Chemical composition and biological activity of propolis from Brazilian meliponinae

Twenty-one propolis samples produced by 12 different Meliponinae species were analyzed by GC-MS. Several chemical types of stingless bees' propolis could be grouped, according to the prevailing type of compounds like: 'gallic acid", "diterpenic" and "triterpenic" types. The results confirm that neither the bee species nor the geographical location determine the chemical composition of Meliponinae propolis and the choice of its plant source, respectively. This could be explained by the fact that Meliponinae forage over short distances (maximum 500 m) and thus use as propolis source the first plant exudate they encounter during their flights. The antibacterial, antifungal and cytotoxic activities of the samples were also investigated. Most samples had weak or no activity against E. coli, weak action against Candida albicans. Some of them showed significant activity against St. aureus., presumably connected to the high concentration of diterpenic acids. Samples rich in diterpenic acids possessed also high cytotoxic activity (Artemia salina test).     

The additive and synergistic antimicrobial effects of select frankincense and myrrh oils--a combination from the pharaonic pharmacopoeia

Aims: The in vitro antimicrobial activity of three essential oil samples of frankincense (Boswellia rivae, Boswellia neglecta and Boswellia papyrifera) and two essential oil samples of myrrh and sweet myrrh (Commiphora guidotti and Commiphora myrrha), collected from different regions of Ethiopia, was investigated independently and in combination to determine their anti‐infective properties. Methods and Results: The microdilution minimum inhibitory concentration (MIC) assay was performed, whereby it was noted that generally Cryptococcus neoformans (MIC values in the range of 0·8–1·4 mg ml^?1) and Pseudomonas aeruginosa (MIC values in the range of 0·5–1·3 mg ml^?1) often appeared to be the most susceptible micro‐organisms against oils of both Boswellia and Commiphora spp. When assayed in various combinations, the frankincense and myrrh oils displayed synergistic, additive and noninteractive properties, with no antagonism noted. When investigating different ratio combinations against Bacillus cereus, the most favourable combination was between B. papyrifera and C. myrrha. The composition of the oils was determined by gas chromatography coupled to mass spectrometry (GC–MS) to document the specific chemotypes used in the study, and the chemical profiles were found to be congruent with previously reported data. Conclusions: The majority of interactions identified synergistic and additive effects, with strong synergism noted between B. papyrifera and C. myrrha. Significance and Impact of the Study: Frankincense and myrrh essential oils have been used in combination since 1500 bc ; however, no antimicrobial investigations have been undertaken to confirm their effect in combination. This study validates the enhanced efficacy when used in combination against a selection of pathogens.     

Frankincense and myrrh

While frankincense and myrrh have been harvested from a multitude of species, certain species have predominated in history.Boswellia carteri andB. frereana are the main sources of frankincense today, whileB. papyrifera was the principal source of antiquity andB. sacra was the principal species of classical times.Commiphora myrrha is the chief source of myrrh today, butC. erythraea was the principal source of ancient and classical times. Each of these oleo-gum-resins has a characteristic odor that is predominately due to a mixture of complex sesquiterpenes.     

Fourier transform IR study of aggregational behavior of N-acetyl-L- and N-butyloxycarbonyl-L-glutamic acid oligomeric benzyl esters in dioxane and benzene: beta-turn --> antiparallel beta-sheet transition

Resins from several different genera are studied using Fourier transform (FT)-Raman spectroscopy. Tree resins can be broadly divided into those that contain diterpenoid components and those that contain triterpenoid components. The diterpenoid resins analyzed are from the genera Pinus, Cedrus, and Agathis (kauri resin) and the triterpenoid resins examined are samples from Pistacia, Boswellia (frankincense), and Commiphora (myrrh) genera. A protocol is developed to nondestructively distinguish diterpenoid and triterpenoid resins and to differentiate the genera within the two types. The effects of oxidation on the discrimination of the FT-Raman spectra are considered.     

Unifloral Honeys: Chemical Conversion and Pollen Reduction

Physico-chemical and pollen analyses were made of unifloral samples of nectar, crop content and honey. The samples were obtained by carrying out cage experiments exclusively with one plant species. The investigations of four plant species are presented: Centaurea montana, Coriandrum sativum. Euphorbia lathyris and Rubus idaeus. During the conversion of nectar to honey its chemical composition changes. The changes in carbohydrate composition cannot be explained by those enzymes described as specific for honey. During the period of time when the bees are working on gathered nectar, the chemical components of the honey, whose origin is the secretion from the bees, is increasing, while the amount of pollen is decreasing. There are negative correlations between the quantity of pollen grains and both enzymatic activity and the amount of proline. With respect to the botanical origin it is suggested that the absolute amount of pollen should be determined and compared with the chemical analysis of the honey.     

