esche,  I. Koelling-Speer,  K. Speer

Prof. Dr. Karl Speer,

Technical University Dresden, Professorship for Special Food Chemistry / Food Production, Bergstr. 66, D-01069 Dresden

GERMANY

 Even though organic acids represent less than 0.5 – 2.0 % of honey [1, 2, 3], they are significantly involved in the organoleptic, physical and chemical properties of honey. Furthermore, aliphatic organic acids were used to characterize honeys according to their botanical and/ or geographical origin [4, 5].

 A solid-phase extraction procedure was applied to isolate organic acids from honey. Acetic, citric, formic, fumaric, galacturonic, gluconic, glucuronic, glutaric, lactic, malic, oxalic, propionic, pyruvic, succinic and tartaric acid were extracted using an anion-exchange cartridge. Different clean-up steps such as the conditioning and the elution parameters which influence the recovery of the acids were studied and consequently optimized. With the final method the average recovery of the 15 organic acids in honey was above 90 %. The organic acids were determined via HPLC-DAD at 210 nm.

As expected, honeydew honeys contained the highest amount of total organic acids with 11.2 g/kg on average. The results of this research will be presented and discussed.

23. SIMULTANEOUS ANALYSIS OF PHENOLIC ACIDS AND FLAVONOIDS IN HONEY

 K. Speer.,  S. Trautvetter,  I. Koelling-Speer

Prof. Dr. Karl Speer, Technical University Dresden, Professorship for Special Food Chemistry / Food Production, Bergstr. 66, D-01069 Dresden

GERMANY

 Certain phenolic acids and flavonoids are suitable as marker substances to characterize several unifloral honeys. First results were presented by Speer und Montag (1984, 1987) and Steeg und Montag (1987, 1988). Several years later, the study-group of Tomás-Barberán (1992, 1995) reported further results concerning the flavonoids in honey. Up to date, a large number of investigations have been published about phenolic acids and flavonoids in honey.

As most of the authors applied different methods for extraction and determination, the obtained data could, frequently, hardly be compared and showed remarkable discrepancies concerning the qualitative and the quantitative data.

 Hence, a fast and easy extraction method for both the phenolic acids as well as the flavonoids had to be developed. The separation and detection was, then, carried out using HPLC-DAD and UPLC-Q/TOF-MS. Thereby, 34 substances were identified in the ethyl acetate extracts of sunflower, lime, clover, rape, and honeydew honey.

24. ANALYSIS OF THE VOLATILE FRACTION COMPOSITION IN THE BEE POLLEN LOADS IN SPAIN

 Ana Rosa Quintana-Edesa, Amelia Virginia González-Porto, Mª Teresa Sancho-Ortíz, Miguel Ángel Fernández-Muiño

Centro Agrario de Marchamalo. Junta de Comunidades de Castilla-La Mancha. Camino San Martín s/n. Marchamalo. 19180 Guadalajara

Departamento de Biotecnología y Ciencia de los Alimentos. Facultad de Ciencias. Universidad de Burgos. Edif. A. 1ª planta Plaza Misael Bañuelos, s/n. 09001 Burgos

SPAIN

 The bee pollen is one of the beehive products that require a complete study to obtain its characterization as a quality product and its identity appreciation on a botanical profile. Thus, the study of volatile fraction has been carried out. It is known that pollen aroma is determined by the floral origin of the pollen, but it can vary after the drying process required for its marketing.

 As a result of these premises, the study of 13 samples of bee pollen was done. Analysis results from several subsamples that were dried out pollen at different temperatures were compared with fresh pollen.

 The aim of this study was the detection of the volatile compounds that can identify its floral origin, as well as those that can reflect the conditions related to the drying process of the pollen. This information was used to establish which dryness temperature was critical for the aroma loss of the pollen.

 Pollen was dissolved in an ethyl acetate solution for the extraction of the volatile compounds and then analyzed by a gas chromatography / mass spectrometry. 146 aromatic compounds have been found and statistically analysed through a discriminant analysis, in order to identify and detect which ones are related to each dryness temperature. Final evaluation reduces the markers compounds to a 17-27 range. Among them, 16 compounds can be used as a tool to identify botanical origin and dryness temperature.  

This work has been financed by INIA-FEDER project RTA2007-00072-C03 and a predoctoral scholarship financed by INIA.

25. ANALYSIS OF FRENCH ROYAL JELLY FOR QUALITY AND AUTHENTICITY CONTROLS

H. Casabianca, G. Daniele

CNRS Service Central d’Analyse - Echangeur de Solaize – Chemin

 du Canal - 69360 Solaize

FRANCE

The aim of our study is to characterize french Royal Jelly (RJ) in order to define a standard composition and evaluate the quality of commercial products.

More than 300 RJs produced recently in France were analyzed. Due to the important heterogeneity of the materials depending on the environment of the hives, the climate, the soil etc, the samples were collected in different french regions during the course of the harvesting season representing various geographical and botanical origins. All the samples analyzed for creating our data base were provided by beekeepers belonging to the GPGR (Groupement des Producteurs de Gelée Royale), a french cooperative that respect a quality charter concerning the production, the sampling procedures and the storage of the RJ.

We have developed and validated analytical methods to quantify various parameters : water, protein, 10-HDA, amino acid, sugar contents. Moreover stable isotope ratios (¹³C/¹²C and ¹⁵N/¹⁴N) were measured in RJ samples by isotope ratio mass spectrometry.

In comparison, around 60 commercially available RJ samples, as representative materials produced and traded worldwide, were analyzed by the same methods.

In addition, around 30 RJ from feeding experiments with artificial sugars were provided by some apiarists in order to evaluate the impact of sugar feeding on the composition of RJ.

This study has allowed to establish ranges of natural variation of different composition parameters in french RJ and to define criteria for genuine ones. Some parameters are essential to distinguish RJ produced in France from abroad RJ and/or from intensive feeding, included isotopic measurements of ¹³C and the content of some particulate sugars.

26. THE CONTENT OF FATTY ACIDS IN MONOFLORAL POLLEN LOADS

 Róbert Chlebo, Martin Melich, Miroslava Kačániová

Slovak University of Agriculture in Nitra, Tr. Andreja Hlinku 2, 949 76 Nitra, Slovakia

Robert.Chlebo@uniag.sk

SLOVAK

 Contect of fatty acids as well as total lipid content in 7 samples of fresh monofloral bee pollen loads was evaluated in this study using GC/MS. Pollen loads were collected from bees using pollen traps when placed on monocultures of rape (Brassica napus), white mustard (Sinapis alba), sunflower (Helianthus annuus), poppy (Papaver somniferum), apple trees (Malus domestica), willow (Salix spp.) and dandelion (Taraxacum officinale).

 Highest total lipid content was detected in pollen loads from rape (13,34%) and dandelion (11,63%). Dominant fatty acids in various pollen loads are  linoleic, palmitic, tridecanic, linolic, oleic and capronic acids.

 Fatty acids was hypothesised to improve honey bee longevity. Different concentrations of various fatty acids mentioned in this study can be used as a floral origin marker of pollen. This work has been supported by project VEGA 1/0074/08.

27. PROTEIN CONTENT AND SUGAR COMPOSITION OF BEE-COLLECTED POLLEN FROM SELECTED BOTANICAL ORIGINS

 Liolios V. , Tananaki C. , Dimou M. , Kanelis D. , Thrasyvoulou A.

March 2009-March 2010

Laboratory of Apiculture-Sericulture,

School of Agriculture, Aristotle University of Thessaloniki,

 GREECE

   The objective of the study was to investigate the protein content and sugar composition of honey bee-collected pollen. The pollen pellets were harvested with pollen traps, separated and classified according to their botanical origin.  In 37 different botanical  species the protein content was determined using the Kjeldahl method. The crude protein content in the examined samples ranged from 10,69% for Smilax pollen to 25,90% for that of Fallopia type and the mean value was 17.84%. Pollen from insect-pollinated species (Castanea sativa 21,42%) was also found to be richer in protein compared to that from anemophilous plants (Pinus 12,17%). Although honeybees posses the ability to discriminate among pollen types they are incapable of discriminating protein content.

 The analysis of sugars was carried out by high-performance liquid chromatography.  Differences were also found in the concentrations of the eight investigated carbohydrates (fructose, maltose, sucrose, trehalose, turanose, melibiose, melezitose, glucose)  among various botanical species.

28. INHIBITION ACTIVITY OF ETHANOL EXTRACT OF PROPOLIS (EEP) RELATION MYCOTOXIGENIC FUNGI 

Karina Grigoryan, Mariam Sargsyan, Lusine Hakobyan, Grigor Badalyan

Yerevan State University, Laboratory of Biocontrol of Food

ARMENIA 

    The contamination of dried fruits by mycotoxigenic fungi  is a  actual problem  with  economic, and health  value.

