Portable NIR Spectroscopy to Simultaneously Trace Honey Botanical and Geographical Origins and Detect Syrup Adulteration.

Fraudulent practices concerning honey are growing fast and involve misrepresentation of origin and adulteration. Simple and feasible methods for honey authentication are needed to ascertain honey compliance and quality. Working on a robust dataset and simultaneously investigating honey traceability and adulterant detection, this study proposed a portable FTNIR fingerprinting approach combined with chemometrics. Multifloral and unifloral honey samples (n = 244) from Spain and Sardinia (Italy) were discriminated by botanical and geographical origin. Qualitative and quantitative methods were developed using linear discriminant analysis (LDA) and partial least squares (PLS) regression to detect adulterated honey with two syrups, consisting of glucose, fructose, and maltose. Botanical and geographical origins were predicted with 90% and 95% accuracy, respectively. LDA models discriminated pure and adulterated honey samples with an accuracy of over 92%, whereas PLS allows for the accurate quantification of over 10% of adulterants in unifloral and 20% in multifloral honey.

Mass Spectrometry Characterization of Honeydew Honey: A Critical Review.

Honeydew honey is produced by bees (Apis mellifera) foraging and collecting secretions produced by certain types of aphids on various parts of plants. In addition to exhibiting organoleptic characteristics that distinguish them from nectar honey, these honeys are known for their functional properties, such as strong antioxidant and anti-inflammatory activities. Despite their importance, they remain poorly characterized in comparison with flower honeys, as most studies on this subject are not only carried out on too few samples but also still focused on traditional chemical-physical parameters, such as specific rotation, major sugars, or melissopalynological information. Since mass spectrometry has consistently been a primary tool for the characterization and authentication of honeys, this review will focus on the application of these methods to the characterization of the minor fraction of honeydew honey. More specifically, this review will attempt to highlight what progress has been made so far in identifying markers of the authenticity of the botanical and/or geographical origin of honeydew honeys by mass spectrometry-based approaches. Furthermore, strategies devoted to the determination of contaminants and toxins in honeydew honeys will be addressed. Such analyses represent a valuable tool for establishing the level of food safety associated with these products. A critical analysis of the presented studies will identify their limitations and critical issues, thereby describing the current state of research on the topic.

Monitoring the Phenolic and Terpenic Profile of Olives, Olive Oils and By-Products throughout the Production Process.

Olive oil is a food of great importance in the Mediterranean diet and culture. However, during its production, the olive oil industry generates a large amount of waste by-products that can be an important source of bioactive compounds, such as phenolic compounds and terpenes, revalorizing them in the context of the circular economy. Therefore, it is of great interest to study the distribution and abundance of these bioactive compounds in the different by-products. This research is a screening focused on phytochemical analysis, with particular emphasis on the identification and quantification of the phenolic and terpenic fractions. Both the main products of the olive industry (olives, olive paste and produced oil) and the by-products generated throughout the oil production process (leaf, "alpeorujo", liquid and solid residues generated during decanting commonly named "borras" and washing water) were analyzed. For this purpose, different optimized extraction procedures were performed for each matrix, followed by high-performance liquid chromatography coupled with electrospray time-of-flight mass spectrometry (HPLC-ESI-TOF/MS) analysis. Although no phenolic alcohols were quantified in the leaf and the presence of secoiridoids was low, this by-product was notable for its flavonoid (720 ± 20 µg/g) and terpene (5000 ± 300 µg/g) contents. "Alpeorujo" presented a complete profile of compounds of interest, being abundant in phenolic alcohols (900 ± 100 µg/g), secoiridoids (4500 ± 500 µg/g) and terpenes (1200 ± 100 µg/g), among others. On the other hand, while the solid residue of the borras was the most abundant in phenolic alcohols (3700 ± 200 µg/g) and secoiridoids (680 ± 20 µg/g), the liquid fraction of this waste was notable for its content of elenolic acid derivatives (1700 ± 100 µg/mL) and phenolic alcohols (3000 ± 300 µg/mL). Furthermore, to our knowledge, this is the first time that the terpene content of this by-product has been monitored, demonstrating that it is an important source of these compounds, especially maslinic acid (120 ± 20 µg/g). Finally, the phytochemical content in wash water was lower than expected, and only elenolic acid derivatives were detected (6 ± 1 µg/mL). The results highlighted the potential of the olive by-products as possible alternative sources of a wide variety of olive bioactive compounds for their revalorization into value-added products.

