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.

Chemical Composition of the Same Brazilian Propolis Sample Analyzed in 1997 and in 2012: No Freezing Effect.

The same propolis sample that was analyzed by GC-MS in 1997 and 2012 was examined again after being frozen for 15 years. No qualitative changes were seen in its composition; artepilin C, prenyl-p-coumaric acid, dihydrocinnamic acid, caffeic acid were the major compounds.

International Regulations of Propolis Quality: Required Assays do not Necessarily Reflect their Polyphenolic-Related In Vitro Activities.

Propolis has been proposed as a polyphenolic-rich natural product potentially able to be used for human consumption or even for medicinal proposes. To guarantee a minimum phenolic and flavonoid content and as consequence of their related-biological activities, international requirements of propolis quality are commonly applied. In this work we assessed phenolic and flavonoid contents of propolis; the antioxidant capacity (toward peroxyl radicals and hypochlorous acid); the ability to generate nitric oxide (NO); and, finally the antimicrobial activity of 6 propolis samples from the VI region of Chile. Our results show that the total phenolic and flavonoid content of propolis samples are not always in agreement with their polyphenolic-associated in vitro activities. For example, P03 and P06 samples showed the lowest (25 ± 4 GAE/g propolis) and the highest (105 ± 3 GAE/g propolis) total phenolic content, respectively. This was in agreement with flavonoid content and their Oxygen Radical Absorbance Capacity (ORAC) activity. However, this dependence was not observed toward HOCl, NO release and antimicrobial activity. Based on our results, we consider that, in order to guarantee the antioxidant or antimicrobial in vitro effects, the international regulations of propolis quality should contemplate the convenience of incorporating other simple analytical test such as ORAC or antimicrobial tests.

Composition, labelling, and safety of food supplements based on bee products in the legislative framework of the European Union - Croatian experiences.

The European Union market is overflown by food supplements and an increasing number of consumers prefer those where bee products play an important part in their composition. This paper deals with complex European Union legislation concerning food supplements based on bee products, placing a special emphasis on their composition, labelling, and safety. Correct labelling of food supplements also represents a great challenge since, in spite of legal regulations in force, there are still open issues regarding the statements on the amount of propolis, which is not clearly defined by the legal framework. One of the key issues are the labels containing health claims from the EU positive list approved by the European Food Safety Authority. Emphasis will also be placed on informing consumers about food, as statements which imply the healing properties of food supplements and their capacity to cure diseases are forbidden. One of the key elements of product safety is HACCP based on the EU Regulations EC 178/02 and 852/2004. Health safety analyses of food supplements with bee products used as raw materials, which are standardised by legal regulations will also be discussed. In the future, attention should also be paid to establishing the European Union "nutrivigilance" system. Croatian experiences in addressing challenges faced by producers, supervisory entities, and regulatory and inspection bodies may serve as an example to countries aspiring to become part of the large European family.

Foreign matter contaminating ethanolic extract of propolis: a filth-test survey comparing products from small beekeeping farms and industrial producers.

Propolis is a resinous material collected by honeybees from the exudates and buds of plants. It has been widely used as a remedy by humans since ancient times, as well as for dietary supplements and cosmetics. European legislation recently focused on the quality and hygiene standards of foods, including beehive products, and extensive efforts have been made to avoid the presence of chemical contaminants, whilst in contrast few studies have investigated the magnitude of contamination by physical ones. We conducted a filth-test survey to evaluate the contamination of ethanolic extract of propolis by foreign materials. We also compared the abundance of contaminants in propolis extracts currently marketed by small beekeepers and industrial producers. We found different foreign materials in the ethanol extract of propolis. Contaminants differed in abundance, with a higher number of carbon particles (small beekeepers: 2.70 ± 0.63; industrial producers: 1.25 ± 0.49; mean (n/30 ml) ± SE) and other inorganic fragments (small beekeepers: 3.50 ± 0.31; industrial producers: 3.88 ± 1.11) than arthropod fragments (small beekeepers: 0.30 ± 0.21; industrial producers: 0.38 ± 0.26) and mammal hairs (small beekeepers: 0.10 ± 0.10; industrial producers: 0.38 ± 0.26). No differences in the abundance of foreign matter between propolis from small beekeepers and industrial producers were found, allowing us to point out an increased awareness by small producers of issues inherent in hygiene management. Contamination of propolis extracts by animal body parts, such as insect fragments, mites and rodent hairs, indicates poor management of hygiene in the production process and low effectiveness of the filtration phase. Animal-borne contaminants can act as pathogen vectors as well as introducing dangerous allergens when ingested or applied to human skin. The filth-test applied to ethanolic propolis extract quality control can be considered a promising tool, also for small beekeeper activities, since it is cheap and allows the quick interpretation of results.

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.

Chemical diversity of propolis and the problem of standardization.

Chemical variability of propolis is discussed with respect to the problem of standardization. Several chemical types of propolis are formulated, based on their plant source. Reliable criteria for chemical standardization of different propolis types are needed but such generally accepted criteria do not yet exist. The chemical profile of "poplar" propolis, typical for the temperate zone, can be characterized by the following parameters: total flavone and flavonol content, total flavanone and dihydroflavonol content, and total phenolics content. These parameters correlate better with the biological activity and are more informative that the quantification of individual components. There is still a lot of work to be done to achieve standardization of other propolis types. Working with standardized material will allow scientists to connect a particular chemical propolis type to a specific type of biological activity and formulate recommendations for mainstream practitioners.