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Ajuga turkestanica preparations are used as anti-aging cosmeceuticals and for medicinal purposes. Herein we describe the characterization and quantification of its metabolites in different organs using UHPLC-MS and NMR spectroscopy. A total of 51 compounds belonging to various phytochemical classes (11 flavonoids, 10 ecdysteroids, 9 diterpenes, 6 fatty acids, 5 iridoids, 3 phenylpropanoids, 3 sugars, 2 phenolics, 1 coumarin, 1 triterpene) were annotated and tentatively identified by UHPLC-ESI-QqTOF-MS/MS of methanolic extracts obtained separately from the organs. 1D and 2D NMR spectroscopy independently confirmed the identity of six major compounds. The abundances of these main constituents in flowers, fruits, leaves, roots, seeds, and stems were compared and quantified using 1 H NMR. The results showed that 8-O-acetylharpagide, 20-hydroxyecdysone (ecdysterone) and ajugachin B were the most abundant constituents in the species. The two major compounds, 8-O-acetylharpagide and 20-hydroxyecdysone, were chosen as the markers for the quality assessment of A. turkestanica material. The methanolic extract of the aerial parts of A. turkestanica showed no noteworthy anthelmintic (antihelmintic), antifungal, or cytotoxic effect in in vitro assays., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Hydrophilic Interaction Liquid Chromatography (HILIC) is an outstanding strategy for the challenging analysis of hydrophilic and polar components. Nevertheless, analysis under HILIC mode typically consumes 70%-95 % acetonitrile with the disadvantage of high analytical costs, being environmentally unfriendly and causing biohazards, which is not in line with the concept of green chromatography. Research has shown that Per Aqueous Liquid Chromatography (PALC) simultaneously emphasizes efficient analytical performance for hydrophilic analytes and green analytical concepts. The development of new PALC stationary phases with superior performance is necessary., Results: In this paper, silanized silica was sequentially subjected to esterification reaction, polymerization reaction and covalent bonding through five steps to obtain SiO 2 -RHP-AuNPs material, which was prepared as a novel stationary phase for PALC. Comprehensive characterization of the materials by means of Fourier transform infrared spectroscopy, Transmission scanning electron microscope, Elemental analysis and Thermogravimetric analysis showed the successful bonding of the functionalized groups on the original silica. The polymeric stationary phase based on Radix hedysari polysaccharide and Au nanoparticles had higher density of hydroxyl and ester functionalized groups. The Au nanoparticles upgraded their mesoporous structure and thermal stability, providing exceptional chromatographic performance and selectivity for chromatographic analysis. The influence of mobile phase water content, salt concentration, pH and column temperature on the retention behavior was evaluated. The novel Column was found to exhibit a dual mechanism of hydrophobic interactions/ion exchange interactions in a mobile phase with high water content., Significance and Novelty: The separation efficiency and selectivity of SiO 2 -RHP-AuNPs columns for synthetic pigments and organic acids in PALC mode were superior to those of commercial HILIC and C 18 columns. In addition, a method for the determination of seven active ingredients in Fructus Ligustri Lucidi by SiO 2 -RHP-AuNPs column in PALC mode was developed. The method had good stability, reproducibility and accuracy, which was capable of realizing the quality evaluation of Chinese Materia Medicas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: Catnip (Nepeta cataria, L.) has well-documented applications in arthropod repellency because of its bioactive iridoids. Long-term stability of nepetalactones and other iridoids in N. cataria are needed to develop as effective pest repellents., Objectives: The present work intends to measure iridoid concentration over time in biomass, plant extracts, and extract solution while identifying degradative byproducts under different storage conditions., Methodology: Samples of desiccated biomass, ethanol extract, and extract in ethanol solution were stored in ambient light or darkness. Through UHPLC-QTOF/MS or UHPLC-QQQ/MS, the concentration of Z,E-nepetalactone, E,Z-nepetalactone, nepetalic acid, and dihydronepetalactone were examined over 2 years and statistically analyzed for determination of best storage practices. Degradation kinetics were applied to each analyte using graphical estimation. With targeted formula searching, degradative byproducts were identified and quantified., Results: Light exposure caused significant decreases in E,Z-nepetalactone concentration in all sample types, while having no effect on Z,E-nepetalactone as it decayed more rapidly. Extract samples lost nepetalactone content faster than biomass or extract solution. Dihydronepetalactone levels were low, but never declined over 2 years. Nepetalic acid increased over some periods, depending on sample type, indicating a relationship between the acid and nepetalactone. Four degradative byproducts-nepetonic acid, dehydronepetalactone, an anhydride, and an ethanolic ester-were identified, with variable responses to light exposure., Conclusions: Protecting catnip products from light is necessary to preserve nepetalactones, and a discernable difference in nepetalactone isomer stability was discovered. Identifying Nepeta chemotypes rich in dihydronepetalactone may provide more resilient botanicals as starting materials for processing., (© 2024 The Author(s). Phytochemical Analysis published by John Wiley & Sons Ltd.)

