Insights into Q-markers of honey-fried licorice in treating spleen deficiency based on substance and energy metabolism regulation.

BACKGROUND: Honey-fried Licorice (HFL) is a dosage form of Glycyrrhizae Radix et Rhizome processed with honey, which has been recorded to exhibit better efficacy in tonifying the spleen compared to the raw product. In contrast, different processing methods of Glycyrrhizae Radix et Rhizome exhibit different efficacies and applications, but their current quality control index components remain consistent. PURPOSE: Based on the discovery and research strategy of traditional Chinese medicine decoction piece quality marker (Q-marker), this study aimed to conduct a multidimensional integration of constituents absorbed into the body and metabolomics based on the tonifying spleen and stomach effects of HFL to effectively identify the Q-marker of HFL. METHODS: In this study, a spleen deficiency rat model was established using the "exhausted swimming + poor diet" method to investigate the pharmacodynamics of tonifying the spleen and stomach by HFL. The constituents absorbed into blood was conducted using UPLC-Q-TOF/MS, correlation analysis between metabolomics and constituents absorbed into blood recognized the Q-Marker of HFL. RESULTS: The pharmacodynamic data demonstrated that HFL exhibited a significant regulatory effect on the disordered levels of PP, trypsin, chymase, PL, α-Glu, MTL, GAS, VIP, IL-2, IFN-γ, and IgA in the spleen deficiency model. Furthermore, HFL was found to improve the pathological changes in the spleen and intestine in the spleen deficiency model, highlighting its significant "tonifying spleen and stomach" effect. In the serum containing HFL, a total of 17 constituents were identified as being absorbed into the blood. Among these, 11 were prototypical components, while 6 were metabolites. Metabolomics data revealed that 9 differentially expressed metabolic markers were observed. Furthermore, the analysis of endogenous metabolic markers indicated that 10 components exhibited significant correlations with these biomarkers. CONCLUSION: The effect of "tonifying spleen and stomach" of HFL is closely related to the regulation of the material and energy metabolism pathway. The Q-Marker of HFL is glycyrrhizic acid and 18ß-glycyrrhetinic acid as the main control standards and liquiritin, isoliquiritin, liquiritin, isoliquiritin, isolicorice flavonol, licorice chalcone C and Formononetin were used as auxiliary standards.

The amount of antioxidants in honey has a strong relationship with the plants selected by honey bees.

As one of the main sources of natural antioxidants, flowering plants play a role in the prevention and treatment of many diseases directly and indirectly. Honey is considered as an important nutrient in the supply of natural antioxidants, the amount of which is directly dependent on the plant origin and geographical location of the bee feeding place. The existence of valuable communities of native and endemic plant species has turned Alborz, Zagros and Azerbaijan into the most important hubs of honey production in Iran. In this study, we collected samples of honey from more than 90 regions in Alborz, Zagros and Azerbaijan during the years 2020 to 2021. We evaluated the samples using melissopalynology method and measuring the amount of antioxidant activity. The rise of antioxidant activity in honey is dependent on the abundance of some plant families as well botanical origins. The abundance of plant families Rosaceae, Amaranthaceae, Fabaceae and Asteraceae showed a higher influence on the amount of antioxidants in honey than other plant families. Also, the abundance of plant families Rosaceae and Fabaceae increased with increasing altitude. In general, the amount of antioxidant activity of honey samples shows a different percentage under the influence of ecological and geographical changes.

Assessing monofloral bee pollens from Türkiye: Palynological verification, phenolic profile, and antioxidant activity.