Elemental Fingerprinting Combined with Machine Learning Techniques as a Powerful Tool for Geographical Discrimination of Honeys from Nearby Regions.

Discrimination of honey based on geographical origin is a common fraudulent practice and is one of the most investigated topics in honey authentication. This research aims to discriminate honeys according to their geographical origin by combining elemental fingerprinting with machine-learning techniques. In particular, the main objective of this study is to distinguish the origin of unifloral and multifloral honeys produced in neighboring regions, such as Sardinia (Italy) and Spain. The elemental compositions of 247 honeys were determined using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The origins of honey were differentiated using Principal Component Analysis (PCA), Linear Discriminant Analysis (LDA), and Random Forest (RF). Compared to LDA, RF demonstrated greater stability and better classification performance. The best classification was based on geographical origin, achieving 90% accuracy using Na, Mg, Mn, Sr, Zn, Ce, Nd, Eu, and Tb as predictors.

Thermal degradation of red cabbage (Brassica oleracea L. var. Capitata f. rubra) anthocyanins in a water model extract under accelerated shelf-life testing.

Red cabbage (RC) represents a source of anthocyanins acylated with hydroxycinnamic acids (HCA) that are described to enhance their stability. Nevertheless, data about their thermal degradation are still controversial. Our aim was to comprehensively analyse the degradation kinetics of individual RC anthocyanins in a model aqueous extract treated at 40 °C × 30 days to simulate severe but realistic storage conditions. Free anthocyanins and radical-scavenging capacity showed different kinetics. The results confirm the high stability of RC anthocyanins (t1/2: 16.4-18.4 days), although HPLC analyses of each molecule displayed distinct kinetics with t1/2 from 12.6 to 35.1 days. In particular, the sinapoyl acylation negatively affected the stability of the anthocyanins, while the forms monoacylated with glycosyl p-coumaric and ferulic acids exhibited higher stability. In conclusion, our results indicate that acylation is not a prerogative of stability, as this is instead more dependent on specific acylation patterns and the glycosylation of HCA.

Multi-Elemental Analysis as a Tool to Ascertain the Safety and the Origin of Beehive Products: Development, Validation, and Application of an ICP-MS Method on Four Unifloral Honeys Produced in Sardinia, Italy.

Despite unifloral honeys from Sardinia, Italy, being appreciated worldwide for their peculiar organoleptic features, their elemental signature has only partly been investigated. Hence, the principal aim of this study was to measure the concentration of trace and toxic elements (i.e., Ag, As, Ba, Be, Bi, Cd, Co, Cr, Cu, Fe, Hg, Li, Mn, Mo, Ni, Pb, Sb, Sn, Sr, Te, Tl, V, and Zn) in four unifloral honeys produced in Sardinia. For this purpose, an original ICP-MS method was developed, fully validated, and applied on unifloral honeys from asphodel, eucalyptus, strawberry tree, and thistle. Particular attention was paid to the method's development: factorial design was applied for the optimization of the acid microwave digestion, whereas the instrumental parameters were tuned to minimize the polyatomic interferences. Most of the analytes' concentration ranged between the relevant LoDs and few mg kg-1, while toxic elements were present in negligible amounts. The elemental signatures of asphodel and thistle honeys were measured for the first time, whereas those of eucalyptus and strawberry tree honeys suggested a geographical differentiation if compared with the literature. Chemometric analysis allowed for the botanical discrimination of honeys through their elemental signature, whereas linear discriminant analysis provided an accuracy level of 87.1%.

Electrochemical Determination of the "Furanic Index" in Honey.