Several processes have been developed in the past to selectively extract oleuropein and its aglycones from olive derived materials. In the present manuscript, we outline a novel approach for processing olive leaves aqueous extracts. This allowed first to select microwave irradiation as the methodology able to provide a large enrichment in oleuropein. Subsequently, the use of lamellar solids led to the selective and high yield concentration of the same. Adsorption on solids also largely contributed to the long term chemical stability of oleuropein. Finally, an eco-friendly, readily available, and reusable catalyst like H 2 SO 4 supported on silica was applied for the hydrolysis of oleuropein into hydroxytyrosol and elenolic acid. This latter was in turn selectively isolated by an acid-base work-up providing its monoaldehydic dihydropyran form (7.8 % extractive yield), that was unequivocally characterized by GC-MS. The isolation of elenolic acid in pure form is described herein for the first time., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Fructus Corni (F. Corni) is the dried mature pulp of Cornus officinalis Sieb. et Zucc.(Cornaceae), which is rich in iridoids. In this study, a simple, sensitive and rapid UPLC-MS/MS method was developed for the simultaneous determination of 13 iridoid glycosides of F. Corni from different areas. Specifically, we included five new compounds (cornusdiridoid C, cornusdiridoid E, cornusdiridoid F, 3'',5''-dehydroxycornuside and 2'-O-p-coumaroyl-kingiside) and isomers (2'-O-p-E-coumaroylloganin and 2'-O-p-Z-coumaroylloganin) for the first time in the quality markers of F. Corni. A total of 13 compounds and two pairs of isomers were well isolated and tested within just 14 min. All calibration curves showed good linear regression (r2 ≥ 0.99) within the tested concentration ranges. The limit of detection and limit of quantification were in the range of 0.19-1.90 and 0.38-3.76 ng/mL, respectively. The intra-day and inter-day precision were <3.21% and 12.49%, the RSD values of repeatability did not exceed 6.81% and the average recoveries were 90.95-113.59% for the analytes. All iridoid glycosides stabilized within 12 h (RSD < 10.99%). This method has been successfully applied to the quality evaluation of crude and processed F. Corni from different areas. The determination of characteristic iridoid glycosides and isomers will provide a more reliable and comprehensive method for the evaluation of F. Corni., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Early changes in sensory quality of phenols-rich virgin olive oil (VOO) and their relationship with the chemical changes are less studied in the literature. Therefore, the objective of this study was to propose a predictive model of dynamics of sensory changes based on specific chemical markers. The evolution of the sensory quality of phenol-rich VOOs from Tuscan cultivars stored under optimal storage conditions (i.e., absence of light, no O 2 exposure, low temperature) was investigated using a multi-step methodological approach combining sensory (official sensory analysis (so-called Panel Test), Descriptive Analysis and Temporal Dominance of Sensation) and chemical measurements. The sensory map from descriptive data was related to the phenolic and volatile profiles, measured using HPLC-DAD and HS-SPME-GC-MS, respectively. A predictive model of the sensory changes over storage based on chemical compounds was developed. Results showed that very early changes involving phenolic and volatile compounds profiles occur in VOOs stored under optimal storage conditions, which turn in changes in sensory properties evaluated by the official panel test, the descriptive analysis and the temporal dominance of sensation. Furthermore, a chemical marker of sensory dynamics of oils during storage was identified as the ratio between two groups of secoiridoids. The proposed model, supported by the mentioned chemical marker, has the potential of improving the control of sensory changes in phenols-rich virgin olive oils during storage in optimal conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

In this study, the phytochemical profile of fifty olive leaves (OL) extracts from Spain, Italy, Greece, Portugal, and Morocco was characterized and their anti-cholinergic, anti-inflammatory, and antioxidant activities were evaluated. Luteolin-7-O-glucoside, isoharmnentin, and apigenin were involved in the acetylcholinesterase (AChE) inhibitory activity, while oleuropein and hydroxytyrosol showed noteworthy potential. Secoiridoids contributed to the cyclooxygenase-2 inhibitory activity and antioxidant capacity. Compounds such as oleuropein, ligstroside and luteolin-7-O-glucoside, may exert an important role in the ferric reducing antioxidant capacity. It should be also highlighted the role of hydroxytyrosol, hydroxycoumarins, and verbascoside concerning the antioxidant activity. This research provides valuable insights and confirms that specific compounds within OL extracts contribute to distinct anti-cholinergic, anti-inflammatory, and anti-oxidative effects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The authentication of ingredients in formulas is crucial yet challenging, particularly for constituents with comparable compositions but vastly divergent efficacy. Rehmanniae Radix and its derivatives are extensively utilized in food supplements, which contain analogous compositions but very distinct effects. Rehmanniae Radix, also a difficult-to-detect herbal ingredient, was chosen as a case to explore a novel HPTLC-QDa MS technique for the identification of herbal ingredients in commercial products. Through systematic condition optimization, including thin layer and mass spectrometry, a stable and reproducible HPTLC-QDa MS method was established, which can simultaneously detect oligosaccharides and iridoids. Rehmannia Radix and its processed products were then analyzed to screen five markers that could distinguish between raw and prepared Rehmannia Radix. An HPTLC-QDa-SIM method was further established for formula detection by using the five markers and validated using homemade prescriptions and negative controls. Finally, this method was applied to detect raw and prepared Rehmannia Radix in 12 commercial functional products and supplements.