Honey bee pollen (HBP) is a hive product produced by worker bees from floral pollen grains agglutination. It is characterized by its excellent nutritional and bioactive composition, making it a superior source of human nutrition. This study aimed to evaluate the monofloral bee pollen samples, including Cistus, Crataegus monogyna, Cyanus, Elaeagnus angustifolia, Papaver somniferum, Quercus, Salix, Sinapis, and Silybum from Türkiye according to palynological analysis, antioxidant activity, phenolic profiles, and color. The phenolic profiles were detected using ultra-high performance liquid chromatography coupled with tandem mass spectrometry. Bee pollens were categorized into monofloral, bifloral, and multifloral, underscoring the significance of confirming the botanical source of them depending on palynological analyses. Total phenolic content (TPC) of bee pollens ranged from 4.5 to 14.4 mg gallic acid/g HBP. The samples exhibited antioxidant activity for 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS â€¢+ ) ranging from 94.9 to 233.5 µmol trolox/g HBP, whereas lower values were seen for 2,2-diphenyl-1-picrylhydrazyl (DPPH•) ranging from 25.86 to 70.81 µmol trolox/g HBP. A yellowish-red tint color was also displayed for whole samples, whereas only E. angustifolia bee pollen indicated a darker color (L* = 31.6). Among the phenolic compounds, luteolin, kaempferol, isorhamnetin, rutin, and genistein were the most abundant, and their profiles varied across the samples. It was also observed that TPC, antioxidant activities, and polyphenol composition were higher in samples containing pollen grains of P. somniferum, Quercus, Plantago, and E. angustifolia species. PRACTICAL APPLICATION: The increasing number of new findings on honey bee pollen is crucial to food science and technology. In this sense, this study offers a robust method for verifying the authenticity and quality of 11 monofloral bee pollens, which is crucial for the food industry. It also identifies potential sources of high-quality pollen, benefiting producers, and consumers seeking superior bee pollen products.

Formal analyses are fundamental for the definition of honey, a product representing specific territories and their changes: the case of North Tyrrhenian dunes (Italy).

Honey is a variegate matrix depending significantly on the floral origin, and it could become an important agri-food product to valorise specific territories. Being so diverse, different analytical techniques are necessary for its description. Herein we characterized the honey produced in one of the Italian sand dunes systems hosting beekeeping activities. In terms of floristic origin, phytochemical characterization, and sensory and colour analysis, honey collected in 2021 and 2022 was comparable. Honey was polyfloral, with several pollens from dune habitat plants classified as minor. The presence of the allochthonous Amorpha fruticosa L. and the ruderal Rubus fruticosus L. pollens in the category of the secondary pollens testifies the alteration of the park vegetation. The phytochemical profile was rich in polyphenols. Other interesting compounds were coumarine derivatives, likely attributable to resin-laden plants as rockroses, long chain hydroxyacids typical of royal jelly and nicotinic acid and its analogues (2-hydroxynicotinic acid and 2-hydroxyquinoline). The above-mentioned honey showed interesting features and was a good representation of the vegetation of this area. Our study pointed out the importance of relying on multiple analytical techniques for the characterization of honey and the advisability of a technical support toward beekeepers to correctly describe and valorise their product.

The Comparison of Honey Enriched with Laboratory Fermented Pollen vs. Natural Bee Bread in Terms of Nutritional and Antioxidant Properties, Protein In Vitro Bioaccessibility, and Its Genoprotective Effect in Yeast Cells.

The aim of the study was to compare the nutritional value and bioactivity of honey enriched with a 10% addition of natural bee bread and its substitutes obtained as a result of laboratory fermentation of bee pollen. Physicochemical parameters, antioxidant properties, as well as the bioaccessibility of proteins using an in vitro static digestion model were analyzed. The bioactivity of the obtained enriched honeys was tested using the yeast model. The research indicates the similarity of honeys with the addition of "artificial bee bread" to honey with natural ones. During in vitro digestion, good bioaccessibility of the protein from the tested products was demonstrated. The ability of the products to protect yeast cells against hydrogen superoxide-induced oxidative stress was demonstrated using a qualitative spot test, which was stronger in the case of enriched honey than in pure rapeseed control honey. Significant inhibition of the growth of both strains of yeast exposed to bee pollen-enriched honeys was also demonstrated. Furthermore, all tested samples showed significant genoprotective activity against the genotoxic effect of zeocin and the reduction of the number of DNA double-strand breaks by a minimum of 70% was observed.

Citrus Honeys from Three Different Regions of Turkey: HPLC-DAD Profiling and in Vitro Enzyme Inhibition, Antioxidant, Anti-Inflammatory and Antimicrobial Properties with Chemometric Study.

