Suppressive effects of ethanolic extracts from propolis and its main botanical origin on dioxin toxicity

Suppressive effects of ethanolic extracts prepared from propolis group 12 and its main botanical origin (leaf bud of Baccharis dracunculifolia) on transformation of the aryl hydrocarbon receptor (AhR), the initial action of dioxin toxicity, were investigated. It was found that suppressive effects of propolis on AhR transformation were relatively higher than those of resins of its botanical origin in cell-free system and in Hepa-1c1c7 cells. When the composition of chemical ingredients was measured, propolis contained slightly higher amounts of flavonoid aglycones as compared with its botanical origin with the same characteristics. Moreover, antiradical activity, one of the typical biological activities of flavonoids, in propolis was also slightly higher than that in its botanical origin. These results indicate that not only propolis but also its botanical origin contains high amounts of flavonoid aglycones and that both of them are useful dietary sources for flavonoids with a potency to prevent dioxin toxicity.     

Botanical origin and chemical composition of Brazilian propolis

Brazilian propolis has been classified into 12 groups based on physicochemical characteristics: five in the southern Brazil group (group 3), one in the southeastern Brazil group (group 12), and six in the northeastern Brazil group (group 6). The plant origins of these groups were investigated using reversed-phase high-performance thin-layer chromatography (RPHPTLC), reversed-phase high-performance liquid chromatography (RPHPLC), and gas chromatography-mass spectrometry (GC-MS). It was concluded that the origins of propolis group 3, group 6, and group 12 are resins of the poplar tree, Hyptis divaricata, and Baccharis dracunculifolia, respectively.     

Cuban and Brazilian red propolis: botanical origin and comparative analysis by high-performance liquid chromatography-photodiode array detection/electrospray ionization tandem mass spectrometry

Chemical composition of propolis depends on the specificity of the local flora at the site of collection and thus on the geographic and climatic characteristics of this place. This paper describes a comparative analysis of Cuban red propolis (CRP), Brazilian red propolis (BRP), and Dalbergia ecastophyllum exudates (DEE) by high-performance liquid chromatography with diode-array detection and tandem mass spectrometry. The aim of this study was to investigate the overall chemical profile and the botanical origin of red propolis and to suggest similarities and differences between samples collected in different tropical regions. Isoliquiritigenin (1), liquiritigenin and naringenin (2 and 17), isoflavones (3-4 and 16), isoflavans (5-7 and 18), and pterocarpans (8-13) were detected in CRP, BRP, and DEE, whereas polyisoprenylated benzophenones (PPBs) guttiferone E/xanthochymol (14a,b) and oblongifolin A (15) were detected only in BRP. Pigments responsible for the red color of DEE and red propolis were also identified as two C30 isoflavans, the new retusapurpurin B (19) and retusapurpurin A (20). PPBs and pigments were isolated and unambiguously characterized by 1D and 2D NMR analysis. These results show that red propolis samples from different tropical zones have a similar chemical composition. DEE is the main red propolis source, but the presence of PPBs in BRP suggests the contribution of different botanical sources for Brazilian samples. This chemical information is important for quality control of red propolis and its commercial products and for biological study.     

Studies of the constituents of Uruguayan propolis

Eighteen flavonoids including two new compounds, four aromatic carboxylic acids, and eleven phenolic acid esters including one new compound were isolated and identified from the ethyl acetate soluble fraction of the 70% ethanol extract of Uruguayan propolis. The new compounds were elucidated as pinobanksin 3-(2-methyl)butyrate (1; recently reported in Usia, T.; Banskota, A. H.; Tezuka, Y.; Midorikawa, K.; Matsushige, K.; Kadota, S. J. Nat. Prod. 2002, 65, 673-676) pinobanksin 3-isobutyrate (2), and 2-methyl-2-butenyl ferulate (24). The constituents isolated from Uruguayan propolis in this study were similar to those of propolis of European and Chinese origin. Thus, it is suggested that the Uruguayan propolis has a plant origin similar to those of propolis from Europe and China.     