This study established an ultrasound-assisted extraction-high performance liquid chromatography method for simulta-neously determinining the content of 11 bioactive compounds including iridoids, phenolic acids, and flavonoids in Lonicera japonica flowers. The flowers at six stages from the rice bud stage(ML) to the golden flower stage(JH) of L. japonica varieties 'Sijuhua' and 'Beihua No.1' in two planting bases in Shandong province were collected. The established method was employed to determine the content of 11 target compounds, on the basis of which the dynamics of active components in L. japonica sampels during different development stages was investigated. The correlation analysis was carried out to reveal the correlations of the content of iridoids, phenolic acids, and flavonoids. Furthermore, the antioxidant activities of samples at different developmental stages were determined, and the relationship between antioxidant activity and chemical components was analyzed by the correlation analysis. The results showed that the total content of the 11 components in 'Sijihua' changed in a &quot;W&quot; pattern from the ML to JH, being the highest at the ML and the second at the slight white stage(EB). The total content of 11 compounds in 'Beihua No.1' was the highest at the ML and decreased gra-dually from the ML to JH. The samples of 'Sijihua' had higher content of iridoids and lower content of phenolic acids than those of 'Beihua No.1'. The content of flavonoids and phenolic acids showed a positive correlation(R~2=0.90, P&lt;0.05) in 'Sijihua' but no obvious correlation in 'Beihua No.1'. The antioxidant activity and phenolic acid content showed positive correlations, with the determination coefficients(R~2) of 0.84(P&lt;0.05) in 'Beihua No.1' and 0.73(P&lt;0.05) in 'Sijihua'. The antioxidant activity of both varieties was the strongest at the ML and the second at the EB. This study revealed that the content dynamics of iridoids, phenolic acids, and flavonoids in 'Sijihua' and 'Beihua No.1' cultivated in Shandong province during different developmental stages. The results indicated that the antioxidant activity of L. japonica flowers was significantly correlated with the content of phenolic acids at different deve-lopmental stages, which provided a basis for determining the optimum harvest time of L. japonica flowers.

Background: Extra virgin olive oil (EVOO), a natural product with a multidisciplinary role, has been and is continuing to be studied from several points of view. Among them, its chemical analysis is of major importance and several methods have been used. Nuclear magnetic resonance (NMR) spectroscopy has inherent advantages, among them monitoring the chemical constituents without the need for a separation technique and without, for instance, possible carry-over effects. Additionally, several magnetic resonance spectroscopic techniques can provide a novel powered insight into the nature and properties of a sample under study. Moreover, -omics procedure can reveal new information and can lead to the classification of populations under study. The main objective of the present work was the possible classification of the EVOO samples based on their aldehyde content using a proposed unreferenced 1 H-NMR spectroscopic quantification method combined with a metabolomic approach. Moreover, the study of the impact of such elevated aldehyde content on several spectra regions of importance in the proton NMR spectra led to the proposal of a possible new isomer indicator., Results: Univariate analysis of 12 EVOO samples showed that oleacein, oleocanthal, elenolic acid, hydroxytyrosol/hydroxytyrosol derivatives and tyrosol/tyrosol derivatives strongly differentiated two classes of EVOO: OEH (for high aldehyde EVOO content) and OE (for non-high aldehyde content). Moreover, we pointed out the 'impact' of such elevated secoiridoid and derivatives content, through their moieties' units, on a range of several resonances of the 1 H-NMR spectrum. The metabolomic approach demonstrated the classification of EVOO samples based on their secoiridoid and derivatives content. Multivariate analysis showed a strong influence on the discrimination of the EVOO classes based on the protons resonating at the aldehyde region of the 1 H-NMR spectrum; the aldehyde protons corresponding to 5S,4R-ligstrodial and 5S,4R-oleuropeindial, oleacein, oleocanthal, elenolic acid, p-HPEA-EA, 3,4-DHPEA-EA, 5S,4R- and 5S,4S-ligstrodial and the proton corresponding to a new compound were reported for the first time. This isomer compound, reported for the first time, could serve as a possible indicator for EVOO classification., Conclusions: An unreferenced quantification method was proposed and EVOO samples were classified into two classes: OEH and OE, according to their aldehyde content, gaining thus probably higher nutrient and possible pharmacological value. Moreover, we point out the 'impact' of such elevated aldehyde content on several spectral regions of the 1 H spectrum. Finally, a new compound was detected in the OEH samples and is reported for the first time. This compound could possibly be an indicator. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Pterocephali Herba (PH), the dried whole plant of Pterocephalus hookeri, is a Tibetan medicine commonly used to treat rheumatoid arthritis (RA). Iridoids, triterpenoids, flavonoids and phenylpropanoids are the major groups of bioactive constituents from PH. However, only ursolic acid and oleanolic acid, two unspecific triterpenoid components, are used as markers for the quality control of PH in Chinese Pharmacopoeia. Herein, an UPLC-TQ-MS/MS integrating SIR and MRM mode method for simultaneously quantifying 18 components, i.e., 9 iridoids, 3 triterpenoids, 3 phenylpropanoids, 2 flavonoids and quinic acid, in PH was developed and validated, and was used to evaluate 10 batches of PH samples from different origins. Hierarchical cluster analysis (HCA) was used to show the clustering of PH samples, while spearman correlation analysis was adopted to evaluate the correlation between ursolic acid/oleanolic acid and other quantified components. It was found that the established method was sensitive, precise, and accurate enough for the simultaneous quantification of 18 analytes in PH samples. Significant differences were found among the contents of 18 components in PH samples, no apparent clustering of the quality of PH samples was found to be related to its origins, and the contents of ursolic acid/oleanolic acid were only significantly correlated to the content of sylvestroside I, dipsanoside B, dipsanoside A in PH. Our results suggested that the newly established multi-components quantitative method is an improved approach for quality evaluation of PH samples. Furthermore, the holistic quality was inconsistent among PH samples, and ursolic acid/oleanolic acid alone could not indicate the holistic quality variation trend of PH., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Valeriana jatamansi Jones is a commonly used traditional Chinese medicine, boasting rich effective compositions with versatile chemical structures and wide polarity, including iridoids, chlorogenic acid, and flavonoids. Previous reports indicate that conventional high-performance liquid chromatography (HPLC) analytical methods have proven inefficient performance in comprehensively characterizing components in Valeriana jatamansi. In the present study, a hybrid online analytical platform combining supercritical fluid extraction with both conventional HPLC separation (reverse phase) and supercritical fluid chromatography (normal phase) has been established and validated. This system can provide online extraction with two different chromatographic separation modes to increase separation ability and has been connected to a mass spectrometer to acquire high-resolution mass spectrometry data. Then, the online platform was applied to screening components in Valeriana jatamansi. A total of 117 compounds were identified, including five lignans, 18 organic acids, six flavonoids, and 88 iridoids. Thirty-three compounds were reported from Valeriana jatamansi for the first time. These results enrich our understanding of the components of Valeriana jatamansi and prove that the developed online platform in this study is a robust approach for accelerating working efficiency in comprehensively analyzing complicated samples., (© 2023 Wiley-VCH GmbH.)