Isolation of iridoid and secoiridoid glycosides and comparative study on Radix gentianae and related adulterants by HPLC analysis

HPLC profile guided study led to the isolation of an acylated secoiridoid glycoside, named gentiotrifloroside (1), together with six known compounds, i.e., loganic acid (2), 6-O-beta-d-glucopyranosylgentiopicroside (3), swertiamarin (4), gentiopicroside (5), sweroside (6) and 2 -(o,m-dihydroxybenzyl)-sweroside (7) from Gentiana triflora and Gentiana rigescens. The structure of 1 was deduced from one- and two-dimensional NMR spectroscopic experiments. Compounds 1-7 were used successfully as chemical markers for the comparison of the four species of Gentiana used as Radix gentianae. Additionally, differentiation of Gentiana species mentioned and those used as adulterants was evaluated. The close similarity of chemical composition among the four genuine Gentiana species explain their popular usage as R. gentianae in Chinese medicine. We have also shown that the variation of chemical composition in R. gentianae and related adulterants agree well with their botanical phylogeny.     

An NMR-based metabolomic approach to identify the botanical origin of honey

NMR can be used in food analysis for origin discrimination and biomarker discovery using a metabolomic approach. Here, we present an example of this strategy to discriminate honey samples of different botanical origins. The NMR spectra of 353 chloroform extracts of selected honey samples were analyzed to detect possible markers of their floral origin. Six monofloral Italian honey types (acacia, linden, orange, eucalyptus, chestnut, and honeydew) were analyzed together with polyfloral samples. Specific markers were identified for each monofloral origin: two markers for acacia (chrysin and pinocembrin), one for chestnut (??-LACT-3-PKA), two for orange (8-hydroxylinalool and caffeine), one for eucalyptus (dehydrovomifoliol), one for honeydew (a diacylglycerilether) and two for linden (4-(1-hydroxy-1-methylethyl)cyclohexa-1,3-diene-carboxylic acid and 4-(1-methylethenyl)cyclohexa-1,3-diene-carboxylic acid). An NMR-based metabolomic approach that used O2PLS-DA multivariate data analysis allowed us to discriminate the different types of honey. Two different classifiers were built based on different multivariate techniques. The high precision of the classification obtained suggests that this approach could be useful to develop generally applicable metabolomic tools to discriminate the origin of honey samples.     

Numerical taxonomic studies of the genus Pyrus using both chemical and botanical characters*

Of three computer-generated classifications of Pyrus species, produced respectively from 29 chemical characters, 22 botanical characters and all 51 characters taken together, the classification based upon both chemical and botanical characters agrees best with the known geographical distributions of the species. Where either chemical or botanical characters are separately employed, the classifications contain seriously misplaced species. The numerical classification based upon all 51 characters lends some support to the division of the genus into four principal groups of species: the East Asian pea pears, the larger-fruited East Asian pears, the North African pears and the European and West Asian pears. P. longipes, P. betulifolia and P. cordata are considered as being connecting links between the East Asian pea pears, the larger-fruited East Asian pears and the European and West Asian pears. Some speculations are made on the phylogenetic relationships among Pyrus species.     

Holy Smoke in Medieval Funerary Rites: Chemical Fingerprints of Frankincense in Southern Belgian Incense Burners

Frankincense, the oleogum resin from Boswellia sp., has been an early luxury good in both Western and Eastern societies and is particularly used in Christian funerary and liturgical rites. The scant grave goods in late medieval burials comprise laterally perforated pottery vessels which are usually filled with charcoal. They occur in most regions of western Europe and are interpreted as incense burners but have never been investigated with advanced analytical techniques. We herein present chemical and anthracological results on perforated funerary pots from 4 Wallonian sites dating to the 12–14^th century AD. Chromatographic and mass spectrometric analysis of lipid extracts of the ancient residues and comparison with extracts from four Boswellia species clearly evidence the presence of degraded frankincense in the former, based on characteristic triterpenoids, viz. boswellic and tirucallic acids, and their myriad dehydrated and oxygenated derivatives. Cembrane-type diterpenoids indicate B. sacra (southern Arabia) and B. serrata (India) as possible botanical origins. Furthermore, traces of juniper and possibly pine tar demonstrate that small amounts of locally available fragrances were mixed with frankincense, most likely to reduce its cost. Additionally, markers of ruminant fats in one sample from a domestic context indicate that this vessel was used for food preparation. Anthracological analysis demonstrates that the charcoal was used as fuel only and that no fragrant wood species were burned. The chars derived from local woody plants and were most likely recovered from domestic fires. Furthermore, vessel recycling is indicated by both contextual and biomarker evidence. The results shed a new light on funerary practices in the Middle Ages and at the same time reveal useful insights into the chemistry of burned frankincense. The discovery of novel biomarkers, namely Δ²-boswellic acids and a series of polyunsaturated and aromatic hydrocarbons, demonstrates the high potential for organic chemical analyses of incense residues.     