The objectives of the present study are to compare the phenolic profiles and biological activities of 15 citrus honey samples from three different locations in Turkey using a chemometric approach. The HPLC-DAD analysis was used to determine phenolic profiles. Nineteen phenolic compounds were identified. Gallic acid (107.14-717.04 µg/g) was recorded as the predominant compound. AF (Antalya-Finike) had the highest antioxidant activity in ABTS⋅+ (IC50 : 18.01±0.69 mg/mL), metal chelating (IC50 : 6.20±0.19 mg/mL) and CUPRAC (A0.50 : 12.05±0.68 mg/mL) assays, while it revealed the best anti-inflammatory activity against COX-2 (17.28±0.22 %) and COX-1 (43.28±0.91 %). AM (Antalya-Manavgat) was the most active in ß-carotene-linoleic acid (IC50 : 10.05±0.19 mg/mL), anti-urease (38.90±0.69 %), anti-quorum sensing and antimicrobial activities. AKO1 (Adana-Kozan-1) in DPPH⋅ (IC50 : 34.25±0.81 mg/mL) assay, AKU1 (Antalya-Kumluca-1) in tyrosinase inhibition activity (37.73±0.38 %) assay, AKU2 (Antalya-Kumluca-2) in AChE (10.55±0.63 %) and BChE (9.18±0.45 %) inhibition activity assays showed the best activity. Chemometric tools were applied to the phenolic compositions and biological properties. PCA and HCA ensured that 15 citrus honey samples were grouped into 3 clusters. The results showed that myricetin, kaempferol, vanillin, protocatechuic acid, rosmarinic acid, rutin, vanillic acid, gallic acid, catechin and p-hydroxyphenyl acetic acid are phenolic compounds that can be used in the classification of citrus honeys.

Assessing the quality of bee honey on the basis of melissopalynology as well as chemical analysis.

Melissopalynological and chemical analysis of honey provide us useful and valuable information about the botanical and geographical origin of honey. The data in question is very important for authentication as well as for testing the quality of honey, so this is considered the main method in honey regulation here, we have used chemical analysis and melissopalynology to evaluate different honey samples from two main Iranian hubs of honey. Sampling was carried out on two important poles in Iran's honey production, the central Alborz region, and the mountainous Zagros ecosystems in the years 2020 to 2021. Therefore, 52 samples from Alborz (Northern Iran), as well as 42 samples from the Zagrosian ecosystems (western Iran) belonging to different ecological habitats, were collected. In addition, samples were taken at 7 altitudes from 0 to 3500 m a.s.l. Furthermore, in this study, various chemical analyses such as the effect of antioxidant activity, the amount of total phenolic content, pH, and moisture content of honey samples were evaluated. Our results showed that all honey samples were classified as polyfloral honey. Based on our findings, 57 honey samples (61%) contained the standard amount of pollen. A total of 42 plant families and 55 genera were identified in the studied samples, with the highest presence of Asteraceae, Fabaceae, Rosaceae, Apocynaceae, and Apiaceae. Finally, an antioxidant activity ratio of 19% to 98%, total phenolic content from 0.08 to 0.51 ppm, pH from 1.90 to 5.21, and moisture content from 13% to 18.40%.

Pyrrolizidine Alkaloids in Food on the Italian Market.

Pyrrolizidine alkaloids (PAs) are secondary metabolites produced by over 6000 plant species worldwide. PAs enter the food chain through accidental co-harvesting of PA-containing weeds and through soil transfer from the living plant to surrounding acceptor plants. In animal studies, 1,2-unsaturated PAs have proven to be genotoxic carcinogens. According to the scientific opinion expressed by the 2017 EFSA, the foods with the highest levels of PA contamination were honey, tea, herbal infusions, and food supplements. Following the EFSA's recommendations, data on the presence of PAs in relevant food were monitored and collected. On 1 July 2022, the Commission Regulation (EU) 2020/2040 came into force, repealed by Commission Regulation (EU) 2023/915, setting maximum levels for the sum of pyrrolizidine alkaloids in certain food. A total of 602 food samples were collected from the Italian market between 2019 and 2022 and were classified as honey, pollen, dried tea, dried herbal infusions, dried herbs, and fresh borage leaves. The food samples were analyzed for their PA content via an in-house LC-MS/MS method that can detect PAs according to Regulation 2023/915. Overall, 42% of the analyzed samples were PA-contaminated, 14% exceeded the EU limits, and the items most frequently contaminated included dried herbs and tea. In conclusion, the number of food items containing considerable amounts of PAs may cause concern because they may contribute to human exposure, especially considering vulnerable populations-most importantly, children and pregnant women.

Identification and quality evaluation of different processed products of Aconitum carmichaelii by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry and ultra-high-performance liquid chromatography-tandem mass spectrometry.