In this study, a sensitive high-performance liquid chromatography detector was established and validated for the simultaneous determination of geniposide, ellagic acid, piperine, costunolide and dehydrocostuslactone in Liuwei Muxiang Capsules. The analysis was achieved on CHANIN 100-5-C18-H column (5μm, 250 mm×4.6 mm) with the temperature of 30 o C. Gradient elution was applied using 0.1% phosphoric acid solution-methanol-acetonitrile (50:50) as mobile phase at the flow rate of 1.0 mL/min. The determination was performed at the wavelength of 225 nm (detecting geniposide), 254 nm (detecting ellagic acid), 343 nm (detecting piperine) and 225 nm (detecting costunolide and dehydrocostuslactone) along with the sample volume of 10μL. The linear ranges of geniposide, ellagic acid, piperine, costunolide and dehydrocostuslactone demonstrated good linear relationships within their respective determination ranges. The average recoveries were 100.04%, 99.86%, 99.79%, 100.17% and 100.41%, respectively. RSD% was 1.3%, 1.2%, 1.2%, 1.2%, 1.5%, respectively. The developed method was proved to be simple, accurate and sensitive, which can provide a quantitative analysis method for the content determination of geniposide, ellagic acid, piperine, costunolide and dehydrocostuslactone in Liuwei Muxiang capsules.

Background: White rice is poor in health-promoting phytochemicals; therefore, the production of a phenol-enriched commodity is highly desirable. Recent findings on its enrichment via cooking in plant extracts are promising, yet studies employing aqueous extracts of olive leaves (OLs), containing well-recognized bioactive phenols (e.g. oleuropein) are absent. In addition, little is known about the levels of phenols that are maintained after rice drying and rehydration, an important aspect for the future design of 'ready-to-eat' functional rice., Results: The examination, for the first time, of white rice adsorption capacity of phenols from OLs upon cooking in infusions containing different levels of phenols, after freeze-drying and rehydration, showed the following: (i) the total phenol content, the antioxidant activity (assessed via 2,2-diphenyl-1-picrylhydrazyl radical and ferric reducing antioxidant power assays), the oleuropein and luteolin-7-O-glucoside levels increased dose dependently; (ii) upon rehydration, the average decrease of total phenol content and antioxidant activity values was significantly lower when an exact volume of water was used compared with an excess (~10% versus 63%). A similar trend was observed for oleuropein (36% versus 83%) and the luteolin-7-O-glucoside (24 versus 82%) levels; (iii) the dried enriched kernels were less bright with a hay-yellow hue (CIELab coordinates)., Conclusion: White rice enrichment with biophenols from OLs, a by-product of olive tree cultivation, was successful using a simple approach. Despite leaching upon freeze-drying/rehydration, sufficient amounts were maintained to obtain a functional rice that could serve as an alternative dietary source of OLs phenols to non-traditional olive tree product consumers or those refraining from sodium and fats. © 2023 Society of Chemical Industry., (© 2023 Society of Chemical Industry.)

