Thin-layer chromatography in the authenticity testing of bee-products.

Quality control, nutritional value and the monitoring of hazardous residues in honey bee- products have become major topics for both producers and consumers. Due to its potential role in human health, bee-products rich in bioactive compounds are becoming increasingly popular. This review aims to provide an overview of thin-layer chromatography methods used in quality control,authenticity testing and chemical profiling of bee-products in order to help scientists engaged in the field of bee-products chemistry to utilize the advantages of this technique in the detection and elimination of fraudulent practices in bee-product manufacturing. Recently, hyphenation of thin-layer chromatography, image analysis and chemometrics support bee-products analysisbysimultaneousdeterminationofanalytes with different detection principles, identification of individual bioactive compounds as well as structure elucidation of compounds. Highlighted opportunities of thin-layer chromatography could encourage further investigations that would lead to improvements in the detection and elimination of marketing fraudulent practices.

Manuka Honey: Feasibility and Safety in Postoperative Neurosurgical Wound Care.

OBJECTIVE: To date, no reports have been published on active Leptospermum manuka honey (ALH) feasibility as a postoperative topical wound supplement in neurosurgical patients. The objective of the study is to present the authors' initial experience with using ALH in postoperative neurosurgical patients. METHODS: A single-surgeon retrospective case series review of cranial and spinal operations between 2018 and 2020 was performed in patients with nonhealing wounds or wounds deemed "at risk" as defined by grade 1 Sandy surgical wound dehiscence grading classification. An ALH gel or ointment was applied to these incisions once a day for 2 to 4 weeks. Patients were followed up in the clinic every 2 weeks until incisions had healed. RESULTS: Twenty-five postoperative patients (12 cranial, 13 spinal) were identified to be at high risk of operative debridement. All 25 patients were prescribed a topical application of ALH, which was easily adopted without patient-related adverse events. Seven (four cranial, three spinal) patients required operative debridement and treatment with long-term antibiotic therapy. CONCLUSIONS: In this small case series of neurosurgical patients who were at risk of poor wound healing, the application of medical-grade ALH was well tolerated without patient-reported adverse events. The ALH may have prevented the need for operative debridement in the majority of patients. Further prospective studies are necessary to establish its efficacy in wound healing in the neurosurgical population.

Quality Assessment, Functional Potentials, and Safety Evaluation of Stored Egyptian Honey as an Environmental Pollution Bioindicator.

Egyptian honeys are distinguished by their variety, properties, and therapeutic applications. The aim of the present study was to evaluate the quality and the functional and safety characteristics of Egyptian honey as a sensitive environmental bioindicator collected from 9 different locations in Egypt that were exposed to various air pollution sources, either stationary, mobile, or area sources, and stored for 6 mo at ambient temperature (22 ± 2 °C) compared with Codex Alimentarius international standards. Physicochemical properties, sugar profile via high performance liquid chromatography, functional potentials (antioxidant and antimicrobial), and safety parameters (microbiological quality, aflatoxins, and heavy metal pollutants) were assessed. The results revealed that honey obtained from Shabshir Hessa, El Gharbia Governorate (GSH-1), showed the best quality with the highest monosaccharide and phenolic and flavonoid content (73.07%, 363.07, and 15.33 mg/g, respectively) and clear of biotic and abiotic contaminants, reflecting good hygienic, environmental conditions and apiarian practices. The other 8 honey samples reflected inferior quality in physicochemical parameters either of low monosaccharide content (40.46-50.25%), high hydroxymethylfurfural (HMF) values as in Borg El-Arab Industrial Zone, Alexandria Governorate (BAI-9), and Nubaria Desert Road, Alexandria Governorate (NDR-4; 50.83 and 48.25 mg/kg), or of high acidity as in Agricultural Road, El Gharbia Governorate (GAR-3), and NDR-4 (74.72 and 68.47 mEq/kg) attributable to contaminated locations (fermentations) or thermal treatment (HMF). Furthermore, safety assessment revealed that coliform counts exceeded 2.5 log10 cfu/g in 6 samples-Chemicals Company, Kafr El Zayyat Governorate (KZC-2); GAR-3; NDR-4; Mansoura Fertilizer Talkha Company, Daqahlia Governorate (MFT-5); Gharbaniyat Cement Company, Beheira Governorate; and BAI-9-and the highest counts of yeast and molds in Abees Ceramic Company, Alexandria Governorate (3.72 log10 cfu/g), which was reflected in its total aflatoxins (679.83 µg/kg). Samples MFT-5; GAR-3; Kafr El-Sheikh Petroleum Company, Kafr El-Sheikh Governorate; and KZC-2 exceeded the lead permissible limit (0.75, 0.61, 0.57, and 0.51 mg/kg, respectively) as a result of inferior hygienic quality and the negative effects of environmental pollutants. The results ring alarm bells about how we should take substantial steps toward stringent standards for quality control practices of honey production operations. Environ Toxicol Chem 2020;39:1894-1907. © 2020 SETAC.

