The Folin-Ciocalteu assay is a widely used method for measuring the total phenolic content (TPC) in honey, but it can be affected by the presence of reducing sugars in honey, which can lead to interference and an over-estimation of its TPC. To optimize the Folin-Ciocalteu assay for honey analysis, the effect of pH on the assay was investigated. A number of pH scenarios were tested using different concentrations of Na₂CO₃ (0.00%, 0.75%, 0.94%, and 7.50%) in order to minimize reducing sugar interference and maximize the reaction of phenolics in the assay. The modified TPC method was then validated in accordance with current International Council on Harmonisation (ICH) guidelines. The findings of this study demonstrate that the traditional Folin-Ciocalteu assay (using 7.50% aqueous Na₂CO₃ solution, pH 10.8) leads to a significant overestimation of the TPC of honey due to the interference of reducing sugars. However, a pH of 7.9, achieved by using a 0.75% aqueous Na₂CO₃ solution, provides suitable conditions to account for most of the phenolic compounds without interference from reducing sugars. This finding was further confirmed by testing various sugar solutions and artificial honey which yielded TPC values below the established limit of detection and quantification of the assay. However, a slight increase in pH, even by a moderate deviation (pH 8.9), leads to significant discrepancies in absorbance readings, indicating that pH control is crucial for the accurate analysis of TPC in honey.

Honey, a naturally sweet and viscous substance is mainly produced by honeybees (Apis mellifera) from flower nectar. Honey exerts a plethora of biological and pharmacological activities, namely, antioxidant, antimicrobial and anti-inflammatory activity, because of the presence of an extensive variety of bioactive compounds. The antibacterial activity is one of the most reported biological properties, with many studies demonstrating that honey is active against clinically important pathogens. As a result, beside honey’s widespread utilization as a common food and flavouring agent, honey is an attractive natural antimicrobial agent. However, the use of neat honey for therapeutic purposes poses some problems, for instance, its stickiness may hamper its appeal to consumers and health care professionals, and the maintenance of an adequate therapeutic concentration over a sufficient timeframe may be challenging due to honey liquidity and leakage. It has motivated researchers to integrate honey into diverse formulations, for example, hydrogels, dressings, ointments, pastes and lozenges. The antibacterial activity of these formulations should be scientifically determined to underscore claims of effectiveness. Some researchers have made efforts to adapt the disc carrier and suspension test to assess the antimicrobial activity of topical products (e.g., silver-based wound dressings). However, there is currently no established and validated method for determining the in vitro antimicrobial potential of natural product-based formulations, including those containing honey as the active principle. Against the backdrop of a brief discussion of the parameters that contribute to its antibacterial activity, this review provides an outline of the methods currently used for investigating the antibacterial activity of neat honey and discusses their limitations for application to honey-based formulations.

Basic fibroblast growth factor (FGF)-2 has been shown to regulate many cellular functions including cell proliferation, migration, and differentiation, as well as angiogenesis in a variety of tissues, including skin, blood vessel, muscle, adipose, tendon/ligament, cartilage, bone, tooth, and nerve. These multiple functions make FGF-2 an attractive component for wound healing and tissue engineering constructs; however, the stability of FGF-2 is widely accepted to be a major concern for the development of useful medicinal products. Many approaches have been reported in the literature for preserving the biological activity of FGF-2 in aqueous solutions. Most of these efforts were directed at sustaining FGF-2 activity for cell culture research, with a smaller number of studies seeking to develop sustained release formulations of FGF-2 for tissue engineering applications. The stabilisation approaches may be classified into the broad classes of ionic interaction modification with excipients, chemical modification, and physical adsorption and encapsulation with carrier materials. This review discusses the underlying causes of FGF-2 instability and provides an overview of the approaches reported in the literature for stabilising FGF-2 that may be relevant for clinical applications. Although efforts have been made to stabilise FGF-2 for both in vitro and in vivo applications with varying degrees of success, the lack of comprehensive published stability data for the final FGF-2 products represents a substantial gap in the current knowledge, which has to be addressed before viable products for wider tissue engineering applications can be developed to meet regulatory authorisation.

Honey, a concentrated natural product, is produced by honeybees (Apis mellifera) from the nectar of flowers. It contains over 200 compounds that exert various biological or pharmacological activities, ranging from antioxidant, anti-inflammatory, antimicrobial, and antihypertensive to hypoglycemic effects. Due to the presence of a plethora of bioactive compounds, as well as unique physicochemical properties, honey has been widely used as medicine throughout human history along with its extensive utilization as common food and flavoring agent. The application of neat honey for therapeutic purpose, however, poses some difficulties such as the maintenance of a required therapeutic concentration over an adequate timeframe due to the problem of liquefaction and leakage. This has driven researchers to incorporate honey into a range of formulations, for example, hydrogels, dressings, ointments, pastes, or lozenges. After a brief discussion of the chemistry and medicinal use of honey, this review focuses on commercial honey-based medicinal formulations as well as in vitro, in vivo, and clinical studies on noncommercial honey formulations for the treatment of various ailments. In addition to this, it also covers the application of honey formulations and the evidence underpinning their use.

Honey adulteration, where a range of sugar syrups is used to increase bulk volume, is a common problem that has significant negative impacts on the honey industry, both economically and from a consumer confidence perspective. This paper investigates High-Performance Thin Layer Chromatography (HPTLC) for the authentication and detection of sugar adulterants in honey. The sugar composition of various Australian honeys (Manuka, Jarrah, Marri, Karri, Peppermint and White Gum) was first determined to illustrate the variance depending on the floral origin. Two of the honeys (Manuka and Jarrah) were then artificially adulterated with six different sugar syrups (rice, corn, golden, treacle, glucose and maple syrup). The findings demonstrate that HPTLC sugar profiles, in combination with organic extract profiles, can easily detect the sugar adulterants. As major sugars found in honey, the quantification of fructose and glucose, and their concentration ratio can be used to authenticate the honeys. Quantifications of sucrose and maltose can be used to identify the type of syrup adulterant, in particular when used in combination with HPTLC fingerprinting of the organic honey extracts.

The aim of this review is to provide a comprehensive overview of the large variety of phenolic compounds that have to date been identified in a wide range of monofloral honeys found globally. The collated information is structured along several themes, including the botanical family and genus of the monofloral honeys for which phenolic constituents have been reported, the chemical classes the phenolic compounds can be attributed to, and the analytical method employed in compound determination as well as countries with a particular research focus on phenolic honey constituents. This review covers 130 research papers that detail the phenolic constituents of a total of 556 monofloral honeys. Based on the findings of this review, it can be concluded that most of these honeys belong to the Myrtaceae and Fabaceae families and that Robinia (Robinia pseudoacacia, Fabaceae), Manuka (Leptospermum scoparium, Myrtaceae), and Chestnut (Castanea sp., Fagaceae) honeys are to date the most studied honeys for phenolic compound determination. China, Italy, and Turkey are the major honey phenolic research hubs. To date, 161 individual phenolic compounds belonging to five major compound groups have been reported, with caffeic acid, gallic acid, ferulic acid and quercetin being the most widely reported among them. HPLC with photodiode array detection appears to be the most popular method for chemical structure identification.