The simultaneous extraction of crocin and geniposide from gardenia fruits (Gardenia jasminoides Ellis) was performed by integrating natural deep eutectic solvents (NADES) and ultrasound-assisted extraction (UAE). Among the eight kinds of NADES screened, choline chloride-1,2-propylene glycol was the most suitable extractant. The probe-type ultrasound-assisted NADES extraction system (pr-UAE-NADES) demonstrated higher extraction efficiency compared with plate-type ultrasound-assisted NADES extraction system (pl-UAE-NADES). Orthogonal experimental design and a modified multi-index synthetic weighted scoring method were adopted to optimize pr-UAE-NADES extraction process. The optimal extraction conditions that had a maximum synthetic weighted score of 29.46 were determined to be 25 °C for extraction temperature, 600 W for ultrasonic power, 20 min for extraction time, and 25% (w/w) for water content in NADES, leading to the maximum yields (7.39 ± 0.20 mg/g and 57.99 ± 0.91 mg/g, respectively) of crocin and geniposide. Thirty-three compounds including iridoids, carotenoids, phenolic acids, flavonoids, and triterpenes in the NADES extract were identified by LC-Q-TOF-MS 2 coupled with a feature-based molecular networking workflow. The kinetics evaluation of the conjugated dienes generation on Cu 2+ -induced low density lipoprotein (LDL) oxidation via the four-parameter logistic regression model showed that crocin increased the lag time of LDL oxidation in a concentration-dependent manner (15 μg/mL, 30 μg/mL, 45 μg/mL) by 12.66%, 35.44%, and 73.42%, respectively. The quantitative determination for fluorescence properties alteration of the apolipoprotein B-100 exhibited that crocin effectively inhibited the fluorescence quenching of tryptophan residues and the modification of lysine residues caused by reactive aldehydes and malondialdehydes. The pr-UAE-NADES showed significant efficiency toward the simultaneous extraction of crocin and geniposide from gardenia fruits. And this study demonstrates the potential utility of gardenia fruits in developing anti-atherogenic functional food., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Olive drupe traits (i.e., ripening index and pericarp water content) and minor components (i.e., phenols and pigments in both fruit and oil) are important for human health and are affected by agronomic background. The aim of this study was to investigate the relationship between fruit traits, phenols, and pigments in samples derived from different soil and water management practices. Chromatographic (UHPLC-MS/MS) and spectroscopic ( 1 HNMR and near UV-Vis spectroscopy) techniques were employed for the characterization of olive fruits and oils. The use of various techniques allowed the identification of interesting trace compounds. We observed that most of the fruit phenols (a total of 29 compounds) were correlated with the degree of ripening: most of the phenolic acids (and their derivatives), phenolic alcohols, and secoiridoids were negatively correlated, whereas the majority of the studied flavonoids were positively correlated. The relationship between the ripening index and fruit phenolic compounds appears to be dependent on the metabolic pathway that controls the synthesis of each individual compound. Conversely, the secoiridoids and pigments in olive oil showed a negative correlation with pulp moisture, probably because of the influence of the water content on the extractability and transfer in the oil phase of these minor components.

Gardenia jasminoides is an important ornamental greening plant with medicinal and edible values. This study investigated volatile constituents, alcoholic components and physiological activities on flowers of G. jasminoides Ellis and its variety. It was found that a total of 56 volatile components were identified, and terpenoids and esters were the main compounds to distinguish these species. Furthermore, the alcohol-soluble extracts of G. jasminoides flowers have the high contents of total phenols and total flavonoids, with potential antioxidant and hypoglycemic activities. In addition, nine compounds were identified, whose distribution in petals and stamens of G. jasminoides were significantly dissimilar. The contents of flavonoids and phenolics were stable after blanching confirmed by our findings, while iridoids were remarkably higher after freeze-drying (FD) and hot-air drying (HD). This research provides evidences that the fragrance, active components and activity of flowers of these species were affected by species, flower parts and processing methods., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Three undescribed glycoside constituents, macrophyllosides E-G and a pair of iridoid glycosides genticrasides A/B, together with eleven known glycoside compounds were isolated from the roots of Gentiana crassicaulis Duthie ex Burk. Their structures were identified by means of spectra analysis and data comparison with previous literatures. Interestingly, the glucose moieties in macrophylloside E and F possess free anomeric hydroxy groups. Genticrasides A/B, identified as a pair of iridoid originated lactones, have not been reported from Gentianaceae family up to now. The anti-inflammatory effects of selected compounds were also evaluated through the nitric oxide (NO) production inhibition in lipopolysaccharides (LPS)-induced RAW264.7 macrophage cells. In which, macrophyllosides G and D showed NO inhibitory activities with rates of 76.14±4.02 % and 52.44±8.29 % at 100 μg/mL., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

In this study, to identify bioactive components of Olea europaea leaves extract (OLE), chemometrics analyses including bivariate correlation analysis and partial least squares regression were used to establish the relationships between the chromatograms and anti-photoaging effect of OLE samples. Firstly, the fingerprint of olive leaves extract was determined by high-performance liquid chromatography (HPLC). Photoaging models of HaCaT cells were established by UVB irradiation. The photoaging resistance of OLE was evaluated by cell viability using the MTT assay. Chemometrics analyses showed that compounds 14, 19, 20, 24, 26, and 28 might be the major anti-photoaging components of OLE. Furthermore, after separation by HSCCC and NMR identification, compound 19 is luteoloside and compound 24 is oleuropein. Oleuropein and luteoloside were docked with collagenase (MMP-1), stromelysin (MMP-3), and gelatinase (MMP-9), respectively. The results showed that oleuropein and luteoloside inhibited their activity by directly interacting with MMP-1, MMP-3, and MMP-9, thereby exhibiting anti-photoaging activity. The current bioassay and spectrum-effect relationships are proper for associating sample quality with the active ingredient, and our finding would provide foundation and further understanding of the quality evaluation and quality control of Olea europaea., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Premna fulva Craib, rich in iridoid glycosides, is widely used to treat periarthritis, osteoproliferation, pain, and other diseases. However, no studies have reported effective purification methods for obtaining iridoid glycosides as active materials. This paper describes an efficient strategy for separating iridoid glycosides from Premna fulva leaves using high-speed counter-current chromatography and preparative high-performance liquid chromatography. A two-phase solvent system, ethyl acetate/n-butanol/water (7.5:2.5:10, v/v), was selected for high-speed counter-current chromatography separation. The proposed method effectively separated and purified four iridoid glycosides and four lignans, including three new iridoid glycosides (4-6) and five known compounds (1-3, 7, 8), from Premna fulva leaves, indicating that high-speed counter-current chromatography combined with prep-HPLC can efficiently isolate catalpol derivatives from the genus Premna. Additionally, the in vitro anti-inflammatory activities of all isolated compounds were analyzed using lipopolysaccharide-stimulated RAW 264.7 cells, and the results indicated that six compounds (1 and 3-7) exhibited potential anti-inflammatory activities., (© 2023 Wiley-VCH GmbH.)