Aconitum carmichaelii is widely used to treat chronic and intractable diseases due to its remarkable curative effect, but it is also a highly toxic herb with severe cardiac and neurotoxicity. It has been combined with honey for thousands of years to reduce toxicity and enhance efficacy, but there has been no study on the chemical constituent changes in the honey-processing so far. In this study, the chemical constituents of A. carmichaelii before and after honey-processing were characterized by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry. The results showed that a total of 118 compounds were identified, of which six compounds disappeared and five compounds were newly produced after honey-processing, and the cleavage pathway of main components was elucidated. At the same time, 25 compounds were found to have significant effects on different products, among which four compounds with the biggest difference were selected for quantitative analysis by ultra-high-performance liquid chromatography-tandem mass spectrometry. This study not only explained the chemical differences between the different products, but also helped to control the quality of the honey-processed products more effectively, and laid a foundation for further elucidating the mechanism of chemical constituent change during the honey-processing of A. carmichaelii.

FTIR-ATR detection method for emerging C3-plants-derivated adulterants in honey: Beet, dates, and carob syrups.

The European Union Publications Office has recently presented a report on the European Union's coordinated action with the Joint Research Centre to determine certain fraudulent practices in the honey sector, in which it has been indicated that 74% of the samples analyzed, imported from China, and 93% of the samples analyzed, imported from Turkey, the two largest honey producers worldwide, presented at least one indicator of exogenous sugar or suspicion of being adulterated. This situation has revealed the critical state of the problem of honey adulteration worldwide and the need to develop analytical techniques for its detection. Even though the adulteration of honey is carried out in a general way with sweetened syrups derived from C4 plants, recent studies have indicated the emerging use of syrups derived from C3 plants for the adulteration of honey. This kind of adulteration makes it impossible to analyze its detection using official analysis techniques. In this work, we have developed a fast, simple, and economical method based on the Fourier transform infrared spectroscopy technique, with attenuated total reflectance, for the qualitative, quantitative, and simultaneous determination of beetroot, date, and carob syrups, derived from of C3 plants; whose available bibliography is very scarce and analytically not very conclusive for its use by the authorities. The proposed method has been based on the establishment of the spectral differences between honey and the mentioned syrups at eight different points in the spectral region between 1200 and 900 cm-1 of the mid-infrared, characteristic of the vibrational modes of carbohydrates in honey, which allows the pre-discrimination of the presence or absence of the syrups studied, and their subsequent quantification, with precision levels lower than 2.0% of the relative standard deviation and relative errors lower than 2.0% (m/m).

Anti-arrhythmia potential of honey-processed licorice in zebrafish model: Antioxidant, histopathological and tissue distribution.

ETHNOPHARMACOLOGICAL RELEVANCE: Honey-processed licorice (HPL) is the roasted product of licorice. It is recorded in the "Shang Han Lun" that licorice has better protection on heart after honey-processed. However, researches regarding its protective effect on the heart and the distribution of HPL in vivo are still limited. AIM OF THE STUDY: To evaluate the cardio-protection of HPL and explore the law of ten main components distribution in vivo under physiological and pathological conditions for an attempt to clarify the pharmacological substance basis of HPL in treating arrhythmia. MATERIALS AND METHODS: The adult zebrafish arrhythmia model was established by doxorubicin (DOX). Electrocardiogram (ECG) was used to detect the heart rate changes of zebrafish. SOD and MDA assays were used to evaluate oxidative stress levels in the myocardium. HE staining was used to observe the morphological change of myocardial tissues after HPL treatment. The UPLC-MS/MS was adapted to detect the content of ten main components of HPL in heart, liver, intestine, and brain under normal and heart injury conditions. RESULTS: Heart rate of zebrafish was decreased, the SOD activity was attenuated and MDA content was increased in myocardium after administration of DOX. Moreover, tissue vacuolation and inflammatory infiltration were detected in zebrafish myocardium induced by DOX. HPL could ameliorate heart injury and bradycardia induced by DOX to a certain extent by increasing SOD activity and reducing MDA content. In addition, the study of tissue distribution revealed that the content of liquiritin, isoliquiritin, and isoliquiritigenin in the heart was higher in the presence of arrhythmias than those in the normal condition. Under pathological conditions, the heart highly exposed to these three components could elicit anti-arrhythmic effects by regulating immunity and oxidation. CONCLUSION: These findings indicate that the HPL is protective against heart injury induced by DOX, and its effect is associated with the alleviation of oxidative stress and tissue injury. And the cardioprotective effect of HPL under pathological conditions may be related to the high distribution of liquiritin, isoliquiritin, and isoliquiritigenin in heart tissue. This study provides an experimental basis for the cardioprotective effects and tissue distribution of HPL.