     Results of mycological analysis of dried fruits and raisin in Armenia , show their high  contamination by ochratoxigenic and afltoxigenic fungi from A. nigri  and A. flavi section. In recent years attention has been focused on the use of propolis as a health supplement suited to consumers in developed  countries. Propolis, a natural honey bee product, has different biological activities.

      The aim, of this study was to determine the effectiveness of ethanol extract of propolis (EEP), from some region and  different concentration , relation samples of raisin with high level contamination by ochratoxigenic fungi and afltoxigenic  fungi from A. nigri  and A.flavi section. Antifungal activity    also was studied, in vitro, by injection of certain quantities  of extracts of propolis in Czapek-Dox agar. After solidifying, the disc (5 mm dia.) of the fungus culture was placed in the center of Petri dish. The inhibition percentage of mycelial growth   of 10 strains of A. carbonarius , A. niger, A. flavus species, was determinated according to Singh (Singh et al., 1999). MIC (minimal inhibition concentration) was defined as the lowest concentration that did not yield visual growth. All experiments were performed in triplicate.

Along with ЕЕР, during raisin processing (treatment of raisin), defined concentrations of mixture of propolis and hydrogen peroxide were used (some concentration). Duration of treatment  varied from 5 to 20 minutes.

The EEP and mix with hydrogen peroxide has shown the stable antifungal effect relation ochratoxin A producing  strains A. carbonarius , is  in experiment in vitro , on pure culture (60%) and treatment  of raisin (95%), which contaminating some species from A. nigri section. The EEP  inhibition percentage of A. flavus mycelial growth was between  40% -60%.

   The results obtained in this work could be of importance in the search of new natural sources bioactive compounds.

29. IS PORTUGAL A COUNTRY OF PROPOLIS DIVERSITY?

S.I. Falcão^a, V. Batista^a, S.M. Cardoso^b, M.R.M. Domingues^c, M. Vilas-Boas^a*

 ^a CIMO - Escola Superior Agrária, Instituto Politécnico de Bragança, Campus de Sta. Apolónia, Apartado 1172, 5301-855 Bragança, Portugal

^b CERNAS - Escola Superior Agrária, Instituto Politécnico de Coimbra, Bencanta 3040-316 Coimbra, Portugal

^c Centro de espectrometria de massa, Departamento de Química, Universidade de Aveiro, 3810-193 Aveiro, Portugal

^*mvboas@fc.up.pt

Portugal geographical situation and therefore their floral diversity points towards the existence of typical propolis from temperate zones, where bud exudates of poplar trees (Populus species) are the main source of this honeybee product. With the exception of Madeira and Azores Islands, where the floral diversity could present significant differences, it would be expected to find a product rich in flavonoid aglycones (flavones and flavanones), phenolic acids and their esters, characteristic of European propolis. In order to clarify this issue, several propolis samples were collected along the country, including islands, and their colour, ashes, content of wax, phenols and flavonoids, as also their phenolic profile, were determined. Albeit this work seems endless, it is clear that as dipper we study each sample, more diversity we find. In fact, the first phenolic profile we analyse by mass spectroscopy,^[1] a propolis sample with a significant poplar source, revealed the presence of 37 phenolic compounds, from which, seven were identified for the first time in propolis. More, the straight comparison of the phenolic profile obtained by HPLC for all samples, allow the identification of five distinct propolis phenolic extracts, with a correspondence diversity found as well in colour and odour. Although some samples show a typical odour of Cistus species, a common resinous plant in the east side of Portugal, the exact botanical origin seems another giant task to overcome.

The bioactive properties of the phenolic extracts were also studied in terms of reducing power and radical scavenging effect. Once again, the differences between propolis samples were clearly observed.

 [1] Falcão, S.I., Vilas-Boas, M., Estevinho, L.M., Barros, C., Domingues M.R.M., Cardoso, S.M. (2010). Anal Bioanal Chem, 396: 887-897. DOI:10.1007/s00216-009-3232-8.

 Acknowledgements: Soraia I. Falcão thanks FCT for a PhD grant (SFRH/BD/44855/2008).

                              Thanks to National Federation of Portuguese Beekeepers for propolis samples.

30. PARAMETERS THAT AFFECT IN PROTEIN AND SUGAR CONTENT IN ROYAL JELLY

Kanelis D., Tananaki Ch., Lazaridou E., Liolios V., Thrasyvoulou A.

Laboratory of Apiculture – Sericulture, School of Agriculture, Aristotle’s University of Thessaloniki,

GREECE .

Several groups of bee colonies were made to produce royal jelly in order to be investigated; the variation among bee colonies, the artificial sugar feeding and the kind of grafted larvae in the quality of collected royal jelly. The produced royal jelly harvested, placed into bottles and stored at -18 ^oC until analyzed. Totally 145 samples were collected. The samples were analyzed using High Performance Liquid Chromatography and Kjeldahl methods for the determination of sugars and proteins respectively.  The proteins in analyzed samples, ranged from 11,0% to 17,1%. Using a mixture of acetonitrile and water as mobile phase totally eight sugars were investigated in royal jelly samples (fructose, glucose, sucrose, melezitose, maltose, trehalose, turanose and melibioze).

31. ANALYSIS OF FRENCH ROYAL JELLY FOR QUALITY AND AUTHENTICITY CONTROLS

H. Casabianca, G. Daniele

CNRS Service Central d’Analyse - Echangeur de Solaize –

Chemin du Canal  69360 Solaize

FRANCE

The aim of our study is to characterize french Royal Jelly (RJ) in order to define a standard composition and evaluate the quality of commercial products.

More than 300 RJs produced recently in France were analyzed. Due to the important heterogeneity of the materials depending on the environment of the hives, the climate, the soil etc, the samples were collected in different french regions during the course of the harvesting season representing various geographical and botanical origins. All the samples analyzed for creating our data base were provided by beekeepers belonging to the GPGR (Groupement des Producteurs de Gelée Royale), a french cooperative that respect a quality charter concerning the production, the sampling procedures and the storage of the RJ.

We have developed and validated analytical methods to quantify various parameters : water, protein, 10-HDA, amino acid, sugar contents. Moreover stable isotope ratios (¹³C/¹²C and ¹⁵N/¹⁴N) were measured in RJ samples by isotope ratio mass spectrometry.

In comparison, around 60 commercially available RJ samples, as representative materials produced and traded worldwide, were analyzed by the same methods.

In addition, around 30 RJ from feeding experiments with artificial sugars were provided by some apiarists in order to evaluate the impact of sugar feeding on the composition of RJ.

This study has allowed to establish ranges of natural variation of different composition parameters in french RJ and to define criteria for genuine ones. Some parameters are essential to distinguish RJ produced in France from abroad RJ and/or from intensive feeding, included isotopic measurements of ¹³C and the content of some particulate sugars.

32. PREDICTION OF 16 PHYSICOCHEMICAL MEASURANDS OF HONEY USING FT-IR ATR

Ligia Bicudo de Almeida-Muradian¹, Werner Luginbühl², Peter Gallmann³, René Badertscher³

¹ Faculdade de Ciências Farmacêuticas da USP, Av. Prof. Lineu Prestes 580, bloco 14, 05508-900, São Paulo, SP, Brazil

² ChemStat, Aarstrasse 98, CH-3005, Bern, Switzerland

³ Swiss Bee Research Centre/Agroscope Liebefeld-Posieux, Schwarzenburgstrasse 161, CH-3003, Bern,

SWITZERLAND

Fourier Transform Infrared spectroscopy with an attenuated total reflection accessory (FT-IR ATR) was used to predict 16 measurands in honey. 416 different honey samples obtained from the Swiss National Honey Quality Program of the years 2006 and 2007 were analyzed by classical physicochemical methods and FT-IR ATR. Partial least squares regression (PLS) was used to develop the calibration models for the measurands studied (electrical conductivity, erlose, free acidity, fructose, fructose/glucose ratio, glucose, glucose/moisture ratio, HMF, isomaltose, , melezitose, moisture, pH, sum of fructose + glucose, , total nitrogen, trehalose and turanose) They were validated using independent samples and proved satisfying accuracies for the determination of electrical conductivity, melezitose, and moisture. Poor predictive quality was found for erlose, fructose, HMF, isomaltose, total nitrogen, trehalose, and turanose while the calibrations for the remaining measurands can be applied for rough screening purposes. The results showed that mid-infrared spectrometry can be used as a screening method for the routine analysis of some quality characteristics of honey with the advantages of being rapid and non-destructive.