Chirayata-the whole dried plant of Swertia chirayita-is an important traditional drug of Indian System of Medicines. A novel reverse-phase high performance liquid chromatography (RP-HPLC) method has been developed for the simultaneous determination and quantification of amaroswerin, amarogentin and andrographolide in a herbal drug "Chirayata," which is oftenly adulterated/substituted with herbal drug Kalmegh. The developed method is in accordance with International Council for Harmonization guidelines and is simple, precise, accurate, rapid, reproducible and specific to determine amarogentin, amaroswerin and andrographolide. Reverse-phase column (Water's X-bridge C18, 5 μm, 4.6 mm × 250 mm) with high resolution for all marker compounds was used with binary gradient elution (methanol:water) with a flow rate of 1 mL/min and detection at 235 nm. The developed method showed good linearity (R2 > 0.999) in a relatively wider range of concentration 2.968-95.00 ppm for amarogentin, amaroswerin and 5.625-180 ppm for andrographolide. The method is important for quality control analysis of drug Chirayata., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The present study investigated the effects of genipin cross-linking on the gelling properties of ginkgo seed protein isolate (GSPI). Cross-linking of GSPI was achieved with different concentrations (0, 0.05, 0.1, 0.2, 0.4, 0.6% w/v) of genipin at pH 6.0. Compared to pure GSPI, genipin treatment led to lower solubility, surface hydrophobicity, and fluorescence intensity, while promoted protein aggregation. Cross-linked GSPI gels exhibited markedly improved gelling properties and water holding capacity (WHC), with up to 2.1-fold increases in gel hardness and 1.3-fold increases in WHC over non-treated GSPI gel. Electrophoresis and Fourier-transform infrared spectroscopy confirmed the cross-linking. Moreover, microstructural examination showed that cross-linking with genipin resulted in protein aggregation and more porous gel matrix. Overall, genipin cross-linking demonstrated great potential for the enhancement of gelling properties of ginkgo seed protein. The current research may expand the utilization of ginkgo seeds in food applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Olea europaea is an economically significant crop native to Mediterranean countries. Its leaves exhibit several biological properties associated to their chemical composition. The aqueous ethanolic extracts of olive leaves from twelve different cultivars were analyzed by high performance liquid chromatography coupled to photodiode array and electrospray ionization mass spectrometry (HPLC/PDA/ESI-MS/MS). A total of 49 phytochemicals were identified in both positive and negative ionization modes. The identified compounds belonged to four classes of secondary metabolites including secoiridoids, flavonoids, pentacyclic triterpenoids and various phenolic compounds. Seasonal variation in chemical composition among the studied cultivars was apparent in autumn and spring. Secologanoside, oleuropein, hydroxy-oleuropein, demethyl oleuropein, gallocatechin, luteolin-O-hexoside, diosmetin, oleanolic acid and maslinic acid were detected in all cultivars in both seasons. Oleuropein-O-deoxyhexoside was tentatively identified for the first time in olive leaf extracts; detected only in the Spanish cultivar Picual (PIC) collected in spring. Also, dihydroxy-oxooleanenoic acid and hydroxy-oxooleanenoic acid, two bioactive pentacyclic triterpenes, were identified. Principle component analysis (PCA) showed good discrimination among the studied cultivars in terms of their botanical origin. This study is considered the first study for non-targeted metabolic profiling of different olive leaf cultivars cultivated in Egypt., (© 2023. The Author(s).)