Photodegradation of Organophosphorus Pesticides in Honey Medium by Solar Light Irradiation.

In this study, the photodegradation of organophosphorus (OPs) pesticides in the honey medium was evaluated under sunlight irradiation. Some of the 22 samples collected at different sites contained OPs pesticides (Methyl parathion, Coumaphos and Fenitrothion) with an average of 8 ng/g. Moreover, three samples were found with pesticide residue levels exceeding the maximum residue limits (MRL ≥ 50 ng/g) imposed by the standard water (WHO). Gas chromatography (GC) combined with a tritium electron capture detector system was used for the analysis of OPs pesticides in honey. Total degradation of the Methyl parathion was obtained in less than 60 min of irradiation. Moreover, the elimination of the other OPs found in the samples was also effective with a rate of 85% for Coumaphos and Fenitrothion after 50 min of sunlight irradiation. The kinetics of the photodegradation reaction of all OPs pesticides studied followed a pseudo-first order model.

Honey Volatiles as a Fingerprint for Botanical Origin-A Review on their Occurrence on Monofloral Honeys.

Honeys have specific organoleptic characteristics, with nutritional and health benefits, being highly appreciated by consumers, not only in food but also in the pharmaceutical and cosmetic industries. Honey composition varies between regions according to the surrounding flora, enabling its characterization by source or type. Monofloral honeys may reach higher market values than multifloral ones. Honey's aroma is very specific, resulting from the combination of volatile compounds present in low concentrations. The authentication of honey's complex matrix, according to its botanical and/or geographical origin, represents a challenge nowadays, due to the different sorts of adulteration that may occur, leading to the search for reliable marker compounds for the different monofloral honeys. The existing information on the volatiles of monofloral honeys is scarce and disperse. In this review, twenty monofloral honeys and honeydews, from acacia, buckwheat, chestnut, clover, cotton, dandelion, eucalyptus, fir tree, heather, lavender, lime tree, orange, pine, rape, raspberry, rhododendron, rosemary, strawberry tree, sunflower and thyme, were selected for volatile comparison purposes. Taking into consideration the country of origin, the technique of isolation and analysis, the five main volatiles from each of the honeys are compared. Whereas some compounds were found in several types of monofloral honey, and thus not considered good volatile markers, some monofloral honeys revealed characteristic volatile compounds independently of their provenance.

Detection and quantification of adulterants in honey by LIBS.

The present work proposes methods for detection and quantification of honey adulterants using laser-induced breakdown spectroscopy (LIBS). The sample set consisted of 6 pure honey from different botanical sources, 2 sweetener syrups and 228 fortified samples. The spectra acquired using a spark discharge coupled to the LIBS system were used for the development of the PLS-DA (classification) and PLS (calibration) models. Several data preprocessing and variable selection methods were evaluated to obtain the best fit. The detection of adulterants was performed with 100% of accuracy. The quantification of adulterants was possible through a PLS model with the variables selected by iPLS. The PLS model was validated with external samples and presented good accuracy, selectivity, sensitivity, and linearity. The proposed methods highlighted the potential of the LIBS technique for honey authenticity certification, providing fast, simple, and clean determinations since no sample pretreatment was required.

Pesticide and trace element residues in honey and beeswax combs from Israel in association with human risk assessment and honey adulteration.

Beehive products are considered sentinels for environmental pollutants. The presence of trace elements and pesticides in honey and beeswax may pose a health hazard to consumers. The study's aim was to determine the profile of pesticides and trace elements in Israeli honey and beeswax samples in relation to human risk assessment. At least two pesticides contaminated the honey and beeswax samples simultaneously, in which, amitraz metabolites and coumaphos were frequently detected. The neonicotinoid insecticides and 2,4-dichlorophenoxyacetic acid, were found only in honey samples, whereas the more lipophilic pesticides were predominantly found in beeswax. In honey, chromium displayed the highest mean concentration, followed by zinc, whereas lead and molybdene occurred only in beeswax. Our findings indicate that the daily consumption of honey and beeswax together may compromise children's health. Sucrose-syrup fed honey could not be distinguished from floral honey based on sugar profile, rather by its trace elements levels.