Acknowlegements: CAPES, CNPq, FAPESP and USP

33. PhYsicochemical and PALYNOLOGICAL ANALYSIS OF BRAZILIAN Apis mellifera BEE AND STINGLESS bee (Tetragonisca.angustula) HONEY

Ligia Bicudo de Almeida-Muradian*, Graziela Leal Sousa*, Alex da Silva de Freitas**, Ortrud Monika Barth**

* Faculdade de Ciências Farmacêuticas da USP, Av. Prof. Lineu Prestes 580, bloco 14, 05508-900, São Paulo, SP, Brazil, E-mail: ligiabi@usp.br

** Laboratório de Morfologia e Morfogênese Viral, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro,

BRAZIL.

 Honey is considered as a food that provides energy, being elaborated from the dehydration and transformation of the nectar of the flowers by the bees. For the human consumption, honey needs to attend the minimum requirements of identity and quality demanded by the regulation. In Brazil beekeepers can be divided in two practical distinct ones: the traditional ones, which use Apis mellifera bees and the Meliponiculture which uses native stingless bees such as “jataí” bee (Tetragonisca angustula).

 There are no identity and quality parameters or regulation for this type of honey. Honey from stingless bees is more expensive compared with the traditional honey; however it is commercialized without a proper regulation. The objective of the present work is to compare the composition and quality of honey from Apis mellifera and from Tetragonisca angustula bee from six cities of the state of Sao Paulo (southeast region of Brazil). The honey samples were obtained from both bees in the same botanical region. From botanical analysis it can be state that the honey from stingless bees present a bigger botanical diversity compared with Apis honey. It was used the methods based on the Brazilian Regulation for quality control of honey from Apis mellifera.  The results for honey from Apis mellifera honey and stingless bee honey were respectively: moisture (15.40 -19.00 ; 23.40 -25.60 %), acidity (16.82-32.47; 21.65 - 63,85 mEq/Kg), reducing sugars (52.98 – 84.24; 44.78 – 67.54 %), apparent sucrose (0,56 –7,64; 0,43 –4,46 %); total sugars  (53.54 – 83.75; 45.57 – 67.68 %),  HMF (2.00 – 21.00; 0.30 – 0.93  mg/Kg); diastase number (2.20 – 11.49; 11.01- 22.45); insoluble solids (0.01 – 0.08; 0.02- 0.10%) and ashes (0.11- 0.26; 0.17 – 0.42 %).   

Acknowlegements: FAPESP, CNPq, CAPES and USP

34. VITAMIN B₂ STABILITY OF DRIED BEE POLLEN DURING STORAGE

Aline Aparecida Santos Pereira, Vanilda Aparecida Soares de Arruda, Ligia Bicudo de Almeida-Muradian*

Faculdade de Ciências Farmacêuticas da USP, Av. Prof. Lineu Prestes 580, bloco 14, 05508-900, São Paulo, SP,

BRAZIL

Bee pollen has been used as a food supplement on account of its importance as a source of essential nutrients, among them, vitamins. Vitamin B₂ (riboflavin) has an important role in cell respiration, metabolism of proteins, fats and carbohydrates and also has participation in vitamins B₆, B₉ and B₁₂ metabolism. Riboflavin is stable during food processing and storage. However, it is very sensitive to light and there is a lack about bee pollen riboflavin values in literature. The aim of this work was to analyze this vitamin in seven samples of dried bee pollen commercialized in Brazil and to study its stability after eight months of storage in the original packages under three different conditions: room temperature (with and without light) and frozen. Riboflavin was analyzed by HPLC with fluorescence detection. The average content of this vitamin in time zero was 2.09 ± 0.32 mg/ 100g and the average contend after storage was 24.40%, 28.23% and 33.50% less in samples stored at freezer, room temperature without light and room temperature exposure to light, respectively. Statistically there is no variation (p<0.05) between the three storages conditions. 

Acknowlegements: FAPESP, CNPq, USP

37. BEE FARMING IN EAST AFRICA.

Mugisha Fred    ,2. Jonathan Kakande

 NAKASONGOLA BEE FARMERS ASSOCIATION   P.o.box 71183 clock tower ampala                            Tel:+256772657714 fax:+256414235858

EAST AFRICA

1.      Uganda

Races of honeybees, including African honey-bees, bee forage plants, activities of honeybees, honey production, pests, diseases of honeybees, honey hunting, traditional beekeeping using traditional hives, modern beekeeping using modern hives, in these 26 countries, are included in this work. History of beekeeping, research activities, and pioneers of beekeeping research are needed

Uganda has a very high potential for honey production, which not yet been fully achieved. Traditional beekeeping are in Nakasongola,Teso and West Nile ar-eas. Trees are the main forage plants, while in the Kigezi area, crops, pasture, weeds and exotic trees. Introduction of modern beekeeping in Uganda was de-scribed.

A promotion programme with CARE, YMCA and Red Cross has been started to introduce more effective modern beekeeping in Uganda. 4 major honey refinery plants in Nakasongola, Nalukolonga, Mbale and Soroti, 14 apiary demonstration firms are being established. "Uganda Beekeeping Association" (UBA) has recently formed in 1986 and 1st edition of their Newsletter was published with CARE-Uganda. "Apiculture Section" was established in Ministry of Animal In-dustry and Fisheries, Kampala. Apiculture Project CARE-Uganda, Kampala, was conducted. In 1990, beekeeping was started in Bunyaruguru County-Bushenyi. People are drinking their local brew mixed with honey. The UBA has embarked upon a "Beekeeping Research Project", which commenced in September 1995, in Luweero District, Kampala. They compared traditional hives with sloping sides, with straight sides, Langstroth hives with traditional hives instead of frames and tradi-tional hives, concerning production, duration and susceptibility to pests and preda-tors.

"Tropical Projects Ltd", is a company that specialises in beekeeping extension, including 70 women, with the aim of collecting honey and wax in large quantities. They have 150 beekeepers and over 2500 colonies. Orders for honey were received from Arabia, France, Germany. Most hives are traditional hives, but they intend to acquire more Tanzanian beehives. So far we  held a seminars  in our country  about marketing of honey and how to use Tanzanian beehives. "Beekeeping and Development Meeting", was conducted in April, 1997, Lira. "Nakasongola bee farmers” Group, is interested to contact beekeepers in other countries. "The Uganda Honey Beekeepers Association" was formed in 1995. Honey and wax production can be more than doubted]. UHA is involved in a broad spectrum of rural beekeepers in Uganda and designed a programme of apiculture development, based on strengthening UHA coordination at national level, down to districts to villages establishing honey refining, for export grade bee products.

 2.Kenya

Keeping bees in traditional hive, movable comb frameless hives, Kenya top-bar hive, and Tanzania transitional hive, in which comb are moved in pairs, were described. Kenya top-bar hive is successfully replacing traditional hive. The "David Hive" is more or less like Kenya top-bar hive, full honey combs are ex-tracted. A planned research programme in bee selection in Kenya was suggested. 80% of Kenya-land, including some arid areas, is suitable for beekeeping. In Kenya, the imported European bees, suitably managed, were less useful than Afri-can honeybees. The foreign bees are confronted with competition in foraging and defence from African honeybees, which is well adapted to the tropical conditions. Traditional beehives are to be found in the Wakamba and Kalenjan, and pro-posed methods for improvement. Traditional beehives are held by various tribes in the Embu District. There are economic life in Dorobo, hive desgins in Kenya, and Kenya-Pilot Project.

. A project for forest conservation and income generation is based in Samburu District, in the semi-arid area of Kenya, has included beekeeping in its research programme. Education, training and working of women as beekeepers, utilization of modern hives in Tropical Africa for honey production is recent, as compared with traditional bee-keeping. Traditional hives consists of a centrally split hollowed-out log, used by Turgen . The upper section (male) is larger than the bottom section (fe-male). During honey harvesting the female sections is detached to expose the upper fixed combs without damage. The multichamber traditional beehive was described and the constraint in transforming traditional hives to modern hives which offer some solutions resulting in increased yields of better quality of honey. In Laikipia District, a frame for Kenya top-bar, and comb honey production, were described. "Ruai Farm" motivates beekeepers have  produced honey since 1977. From Euphorbia candelabrum and Acacia mellifera, it is possible to harvest "bitter honey" from traditional hives in September, and "sweet honey" in October, then in December there is mixed floral "grass" honey. The brood nest arrangement in Kenya top-bar hives was described. The communication dances performed by African honeybees clusters after vacating a nest was de-scribed. Exposed clustered colonies are more open to weather and predators, than colonies in hives or closed nests.