Fingerprinting Chemical Markers in the Mediterranean Orange Blossom Honey: UHPLC-HRMS Metabolomics Study Integrating Melissopalynological Analysis, GC-MS and HPLC-PDA-ESI/MS.

(1) Background: Citrus honey constitutes a unique monofloral honey characterized by a distinctive aroma and unique taste. The non-targeted chemical analysis can provide pivotal information on chemical markers that differentiate honey based on its geographical and botanical origin. (2) Methods: Within the PRIMA project "PLANT-B", a metabolomics workflow was established to unveil potential chemical markers of orange blossom honey produced in case study areas of Egypt, Italy, and Greece. In some of these areas, aromatic medicinal plants were cultivated to enhance biodiversity and attract pollinators. The non-targeted chemical analysis and metabolomics were conducted using ultra-high-performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS). (3) Results: Forty compounds were disclosed as potential chemical markers, enabling the differentiation of the three orange blossom honeys according to geographical origin. Italian honey showed a preponderance of flavonoids, while in Greek honey, terpenoids and iridoids were more abundant than flavonoids, except for hesperidin. In Egyptian honey, suberic acid and a fatty acid ester derivative emerged as chemical markers. New, for honey, furan derivatives were identified using GC-MS in Greek samples. (4) Conclusions: The application of UHPLC-HRMS metabolomics combined with an elaborate melissopalynological analysis managed to unveil several potential markers of Mediterranean citrus honey potentially associated with citrus crop varieties and the local indigenous flora.

A novel strategy on the study of whole intestinal metabolic profiles for Polygalae Radix before and after processing.

INTRODUCTION: Relieving toxicity and enhancing a calming effect after processing Polygalae Radix (PR) are widely known. Aromatic carboxylic acids (ACAs) may be crucial processed products. However, due to the limited detection methods for ACAs, the whole metabolic profiles via intestinal bacteria are still not very clear. OBJECTIVE: Designing a novel strategy for the detection of ACAs and tracking the whole metabolic profiles before and after processing PR. MATERIALS AND METHODS: The stable-isotope labelling derivatisation (SILD) method based on multidimensional ultra-high performance liquid chromatography coupled with a mass spectrometer (UHPLC-MS) technology and UNIFI-pathway mode was firstly designed to systematically study the metabolisms of all the drug-derived ingredients ranging from m/z 100 to 2000 in processing PR via intestinal bacteria. Firstly, the SILD with UHPLC coupled with a triple-quadrupole MS technology was designed to trace eight ACA metabolites of the processed PR with intestinal bacteria. Additionally, the UHPLC coupled with a quadrupole time-of-flight MS with UNIFI-pathway mode was adopted to monitor relatively big metabolites. RESULTS: The metabolism mechanism of ACAs (eight kinds) and the relatively big molecular metabolites (98 kinds) were deeply traced in PR, PR with refined honey (HP), and PR with licorice (LP) via the intestinal bacteria. Totally 106 intact metabolic profiles of drug-derived ingredients were presented. Importantly, the influence of LP on the metabolism of compounds after incubation of intestinal bacteria was greater than that of HP. CONCLUSION: This research provides a comprehensive and systematic guidance for further study on in vivo metabolisms of the processed PR.

Target screening method for the quantitative determination of 118 pyrrolizidine alkaloids in food supplements, herbal infusions, honey and teas by liquid chromatography coupled to quadrupole orbitrap mass spectrometry.

An analytical procedure for the screening of 118 pyrrolizidine alkaloids (PAs) was successfully validated and applied to their quantitative determination in food supplements, herbal infusions, honey, and teas. It provides the reliable analyte identification by high-resolution tandem mass spectrometry (HRMS/MS), the accurate determination of 21 regulated PAs, and broad contamination profiles. 10% of 281 analyzed samples resulted contaminated at levels above the maximum levels (MLs) of European legislation. The contamination of herbal infusions of mixed plants can represent a possible health concern (23%; mean of PA sum above ML). A high number of PAs not included in the regulation was detected in honey and herbal food supplements, but their contribution was only relevant to the overall level in honey. The results indicate the need to continue collecting contamination data in food supplements and infusions of mixed herbs and to expand the PA-pool to be monitored in honey and related products.