5-(hydroxymethyl)furan-2-carbaldehyde, better known as hydroxymethylfurfural (HMF), is a well-known freshness parameter of honey: although mostly absent in fresh samples, its concentration tends to increase naturally with aging. However, high quantities of HMF are also found in fresh but adulterated samples or honey subjected to thermal or photochemical stresses. In addition, HMF deserves further consideration due to its potential toxic effects on human health. The processes at the origin of HMF formation in honey and in other foods, containing saccharides and proteins-mainly non-enzymatic browning reactions-can also produce other furanic compounds. Among others, 2-furaldehyde (2F) and 2-furoic acid (2FA) are the most abundant in honey, but also their isomers (i.e., 3-furaldehyde, 3F, and 3-furoic acid, 3FA) have been found in it, although in small quantities. A preliminary characterization of HMF, 2F, 2FA, 3F, and 3FA by cyclic voltammetry (CV) led to hypothesizing the possibility of a comprehensive quantitative determination of all these compounds using a simple and accurate square wave voltammetry (SWV) method. Therefore, a new parameter able to provide indications on quality of honey, named "Furanic Index" (FI), was proposed in this contribution, which is based on the simultaneous reduction of all analytes on an Hg electrode to ca. -1.50 V vs. Saturated Calomel Electrode (SCE). The proposed method, validated, and tested on 10 samples of honeys of different botanical origin and age, is fast and accurate, and, in the case of strawberry tree honey (Arbutus unedo), it highlighted the contribution to the FI of the homogentisic acid (HA), i.e., the chemical marker of the floral origin of this honey, which was quantitatively reduced in the working conditions. Excellent agreement between the SWV and Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) data was observed in all samples considered.

Early Immunocastration of Pigs: From Farming to Meat Quality.

The study aimed to test a very early immunization of pigs to prevent boar taint with regard to its practicability and influence on production performance, its reliability in ensuring good meat and fat quality, and animal welfare. Immunization was already conducted at piglet production stage and could be easily integrated into routine vaccination (week 3) and weaning practices (week 7). The fattening and slaughter performance of the animals was not affected by the immunization regime and was within the usual range. In addition, there were no abnormalities in animal behavior and the prevalence of injuries caused by aggressive interactions. All animals were classified as infertile on the basis of the histological examination of the testicles. However, the testosterone levels measured at slaughter were significantly higher in animals of the early immunization regime than in animals subjected to the standard immunization regime. Androstenone and skatole levels as the main components of boar taint were, on average, higher and varied to a greater extent in early immunized animals. Furthermore, the comparison of the immunization scheme did not result in significant differences for meat quality and for fatty acid composition.

Classification of Unifloral Honeys from SARDINIA (Italy) by ATR-FTIR Spectroscopy and Random Forest.

Nowadays, the mislabeling of honey floral origin is a very common fraudulent practice. The scientific community is intensifying its efforts to provide the bodies responsible for controlling the authenticity of honey with fast and reliable analytical protocols. In this study, the classification of various monofloral honeys from Sardinia, Italy, was attempted by means of ATR-FTIR spectroscopy and random forest. Four different floral origins were considered: strawberry-tree (Arbutus Unedo L.), asphodel (Asphodelus microcarpus), thistle (Galactites tormentosa), and eucalyptus (Eucalyptus calmadulensis). Training a random forest on the infrared spectra allowed achieving an average accuracy of 87% in a cross-validation setting. The identification of the significant wavenumbers revealed the important role played by the region 1540-1175 cm-1 and, to a lesser extent, the region 1700-1600 cm-1. The contribution of the phenolic fraction was identified as the main responsible for this observation.

Effect of the irrigation method and genotype on the bioaccumulation of toxic and trace elements in rice.