Quantitation of the main constituents of some authentic grape-seed oils of different origin

This paper describes the composition of 30 grape-seed oils obtained from France, Italy, and Spain during 2002-2003. Oils were extracted from the seeds using small-scale industrial solvent extraction equipment and analyzed in their unrefined state using standard methods for fatty acids, fatty acids in the triacylglycerol 2-position, tocopherols and tocotrienols, triglycerides, sterols, steradienes, and iodine value. Values for the composition of the sterols, triglycerides, fatty acids, iodine value, and tocopherol composition were generally in good agreement with the results of previous similar surveys. Steradienes (stigmastadiene, campestadiene, stigmastatriene, and campestatriene) were detected in the oil and were probably formed from sterols during the extraction process.     

Quantitation of the main constituents of some authentic sesame seed oils of different origin

This paper describes the composition of sesame seed oils obtained from seeds collected from five countries that are major suppliers of traded sesame seed oil. Oils were extracted from the seeds using small-scale industry pressing equipment and analyzed using standard methods for fatty acids, fatty acids in the triglyceride 2-position, tocopherols and tocotrienols, triglycerides, sterols, steradienes, and iodine value. Values for the composition of the sterols, triglycerides, fatty acids, iodine value, and tocopherol composition were generally in good agreement with the results published elsewhere. All of the oils from roasted seeds contained low levels of the sterol degradation products stigmasta-3,5-diene and campesta-3,5-diene, which were probably formed by dehydration of the parent sterols during roasting.     

Authentication of the botanical origin of honey by near-infrared spectroscopy

Fourier transform near-infrared spectroscopy (FT-NIR) was evaluated for the authentication of eight unifloral and polyfloral honey types (n = 364 samples) previously classified using traditional methods such as chemical, pollen, and sensory analysis. Chemometric evaluation of the spectra was carried out by applying principal component analysis and linear discriminant analysis. The corresponding error rates were calculated according to Bayes' theorem. NIR spectroscopy enabled a reliable discrimination of acacia, chestnut, and fir honeydew honey from the other unifloral and polyfloral honey types studied. The error rates ranged from <0.1 to 6.3% depending on the honey type. NIR proved also to be useful for the classification of blossom and honeydew honeys. The results demonstrate that near-infrared spectrometry is a valuable, rapid, and nondestructive tool for the authentication of the above-mentioned honeys, but not for all varieties studied.     

Identification of flavonoid markers for the botanical origin of Eucalyptus honey

European Eucalyptus honeys showed a common and characteristic HPLC profile in which the flavonoids myricetin (3,5,7,3',4', 5'-hexahydroxyflavone), tricetin (5,7,3',4',5'-pentahydroxyflavone), quercetin (3,5,7,3',4'-pentahydroxyflavone), luteolin (5,7,3', 4'-tetrahydroxyflavone), and kaempferol (3,5,7, 4'-tetrahydroxyflavone) were identified. Their contents, and relative amounts, in the analyzed honey samples were quite constant and supported their floral origin. In addition, ellagic acid and the propolis-derived flavonoids pinobanksin, pinocembrin, and chrysin were detected in most samples. The contents of these nonfloral phenolics were much more variable as could be expected for their propolis origin. Myricetin, tricetin, and luteolin had not been identified as floral markers in any other honey sample previously analyzed in our laboratory (chestnut, citrus, rosemary, lavender, acacia, rapeseed, sunflower, heather, lime tree, etc.) or reported in the literature, suggesting that these could be useful markers. Only in some individual heather samples produced in Portugal has tricetin previously been detected in minor amounts. These samples, however, were contaminated with Eucalyptus as revealed by their pollen analysis and the lack of tricetin or their glycosides in heather floral nectar. It remains to be established if myricetin, tricetin, and luteolin originate from Eucalyptus floral nectar where the corresponding glycosides should be present.     