Towards DNA-Based Methods Analysis for Honey: An Update.

Honey is a natural product widely consumed all over the world due to its relationship with healthy benefits. Additionally, environmental and ethical issues have a higher weight in the consumer's choice to buy honey as a natural product. Following the high demand of this product, several approaches have been suggested and developed aiming at the assessment of honey's quality and authenticity. Target approaches, such as pollen analysis, phenolic compounds, sugars, volatile compounds, organic acids, proteins, amino acids, minerals, and trace elements, showed an efficacy, particularly concerning the honey origin. However, a special highlight is given to DNA markers, due to their useful applicability in environmental and biodiversity studies, besides the geographical, botanical, and entomological origins. Different DNA target genes were already explored for addressing diverse sources of honey DNA, with DNA metabarcoding attaining a relevant importance. This review aims to describe the latest advances on DNA-based methods applied in honey related studies, identifying the research needs for the development of new and additional required methodologies, and to select the most adequate tools for future research projects.

Quality Assessment of Bee Pollen-Honey Mixtures Using Thin-Layer Chromatography in Combination with Chemometrics.

The aim of this study is to develop a rapid, effect-directed screening method for quality assessment of bee pollen-honey mixtures. The comparative antioxidant potential and phenolic content of honey, bee pollen, and the bee pollen-honey mixtures, was performed using spectrophotometry. The total phenolic content and antioxidative activity of bee pollen-honey mixtures with 20 % bee pollen share were in the range 3.03-3.11 mg GAE/g, and 6.02-6.96 mmol TE/kg, respectively, while mixtures with 30 % bee pollen share contained 3.92-4.18 mg GAE/g, and 9.69-10.11 mmol TE/kg. Chromatographic fingerprint of bee pollen-honey mixtures was performed by high-performance thin-layer chromatography with conditions developed by authors and reported for the first time. Fingerprint analysis hyphenated with chemometrics enabled authenticity assessments of honey in mixtures. Results indicate that bee pollen-honey mixtures represent a food with highly, both, nutritious characteristics and health-promoting effect.

Protective Effects of Arbutus unedo L. Honey in the Alleviation of Irinotecan-Induced Cytogenetic Damage in Human Lymphocytes-An In Vitro Study.

Strawberry tree (Arbutus unedo L.) honey (STH) has been used since ancient times as a folk medicine remedy, especially in certain Mediterranean countries. This honey, rich in phenolic content, is well recognized for its antioxidant, anti-inflammatory, and antimicrobial activities, and is used for the treatment of skin lesions as well as gastrointestinal and respiratory disorders. This study investigated whether STH alleviates genome damage in human peripheral blood lymphocytes produced by the cytotoxic drug irinotecan. The phenolic profile of STH was previously estimated by ultra-high-performance liquid chromatography coupled to a linear ion trap-Orbitrap hybrid mass spectrometer. The effects of STH were evaluated at three concentrations (1×, 5×, and 10×), based on the daily consumption of the honey by an adult person. After 2 h of in vitro exposure, standard lymphocyte cultures for the analysis of chromosome aberrations and the cytokinesis-block micronucleus cytome assay were established. Our results demonstrate that STH offered remarkable geno- and cytoprotection when administered with irinotecan. These findings are relevant for drawing preliminary conclusions regarding the in vitro safety of the tested honey. However, further studies are needed with the application of more complex experimental models.

Membrane-processed honey samples for pollen characterization with health benefits.