The total concentration of three toxic elements (As, Cd and Pb) and five oligoelements (Cu, Mn, Mo, Ni and Se) has been determined using an original and completely validated ICP-MS method. This was applied to rice grains from 26 different genotypes cultivated in the same soil and irrigated with the same water in three different ways: by the traditional continuous flooding (CF) and by two intermittent methods, the sprinkler irrigation (SP) and the periodical saturation of the soil (SA). The adoption of SP hugely minimizes the average amounts of almost all elements in kernels (-98% for As, -90% for Se and Mn, -60% for Mo, -50% for Cd and Pb), with the only exception of Ni, whose concentration increases the average amount found in the CF rice by 7.5 times. Also SA irrigation is able to reduce the amounts of As, Mo and Pb in kernels but it significantly increases the amounts of Mn, Ni and - mainly - Cd. Also the nature of the genotype determined a wide variability of data within each irrigation method. Genotypes belonging to Indica subspecies are the best bioaccumulators of elements in both CF and SP methods and, never, the worst bioaccumulators for any element/irrigation method combination. In the principal component analysis, PC1 can differentiate samples irrigated by SP by those irrigated by CF and SA, whereas PC2 provides differentiation of CF samples by SA samples. When looking at the loading plot Ni is negatively correlated to the majority of the other elements, except Cu and Cd having negative loadings on PC2. These results allow to envisage that a proper combination of the irrigation method and the nature of rice genotype might be a very valuable tool in order to successfully achieve specific objectives of food safety or the attainment of functional properties.

The Effect of Algae or Insect Supplementation as Alternative Protein Sources on the Volatile Profile of Chicken Meat.

The aim of this study was to investigate the differences in the volatile profile of meat from chickens fed with alternative protein diets (such as algae or insect) through two different trials. In Trial 1, broiler chicken at one day of age were randomly allocated to three experimental groups: a basal control diet (C) and two groups in which the soybean meal was replaced at 75% (in the starter phase) and 50% (in the grower phase) with partially defatted Hermetia illucens (HI) larvae or Arthrospira platensis (SP). In Trial 2, broiler chickens were housed and reared similar to Trial 1, with the exception that the experimental diets replaced soybean meal with either 100% partially defatted HI or 100% SP. In both trials, chickens were slaughtered at day 35. Per group, 10 chickens were submitted to volatile analysis by using solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS) analysis. Results in both trials showed that levels of several lipid-derived compounds were found to be lower in chickens fed an HI diet, which could be linked to a possibly lower level of polyunsaturated fatty acid content in HI-fed chicken. In addition, the dietary treatments could be discriminated based on the volatile profile, i.e., the substitution of soy with HI or SP distinctively affected the levels of flavor compounds.

Meat Quality Parameters and Sensory Properties of One High-Performing and Two Local Chicken Breeds Fed with Vicia faba.

The current practices of the poultry industry have raised concerns among consumers. Among these is the culling of day-old male chicks of laying hybrids; a suitable alternative for this could be the use of dual-purpose breeds where both sexes are used. Another practice that causes concern is the import of large quantities of soybeans for feedstuff production. Substitutes for these soybean-based products are regional protein crops, such as faba beans (Vicia faba L.; FBs). The objective of this study was to test the suitability of FB as a locally produced soybean meal replacement for two local dual-purpose chicken breeds and one high-performing layer line. The breast and leg meat of male Bresse Gauloise (BG), Vorwerkhuhn (VH), and White Rock (WR) animals was evaluated for different meat quality parameters: pH, color, water holding capacity, and tenderness. Sensory properties of the samples were evaluated by a trained panel with a conventional descriptive analysis. Results show different effects of FB diets on meat quality parameters in the different breeds. The attributes mostly affected by the diet are related to aroma, flavor, and texture, particularly in VH and WR. Overall, faba beans appear to be an acceptable dietary protein source for rearing these breeds for meat production.

The effect of insect or microalga alternative protein feeds on broiler meat quality.

BACKGROUND: In order to combat environmental and food security concerns associated with the increasing demand for soymeal related to increasing meat consumption, this study determines the chicken meat quality derived when soymeal is substituted for either partially de-fatted Hermetia illucens larval meal or spirulina (Arthrospira platensis) in broiler diets. Physicochemical parameters, sensory traits, and fatty acid composition of the meat are investigated, as well as an experiment to evaluate the impact of highly oxygenated atmosphere versus vacuum-bag packaging on shelf life was conducted. RESULTS: Hermetia illucens did not compromise quality; meat was slightly more yellow (higher b*), had a slightly decreased pH, and was less adhesive during chewing compared to the soy-fed control. Furthermore, Hermetia illucens resulted in higher saturated fatty acids proportions in thigh meat. Spirulina resulted in redder (higher a*) and more yellow (higher b*) meat with a slightly increased umami and chicken flavour. Spirulina-fed chicken meat had higher lipid oxidation levels compared to the control after being packaged in a highly oxygenated atmosphere; although, differences between the spirulina-fed and control fatty acid composition in thigh meat were minor. CONCLUSION: Both alternative protein feeds show potential to replace soymeal in broiler diets; however, they do result in moderately altered products.