3. Tanzania

In Tanzania we have discovered that Classical work on identification of pollen grains from 236 plant species, scale-hive records, discovery of European foulbrood, African "Dadant Hive", reasons for advocating it for production of high quality honey are seen as the compititve Idea.. A report covers improvement of beekeeping in Ujamma villages, honey and wax production, hives and management methods for African honeybees, diseases .
Beeswax is a very important by-product of traditional beekeeping. Tanzania has been one of largest exporters of wax in the world. In 1973, 275 tons were exported. In Handern District, mean yield/traditional hive, is 15 kg honey. Assuming that 1/2 to 2/3 of harvested wax is obtained for export, the number of colonies must be between 800 thousands to a million. Modern hives were used in “Tanga Inte-grated Rural Development Programme”, in north-eastern Tanzania and in Handeni, A beer, “pombe”, are prepared from honey. A ratio of 1:15 between wax and honey provides a basis for calculation. Tanzanian-commercial hive and Tanzanian tradi-tional hive are the two recommended blank hives. In order to avoid overcrowding, the carrying capacity of different areas must be investigated.

Problems of beekeeping programme are associated with man and his traditions, and with bees, and its enemies. Financial support is needed. Train-ing in Tropical beekeeping is conducted in "NJIRO Wildlife Research Centre, Arusha". They produce a "Newsletter" in beekeeping. A "Beekeeping Division" exists in Ministry of Natural Resources and Tourism.

1st East Africa Workshop was held on "Tabora Natural Organic Honey", gathered without smoke. Nearly all honey is exported. Stored honey for a long time which means high HMF can sell as industrial honey with lower prices. In Njiro Centre, the best time of day to harvest honey combs from TBH is in the evening before dusk. When procedures are fol-lowed well, the best honey, with no venom can be obtained, with a few dead bees. El-Niño rains affected honey and wax production in June and August 1997 and 1998. Members of TBCS, reported that many colonies absconded and again occu-pied.

Traditional beekeeping among Wameru people of northern Tanzania. Social aspects of Ngindo beekeeping, "Tanzania-Canada Beekeeping Project" with CIDA, honey hunting and beekeeping situation in Tanga region, use of modern hives in development programme were summarized. CLAUSS's book "The Bee-keeping Handbook" was translated to Swahili for Tanzania, in 1987.

Traditional beekeeping in Tanzania, is done side by side with other socio-economic activities. Stocking of traditional hive is left to chance, excepting baited hives. Maximum occupancy in: Lindi, Mtwara, Rungwa Game Reserve, where Tanzanian traditional hives and Tanzanian commercial hives are used with: baiting, transferring swarms, dividing established colonies, and using of emergency queen rearing during April and June, in lower plains and mountains. Beekeeping industry plays an important part of the economy of arid areas. Twelve African honeybee races were described. A.m. scutellata, is superior to European races. Most African honeybee races abandon hives by reproductive swarming, migration and absconding. They defend themselves against intruders. With selective breeding we can have gentle bees. Foraging activity was studies at Njiro Centre. Beekeeping in the "Tanzanian Tropical Forestry Action Plan", was described. Aid was re-quested from "Overseas Development Administration", . Teaching beekeeping, research on African honeybee, using of propolis in medicine, in Kilimanjaro region, are summarized.

"Arusha Beekeepers Association", held a meeting in October 1993. At Ta-bora, 1994, was a poor year for honey production. In this honey area, the esti-mated harvest of 360 tonnes could not be obtained due to lack of rain, and less than a tonne was harvested. Training in "Tropical Beekeeping" was conducted in Njiro Centre, in 1996. Fair rains, in 1999, were giving hope that the harvest would be good. TBCS were awaiting June for harvesting honey from Madaha's apiary in Malongwe Forest. The "let alone" method is discouraged by many beekeepers, but still the easiest way for catching swarms. In August 1998 and August 2000 "Bee-keeping in Rural Development" courses were conducted in Njiro Centre and at the Cardiff University, UK. The disadvantages of using modern hives with African hon-eybees, were discussed. A seminar was organized by FAIDERS, in Biharamulo, during 1997, to discuss modern beekeeping and tools to produce good quality bee products in increased quantities. "Arumeru Beekeepers Society", Usa River, and "Tree Planting Foundation" are promoting beekeeping in Usa River, Arusha.

In Tarangaire and Manyra National Parks, bees nests are located in hollow trees. The Gorowa and Iraqw beekeepers use a strongly scented plant, Ocimeras suave, to bait hives. Honey badger caused the decline of traditional beekeeping in some areas of bambati District. Barbaigs call honey beer "gesuda". Beekeeping is capable of further development. Beekeeping camps of 6 to 15 persons are used over generations along Ugalla River. Individual beekeepers of TBCS can produce large quantities of organic honey, which is sold on international markets. In Tan-zania, 3 types of smokers are used: traditional locally made smokers, modern style smokers, and imported smokers. Ideal was a wide smoker with 2.1 litre volume, using elephant dung or papaya fibre. Low cost gloves are made from available plastic tubes in Tanzania.

Honey production in Tanzania is dependant on small holders beekeepers, using traditional hives for African honeybees. Over 95% of beekeeping is practised in Savannah Forests "Miombo Woodlands", the rest is carried out in banana and coffee plantations, and where trees are used for hanging hives. Average productiv-ity A. m. scutellata colony/year, using traditional hives, was 15 kg honey and 1 kg wax. Suggested formula for calculation of honey production using wax figures was discussed. Traditional uses of honey and wax in making remedies used to cure various diseases were summarized. A floral calender for beekeeping in North Tanzania was worked-up. Traditional beekeepers use various indicators to deter-mine when it is time to harvest honey. The success of improved traditional hives beehive in field trials was discussed. A slight slope of the longside wall is necessary in order to reduce the degree of comb attachment. Traditional hives beehive, with 20 bars is recommended in North Tanzania. Entrance hole of 8 mm diameter, is ideal.

B. POSTERS

38. Valorisation of the honey food chain: traceability and botanical origin

Morcia C. ⁽¹⁾, Cafiero C. ⁽³⁾, Faccioli P. ⁽¹⁾, Gardini S. ⁽²⁾, Marcazzan G.L. ⁽²⁾  Savoldelli J. ⁽¹⁾, Spini M. ⁽¹⁾, Terzi V.⁽¹⁾, Valentini M. ⁽³⁾

(1)                                                                                                                                                                                                                                                   CRA-GPG, Genomic Research Centre,  Via S.Protaso 302, 29017-Fiorenzuola d’Arda (PC), ⁽²⁾ CRA-API, Research  Unit for Apicolture and Sericolture, Via di Saliceto 80, 40128-Bologna, ⁽³⁾ CRA-RPS, Research Centre for the Soil-Plant System, Instrumental Centre of Tor Mancina, Strada della Neve km. 1, 00015 Monterotondo

ITALY

Traceability, Quality, Honey, Botanical and Geographical origin

 Quality and safety of food products has always been a primary need in the food chain, in order to guarantee the consumer not only microbiological healthiness, but different qualitative aspects of  all food. Quality is a multifactorial parameter linked to species and variety, agricultural methods and geographical origin.

 For honey, one of the fundamental aspects of quality that affects its commercial value, is its botanical and geographical declaration of origin, as in its food chain, from harvesting to packing, the product should not have been submitted to any treatment that has modified its composition. Nowadays the diagnostic methods to determine the botanical and geographical origin are based on the sensory, microscopic and physico-chemical analysis.    

In this work, analytical approaches of molecular traceability have been developed with the aim of establishing the botanical species in products from bees.

 The use of methods of analysis based on nucleic acids allows a greater flexibility, due to the variety of the methodological approaches, as well as the robustness, speed and reproducibility of the process. DNA can be extracted from a variety of matrixes, is stable to technological treatments and therefore is particularly suitable for traceability in the process chain.

 The Real-Time PCR tests allow for the identification, in a precise manner, of the species, variety and the place of origin of the food product.

Furthermore, the metabolic profile of honey has been defined by NMR spectroscopy. The aim was to identify the low molecular weight components, i.e. metabolites, responsible of specific quality features, such as the botanic species, any treatments undergone and the area of origin.

39. THE BEE PERGA-ECOLOGICAL FOOD AND VALUABLE PROTEIN CONCENTRATE OF PATIENTS SUFFERING  FROM  ANDROID OBESITY AND METABOLIC SYNDROME

Dr.Evgeni Petkov,

 Apitherapy Union in

E-mail: apispharma@abv.bg 

BULGARIA

Bee pollen is male sexual cells of blooming plants. Pollen seeds are invisible with the naked eye. Their size varies from 2 to 250 micrones. Pollen seeds are living cells preserved from the hostile environment by a cover called exzine.

 Perga is a bee product which is stored in the cells of honey-combs. It is used in medical practice as food, medicine, for disease prevention .  