Better processing techniques must be utilized widely due to the rising demand for honey. The most common honey processing techniques are applied to melissopalynomorphs to check the quality and quantity of valuable honey using microporous ultrafiltration membranes. It is essential to have the ability to selectively filter out sugars from honey using ultrafiltration. This study authenticated 24 honey samples using membrane reactors ultrafiltration protocol to describe the pollen spectrum of dominant vegetation. The purpose of this study was also to explore nutritional benefits as well as the active phytochemical constituents of honey samples. Honey samples were collected and labeled Acacia, Eucalyptus, and Ziziphus species based on plant resources provided by local beekeepers. A variety of honeybee flora was collected around the apiaries between 2020 and 2021. Honey analysis revealed that the pollen extraction of 24 bee foraging species belonging to 14 families. The honey membrane technology verified the identities of honey and nectar sources. Also, pollen identified using honey ultrafiltration membranes revealed dominant resources: Acacia spp. (69%), Eucalyptus spp. (52%) and Ziziphus spp. Honey filtration using a membrane technology classified 14 samples as unifloral, represented by six dominant pollen types. The absolute pollen count in the honey sample revealed that 58.33% (n = 14) belong to Maurizio's class I. Scanning ultrasculpturing showed diverse exine patterns: reticulate, psilate, scabrate-verrucate, scabrate-gemmate, granulate, perforate, microechinate, microreticulate, and regulate to fossulate for correct identification of honey pollen types. Honey ultrafiltration should be utilized to validate the botanical sources of honey and trace their biogeographic authenticity. Thus, it is imperative to look at the alternative useful method to identify the botanical origin of filtered honey. It is critical to separate honey from adulteration by a standardized protocol. Membrane technology has yielded significant outcomes in the purification of honey.

Chemical and pharmacological difference between honey-fried licorice and fried licorice.

ETHNOPHARMACOLOGICAL RELEVANCE: According to textual research of books from ancient times till now, there are three main preparation methods of "fried licorice", including frying licorice without excipients (F), frying licorice after dipping with water (W), and frying licorice with honey (H). However, with the development over many successive generations, honey frying has gradually become the main processing form of licorice, whereas the fried licorice is nowadays rarely used. AIM OF THE STUDY: The objectives of this study were to clarify the differences of the three forms of "fried licorice" in chemical composition and pharmacological activities, and to screen quality markers for differently processed licorice. It is expected to provide a scientific basis for the rational choice of "fried licorice" as medicine. MATERIALS AND METHODS: Non-target metabolomic analysis based on UHPLC-QE-Orbitrap-MS was conducted to compare the chemical differences between the differently processed licorice material. Pharmacodynamically, the differences in immunomodulatory activity (including intestinal flora experiment), anti-inflammatory activity, and hepatoprotective activity of the differently processed licorice were evaluated. Furthermore, multivariate statistical analysis was performed to screen potential quality markers of honey-fried licorice. The serum concentration of selected markers was determined by UHPLC-QqQ-MS. RESULTS: Metabolomic analysis showed no difference in the chemical composition of F and W, whereas the chemical composition of H was significantly different from that of F and R. The immunomodulatory activity, anti-acute inflammatory effect, and hepatoprotective effect of licorice were significantly improved after frying with honey; No significant differences were observed between F and H in term of immunomodulatory activity and anti-acute inflammatory effect, whereas, H is better than F in terms of liver-protective activity. The intestinal flora experiment confirmed that H does have immunomodulatory activity, while F may induce an increased abundance of certain pathogenic bacteria in the intestine. Multivariate statistical analysis suggests that the content of liquiritin (2), liquiritigenin (3), isoliquiritin (5), isoliquiritigenin (6) and glycyrrhizic acid (7) plusing glycyrrhetinic acid (8) in H group is closely correlated with its improved effects. CONCLUSIONS: This study provides a scientific rational for the selection of "fried licorice" processing methods. In addition, it provides a scientific basis for the selection of quality markers of differently processed licorice.

An analytical platform for the screening and identification of pyrrolizidine alkaloids in food matrices with high risk of contamination.

An analytical platform for the detection of pyrrolizidine alkaloids (PAs) in honey, pollen, teas, herbal infusions, and dietary supplements is proposed; it includes a wide-scope suspect screening method, based on a diagnostic product ion filtering strategy for the characterization of PAs, and a target screening and identification method for the high-throughput detection of 118 PAs of a high-resolution mass spectral library. Salting-out assisted liquid-liquid extraction of aqueous extracts combined to ultra-high performance liquid chromatography-high-resolution tandem mass spectrometry was employed. The limit of identification (0.6-30 µg kg-1) of 28 standards were fit-for-purpose in PA-monitoring applications, with a false negative rate <1.3 % at 4 µg L-1. The wide-scope suspect screening method allowed the tentative identification of 88 compounds. The screening of 282 commercial samples revealed a broad contamination of the studied matrices, demonstrating the effectiveness of the platform in detecting and identifying both target and untarget PAs.