Antioxidant activity and constituents of propolis collected in various areas of Korea

Propolis is a resinous substance collected by honeybees from various plant sources. The composition of propolis depends on time, vegetation, and the area of collection. This study examined the antioxidant activity of propolis from various areas of Korea: Chilgok, Cheongju, Geochang, Muju, Pocheon, and Sangju. Ethanol extracts of propolis (EEP) were prepared and evaluated for their antioxidant activity by beta-carotene bleaching, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization assays. Furthermore, the major constituents in EEP were identified by high-performance liquid chromatography analysis with a photodiode array and mass spectrometric detection, and each component was quantitatively analyzed. EEP from Cheongju and Muju had relatively strong antioxidant activity accompanied by high total polyphenol contents. Propolis from Cheongju contained large amounts of antioxidative compounds, such as caffeic acid, kaempferol, and phenethyl caffeate. On the other hand, propolis from Pocheon had compounds not seen in propolis from other areas.     

Authentication of the botanical and geographical origin of honey by mid-infrared spectroscopy

The potential of Fourier transform mid-infrared spectroscopy (FT-MIR) using an attenuated total reflectance (ATR) cell was evaluated for the authentication of 11 unifloral (acacia, alpine rose, chestnut, dandelion, heather, lime, rape, fir honeydew, metcalfa honeydew, oak honeydew) and polyfloral honey types (n = 411 samples) previously classified with traditional methods such as chemical, pollen, and sensory analysis. Chemometric evaluation of the spectra was carried out by applying principal component analysis and linear discriminant analysis, the error rates of the discriminant models being calculated by using Bayes' theorem. The error rates ranged from <0.1% (polyfloral and heather honeys as well as honeydew honeys from metcalfa, oak, and fir) to 8.3% (alpine rose honey) in both jackknife classification and validation, depending on the honey type considered. This study indicates that ATR-MIR spectroscopy is a valuable tool for the authentication of the botanical origin and quality control and may also be useful for the determination of the geographical origin of honey.     

Authentication of the botanical and geographical origin of honey by front-face fluorescence spectroscopy

Front-face fluorescence spectroscopy, directly applied on honey samples, was used for the authentication of 11 unifloral and polyfloral honey types (n = 371 samples) previously classified using traditional methods such as chemical, pollen, and sensory analysis. Excitation spectra (220-400 nm) were recorded with the emission measured at 420 nm. In addition, emission spectra were recorded between 290 and 500 nm (excitation at 270 nm) as well as between 330 and 550 nm (excitation at 310 nm). A total of four different spectral data sets were considered for data analysis. Chemometric evaluation of the spectra included principal component analysis and linear discriminant analysis; the error rates of the discriminant models were calculated by using Bayes' theorem. They ranged from <0.1% (polyfloral and chestnut honeys) to 9.9% (fir honeydew honey) by using single spectral data sets and from <0.1% (metcalfa honeydew, polyfloral, and chestnut honeys) to 7.5% (lime honey) by combining two data sets. This study indicates that front-face fluorescence spectroscopy is a promising technique for the authentication of the botanical origin of honey and may also be useful for the determination of the geographical origin within the same unifloral honey type.     

Rapid screening of polar compounds in Brazilian propolis by high-temperature high-resolution gas chromatography-mass spectrometry

Methanol extracts of propolis from six different places, five in Rio de Janeiro state and one in S?o Paulo state, both in the Southeast of Brazil, were investigated using high-temperature high-resolution gas chromatography (HT-HRGC) and HT-HRGC-mass spectometry. The main purpose of the study was to establish the applicability of HT-HRGC as an analytical method for systematic studies of polar propolis fractions. Several compounds, including carbohydrates, phenolic acid derivatives, and high molecular weight compounds (e.g., wax esters of long chain fatty alcohols) could be readily characterized in the crude extracts by HT-HRGC-MS. HT-HRGC and HT-HRGC-MS were shown to be quick and informative tools for rapid analysis of crude polar extracts without cleanup.     

