Open in a separate window 2. Folin-Ciocalteu reagent was supplied by the Merck Darmstadt, Germany. Acetonitrile Sodium carbonate and acetic acid Bratislava, Slovakia. Honey 2 g or royal jelly 2 g were dissolved in 15 mL of bidistilled water before analysis. Bee pollen, beebread and propolis samples each 2 g were extracted with 15 mL of bidistilled water for 24 h at room temperature.
Prepared solutions were filtered through a 0. Calibration of pH-meter was performed with three different buffer solutions having pH values of 4, 7 and 10 [ 21 ]. Refractometry Bee product under test 2 legjobb anti aging éjszakai krém termékek was mixed with 8 mL of bidistilled water and macerated for 24 h.
The prepared solutions were centrifuged at 10,× g for 15 min and filtered through a 0. Content of soluble solids and amount of NaCl were expressed as a percentage [ 22 ].
Evaluation of Oxidation-Reduction Potential An amount of 1. Honey and royal jelly were filtered with a 0.
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The results were expressed in mV. Ultraviolet-Visible Scanning Spectrophotometry Tested bee product 0.
The extracts were filtered through a paper filter Labbox, Barcelona, Spain and then through a 0. The UV-Vis spectra of bee products were performed using different dilution levels: bee pollen samples were diluted 11 times, beebread 18 times, royal jelly 10 times, propolis 19 times and dilution of three times was used for honey solutions. For all absorbance measurements Quartz cells 1 cm were used [ 25 ].
Spectrophotometric Evaluation Total phenolic compound content, total best anti aging cream in india for 30s content and radical scavenging activity were determined spectrophotometrically in bee products using methodology described in Kaškonienė et al. For these tests an amount of 1 g of bee product was suspended in 10 mL of bidistilled water.
The insoluble products, namely, bee pollen, beebread and propolis, were subjected to traditional maceration extraction for 24 h. Obtained extracts and solutions were filtered with a 0. Total phenolic content was estimated by the Folin-Ciocalteu method.
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Extracts 8 µL were mixed with µL of 3. A calibration curve of rutin was prepared 0.
Results are expressed as mg rutin equivalent RUE per 10 g of raw sample. Each prepared extract 10 µL was mixed with µL of the stock solution and left for 30 min at 4 °C temperature.
After incubation, samples were measured at nm wavelength.
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Total flavonoid content was evaluated using standard curve of rutin 0. Radical scavenging activity was determined according to colorimetric reaction using 1,1-diphenylpicrylhydrazyl DPPH free radical.
Bee product extract 5. All prepared samples were kept for 15 min at 22 ± 2°C temperature in the dark room and then measured at nm wavelength. Radical scavenging activity was evaluated using a standard curve of rutin 0.
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Homogenised sample 0. Prior to digestion, the samples were soaked in acid solution for 30 min at room temperature.
Digestion was performed under following conditions: temperature °C, pressure psi, ramp time 20 min, hold time 20 min, microwave power W. Then, the digested sample was cooled down, thoroughly transferred into a mL volumetric flask and diluted using bidistilled water to the mark. Each sample was prepared in triplicate and a blank sample was included in each digestion run.
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Qualitative and quantitative evaluation of microelements was performed using inductively coupled plasma mass spectrometer ICP-MS. Inductively coupled plasma mass spectrometry was performed under helium collision-cell He-cell with kinetic energy discrimination mode to remove polyatomic interferences.
The data set representing physicochemical properties was composed of 53 samples best anti aging cream in india for 30s bee pollen, 10 propolis, 11 honey, eight beebread and six royal jelly sampleswhen each of them was described by nine variables pH, electrical conductance, oxidation-reduction potential, amount of NaCl, refraction index, Brix value, total phenolic compound content, total flavonoid content and antiradical activity measured five times.
Data standardization procedure by centering each variable around zero i.
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Hierarchical clustering analysis allowed to present similarity among underlying groups of data using multilevel hierarchical structure, dendrogram, was applied on both prepared data sets—the data set representing physicochemical properties of the samples and the data set built from mineral analysis results. These metrics were used trying to maximise a cophetetic correlation coefficient.
The cophenetic correlation for a dendrogram tree is defined as the linear correlation coefficient between the cophenetic distances obtained from the tree, and the original distances dissimilarities used to construct this tree. Therefore, the cophenetic correlation reveals the adequacy of the built dendrogram representing the dissimilarities among observations.
The cut-off level of the tree to set data into clusters was chosen as a percentage of the maximum observed distance.
Results and Discussion 3. Physicochemical Properties of Bee Pollen and Other Bee Products Bee pollen and four others investigated in the research bee products honey, beebread, propolis and royal jelly were analyzed according to their physicochemical properties Table 2.
Bee pollen samples distinguished by the highest pH values 4. Other studies in literature showed that the pH values of bee pollen varied from 4. As it can be seen, the results of this research coincide with the data published by other authors.
Table 2 Physicochemical properties of bee pollen and other bee products sample codes see in Table 1.