Análise da qualidade físico-química com ênfase na detecção de fraudes em méis comercializados no Estado do Rio de Janeiro / Analysis of the physical-chemical quality with an emphasis in the detection of frauds in means marketed in the State of Rio de Janeiro

O presente estudo avaliou a qualidade e a possível ocorrência de fraudes em méis comercializados no Estado do Rio de Janeiro. Dez amostras de méis foram adquiridas em comércios da região, onde foram submetidas às analises de qualidade (HMF, Açucares Redutores, Sacarose aparente e Umidade) estabelecidas pela Normativa nº11 de outubro de 2000 do MAPA (BRASIL, 2000), bem como a analises específicas para detecção de fraudes (Reação de Fiehe, Reação de Lund e Teste de Lugol). Os resultados demonstraram que os níveis de sacarose aparente e umidade apresentaram-se acima do limite estabelecido pela normativa. Os outros parâmetros de qualidade avaliados estavam dentro do permitido. Nos testes específicos de adulteração apenas uma amostra (10%) foi positiva, tanto para o Teste de Lugol, quanto para Reação de Fiehe, confirmando a presença de fraude e a necessidade de maior fiscalização desses produtos.

Análise físico-quimica e microbiológica de amostras de mel comercializadas no município de Juiz de Fora-MG / Physical-chemical and microbiological analysis of honey samples marketed in the municipality of Juiz de Fora-MG

O mel é um produto obtido das abelhas melíferas contem altas quantidades de monossacarídeos, como a glicose e a frutose, sendo considerado um alimento de alto valor nutricional. Devido seu alto valor comercial, grande parte do mel comercializado não tem sua qualidade preservada. O trabalho teve o objetivo avaliar amostras de mel comercializadas na cidade de Juiz de Fora-MG através de análises físico-química e microbiológica. Das onze amostras analisadas: três apresentaram umidade acima de 20% permitido pela legislação; cinco amostras foram positivas para o Teste de Lugol; cinco foram reprovadas no teste de Lund por apresentarem baixa quantidade de albuminóides e duas amostras apresentaram positividade na análise de Fiehe. Na análise microbiológica não ocorreu crescimento de coliformes totais, termotolerantes, bolores e leveduras. O consumidor deseja um produto seguro e de boa qualidade. Os resultados reforçam a necessidade do controle do comércio e a conscientização dos consumidores.

Avaliação da qualidade de méis artesanais e méis oriundos de entrepostos registrados / Evaluation of the honey quality of beekeepers and honey from registered places

O presente estudo analisa as propriedades físico-químicas de méis oriundos de entrepostos registrados e de méis de apicultores não registrados. Dez amostras de méis da região Sudeste do Brasil foram analisadas. Os resultados demonstraram que em méis registrados a umidade variou de 17,8 a 19,9%, o hidroximetilfurfural (HMF) de 0,5 a 41,3 meq/kg, os açúcares redutores de 73,8 a 77%, a sacarose aparente de 2,25 a 6% e a acidez de 37,5 a 52,7 meq/kg. Já em méis não registrados, a umidade variou de 18,4 a 22,6%, o HMF de 2 a 25 meq/kg, os açúcares redutores de 67 a 77%, a sacarose aparente de 2 a 11% e a acidez de 31,2 a 140,5 meq/kg. Os resultados sugerem que a grande variação na qualidade das amostras pode ser diretamente influenciada pela forma de manejo de coleta, transporte, beneficiamento e armazenamento do produto.

Características físico-químicas e microbiológicas do mel de abelhas sem ferrão produzido em comunidades tradicionais do Amazonas / Physico-chemical and microbiological characteristics of the honey of stingerless bees produced in tradicional communities of the Amazon

Este trabalho foi desenvolvido com dois objetivos, (1) verificar se parâmetros físico-químicos e microbiológicos do mel de abelhas sem ferrão produzido em comunidades tradicionais do município de Boa Vista do Ramos atendem a legislação vigente, (2) comparar parâmetros físico-químicos e microbiológicos de amostras de mel de abelhas sem ferrão coletadas diretamente das colônias, de amostras recebidas na agroindústria para posterior processamento. Foram coletas nove amostras de mel de cinco comunidades ribeirinhas e oito amostras da agroindústria. As características físico-químicas e microbiológicas avaliadas foram: pH, umidade, acidez titulável total, cinzas totais, sólidos insolúveis em água, coliformes totais, coliformes termotolerantes, bolores e levedura. Os resultados médios e devios padrões do pH, umidade, acidez, cinzas e sólidos insolúveis das amostras de mel coletadas diretamente dos potes de mel foram: 3,2±0,17; 26,9±1,22; 27,02± 13,57; 0,49±0,04; 0,06±0,02, respectivamente. Os mesmos parâmetros avaliados de amostras coletadas da agroindústria foram: 3,26 ±0,14; 26,34±1,00; 28,59±7,51; 0,49±0,05 e 0,07±0,01, respectivamente. Não houve crescimento de coliformes a 35ºC e 45ºC. A média de bolores e leveduras das amostras coletadas diretamente das colônias foi de 3,2 x 10³ est. e da agroindústria de 3,4 X 10³ est. Estatisticamente essas contagens não apresentam diferença. Portanto, conclui-se que as amostras de mel de abelha sem ferrão coletadas atenderam a legislação vigente, e a forma de acondicionamento e transporte do mel não afetou os parâmetros analisados.