Study of the main constituents of some authentic walnut oils

This paper describes the composition of walnut oils obtained from nuts collected from seven countries that are major suppliers of walnut oil. Oils were extracted from the nuts using small-scale industry pressing equipment and analyzed using standard methods for fatty acids, fatty acids in the triacylglycerol 2-position, tocopherols and tocotrienols, triacylglycerols, sterols, steradienes, and iodine value. Values for the composition of the sterols, triacylglycerols, fatty acids, iodine value, and tocopherol composition were generally in good agreement with the results of previous similar surveys. Tocotrienols were not detected in any sample. Steradienes (stigmastadiene, campestadiene, stigmastatriene, and campestatriene) were not detected in any oil.     

Study of the main constituents of some authentic hazelnut oils

This paper describes the composition of authentic hazelnut oils obtained from nuts collected from five countries that are major suppliers of hazelnut oil. Oils were analyzed using standard methods for fatty acids, fatty acids in the triacylglycerol 2-position, tocopherols and tocotrienols, triacylglycerols, sterols, steradienes, and iodine value. The results were generally in good agreement with those of other publications. Tocotrienols, previously unreported in hazelnut oil, were detected in one sample. There were no major differences in the composition of oils from different countries. Roasting the nuts prior to pressing had little effect on oil composition.     

Cyclic polyalcohols: fingerprints to identify the botanical origin of natural woods used in wine aging

Cyclic polyalcohol composition of 80 natural wood samples from different botanical species, with the majority of them used in the oenology industry for aging purposes, has been studied by gas chromatography-mass spectrometry (GC-MS) after its conversion into their trimethylsilyloxime derivatives. Each botanical species showed a different and specific cyclic polyalcohol profile. Oak wood samples were characterized by the richness in deoxyinositols, especially proto-quercitol. Meanwhile, other botanical species showed a very low content of cyclic polyalcohols. The qualitative and quantitative study of cyclic polyalcohols was a useful tool to characterize and differentiate woods of different botanical origin to guarantee the authenticity of chips used in the wine-aging process. Monosaccharide composition was also analyzed, showing some quantitative differences among species, but cyclic polyalcohols were the compounds that revealed the main differentiation power.     

Determination by near-infrared spectroscopy of perseitol used as a marker for the botanical origin of avocado (Persea americana Mill.) honey

This paper reports the application of near-infrared (NIR) reflectance spectroscopy to determine the concentration in honey of perseitol, a sugar that is specific to avocado honey. Reference values for perseitol were obtained by high-performance liquid chromatography analysis in 109 honey samples. Although the average concentration of perseitol in honey samples was only 0.48%, accurate prediction equations were successfully developed. The regression model of modified partial least squares was superior to that of principal component regressions. Calibrations based on the first or second derivative of Log(1/R) were equally good (R(2) > 0.95). Using half of the samples for calibration and the second half for validation, the correlation between actual and predicted values of the second half was satisfactory (R(2) = 0.87), the slope did not differ from 1, bias was low (0.005%), and the standard error of prediction was relatively low (0.13%). It was concluded that NIRS analysis may be used to detect to what extent honeybees have harvested avocado nectar but not to authenticate avocado honey as unifloral.     

Authentication of the botanical origin of honey by front-face fluorescence spectroscopy. A preliminary study

The potential of front-face fluorescence spectroscopy for the authentication of unifloral and polyfloral honey types (n = 57 samples) previously classified using traditional methods such as chemical, pollen, and sensory analysis was evaluated. Emission spectra were recorded between 280 and 480 nm (excit: 250 nm), 305 and 500 nm (excit: 290 nm), and 380 and 600 nm (excit: 373 nm) directly on honey samples. In addition, excitation spectra (290-440 nm) were recorded with the emission measured at 450 nm. A total of four different spectral data sets were considered for data analysis. After normalization of the spectra, chemometric evaluation of the spectral data was carried out using principal component analysis (PCA) and linear discriminant analysis (LDA). The rate of correct classification ranged from 36% to 100% by using single spectral data sets (250, 290, 373, 450 nm) and from 73% to 100% by combining these four data sets. For alpine polyfloral honey and the unifloral varieties investigated (acacia, alpine rose, honeydew, chestnut, and rape), correct classification ranged from 96% to 100%. This preliminary study indicates that front-face fluorescence spectroscopy is a promising technique for the authentication of the botanical origin of honey. It is nondestructive, rapid, easy to use, and inexpensive. The use of additional excitation wavelengths between 320 and 440 nm could increase the correct classification of the less characteristic fluorescent varieties.     

