S u m m a r yConiferous honeydew honey, mainly Abies alba was characterised. Samples chosen for the study had organoleptic traits characteristic for the variety: greenish, opalescence tone of brown colour, mild, sweet flavour with pleasant, slightly resinous aftertaste and aroma as well as electrical conductivity over 0.95 mS/cm. To define composition and physicochemical parameters of the variety, contents of water and total sugars were determined. In addition various carbohydrates were identified and their contents were assessed as well. These were: fructose, glucose, sucrose, maltose, turanose, trehalose, isomaltose, malezitose. Other examined parameters related to honey quality were: free acidity, pH, the content of 5-hydroxymetylofurfural (HMF), the main amino acid -proline, and the activity of α-amylase enzyme (Diastase Number). The following properties were proven to be characteristic for this variety: high electrical conductivity with the average value of 1.14 mS/cm, ranging from 0.96 to 1.32 mS/cm; content of monosaccharides lower by few percent in relation to other honey varieties (from 58.2 to 67.4 g/100 g; on average 62.0 g/100 g) and a higher content of disaccharides and trisaccharide -melezitose. The presence of this sugar confirms that a considerable part of the honey was produced from honeydew. The average value of melezitose was 3.2 g/100 g, ranging from 0.9 to 5.9 g/100 g. Also, the results of the pH measurements were slightly higher than in other honey varieties (from 4.23 to 4.99; on average 4.63). The colour value in mm Pfund ranged from 74 to 105, with the average of 93.

a b s t r a c t here we describe a method of hydrocarbon (alkanes, alkenes, dienes) identification and quantitative determination of linear saturated hydrocarbons (n-alkanes) in beeswax using gas chromatography with a mass detector technique (gc-ms). beeswax hydrocarbons were isolated using a solid-phase extraction (spe) technique with neutral aluminum oxide (alumina -n, 1000 mg, 6 mL), then were separated on a non-polar gas chromatography column Zb-5ht inferno (20 m×0.18 mm×0.18 µm). qquantitative analysis of n-alkanes was conducted by the method of internal standard with squalane used as the internal standard. the basic parameters of validation (linearity and working range, limit of determination, repeatability and reproducibility, recovery) were determined. for all of the identified compounds, satisfactory (≥0.997) coefficients of correlation in the working ranges of the method (from 0.005 to 5.0 g/100 g) were obtained. the elaborated method was characterized by satisfactory repeatability and within-laboratory reproducibility. the average coefficients of variation for the total n-alkanes did not exceed 2% under conditions of repeatability or 4% under conditions of reproducibility. the recovery for individual n-alkanes was above 94%; for their total content, it was 100.5%. in beeswax originating from apis mellifera, n-alkanes containing from 20 to 35 carbon atoms in their molecules were determined. the total content of these alkanes was between 9.08 g and 10.86 g/100 g (on average, 9.81 g/100 g). additionally, apart from the saturated hydrocarbons, unsaturated hydrocarbons and dienes were identified.