Two studies over patients suffering from android obesity were conducted. A three-month course of perga was carried out. The patients reduced their food taking to 1200ccal per day. The two main indicators-waist/hip ratio and BMI/body mass index/were observed. Twenty grams of perga, equalled to 49, 20 ccal, was added to the breakfast at ten a.m. Waist/hip ratio and BMI was measured every two weeks.

  The aim of the second study was to estimate the prega effect on BMI and the cholesterol.One-month course was carried out. Twenty grams of perga were taken by patients two times a day in the morning and in the night.   

 During both of the experiments the patients haven`t taken any fibrates or statins. Patients suffering from type 2 diabetes were on their normal therapy.

 The study showed decrease in cholesterol, triglyceride and basal glucemia. 

 Obesity is a complex, multifactor disease. It has connection with a series of factors such as social, psychological, behavioral and metabolic. Android obesity leads to metabolic and blood-vessel disoreder –insulin resistance and following diabetes, hyperprieteinemia, hypercholesterolemia, high blood pressure, metabolic syndrome, /syndrome’’X’’/ and following aterosclerosis. So reducing body weight to normal BMI is the way to overcome metabolic and blood-vessel complications in this specific type of obesity. The treatment with bee pollen and perga shows decrease in body mass to the normal values of BMI. This is a basic therapeutic method of prediabetes diabetes treatement.. A significant correlation between improvement of lipid profile and reduction of the risk of blood-vessel diseases has been found.  

 Perga with its quality to decrease lipid metabolism disorder is a vital part from food regime for improving the standard of life for these people. This is the way to broaden the boundaries of the complex treatment of android obesity and metabolic syndrome.

40. NON-AUTHENTIC ENZYMES IN HONEY

Michal Bednar, Dalibor Titera

Bee Research Institute at Dol

Libcice nad Vltavou, 252 66,

Bednar@beedol.cz

CZECH REPUBLIC

There are present several authentic enzymes in honey. Natural origin of these enzymes may be nectar, pollen, bees, or even producers of honeydew.

We have met unusual property of the enzyme activity in some honey samples from markets recent years.

We have found problems with determination of diastase activity in some market honey samples suspected for adulteration. Results obtained using Phadebas method were significantly different to results of Shade method.

The enzyme activity of several honey samples exceeds markedly natural range of enzyme activity in an authentic honey.

The other unusual enzyme in honey is ß-fructofuranosidase. This enzyme is absent in authentic honey, but is present very often in market products, declared as honey. The amount of ß-fructofuranosidase in this suspicious samples is probably huge, because the activity of this enzyme is over limits of quantification.

The enzyme ß-fructofuranosidase is used for industrial preparation of invert syrups. This enzyme is not authentic in honey. We suppose that market products declared as honey with such high activity of ß-fructofuranosidase is adulterated by industrially inverted syrup.

41. IDENTIFICATION OF AROMATIC PATTERNS BY ELECTRONIC NOSE AND MULTIVARIATE ANALYSIS OF COLOMBIAN STINGLESS BEE HONEY

Consuelo Díaz M, Carlos M. Zuluaga D, Martha C. Quicazán de C

Instituto de Ciencia y Tecnología de Alimentos

Universidad Nacional de Colombia

COLOMBIA

Fifty three samples of Colombian stingless bees honey corresponding to genus Mellipona and Trigona from 2 different geographical regions were analyzed with an electronic nose. Stingless bees are known traditionally by indigenous population and recognized by sensorial and therapeutically properties of their products. The electronic nose was used to generate a pattern of volatile compounds present in honey samples in order to identify aromatic patterns which allow assessing the suitability of this instrument for discriminating among honey from different geographical origins. Data were evaluated by Principal Component Analysis (PCA); Discriminant Function Analysis (DFA) and Hierarchical Cluster Analysis (HCA). It was found that the main groups of volatile compounds responsible of aroma are those with a ring structure, short chain aliphatics, alcohols and organic sulfur substances. The results were also compared to data obtained from the volatile analysis of 53 samples of honey of the specie Apis mellifera of the same geographical regions; the multivariate analysis allowed to differentiate the samples of stingless bee honey and Apis mellifera. The obtained results suggest that electronic nose is a useful tool for the characterization of honey, which would allow searching differences that permit the establishment of Designation of Origin in order to add value to this product and enhance its popularity and demand.

42. HONEYDEW HONEYS CHARACTERIZED BY THEIR ANTIOXIDANT ACTIVITY, FLAVONOID AND POLYPHENOL CONTENTS.

Rodríguez-Malaver AJ¹, Sancho MT², Gutiérrez MG¹,Thrasyvoulou A³, Vit P¹.

¹Universidad de Los Andes, Venezuela; ²Universidad de Burgos, Spain; ³Aristotle Univesity Thessaloniki , Greece.

VENEZUELA

European consumers can choose from nectar and honeydew honey, based on sensory perceptions. Honeydew honeys have also been characterised by chemical and microscopic analysis, to differentiate the floral or non floral origin of honey. In this work, a collection of honeydew honeys from all over the world was provided by the International Honey Commission to test bioactive properties such as the antioxidant activity and  also the flavonoid and polyphenol content. We followed spectrophotometric methods to measure the antioxidant activity AA (210.16 ± 154.34) mmol Trolox equivalents/100 g honey, flavonoid contents F (3.89 ± 1.37) mg quercetine equivalents/100 g honey, and polyphenol contents P (73.32 ± 31.51) mg gallic acid equivalents/100 g honey. According to the geographical origin of the honeydew, the following values were achieved in mediterranean countries including Brasil (Brasil, Croatia, France, Greece, Slovenia, Spain): A (247.58 ± 170.07), F (3.95 ± 1.39), P (83.99 ± 36.42) and others (Bulgaria, Czech Republic, Germany, Rumania, Slovak Republic, Switzerland): A (168.81 ± 124.36), F (3.84 ± 1.37), P (61,83 ± 19.95). This output shows that higher antioxidant activity is not explained by flavonoid and polyphenol contents.

43. EFFECT OF HEAT ON VOLATILE COMPOUNDS OF HONEY

Allisandrakis E and P. Xarizanis

Agriculture University Athens,

GREECE

44. VOLATILE COMPOSITION OF HONEYDEW HONEY FROM DIFFERENT GEOGRAPHICAL ORIGINS BY HS-SPME-GC-MS ANALYSIS

Silvia Sponza¹, Lanfranco Conte², Mojca Korošec³, Dražen Lušić⁴, Chlodwig Franz¹, Johannes Novak¹

¹Institute for Applied Botany, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, AUSTRIA

²Department of Food Science, University of Udine, Via Sondrio 2A, I-33100 Udine, Italy

³Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101,SI-100 Ljubljana, Slovenia

⁴Department of Food Technology & Control, Faculty of Medicine, University of Rijeka, Braće Branchetta 20, 51000 Rijeka, CROATIA

Honeydew honey is increasingly valued by consumers because of its strong and characteristic flavor that is different from floral honeys. Such importance notwithstanding, the honeydew honey is so far not well investigated.

The volatile profile is one of the typical features of honey. The volatile’s composition is specific for each type of honey and closely related to the geographical region of production. In order to characterize the volatile fraction of honeydew honey, HS-SPME-GC-MS and HS SPME-GC-FID analysis were performed. The identification of the volatile compounds were done with the aim to find marker compounds useful for the quality and authenticity control of the honeydew honeys. The studied samples were coming from three different geographical regions: Croatia, Slovenia and Austria.

The results show a high number of volatile compounds. The compound’s mean values range between 0.1-10.0 percent. Some of the compounds present in higher percentage are: benzaldehyde, n-nonane, α and γ-terpinene, benzeneacetaldehyde, trans- and cis-linalool oxide, n-nonanal, phenyl ethyl alcohol, 1-nonanol, n-decanal. Unfortunately it is not possible to highlight single volatile compounds that can be used as geographical markers, but all three groups of samples present characteristic volatile profiles. Due to particular geographical and botanical characteristics as well as climate conditions, a canonical discriminat function analysis (CDA) was applied to volatile compositions to distinguish honeys from Croatia, Slovenia and Austria. The territorial map obtained with CDA shows good discrimination between the honeydew honeys of the three regions. The two functions allowed a good classification of unknown sample (cross-validated). The high classification percentage indicates the possibility to apply the CDA using the composition of the volatile profile of honeys as a quality assurance tool enabling authenticity control.

Keywords: honeydew honey, volatile profiles, geographical markers, HS-SPME-GC-MS, CDA

 Corresponding author: Silvia Sponza, rovigno_2000@yahoo.it

45.  TOTAL FLAVONOID CONTENT OF HONEYS FROM BURGOS (SPAIN) AND ITS RELATIONSHIP WITH ELECTRICAL CONDUCTIVITY

 Fernández-Muiño MA*, Sánchez-Susinos E, Cavia MM, Alonso-Torre SR, and Sancho, MT.