The European Food Safety Authority highlights the beneficial effects of olive oil phenols, mainly, secoiridoids. Nevertheless, the metabolism of secoiridoids in humans has not been fully elucidated. This research evaluated the metabolism of secoiridoids in humans after intake of olive oils with diverse phenolic profiles. For this purpose, three extra virgin olive oils (EVOOs) were ingested by six volunteers at scheduled meals, and urine samples were collected the following morning for subsequent LC-MS/MS analysis. Using untargeted analysis, urinary metabolites revealed representative patterns associated with the various olive oil phenolic contents in absolute and relative terms. We were able to identify metabolites obtained through phase I, phase II, and microbial metabolism with discrimination between tyrosol and hydroxytyrosol derivatives. Metabolism of phenols is differentially activated as a function of the olive oil secoiridoids content, and this proof-of-concept study shows how urinary metabolites represent olive oil phenolic content., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

As a traditional Chinese medicine, Eucommia ulmoides Oliver ( E. ulmoides Oliv.) is an important medicinal plant, and its barks, male flowers, leaves, and fruits have high value of utilization. The seed meal of E. ulmoides Oliv. is the waste residue produced after oil extraction from seeds of E. ulmoides Oliv. Though the seed meal of E. ulmoides Oliv. is an ideal feed additive, its medicinal value is far from being developed and utilized. We identified six natural iridoid compounds from the seed meal of E. ulmoides Oliv., namely geniposidic acid (GPA), scyphiphin D (SD), ulmoidoside A (UA), ulmoidoside B (UB), ulmoidoside C (UC), and ulmoidoside D (UD). Six natural iridoid compounds were validated to have anti-inflammatory activities. Hence, six compounds were quantified at the optimum extracting conditions in the seed meal of E. ulmoides Oliv. by an established ultra-performance liquid chromatography (UPLC) method. Some interesting conversion phenomena of six tested compounds were uncovered by a systematic study of stability performed under different temperatures and pH levels. GPA was certified to be stable. SD, UA, and UC were only hydrolyzed under strong alkaline solution. UB and UD were affected by high temperature, alkaline, and strong acid conditions. Our findings reveal the active compounds and explore the quantitative analysis of the tested compounds, contributing to rational utilization for the seeds residues of E. ulmoides Oliv.

This study aimed to explore the correlation of the content of 15 non-crocin components of Gardeniae Fructus with its external properties(shape and color). The fruit shape was quantified according to the length/diameter measured by ruler and vernier calliper and the chromaticity values L~*, a~*, b~*, and ΔE~* of all samples were determined by chroma meter. Chromatographic separation was conducted on a Welch Ultimate XB C&#95;(18) column(4.6 mm×250 mm, 5 μm) under gradient elution with acetonitrile solution(A) and 0.1% formic acid aqueous solution(B) as the mobile phase at a flow rate of 1.0 mL·min~(-1). The column temperature was 30 ℃ and the detection wavelength was 238 nm. The high-performance liquid chromatography(HPLC) method was established for simultaneous determination of the content of eight iridoid glycosides, six phenolic acids, and one flavonoid in 21 batches of Gardeniae Fructus samples. The correlation of the content of the 15 components with shapes and chromaticity values in each sample was analyzed by multivariate statistical analysis. According to the circulation situation and traditional experience, 21 batches of Gardeniae Fructus samples were divided into three categories, namely 14 batches of Jiangxi products(small and round, red and yellow), 4 batches of Fujian products(oval, red) and 3 batches of Shuizhizi(Gardenia jasminoides, longest, reddest). The Gardeniae Fructus samples were sequenced as Jiangxi products(1.71) &lt; Fujian products(1.99) &lt; Shuizhizi(2.55) in terms of the length/diameter average, Jiangxi products(17.7) &lt; Fujian products(19.7) ≈ Shuizhizi(19.6) in terms of average value of a~*(red and green), Jiangxi products(24.4) &gt; Fujian products(19.2) ≈ Shuizhizi(19.3) in terms of b~*(yellow and blue), and Jiangxi products(49.8) &gt; Fujian products(48.0) ≈ Shuizhizi(47.8) in terms of L~*(brightness). The total content of the 15 components, 8 iridoid glycosides, 6 phenolic acids, and rutin in Jiangxi products was in the ranges of 65.53-99.64, 52.15-89.16, 6.10-11.83, and 0.145-1.81 mg·g~(-1), respectively. The total amount of the 15 components, 8 iridoid glycosides, 6 phenolic acids, and rutin in Fujian products was in the ranges of 69.33-94.35, 63.52-85.19, 5.39-8.41, and 0.333-0.757 mg·g~(-1), respectively. In Shuizhizi, the total content of the 15 components, 8 iridoid glycosides, 6 phenolic acids, and rutin was in the ranges of 77.35-85.98, 68.69-76.56, 7.30-9.05, and 0.368-0.697 mg·g~(-1), respectively. Pearson correlation analysis revealed that Gardeniae Fructus with leaner and longer fruit shape possessed lower content of total phenolic acids(the sum of the six phenolic acids) and rutin, but the correlation with iridoid glycosides was not high. Additionally, the higher content of total phenolic acids and rutin denoted the yellow coloration of Gardeniae Fructus, and the higher content of cryptochlorogenic acid, chlorogenic acid, and rutin meant the brighter color of Gardeniae Fructus. However, the higher content of geniposide and neochlorogenic acid and the lower content of deacetyl asperulosidic acid methyl ester led to the red coloration of Gardeniae Fructus. The results indicated that the morphological characters of Gardeniae Fructus were closely related to its chemical components. The more round shape and the yellower color reflected the higher content of phenolic acids and flavonoid, and Gardeniae Fructus with redder color had higher content of geniposide. OPLA-DA showed that the length/diameter and the content of six iridoid glycosides(gardoside, shanzhiside, gardenoside, genipin 1-gentiobioside, 6β-hydroxy geniposide, and deacetyl asperulosidic acid methyl ester), two phenolic acids(neochlorogenic acid and cryptochlorogenic acid) and rutin could be used as markers to distinguish three types of samples. This study provided experimental data for the scientific connotation of &quot;quality evaluation through morphological identification&quot; of Gardeniae Fructus.