A b s t r a c t To detect beeswax adulteration with hydrocarbons of alien origin (e.g. paraffi n), gas chromatography with mass detector (GC-MS) technique was used. The method has been verifi ed here on beeswax samples with different addition (3, 5, 10, 30, and 50%) of paraffi n and validated under the conditions of repeatability and within -laboratory reproducibility. The addition of paraffi n to beeswax can already be detected on the basis of an analysis of the chromatograms. The intensity of individual alkane peaks increased with the increase of the amount of paraffi n added to the beeswax. This increase was the mostly visible for the alkanes with even numbers of carbon atoms in the molecule: C 24 H 50 , C 26 H 54 , C 28 H 58 , C 30 H 62 , C 32 H 66 , and C 34 H 70 . These observations have also been proven by quantitative analysis performed using the internal standard method. Adding paraffi n to beeswax resulted in an increase in the total contents of n-alkanes as well as individual alkanes, and in particular, of the even-numbered alkanes. The addition of paraffi n to beeswax also resulted in the appearance of alkanes containing over 35 carbon atoms in the molecule, which were not detected in beeswax. The method for determination of beeswax hydrocarbons with the GC-MS technique is characterised by satisfactory repeatability and within-laboratory reproducibility. This method can be used for the detection of beeswax adulteration with hydrocarbons of alien origin (e.g. paraffi n).Keywords: adulteration, beeswax, GC-MS, hydrocarbons, n-alkanes, paraffi n.Research Institute of Horticulture, Apiculture Department, Puławy, Poland INTRODUDCTIONDue to the insuffi cient domestic production of beeswax and the lack of obligatory legal regulations concerning beeswax quality, instances of its adulteration are quite frequent. The addition of alien substances such as paraffi n, decreases beeswax quality as well as the quality of the foundation made of such adulterated beeswax. As a consequence, it may no longer be possible to use this product in beekeeping. It may also not be possible to use such a product in some branches of industry, for example, the cosmetic, pharmaceutical, and food industry. The sensory assessment and physico-chemical characterisation are only approximate and do not allow to detect the adulteration of beeswax unambiguously. Beeswax which has paraffi n added, becomes chinawhite and pliable when kneaded with the fi ngers. What is more, the addition of paraffi n causes the beeswax to become smooth and shiny where rubbed (Curyło, 1983). As we mentioned previously (Waś et al., 2014a), a lot of methods so far used for the determination of hydrocarbons in beeswax and for the detection of beeswax adulteration were based on the physico-chemical parameters. These parameters include: melting point, density, solubility, and saponifi cation, acid and ester numbers, and iodine index (Tulloch, 1973;Serra Bonvehi et al., 1989;Serra Bonvehi, 1990;Vit et al., 1992;Poncini et al., 1993). Some interrelations were found be...

A b s t r a c t The hydrocarbon composition of beeswax secreted by Apis mellifera was characterised. In the studies, analyses were made of virgin beeswax (obtained from light combs, socalled "wild-built combs") that was collected at different dates, and beeswax obtained from dark combs ("brood combs"). A qualitative analysis did not show any differences in the hydrocarbon composition of beeswax originating from light and dark coloured combs. The same hydrocarbons (n-alkanes, alkenes, and dienes) were identified in virgin beeswax and beeswax collected from brood combs. However, the studies showed differences in the content of n-alkanes in the beeswax obtained from light and dark coloured combs. In comparison to the virgin beeswax, the beeswax obtained from dark combs had higher content of the total n-alkanes, higher total contents of even-numbered alkanes and odd-numbered alkanes, and higher contents of certain alkanes. Furthermore, it has been found that the hydrocarbon composition of beeswax did not depend on the collection period.