HPLC/HR-MS-Based Metabolite Profiling and Chemometrics: A Powerful Approach to Identify Bioactive Compounds from Abarema cochliacarpos

Despite its importance as a medicinal plant, there is a lack of studies that assessed the chemical composition of A. cochliacarpos extracts. Herein, we used a metabolite profiling approach and chemometrics as a powerful strategy to correlate the chemical composition with the antioxidant activity of A. cochliacarpos extracts. Extracts obtained with ethyl acetate showed greater antioxidant activity and higher total phenolic content than extracts obtained with hexane. The chemical composition was assessed by HPLC/HR-MS and it encompassed fatty alcohols, terpenoids, phenolic derivatives, lipids, carotenoid-like compounds, alkaloids, flavonoids, polyketides, and glycerophospholipids. Chemometrics successfully differentiated not only the chemical composition of extracts in response to the nature of the extraction solvent and the botanical part used during extraction but also it allowed us to associate the chemical composition with the antioxidant activity of the extracts, which might be particularly helpful for drug discovery and development programs.     

Floral Pollen Bioactive Properties and Their Synergy in Honeybee Pollen

Honeybee pollen (HBP) is a mixture of floral pollen collected by honeybees near the hive. It is characterized by a composition rich in phenolic compounds, carotenoids and vitamins that act as free radicals scavengers, conferring antioxidant and antibacterial capacity to the matrix. These bioactive properties are related to the botanical origin of the honeybee pollen. Honeybee pollen samples were collected from different geographical locations in central Chile, and their total carotenoid content, polyphenols profile by HPLC/MS/MS, DPPH radical scavenging capacity, and antimicrobial capacity against S. pyogenes, E. coli, S. aureus, and P. auriginosa strains were evaluated. Our results showed a good carotenoids content and polyphenols composition, while antioxidant capacity presented values between 0–95 % for the scavenging effect related to the botanical origin of the samples. Inhibition diameter for the different strains presented less variability among the samples, Furthermore, binary mixtures representing the two most abundant species in each HBP were prepared to assess the synergy effect of the floral pollen (FP) present in the samples. Data shows an antagonist effect was observed when assessing the carotenoid content, and a synergy effect often presents for antimicrobial and antioxidant capacity for bee pollen samples. The bioactive capacities of the honeybee pollen and their synergy effect could apply to develop new functional ingredients for the food industry.     

Frankincense Revisited,Part I: Comparative Analysis of Volatiles in Commercially Relevant Boswellia Species

The genus Boswellia comprises many species which are famous for their production of frankincense, a fragrant gum resin. In the published literature, manifold studies on the volatiles and semivolatiles in individual samples of these gum resins exist, yet very few studies have investigated multiple samples. Contradictory results with regard to the volatile composition exist in literature. In this first part of the study, a large sample set (n = 46) of mostly commercially obtained gum resins and essential oils was investigated by solid‐phase microextraction gas chromatography/mass spectrometry. A total of 216 compounds were identified or tentatively identified from the four commercially relevant species, namely B. sacra, B. serrata, B. papyrifera, and B. frereana, and the statistical evaluation of the resulting chemical profiles allowed a clear distinction between the species by their volatile profile. With only few exceptions, the designated species was found to be in accordance with the composition reported in reliable literature sources and detected in botanically identified samples. Chemotaxonomic marker substances were suggested to facilitate the differentiation of commercial gum resins or essential oils based on their volatile profile.     