Caracterização físico-química do mel de abelhas sem ferrão / Physical-chemical characterization of honey stingless bees

O estudo relata as análises físico-químicas do mel de espécies de abelhas sem ferrão (Meliponini), avaliando os resultados conforme a legislação. Foram obtidas 25 amostras de méis de seis espécies de abelhas sem ferrão de meliponicultores do Vale de Taquari, na Região Central do RS. Foi analisado acidez, açúcares redutores, atividade diastásica, hidroximetilfurfural (HMF), sólidos insolúveis, resíduos minerais e umidade. As análises físico-químicas foram realizadas pelo Laboratório de Apicultura da UFRGS, atendendo as exigências do Laboratório Nacional Agropecuário do Rio Grande do Sul (LANAGRO), baseadas no Manual de Métodos Oficiais para Análise de Alimentos de Origem Animal publicado em 2017. Foi observado que os méis de abelha sem ferrão apresentaram apenas uma não conformidade na atividade diastásica. Salienta-se a necessidade da adequação ou da elaboração de uma legislação que determine as características específicas de identidade e qualidade do mel de abelhas sem ferrão.

Qualidade físico-química do mel produzido por melíponas no Estado de Alagoas, Brasil / Physical-chemical quality of honey produced by melíponas in the State of Alagoas, Brazil

O mel de abelhas sem ferrão possui um grande valor agregado e estas abelhas são de suma importância na manutenção de seus ecossistemas, contribuindo para a conservação da flora e da fauna. Este trabalho possui o intuito de contribuir para a regulamentação da meliponicultura, através da especificação da qualidade físico-química de méis de meliponídeos do Estado de Alagoas. Desta forma, foram analisadas quatro amostras de méis, sendo todas elas obtidas através da espécie Melipona scutellaris. Estas amostras obtiveram resultados consoantes com a legislação relativa a abelha do gênero Apis, com exceção do parâmetro da umidade que se revelou acima do máximo, ou seja, maior que 20%. Então, apesar de alguns parâmetros se adequarem as normas vigentes, há a necessidade de uma legislação própria que leve em conta as suas particularidades.

Análise de possíveis fraudes em méis comercializados no município de Seropédica - RJ e arredores / Analysis of possible frauds in honey commercialized in the municipality of Seropédica-RJ and surroundings

O mel é um produto elaborado pelas abelhas melíferas a partir do néctar e secreções das plantas, sendo proibido por Lei adição de qualquer substância que altere a sua composição. O mel brasileiro é considerado entre os mais puros do mundo, e com grande aceitação nos mercados europeus e norte americano, porém é um produto passível de fraudes, e muitas vezes encontrado sendo comercializado de maneira informal, sem nenhum tipo de fiscalização ou registro. As fraudes e adulterações podem ser detectadas por meio de análises físico-químicas. No presente estudo foram avaliadas dez amostras de méis adquiridas no comércio do Município de Seropédica e seus arredores, com relação a presença de açúcar comercial, para tal foram realizadas as análises de Fiehe e de Lugol, foram avaliadas também as informações na rotulagem do produto. Foi detectado resultado positivo para adição de açúcar comercial em duas das amostras, apresentando resultado positivo da reação do Lugol e no teste de Fiehe, e nas amostras com rótulo, uma não apresentava a indicação da florada, e três não possuíam rótulo.

Caracterização microbiológica do mel produzido por melíponas no Estado de Alagoas / Microbiological characterization of honey produced by melíponas in the State of Alagoas

O mel de abelhas sem ferrão possui um grande valor agregado, chegando a ultrapassar o preço do mel das abelhas com ferrão (gênero Apis) em até quinze vezes. As melíponas são de suma importância na manutenção de seus ecossistemas de origem, contribuindo de forma para a conservação e a sobrevivência da flora e da fauna. A pesquisa objetivou caracterizar os méis de meliponídeos nativos do Estado de Alagoas. Foram coletadas e analisadas quatro amostras de méis, sendo todas elas obtidas da espécie Melipona scutellaris (mais conhecida como Uruçu) em diferentes municipios, condições e momentos. As amostras estavam próprias para consumo humano e de acordo com a legislação vigente verificou-se que os padrões adotados para o mel produzido por Apis melífera podem ser aplicados para o mel produzido por Meliponas.