A b s t r a c tThe efficiency of the gas chromatography -mass detector (GC-MS) technique for the detection of beeswax adulterated with paraffin, was evaluated. For this purpose, beeswax samples with paraffin additions (3, 5, 10, 30, 50%) were analysed. Since not enough is known about paraffin compositions, and since it is difficult to detect paraffin in beeswax, the aim of our research was also to compare the hydrocarbon composition of different types of paraffin. The analysis showed that the types of paraffin available on the market, differ qualitatively and quantitatively as far as their hydrocarbon compositions are concerned. In all kinds of paraffin, we found homologous series of n-alkanes that were much longer than those in beeswax. In beeswax, the amount of added paraffin that is possible to detect, differs and depends on the kind of paraffin used for adulteration. In this study, the minimum estimated percent that was detected using the GC-MS technique, was 3%. The adulteration is indicated by the presence of hydrocarbons containing over 35 carbon atoms in the molecule, and by the higher contents of n-alkanes (C 20 H 42 -C 35 H 72 ), in comparison to the concentration of these compounds determined in pure beeswax. We also presented the results of the quality control of commercial beeswax. Based on our results, it can be stated that beeswax adulteration is currently a problem. Keywords: adulteration, beeswax, detection, efficiency, GC-MS, paraffinResearch Institute of Horticulture, Apiculture Division in Puławy, Kazimierska 2, 24-100 Puławy, Poland INTRODUCTIONThe recent increased interest in the testing of beeswax composition and in the quality assessment of beeswax, is related to the numerous cases of beeswax adulteration. Substances such as ceresin, stearin, natural plant waxes (e.g. Candelilla wax, Carnauba wax), animal waxes and fats (e.g. lanolin, tallow), and mineral waxes (Montan, ozocerite) have been added to beeswax. It is paraffin, though, that is the product most frequently used for beeswax adulteration. Due to having a similar composition and similar physico-chemical properties, the addition of paraffin to beeswax, especially when added in small amounts, is difficult to detect (Bogdanov, 2009;Jimenez et al., 2009;Serra Bonvehi & Ornantes Bermejo, 2012;Maia, Barros, & Nunes, 2013;Svečnjak et al., 2015). The chemical composition of paraffin has not yet been sufficiently analysed. The information found in literature mostly concerned physicochemical properties (Bernal et al., 2005) or concerned the estimated data on the paraffin composition stated by the producers. From these data, it follows that the group of longstraight-chained saturated hydrocarbons (nalkanes) dominate in paraffin. The content of these hydrocarbons is estimated to be at different levels, and even at levels of up to 90%. Depending on the kind of paraffin (in particular the melting point of paraffin), and the purification level of the obtained product, the paraffin can also contain other hydrocarbons (e.g. branched, cyclic, unsa...

A total of 102 honey samples collected from small apiaries (≤ 20 hives) in Poland were analysed for the presence of Clostridium botulinum spores. The samples were prepared using the dilution centrifugation method and cultured in parallel in cooked meat medium (CMM) and tripticase peptone glucose yeast (TPGY) enrichment broths. Identification of toxin types A, B, and E of Clostridium botulinum strains was performed with the use of the multiplex PCR method. Positive samples were also subjected to quantitative analysis with the use of Clostridium botulinum Isolation Agar Base (CBAB). The prevalence analysis showed 22 (21.6%) samples contaminated with C. botulinum spores. The major serotype detected was botulin neurotoxin type A – 16 (72.7%) whereas type B was found in 3 (13.6%) honey samples and type E also only in 3 (13.6%) honey samples. Dual-toxin-producing strains were noted. The average quantity of spores in PCR - C. botulinum positive samples was 190 in 1 gram of honey.

A b s t r a c t A method was elaborated to determine phenolic compounds (vanillin, caffeic, p-coumaric and salicylic acids, and flavonoids: rutin, hesperetin, quercetin, pinocembrin, apigenin, kaempferol, isorhamnetin, chrysin, and acacetin) in bee pollen loads using highperformance liquid chromatography with a diode array detector (HPLC-DAD). Phenolic compounds from bee pollen were isolated on Cleanert C18-SPE columns (500 mg/6 mL, Agela Technologies). Polyphenols were identified by comparing the retention times and spectra of compounds found in pollen load samples with the ones of the standard mixture. Quantitative analysis was conducted using the external standard method. In addition, basic validation parameters for the method were determined. For the identified compounds (except for the salicylic acid), satisfactory (≥0.997) linear correlations were obtained. The elaborated method showed high repeatability and inter-laboratory reproducibility. Variability coeffcients of the majority of phenolic compounds did not exceed 10% in conditions of repeatability and inter-laboratory reproducibility, and for the total polyphenolic content they were 1.7 and 5.1%, respectively. The pollen load samples (n = 15) differed in qualitative and quantitative composition of the phenolic compounds. In all the samples, we identified the p-coumaric and salicylic acids and flavonoids rutin, hesperetin, and apigenin nevertheless, these compounds' contents significantly differed among individual samples. The total phenolic content in the tested samples of pollen loads ranged from 0.653 to 5.966 mg/100 g (on average 2.737 mg/100 g).