Foraging behaviour of the stingless bee Melipona orbignyi (Hymenoptera: Apidae: Meliponini) in a dry forest assessed by multivariate analysis from palynological data

Pollen analysis was applied to study the botanical composition of honey and pollen stored by Melipona orbignyi in its southernmost distributional range. Many studies have revealed that robust buzzing bees of this genus are associated with diverse plant families, some of which are different to those exploited by the remaining genera of Meliponini. Cluster analysis was performed to assess similarity in botanical composition between pot-pollen and pot-honey provisions from different colonies. The results indicated that each colony was capable to access to different plant species for obtaining nectar and pollen, a fact observed when discarding the contaminated honey samples. As honey samples were not clustered together with other ones but with pollen samples indicating a similar botanical composition, it could be assumed that a same plant species provided nectar and pollen to bees. Resource partitioning was not clearly observed among a subset of late spring colonies, as two families of high richness of species and abundant in this dry forest (the mimosoid clade in Fabaceae and Capparaceae) were foraged. A higher number of synchronously sampled colonies would be necessary to detect this ecological aspect. The botanical composition of samples of M. orbignyi was governed by both random factors such as local and temporary flower availability plus preferences for particular plant species such as those from Solanaceae and Ximeniaceae. The Capparaceae and Ximeniaceae are here reported for the first time as intensively foraged pollen resources by the genus Melipona.     

Propolis from the Mediterranean region: chemical composition and antimicrobial activity

The chemical composition of propolis from Bulgaria, Turkey, Greece and Algeria was investigated by GC-MS. All of them contained mainly flavonoids and esters of caffeic and ferulic acids, which indicated that their main source are buds of poplars of the taxonomic section Aegieros. Some Turkish samples contained a low percent of diterpenic acids, while in Algerian samples significant amounts of a hydroxyditerpenic acid (M=322, its structure not determined by its MS) were found. All samples showed significant antibacterial and weak to moderate antifungal activity.     

Distribution of steroidal saponins in Tribulus terrestris from different geographical regions

The steroidal saponins of Tribulus terrestris L. (Zygophyllaceae) are considered to be the factor responsible for biological activity of products derived from this plant. The activity depends on the concentration and the composition of active saponins, which in turn is influenced by the geographical origin of plant material. Samples of T. terrestris collected in Bulgaria, Greece, Serbia, Macedonia, Turkey, Georgia, Iran, Vietnam and India were analyzed by LC-ESI/MS/MS for the presence and the concentration of protodioscin (1), prototribestin (2), pseudoprotodioscin (3), dioscin (4), tribestin (5) and tribulosin (6). The flavonoid rutin (7) was also included in the comparison. The results revealed distinct differences in the content of these compounds depending on region of sample collection, plant part studied and stage of plant development. The samples from Bulgaria, Turkey, Greece, Serbia, Macedonia, Georgia and Iran exhibited similar chemical profile and only some quantitative difference in the content of 1-7 with protodioscin (1) and prototribestin (2) as main components. The Vietnamese and Indian samples exhibit totally different chemical profile. They lack 2 and 5, while tribulosin (6) is present in high amounts. Compounds different from 1 to 7 are dominating in these 3 samples. The presented results suggested the existence of one chemotype common to the East South European and West Asian regions. Most probably, the Vietnamese and Indian samples belong to other chemotypes which are still to be studied and characterized. No clear correlation between the burrs morphology and the chemical composition of the samples has been found.     

Bosellia serrata-induced apoptosis is related with ER stress and calcium release

It has been reported that the gum resin of Boswellia serrata (BS), which has been shown to have antiinflammatory properties, might also have anticancer effects. This study examined the potential of BS as an anticancer agent. The BS extract induces apoptosis in HeLa human cervical carcinoma cells, as confirmed by two apoptosis analyses, Hoechst staining and Annexin V/PI assay. Among the apoptosis pathways, the ER stress-associated mechanism was examined to determine its role in BS-induced apoptosis. The expression of GRP78 and CHOP, which are representatives of the ER stress proteins, and the calcium-binding protein-calpain were determined. The results showed significantly higher levels of both GRP78 and CHOP, and stronger calpain activity in the BS-treated cells than in the control cells. This shows that there is a correlation between ER stress signaling and apoptosis, which suggests the possibility of the BS-ER stress initiator as an anticancer therapeutic agent in human cervical carcinoma.     

Extract of gum resins of Boswellia serrata L. inhibits lipopolysaccharide induced nitric oxide production in rat macrophages along with hypolipidemic property