Physicochemical characteristics of bracatinga honeydew honey and blossom honey produced in the state of Santa Catarina: An approach to honey differentiation.

Due to the increasing demand and interest of the consumers of bracatinga honeydew honey, it becomes important to know its characteristics in order to guarantee the quality of this dark honey. It is also necessary to investigate possible parameters that easily determine their differentiation from other honeys, thus guaranteeing their authenticity. Therefore, in the present study, the physicochemical characteristics and the reducing/scavenging proprieties of 16 bracatinga (Mimosa scabrella Bentham) honeydew honey samples and 25 blossom honey (different botanical origin) samples from Santa Catarina, Brazil, were investigated. Bracatinga honeydew honeys were defined by its high free acidity, electrical conductivity and reducing/scavenging capacity, and low glucose content.Still, these specific parameters allowed the differentiation between bracatinga honeydew honeys and blossom honeys, from the of principal component analysis, contributing to the differentiation of these honeys.

Authentication of Apis cerana Honey and Apis mellifera Honey Based on Major Royal Jelly Protein 2 Gene.

In Asia, honey is mainly produced by Apis mellifera and Apis cerana. However, the price of A. cerana honey is usually much higher than A. mellifera honey. Seeing considerable profits, some dishonest companies and beekeepers mislabel A. mellifera honey as A. cerana honey or incorporate A. mellifera honey into A. cerana honey. In the present study, we developed methods to discriminate A. cerana honey from A. mellifera honey based on the MRJP2 (major royal jelly protein 2) gene. Two pairs of species-specific primers were designed. The amplification products of A. cerana and A. mellifera were 212 and 560 bp, respectively. As little as one percent incorporation of A. mellifera honey in the mixture can be detected by duplex PCR. Additionally, another method based on the melt curve analysis using the same primers was also developed, allowing a rapid discrimination of real-time PCR product of different species. Our study shows that the entomological authentication of honey samples can be identified by nuclear genes other than mitochondrial genes and this extends the possibility of gene selection in identification. The authentication system we proposed could be a useful tool for discriminating A. cerana honey from A. mellifera honey.

Physicochemical characterization and determination of chloramphenicol residues and heavy metals in Algerian honeys.

The concentration of certain heavy metals in various foods (fruits, cereals, legumes, and bee products) produced in industrial and urban cities is increasing each year following industrial development. Quality of honey and its contamination by different polluting agents are related essentially to its production environment, or it can arise from beekeeping practices. In the present study, the determination of physicochemical properties: moisture, pH, total acidity, electric conductivity, hydroxymethylfurfural (HMF), sugars, and chloramphenicol (CAP) residues; the metal content by determination of two toxic metals levels: lead (Pb) and cadmium (Cd); and other trace elements: magnesium (Mg), iron (Fe), zinc (Zn), copper (Cu), and nickel (Ni) in 23 different honey samples collected from North regions of Algeria were investigated. The physicochemical properties and the metal contents were found within the ranges established by the international standards. For the antibiotic residues, only four honey samples are contaminated by CAP. Metals were found in non-significant values and are in safety baseline levels for human consumption except Mg which exceed the limits. These results suggested that honey could be used as an indicator to detect contaminating agents from the environment since bees are excellent sentinels for assessing environmental contamination because of their physiological and biological characteristics.

HPTLC Fingerprinting-Rapid Method for the Differentiation of Honeys of Different Botanical Origin Based on the Composition of the Lipophilic Fractions.

Bee honey possess various nutritional and medicinal functions, which are the result of its diverse chemical composition. The numerous bioactive compounds in honey come from flower nectar; thus, the identification of the specific chemical profiles of honey samples is of great importance. The lipophilic compounds from eight monofloral honeys (rape, buckwheat, clover, willow, milk thistle, dandelion, raspberry and sweet yellow clover) were investigated. Analyses of the lipophilic fractions obtained by UAE (ultrasound assisted extraction) and SPE (solid phase extraction) extractions were performed using high-performance thin layer chromatography (HPTLC). Chromatographic and cluster analyses allowed the identification of a unique, colorful pattern of separated compounds with specific Rf values on the HPTLC plate for each type of monofloral honey. HPTLC is a simple and effective visual method of analysis, and it can serve as a basis for authenticating different types of honey.