Effect of Alternative Protein Feeds on the Content of Selected Endogenous Bioactive and Flavour-Related Compounds in Chicken Breast Meat.

Currently, soybean meal constitutes the main protein source for poultry production. However, the environmental and social issues related to soybean production are calling for more sustainable protein sources that can offset soybean requirements in animal production. Hermetia illucens larvae and the microalga spirulina (Arthrospira platensis) have proven to be effective alternatives to soybean meal for poultry production. In this study, the effect of 100% replacement of soy with partially defatted Hermetia illucens larvae and spirulina on the contents of selected endogenous bioactive (anserine, creatine and carnosine) and flavour-related (inosine and inosine-5´-monophosphate, IMP) compounds in chicken breast meat was evaluated. The results showed that the spirulina-based diet lowered the levels of anserine, carnosine and creatine compared to the control diet (3.3 vs. 4.1 mg/g, 0.15 vs. 0.72 mg/g and 1.49 vs. 2.49 mg/g, respectively) while IMP levels tended to be higher in spirulina-fed samples. Compared to the control group, Hermetia illucens-fed samples showed a lower content of bioactive peptides (anserine: 3.6 vs. 4.1 mg/g; carnosine: 0.39 vs. 0.72 mg/g; creatine: 2.03 vs. 2.49 mg/g), albeit to a lesser extent than the spirulina treatment group.

Extraction and Analysis of Phenolic Compounds in Rice: A Review.

Rice represents the main source of calorie intake in many world countries and about 60% of the world population include rice in their staple diet. Whole grain rice, also called brown rice, represent the unpolished version of the more common white rice including bran, germ, and endosperm. Many health-promoting properties have been associated to the consumption of whole grain rice and, for this reason, great attention has been paid by the scientific community towards the identification and the quantification of bioactive compounds in this food item. In this contribution, the last five years progresses in the quali-quantitative determination of phenolic compounds in rice have been highlighted. Special attention has been devoted to the most recent strategies for the extraction of the target compounds from rice along with the analytical approaches adopted for the separation, identification and quantification of phenolic acids, flavonoids, anthocyanins, and proanthocyanidins. More specifically, the main features of the "traditional" extraction methods (i.e., maceration, ultrasound-assisted extraction) have been described, as well as the more innovative protocols involving advanced extraction techniques, such as MAE (microwave-assisted extraction). The predominant role of HPLC in the definition of the phenolic profile has been examined also presenting the most recent results obtained by using mass spectrometry-based detection systems. In addition, the most common procedures aimed to the quantification of the total amount of the cited classes of phenolic compounds have been described together with the spectrophotometric protocols aimed to the evaluation of the antioxidant properties of rice phenolic extracts (i.e., FRAP, DPPH, ABTS and ORAC).

Chemometric treatment of simple physical and chemical data for the discrimination of unifloral honeys.

In this work, nonspecific physico-chemical parameters were determined in 160 honey samples belonging to the four main botanical categories present in Sardinia Island, Italy (strawberry tree, thistle, asphodel and eucalyptus) in order to develop a discriminant method for determining the botanical origin of honey. All the possible combinations of the seven physico-chemical parameters (pH, free acidity, electrical conductivity, color, total phenolic compounds, FRAP activity, and DPPH activity) measured in the honey samples were evaluated by Linear Discriminant Analysis (LDA). LDA models led to the prediction of each botanical origin with a very low level of misclassification (typically less than 5%). Since very high levels of correct prediction in cross validation (98.3%) and external validation (100%) were obtained considering only four parameters (i.e. pH, acidity, conductivity and DPPH), these results might allow a fast and easy control of the botanical origin of honeys.

Gas chromatography analysis of major free mono- and disaccharides in milk: Method assessment, validation, and application to real samples.