University of Burgos (Spain). Department of Biotechnology and Food Science. Faculty of Sciences. Plaza de Misael Bañuelos s/n. 09001 Burgos.

*Presenting author. E-mail: mafernan@ubu.es

SPAIN.

Flavonoids represent the most important group of honey phenolic compounds. They are currently receiving much attention because they can play a role as natural antioxidants, antibacterial, anti-inflammatory and vasodilator agents. Several relationships were described between flavonoid content and colour of honey. As colour is also related with electrical conductivity, because the darker the colour, the higher the electrical conductivity, the purpose of this work was to research if there was any relationship between total flavonoid content and electrical conductivity of honey. The analyses were carried out on 56 honeys from the province of Burgos (Spain). Total flavonoid content was measured by the Dowd colorimetric procedure. Flavonoids react with AlCl3 resulting in a yellow compound, whose intensity of colour varies depending on the flavonoid content of the sample. Total flavonoid content was very variable. Values ranged from 3.1 to 78.3 mg quercetin/100 g, with a mean value of 17.1 mg quercetin/100 g. 23% of samples showed values of total flavonoid content higher than 20 mg quercetin /100 g. In respect of electrical conductivity, 75% of honeys showed values lower than 0.8 mS/cm. At a 99% confidence level, we found a significant relationship between total flavonoid content and electrical conductivity (r= 0.7576) that would be worth studying in depth.

Acknowledgement: Authors thank the Regional Government “Junta de Castilla y León” (Spain) that supported this work under the project BU007A08

46. COMPARISON OF ULTRASONIC SOLVENT EXTRACTS OF SALIX SPP. HONEYDEW AND NECTAR HONEY

Igor Jerković¹, Zvonimir Marijanović², Dragan Bubalo³, Nikola Kezić³

¹ Faculty of Chemistry and Technology, Teslina 10/V, Split, Croatia

² Marko Marulić Polytechnic in Knin, P. Krešimira IV 30, 22300 Knin, Croatia

³ Faculty of Agriculture, University of Zagreb, Svetošimunska 25, 10000 Zagreb, CROATIA

Among the compounds identified (ultrasonic solvent extraction followed by GC-MS), markers of the Salix spp. nectar honey botanical origin were phenylacetic acid, benzoic acid, 2-hydroxybenzoic acid, 4-hydroxybenzoic acid, 4-hydroxybenzyl alcohol and other compounds that originated from shikimate biosynthetic pathway, similar as for Salix spp. honeydew honey. All these compounds probably originated from Salix spp., a well known source of salicylic acid and its derivatives. High abundance of 3-methylpropanoic acid, 3-methylbutanoic acid, 2-methylpentanoic acid and 3-methylpentan-1-ol can be noted as specific for the nectar honey and several of them were found with lower percentages in the honeydew honey. The presence of pinocembrin and 8-hydroxy-4,7-dimethylcoumarin can be emphasized, since they were not identified in the honeydew honey. Phenylacetonitrile was present only in the nectar honey. Another group of abundant compounds were terepens, particularly linalool derived compounds. In comparison with the honeydew honey much more pronounced qualitative and quantitative composition of lilac aldehydes and alcohols were found. Norisoprenoids can be additionally considered as characteristic for the nectar honey, particularly 3-hydroxy-trans-β-damascenone, trans-β-damascone, 4-ketosiophorone, α-isophorone and others.

Keywords: Salix spp. honeydew and nectar honey, Volatile compounds, Ultrasonic solvent extraction, Gas chromatography–mass spectrometry

47.  ANTIOXIDANT ACTIVITY OF HONEYS FROM BURGOS (SPAIN) AND ITS RELATIONSHIP WITH TOTAL PHENOLIC CONTENT

Sancho MT*, Sánchez-Susinos E, Alonso-Torre SR, Cavia MM, and Fernandez-Muiño MA.

University of Burgos (Spain). Department of Biotechnology and Food Science. Faculty of Sciences. Plaza de Misael Bañuelos s/n. 09001 Burgos.

*Presenting author. E-mail: mtsancho@ubu.es

SPAIN.

Nowadays the determination of the antioxidant activity of honeys is a matter of significance. On the one hand, natural antioxidants are effective in preventing several diseases. On the other hand, honey can minimize deteriorative oxidation reactions in food commodities. Several researchers found relationships between antioxidant activity of honeys and total phenolic contents. Nevertheless, depending on the method employed to measure the antioxidant activity, the correlations were considerably different. The purposes of this work were first, to determine in 56 honeys from the province of Burgos (Spain), the antioxidant activity by ORAC (oxygen radical absorbance capacity) method, as well as the total phenolic content by Folin Ciocalteu procedure, and then, to find a possible relationship between these parameters. Almost the half of the honeys analyzed showed an antioxidant activity between 12 and 24 ORAC units (μmol TE/g). With regard to the total phenolic content, more than 90% of samples exceeded the value of 20 mg gallic acid/100 g honey, whereas 30 honeys exceeded the value of 80 mg gallic acid/100 g honey. A statistically significant relationship (99% confidence level) between antioxidant activity and total phenolic content was found (r= 0.7587). After applying t-test, F-test, Mann-Whitney W-test, and Kolmogorov-Smirnov-test no significant differences were found between true and calculated antioxidant activities at a confidence level of 95%. If this correlation keeps in honeys from other geographical origins, the antioxidant activity of a given honey, whose measure is tedious and time consuming, could be calculated from its total phenolic content, therefore drastically reducing the experimental effort, with a significant saving of time and money.

Acknowledgement: Authors thank the Regional Government “Junta de Castilla y León” (Spain) that supported this work under the project BU007A08.

48 CHARACTERIZATION OF SIERRA MORENA (SOUTH SPAIN) EUCALYPTUS HONEY

Rodríguez¹, I; Serrano¹, S.; Jodral¹, M.; Galán¹, H.; Ubera², J.L.; Vicente²,M.; and Carmona², R.

¹Department of Food Hygiene and Technology. University of Cordoba. Campus Rabanales, Edif. Darwin.14071, Cordoba, Spain.

²Department of Botanical and Vegetal Biology. University of Cordoba. Campus Rabanales, Edif. Celestino Mutis.14071, Cordoba,

SPAIN.

Characterization based on botanical, physicho-chemical and sensory analysis was carried out with the aim of establishing criteria for a future Protected Designation of Origin that includes Eucalyptus and other monofloral origins produced in Sierra Morena (geographical region in south Spain that covers the north of Andalusia). After the analysis of 41 samples provided from beekeepers as Eucalyptus honey, just 13 of these samples were set as homogeneous criteria probing their authenticable origin. Physicochemical parameters analyzed and criteria established were: water (≤17.5 %), pH (4.1-5.0), free acidity (≤35 meq/kg), electrical conductivity (0.38-0.58 mS/cm), color (40-60 mm Pfund), diastase (8.0-29.0 D.N.), glucoseoxidase (2.5-17.2-5 μg peroxide/g and hour) and hydroximethylfurfural (≤28 mg/kg). The botanical analysis confirmed minimum Eucalyptus pollen of 87% and Maurizio class III. Others pollens isolated were Reseda, Echium, Trifolium and Cistus. They were found two sensory profiles (five points scale) set as:

•         Pale amber colour, a medium smell and taste intensity,  (3) with hints of wood and vanilla and animal touches, low pastiness (1-2) and a very fine crystallization (1-2), a slightly salty taste (2), freshness, and a medium persistence (3).

•         Yellow,(3), with a medium to high smell and taste intensity (3-4) with whiffs of liquorice, coffee and chocolate and a fruity taste of crystallized orange,  a medium pastiness (3) and rough crystallization (4), a salty taste (2-4), freshness, spicy and a medium persistence (3).

49 CHARACTERIZATION OF SIERRA MORENA (SOUTH SPAIN) ECHIUM HONEY.

Rodríguez¹, I; Serrano¹, S.; Jodral¹, M.; Galán¹, H.; Ubera², J.L.; Vicente²,M.; and Carmona², R.

¹Department of Food Hygiene and Technology. University of Cordoba. Campus Rabanales, Edif. Darwin.14071, Cordoba, Spain.

²Department of Botanical and Vegetal Biology. University of Cordoba. Campus Rabanales, Edif. Celestino Mutis.14071, Cordoba,

SPAIN.