Improved characterization of the botanical origin of sugar by carbon-13 SNIF-NMR applied to ethanol

Until now, no analytical method, not even isotopic ones, had been able to differentiate between sugars coming from C4-metabolism plants (cane, maize, etc.) and some crassulacean acid metabolism plants (e.g., pineapple, agave) because in both cases the isotope distributions of the overall carbon-13/carbon-12 and site-specific deuterium/hydrogen isotope ratios are very similar. Following recent advances in the field of quantitative isotopic carbon-13 NMR measurements, a procedure for the analysis of the positional carbon-13/carbon-12 isotope ratios of ethanol derived from the sugars of pineapples and agave using the site-specific natural isotopic fractionation-nuclear magnetic resonance (SNIF-NMR) method is presented. It is shown that reproducible results can be obtained when appropriate analytical conditions are used. When applied to pineapple juice, this new method demonstrates a unique ability to detect cane and maize sugar, which are major potential adulterants, with a detection limit in the order of 15% of the total sugars, which provides an efficient mean of controlling the authenticity of juices made from this specific fruit. When applied to tequila products, this new method demonstrates a unique ability to unambiguously differentiate authentic 100% agave tequila, as well as misto tequila (made from at least 51% agave), from products made from a larger proportion of cane or maize sugar and therefore not complying with the legal definition of tequila.     

Evaluation of the botanical origin of estonian uni- and polyfloral honeys by amino acid content

The free amino acid content of 61 honey samples from Estonia has been determined by HPLC-UV with precolumn derivatization with diethyl ethoxymethylenemalonate. Analyzed samples were seven types of unifloral honeys and polyfloral honeys. The main amino acids found in Estonian honeys were proline and phenylalanine. The resulting data have been analyzed by t test and principal component analysis (PCA). t Test revealed that some amino acids (alpha-alanine, beta-alanine, asparagine, gamma-aminobutyric acid, glutamine, glycine, histidine, ornithine, phenylalanine, proline, serine, and tryptophan) are more potent for assigning honey botanical origin than others. PCA enabled differentiation of some honey types by their botanical origin. In the space of the two first principal components, heather honeys form a cluster that is clearly separable from, for example, polyfloral honeys. It is concluded that analysis of the free amino acid profile may serve as a useful tool to assess the botanical origin of Estonian honeys.     

Chemical characterization of Cuban propolis by HPLC-PDA, HPLC-MS, and NMR: the brown, red, and yellow Cuban varieties of propolis

Sixty-five samples of propolis were collected from eleven regions of Cuba; methanolic extracts of propolis were prepared from all samples, and a classification method was developed using a combination of NMR, HPLC-PDA, and HPLC-ESI/MS techniques. The analysis of (1)H and (13)C NMR spectra and chromatographic profiles of all propolis extracts allowed the definition of three main types of Cuban propolis directly related to their secondary metabolite classes: brown Cuban propolis (BCP), rich in polyisoprenylated benzophenones, red Cuban propolis (RCP), containing isoflavonoids as the main constituents, and yellow Cuban propolis (YCP), probably with aliphatic compounds. Subsequently, the principal compounds of the brown and red types were characterized by HPLC-ESI/MS analysis. Instrumental techniques used are complementary; NMR was shown to be a quick and informative tool for the rapid analysis of crude propolis polar extracts and allowed the identification of the main class of secondary metabolites, while LC-PDA and LC-MS techniques were useful tools for qualitative and quantitative analysis of marker compounds of Cuban propolis.