Saccharides are functional constituents of milk. Although d-lactose represents almost the totality of the saccharides in the milk, minor species, like d-glucose, d-galactose, myo-inositol and, as a result of severe thermal treatments, monosaccharides like d-tagatose, are also detectable. Although chromatography has been the main analytical approach used for accomplishing this task, quite surprisingly a validated gas chromatographic method aimed at the simultaneous determination of these compounds is still needed. Hence, our contribution is devoted to fill this gap. After the optimization of clean-up and derivatization (conversion of saccharides in their trimethyl silyl ethers) steps, the adoption of a highly cross-linked silphenylene stationary phase permitted to obtain high resolution and a fast chromatographic run. Validation was accomplished in terms of limit of detection, limit of quantification, linearity, precision, and trueness. The accuracy of the method was successfully tested on a number of partially skimmed milk samples. Excellent limits of detection for all analytes make this method eligible, also with respect to a gas chromatographic/mass spectrometry approach, for routine analysis and quality control in the dairy industries.

Recent Advances in the Analysis of Phenolic Compounds in Unifloral Honeys.

Honey is one of the most renowned natural foods. Its composition is extremely variable, depending on its botanical and geographical origins, and the abundant presence of functional compounds has contributed to the increased worldwide interest is this foodstuff. In particular, great attention has been paid by the scientific community towards classes of compounds like phenolic compounds, due to their capability to act as markers of unifloral honey origin. In this contribution the most recent progress in the assessment of new analytical procedures aimed at the definition of the qualitative and quantitative profile of phenolic compounds of honey have been highlighted. A special emphasis has been placed on the innovative aspects concerning the extraction procedures, along with the most recent strategies proposed for the analysis of phenolic compounds. Moreover, the centrality of validation procedures has been claimed and extensively discussed in order to ensure the fitness-for-purpose of the proposed analytical methods. In addition, the exploitation of the phenolic profile as a tool for the classification of the botanical and geographical origin has been described, pointing out the usefulness of chemometrics in the interpretation of data sets originating from the analysis of polyphenols. Finally, recent results in concerning the evaluation of the antioxidant properties of unifloral honeys and the development of new analytical approaches aimed at measuring this parameter have been reviewed.

A Possible Freshness Marker for Royal Jelly: Formation of 5-Hydroxymethyl-2-furaldehyde as a Function of Storage Temperature and Time.

In this article we present a study of the variability of the concentration of 5-hydroxymethyl-2-furaldehyde (HMF) in natural royal jelly (RJ) as a function of its storage temperature (-18, 4, and 25 °C) and time (up to 9 months after harvesting). For this work HMF is evaluated using an RP-HPLC method we previously assessed. While all RJ samples stored at 4 and -18 °C always showed levels of HMF under the limit of detection (0.13 mg kg(-1)), samples kept at 25 °C showed an exponential increase in the concentration of HMF as a function of the storage time. This behavior and a number of desirable features of the analytical method used (ease of use in routine laboratories, availability of a complete validation protocol specifically developed for RJ, based on consolidated chemical knowledge) allow us to hypothesize the use of HMF as a possible, reliable freshness marker for RJ.

RP-HPLC determination of water-soluble vitamins in honey.

The assessment and validation of reliable analytical methods for the determination of vitamins in sugar-based matrices (e.g. honey) are still scarcely explored fields of research. This study proposes and fully validates a simple and fast RP-HPLC method for the simultaneous determination of five water-soluble vitamins (vitamin B(2), riboflavin; vitamin B(3), nicotinic acid; vitamin B(5), pantothenic acid; vitamin B(9), folic acid; and vitamin C, ascorbic acid) in honey. The method provides low detection and quantification limits, very good linearity in a large concentration interval, very good precision, and the absence of any bias. It has been successfully applied to 28 honey samples (mainly from Sardinia, Italy) of 12 different botanical origins. While the overall amount of the analytes in the samples is quite low (always below 40 mg kg(-1)), we have observed a marked dependence of some of their concentrations (i.e. vitamin B(3) and vitamin B(5)) and the botanical origin of the honey. This insight might lead to important characterization features for this food item.