Characterization based on botanical, physicho-chemical and sensory analysis was carried out with the aim of establishing criteria for a future Protected Designation of Origin that includes Echium and other monofloral origins produced in Sierra Morena (geographical region in south Spain that covers the north of Andalusia). 57 samples provided from beekeepers as Echium honey were set as homogeneous criteria probing their authenticable origin. Physicochemical parameters analyzed and criteria established were: water (≤17.5 %), pH (3.6-5.9), free acidity (≤35 meq/kg), electrical conductivity (≤0.5 mS/cm), color (≤60 mm Pfund), diastase (8.0-33.0 D.N.), glucoseoxidase (2.5-17.2-5 μg peroxide/g and hour) and hydroximethylfurfural (≤28 mg/kg). The botanical analysis confirmed minimum Echium plantagineum pollen of 85% and Maurizio class III or IV. Other pollen isolate was Myrtus communis. It was found a sensory profile (five points scale) set as:

White-very pale yellow colour; low smell and flavour intensity(2-3) clearly vegetal with balsamic touches of camphor and resin, woody hints, cooked vegetable and cabbage; sweet taste (3-4); low pastiness (1-2) and fine crystallization (1-2) and low-medium persistence (2-3).

, 

50 SPANISH LAVENDER (LAVANDULA STOECHAS) HONEY FEATURES IN SOUTH SPAIN.

Ubera, J.L, Granados,C.; Royo, M.; Rodríguez, I; Serrano, S.; Jodral, M. and Galán, H.

¹Department of Botanical and Vegetal Biology. University of Córdoba. Campus Rabanales, Edif. Celestino Mutis.14071, Córdoba, Spain.

²MAZARA Botanical Analysis S. L. Córdoba. Spain.

³Department of Food Hygiene and Technology. University of Córdoba. Campus Rabanales, Edif. Darwin.14071, Córdoba,

SPAIN.

Honey formed from Spanish lavender (Lavandula stoechas), known in Spain as cantueso, is one of the uniflorals produced in Sierra Morena, South Spain. Its palynologycal, physicochemical and sensory characteristics were defined after a whole analysis process of 204 samples of honey with the purpose of establishing the criteria for a future Protected Designation of Origin (PDO). In this PDO, 7 different types of honey were described. Up to 10 of those samples were characterized as Spanish lavender honey, which features are described here.

From the palynologycal analysis it was concluded that the minimum Lavandula stoechas pollen percentage for this honey to be considered unifloral according to this PDO must be 12%. Echium, Myrtus or Reseda can appear as the dominant pollen type. Taking into account its PK_10, this kind of honey can be included either in II or III Maurizio class.

As regards the physicochemical characteristics, the colour in mm Pfund can vary from 35 to 55. The electrical conductivity ranges from 0.25 to 0.45 mS/cm. The diastatic activity varies between13 and 25 Gothe scale units.

Regarding its sensory characteristics it is established (five points scale) that it has yellow colour, low fluidity (1-2), low odour/aroma intensities (2-3) associated with floral and fruit notes respectively; medium sweet taste (3) and low persistence.

51. CHEMICAL COMPOSITION OF SEVERAL MONOFLORAL CUBAN HONEYS AND ITS CORRELATION WITH COLOR AND THEIR ANTIOXIDANT CAPACITY.

Alvarez-Suarez J.M ¹, , Romandini S ¹, Giampieri F ¹, Vidal-Novoa A ², Battino M¹

¹ Department of Biochemistry, Biology & Genetics, Faculty of Medicine. Marche Polytechnic University, Italy, ² Department of Biochemistry, Faculty of Biology, Havana University, CUBA.

Address correspondence to this author: Dr. Jose Miguel Alvarez Suarez. Dept of Biochemistry, Biology & Genetics, Faculty of Medicine, Marche Polytechnic University, Via Ranieri 65, 60100 Ancona, Italy. phone:+39 071 2204136, fax: +39 071 2204136,  e-mail: jmals2000@yahoo.es

CUBA.

Several monofloral Cuban honey (83) were analyzed to determine their amino acid and total protein content. The total phenolic, flavonoid and carotenoid contents as well their total antioxidant capacity using the ferric reducing antioxidant power (FRAP) also were studied. The honey samples coming from Christmas vine flowers (Turbina corymbosa,(L.) Raf), morning glory (Ipomoea triloba L.), black mangrove (Avicennia germinans Jacq.), linen vine (Govania polygama (Jack) Urb) and singing bean (Lysiloma latisiquum (L.) Benth). Free amino acids content, determinate by the Cd-ninhydrin method, and expressed as mg of Leucine equivalent (LE)/100g of honey, report the high values in morning glory honey (146.42 mg LE/100g) and the low content in christmas vine (44.64 mg LE/100g). Total protein content, by the Bradford assay, and expressed as mg of bovine serum albumin (BSA)/100g of honey report the high values in linen vine honey (92.38 mg BSA/100g) and the low result in christmas vine (12.06 mg BSA/100g). The results of the study showed that total phenolic, flavonoid and carotenoid content as well their antioxidant activity differ widely among different honey types. Phenolic content, expressed as gallic acid equivalent, range from 213.6 mg GAE/ Kg in christmas vine to 493.6 mg GAE/Kg in linen vine honey, flavonoid content, expressed as catechin equivalent, range from 10.9 mg CE/Kg in Christmas vine to 29.4 mg CE/Kg in linen vine honey and the carotenoid content, expressed as β-carotene equivalent, varied from 5.57 mg β-CE/Kg honey (linen vine, Ambar color) to 0.35 mg β-CE/Kg honey (christmas vine, Light honey). The antioxidant activity was the lowest in christmas vine honey (27.09 μmol TE/100g) and the highest in linen vine honey (96.96 μmol TE/100g). A high correlation was found between phenolic and total flavonoid content (r= 0.8346) and total poliphenolic content and total antioxidant capacity (r= 0.8268).

52 TOTAL ANTIOXIDANT CAPACITY AND FREE RADICAL FORMATION OF SEVERAL CUBAN HONEYS FROM DIFFERENT FLORAL SOURCES WITH POSSIBLE ANTIBACTERIAL ACTIVITY.

Alvarez-Suarez J.M ¹, , Romandini S ¹, Giampieri F ¹, Vidal-Novoa A ², Battino M¹

¹ Department of Biochemistry, Biology & Genetics, Faculty of Medicine. Marche Polytechnic University, Italy, ² Department of Biochemistry, Faculty of Biology, Havana University, CUBA.

Address correspondence to this author: Dr. Jose Miguel Alvarez Suarez. Dept of Biochemistry, Biology & Genetics, Faculty of Medicine, Marche Polytechnic University, Via Ranieri 65, 60100 Ancona, Italy. phone:+39 071 2204136, fax: +39 071 2204136,  e-mail: jmals2000@yahoo.es

CUBA

53. CHEMOMETRICS AND MINERAL CONTENT FOR THE IDENTIFICATION OF GEOGRAPHICAL ORIGINS OF SPANISH HONEYS .                                    

Suárez-Luque S., Mato I, Huidobro JF, Simal-Lozano J, and Sancho MT

1University of Santiago de Compostela. Department of Analytical Chemistry, Nutrition and Food Science. Faculty of Pharmacy. Campus Universitario Sur. 15782 Santiago de Compostela. Spain.

2University of Burgos (Spain). Department of Biotechnology and Food Science. Faculty of Sciences. Plaza de Misael Bañuelos s/n. 09001 Burgos.

*Presenting author. E-mail: mtsancho@ubu.es

SPAIN.

The mineral content of honey samples could give an indication of environmental pollution and herewith also an indication of the geographical origin of honey. Therefore, the objective of this work is to obtain a multivariate statistical model able to classify honeys according to their geographical origin with using the mineral composition and some of its physicochemical properties. Nineteen parameters were determined (colour parameters such us x and y trichromatic coordinates, dominant wavelength, absorbance net, turbidity electrical conductivity, pH, content of potassium, calcium, sodium, magnesium, manganese, nickel, lithium, chloride, nitrate, sulphate, formic acid and phosphate) in 125 samples of honey from different regions of Spain. The results were analyzed using a multivariate statistical study consisting of a principal component analysis and linear discriminant analysis resulting in a model that differentiates the honeys according to their geographical origin with a percentage of correct classification of 87.3%.

54.  IDENTIFICATION OF BOTANICAL ORIGIN OF HONEYS BY THEIR PHYSICOCHEMICAL PROPERTIES AND MINERAL CONTENT

Suárez-Luque S.1, Mato I1, Huidobro JF1, Simal-Lozano J1, and Sancho MT2*

1University of Santiago de Compostela. Department of Analytical Chemistry, Nutrition and Food Science. Faculty of Pharmacy. Campus Universitario Sur. 15782 Santiago de Compostela. Spain.