Boswellia serrata, Linn F (Burseraceae) is commonly used in Indian system of medicine (Ayurvedic) as an anti-inflammatory, analgesic, anti-arthritic and anti-proliferative agent. This study was planned to investigate the water-soluble fraction of the oleoresin gum of Boswellia serrata (BS extract) on lipopolysaccharide (LPS) induced nitric oxide (NO) production by macrophages under in vivo and in vitro conditions. In the previous condition, rats were fed on atherogenic diet (2.5% cholesterol, 1% cholic acid, 15.7 % saturated fat) along with the BS extract for 90 days. Blood was collected for lipid profile and toxicological safety parameters. Peritoneal macrophages were isolated and cultured to see the LPS induced NO production. Under in vivo experiment, BS extract significantly reduced serum total cholesterol (38-48 %), increased serum high-density lipoprotein- cholesterol (HDL-cholesterol, 22-30%). Under in vitro experiments with thioglycolate activated macrophages, it inhibited LPS induced (NO) production with IC 50 value at 662 ng /ml. Further, this fraction, in the dose of 15 mg/100 g body wt for 90 days, did not show any increase in serum glutamate-pyruvate transaminase (SGPT) and blood urea, in normal control animals. However, it significantly reversed the raised SGPT and blood urea in the atherogenic diet-fed animals. Transverse section of liver and kidney also supported its protective effect. Thus it may be concluded that water extract of Boswellia serrata possesses strong hypocholesterolemic property along with increase in serum HDL. It inhibits the LPS induced NO production by the activated rat peritoneal macrophages and show hepato-protective and reno-protective property.     

六道木属六道木组种间关系的AFLP分析和黄花六道木中国分布新记录的认定

用荧光标记AFLP技术对东亚产六道木属六道木组作了系统关系分析。我们采用 5对AFLP引物 ,对六道木属 9个物种 ,锦带花属和猬实属各 1个物种共 16份样品进行分析 ,共获得 988个位点。不同引物所获得的结果具有很高的相容性 ,Mantel相关系数为 0 72 5～0 919,说明AFLP技术适用于六道木属的种间关系分析。用邻接法 (neighbor joining)对 16个样品 988个位点的分析生成的系统树获得很高的自展分析 (bootstrap)支持率。树系图表明 ,猬实属与六道木属管花六道木组有密切的关系。在六道啪组内 ,温州六道木与糯米条的关系密切 ,单花六道木与大花六道木差异很小 ,黄花六道木与日本产的近缘种组成一单系分支。本文认定 ,根据形态特征鉴定浙江永嘉的标本为黄花六道木 (AbeliaserrataSieb .etZucc .)是正确的 ,属中国分布新记录。文章最后补充描述了中国产的黄花六道木。黄花六道木在中国的发现进一步证明中国植物区系与日本植物区系的密切关系。     

Fast classification and compositional analysis of cornstover fractions using Fourier transform near-infrared techniques

The objectives of this research were to determine the variation of chemical composition across botanical fractions of cornstover, and to probe the potential of Fourier transform near-infrared (FT-NIR) techniques in qualitatively classifying separated cornstover fractions and in quantitatively analyzing chemical compositions of cornstover by developing calibration models to predict chemical compositions of cornstover based on FT-NIR spectra. Large variations of cornstover chemical composition for wide calibration ranges, which is required by a reliable calibration model, were achieved by manually separating the cornstover samples into six botanical fractions, and their chemical compositions were determined by conventional wet chemical analyses, which proved that chemical composition varies significantly among different botanical fractions of cornstover. Different botanic fractions, having total saccharide content in descending order, are husk, sheath, pith, rind, leaf, and node. Based on FT-NIR spectra acquired on the biomass, classification by Soft Independent Modeling of Class Analogy (SIMCA) was employed to conduct qualitative classification of cornstover fractions, and partial least square (PLS) regression was used for quantitative chemical composition analysis. SIMCA was successfully demonstrated in classifying botanical fractions of cornstover. The developed PLS model yielded root mean square error of prediction (RMSEP %w/w) of 0.92, 1.03, 0.17, 0.27, 0.21, 1.12, and 0.57 for glucan, xylan, galactan, arabinan, mannan, lignin, and ash, respectively. The results showed the potential of FT-NIR techniques in combination with multivariate analysis to be utilized by biomass feedstock suppliers, bioethanol manufacturers, and bio-power producers in order to better manage bioenergy feedstocks and enhance bioconversion.     

Analysis of pentacyclic triterpenic acids from frankincense gum resins and related phytopharmaceuticals by high-performance liquid chromatography. Identification of lupeolic acid,a novel pentacyclic triterpene

An HPLC gradient method with photodiode array detection was developed for the simultaneous analysis of 12 different pentacyclic triterpenic acids in Indian and African frankincense gum resins as well as in related phytopharmaceuticals. The triterpenic acids were obtained by an exhaustive extraction procedure. Identification of the compounds was based on retention times, UV-spectra and add on technique with standards isolated from African frankincense. The method allows differentiation of frankincense of different origin and standardization of frankincense-based phytopharmaceuticals. Further, this is the first report identifying a novel pentacyclic triterpene, lupeolic acid, as a constituent of frankincense gum resins.