2University of Burgos (Spain). Department of Biotechnology and Food Science. Faculty of Sciences. Plaza de Misael Bañuelos s/n. 09001 Burgos.

*Presenting author. E-mail: mtsancho@ubu.es

SPAIN.

The mineral content of honey is in a ratio less than 1%. It influences in honey such properties as colour, electrical conductivity as well as pH, and it is important because mineral content could give an indication of environmental pollution and herewith also an indication of the geographical and botanical origin of a given honey sample. In this study, colour parameters, electrical conductivity, pH and the mineral content of 125 honey samples from 12 different botanical origins (Castanea sativa Miller, Citrus sp., Echium sp., Erica sp., Eucalyptus sp., Lavandula sp., Rosmarinus sp., Rosmarinus and Trifolium sp., Rubus sp., Trifolium L. sp., multifloral, and blends of multifloral and honeydew honey) were studied. Results were submitted to unsupervised methods such as principal components analysis and supervised learning methods like linear discriminant analysis in order to evaluate possible data patterns, and the possibility of differentiating Spanish honeys according to their botanical origins. Principal components analysis explained 82.3% of the variance with the first six principal compontent variables. The linear discriminant analysis allowed correct botanical classification of 73.2% of the samples.

55. DETERMINATION OF PHYSICOCHEMICAL CHARACTERISTICS OF HONEYS COMPARING FOURIER TRANSFORM INFRARED SPECTROSCOPY AND REFERENCE METHODS.

Christina Kast and Verena Kilchenmann

Swiss Bee Research Centre, Agroscope Liebefeld-Posieux Research Station ALP, Schwarzenburgstrasse 161, 3003 Bern,

SWITZERLAND

Differentiation between nectar and honeydew honeys is based on sensory and microscopic characteristics and also includes physicochemical characteristics of the honeys, such as the electrical conductivity, the free acidity and the pH. Fourier Transform Infrared Spectroscopy (FT-IR) is faster and less laborious than the reference methods using a conductimeter, a pH meter or the reference method by equivalence point titration. Therefore we evaluated the suitability of the FT-IR method for the determination of some physicochemical measurands distinguishing floral and honeydew honeys.

Three different honey samples with low, intermediate and high electrical conductivities and free acidities were measured 10 times each at 10 different days using the FT-IR and the reference methods. The means and repeatability limits of the methods were compared. Additionally rape and honeydew honeys with low and high electrical conductivities were included in the comparison.

The reference - and FT-IR methods for the pH and the water determination presented similar means and repeatability limits (r=2.83Sr) within the range that was tested, suggesting that both methods are equivalent.

The repeatability limits of the electrical conductivities and the free acidities determined by the FT-IR method were wider than those of the reference methods, but they were still in a range that is not critical for most analyses.

The means of the electrical conductivity of both methods were similar in the intermediate range of the calibration curve, suggesting that the FT-IR method is suitable for the discrimination of honeydew and floral honeys at 0.8mS/cm. However, the FT-IR method gave significantly higher values at the low range of the calibration curve. Therefore, the FT-IR method is not equivalent to the reference method in this range and not suitable for reliable discrimination of some monofloral honeys from polyfloral honeys at the low end of the calibration curve around 0.25-0.28 mS/cm. It may be necessary to recalibrate the FT-IR method separately for the low and the high range.

While the FT-IR methods for the pH and the water determination were equal to the reference methods, the conductimetric reference method for electrical conductivity was superior to the FT-IR method.

56.  DETERMINATION OF BENZO(A)PYRENE IN BEESWAX .

 Corredera, L., Bayarri, S., Pérez, C., Lázaro, R., Herrera, A.

Department of Animal Production and Food Science, Faculty of Veterinary, University of Zaragoza

SPAIN

Polycyclic aromatic hydrocarbons (PAHs) are one of the most important groups of environmental pollutants, some of which are known or suspected carcinogens or mutagens to humans. They are widely present in the environment due to their lipophilic properties, which might allow their adsorption onto beeswax and the use of this bee product as a biological indicator.

 Benzo(a)pyrene (BaP) is one of the most carcinogenic of the PAHs and it has been extensively studied, however, little information is available about this type of pollutants in beeswax and other bee products.

 In this work, an analytical method for the identification and quantification of BaP in beeswax has been developed and evaluated. The method consists of a sample preparation with methanol, followed by a solid phase extraction (SPE) and a quantification performed using high-performance liquid chromatography (HPLC) with fluorescence detection. Analytical performance of the proposed method, including sensitivity, accuracy and precision was satisfactory.

Keywords: Benzo(a)pyrene; PAHs; beeswax; HPLC.

Acknowledgements .The authors wish to thank the Government of Aragón for giving a scholarship to Lourdes Corredera, and for its financial support (Grupo de Investigación Consolidado A01/2009 and Ayudas Apícolas).

57 IDENTIFICATION  OF  UNKNOWN  COLOUR  IN  HONEY  USING  UPLC/TOF AND  HPLC/MS/MS

Robert Germuska, Eva Hrnciarikova, Michaela Vierikova

State Veterinary and Food Institute Dolny Kubin

Janoskova 1611/58, Dolny Kubin,

E-mail: germuska@svpudk.sk

SLOVAKIA

Customers in the Central Europe think that dark honey has better quality than light honey. Forest honey is usually darker than others. It is very easy to add unauthorized colour mixture to light honey or more often to honey diluted with inverted sugar syrups. Effect is that adulterated honey looks like forest honey. It is fraud, because according to the Council Directive 2001/110/EC is banned to add any food ingredients to honey. Unusual cheap pitch dark honey from market is therefore suspected.  

This paper describes analytical strategy for identification this type of adulteration. At the very beginning of study was investigated what can be used as colorant. After asking from beekeepers we received to the laboratory unknown brown syrup. Next investigation confirmed that this unknown syrup is probably food additive E-150d using for colouring of beverages, breads etc. This syrup is mixture of several substances. We found then low amount of this syrup added to light honey has colorific effect. Therefore main goal of this study was to develop sensitive and selective analytical method applicable for identification of this syrup on lowest concentration in honey as soon as possible, the best around 50 mg.kg^-1. The first part of the analytical step was to identify major markers from this unknown solution. Diluted unknown brown syrup was infused using positive ESI to MicrOTOF Focus II, (Bruker Daltonic) TOF (Time of Flight) mass spectrometer. Average high resolution mass spectrum within range 100 – 1000 Da was acquired. After evaluation of data two candidate exact masses as possible markers were identified. Then separation conditions were developed using reverse phase liquid chromatography (UPLC) coupled with TOF. Extracted ion chromatograms showed that two selected exact masses are truly present in brown syrup but not in any type of pure honey. In general TOF is less sensitive than low resolution mass spectrometers worked under MS/MS conditions. Therefore was developed MS/MS fragmentation of one selected exact candidate mass using ion trap mass spectrometer (Gemini, Bruker Daltonic). Both analytical detection approaches showed good agreement but MS/MS is slightly more sensitive like TOF.

In last part of this study were analyzed 26 suspected forest honeys from market. In 11 honeys were detected markers comes from brown syrup what was surprised.

58. OCCURRENCE OF MICROSCOPIC ELEMENTS AND THEIR RELATION WITH THE ORIGIN OF HONEY.

 Escuredo O., Seijo M C. & Chouza M

Dpt. Vegetal Biology and Soil Sciences. Faculty of Sciences. University of Vigo. As Lagoas. 32004 Ourense.

E-mail: oescuredo@uvigo.es

SPAIN.

The microscopic analysis of honeys let to recognize different structures. Pollen grains have been the best studied elements, however others like spores of fungus or yeast are more unknown. Some of them come from primary sources and are associated at the presence of honeydew such Alternaria, Stemphylium or Leptosphaeria. In the case of yeast a variety occurs regularly in the floral nectar of many plant species, where they frequently reach extraordinarily high densities. This is the case of some species of Metschnikowia or Candida. 

The aim of this work is to study some of these microscopic elements and their relation with the source of honey. The samples are from NW Spain and were provided for the beekeepers. We realize a physicochemical analysis includes: humidity, HMF, diastase and invertase content, sugars pattern, mineral content, pH, electrical conductivity, phenols content, flavonoids content and percentage of RSA; a melissopalynological analysis and a sensorial study.

It has been analyzed the relations between microscopic elements traditionally considered as honeydew elements, yeast and algae and physicochemical, sensorial and palynological parameters used for honey typification. The results have allowed to establish relationships on the honeydew contribution to the honeys and their botanical origin.

Acknowledgements

This study is financed by Conselleria de Medio Rural, Ministerio de Medio Ambiente, Medio Rural y Marino and FEADER. Research project FEADER 2008-5.