Sanzi San, a Mongolian medicine, comprises three herbs: Terminalia chebula , Melia toosendan , and Gardenia jasminoides . Clinically, Sanzi San is administered orally and distributed via blood to the action site, which implies that the absorption, distribution, metabolism, and excretion (ADME) are closely related to the pharmacological action and curative effect. Therefore, possible explanations for the material basis of Sanzi San were explored in this study preliminarily. A strategy based on serum pharmacochemistry was first applied to explore the absorbed bioactive components and metabolites of Sanzi San. Wistar rats were randomly divided into normal and dosing groups, which were provided with the Sanzi San's water extract for three days. Then, the rat's blood samples were obtained from their abdomiral aorta using a sterile blood collection tube after administering the medicine. The blood samples were then centrifuged at 3500 r/min for 10 min to obtain the serum samples. A practical method based on high performance liquid chromatography coupled with quadrupole and electrostatic field orbitrap high resolution mass spectrometry (HPLC-Q/Orbitrap HRMS) was developed to screen and analyze numerous bioactive components and metabolites adsorbed in the serum of the dosing rats after oral administration of the Sanzi San's water extract. Chromatographic separation was achieved on a SHIMADZU GIST C 18 chromatographic column (150 mm×4.6 mm, 5 μm). The temperature of the column was maintained at 30 ℃. The flow rate was 0.5 mL/min, and the injection volume was 10 μL. The mobile phase comprised an aqueous solution of 0.1% formic acid and methanol under gradient elution. A heated electrospray ion (HESI) source was used with positive and negative ion scanning modes. To rapidly screen out and identify the absorbed bioactive components and metabolites of Sanzi San in the rat serum samples, a simple three-step approach was developed. First, the known components in Sanzi San were listed systematically by exploring various databases, such as the Web of Science, PubMed, and Chinese National Knowledge Infrastructure. In addition, relevant information on drug biotransformation and the characteristic fragmentation patterns of parent compounds were summarized. Second, the absorbed components and metabolites were ascertained using the Xcalibur 3.0 software. Based on the information related to the parent compound's structure, the software could be used to identify the unique peaks by comparing the chromatograms of the normal and dosing samples. Consequently, the total ion chromatograms of serum samples were established. Finally, the Compound Discover 3.0 software was used to predict the metabolic pathways and fragmentation of the absorbed compounds. Using this approach, 55 compounds were characterized, including 41 prototype components and 14 metabolites. The main prototype components in the serum sample were tannins, iridoids, and phenolic acids. The details of these compounds have been summarized and presented. Regarding the absorbed bioactive components and metabolites in the serum samples of rats administered with Sanzi San, phase Ⅰ and phase Ⅱ biochemical reactions were involved in the biotransformation pathways. The phase Ⅰ reaction modified the components and created sites for the phase Ⅱ reaction, involving reduction and hydrolysis. The phase Ⅱ reaction coupled groups to existing conjugation sites, including glucuronide to glucuronic acid, sulfate, and methyl. MS/MS spectra indicated that methylation, demethylation, and dehydroxylation are the metabolic pathways of procyanidins. Additionally, glucuronidation, deglucosidation, hydration, and demethylation are the metabolic pathways of iridoids in Sanzi San. This study comprehensively analyzed the components of the Sanzi San's water extract absorbed in the rat's serum. Our results revealed information regarding the pharmacodynamic substances and the major pathways involved in the ADME of Sanzi San. Further, potential medicinal ingredients for the pharmacological effects and clinical use of Sanzi San were explored at the serum pharmacochemistry level.

Crocins are valuable bioactive components of gardenia fruit, and their biosynthesis and accumulation have attracted widespread interest. Studies have investigated the biosynthesis and accumulation of crocin based on Illumina sequencing, but there is a lack of reports based on full-length transcriptome sequencing. Utilising SMRT sequencing and high-performance liquid chromatography (HPLC), we explored crocin biosynthesis and accumulation in the fruit of Gardenia jasminoides . HPLC analysis showed that crocins specifically exist in fruit and that the content of crocins increases gradually during fruit development. SMRT sequencing generated 46,715 high-quality full-length isoforms, including 5230 novel isoforms that are not present in the G. jasminoides genome. Furthermore, a total of 46 genes and 91 lncRNAs were involved in the biosynthesis and accumulation of crocin. The qRT-PCR indicated that genes involved in crocin biosynthesis reached a peak in the NOV stage. These findings contributed to our understanding of crocin biosynthesis and accumulation.

Tissue cross-linking represents an important and often used technique to enhance the mechanical properties of biomaterials. For the first time, we investigated biochemical and structural properties of genipin (GE) cross-linked equine pericardium (EP) using optical imaging techniques in tandem with quantitative atomic force microscopy (AFM). EP was cross-linked with GE at 37 °C, and its biochemical and biomechanical properties were observed at various time points up to 24 h. GE cross-linked EP was monitored by the normalized ratio between its second-harmonic generation (SHG) and two-photon autofluorescence emissions and remained unchanged for untreated EP; however, a decreasing ratio due to depleted SHG and elevated autofluorescence and a fluorescence band at 625 nm were found for GE cross-linked EP. The mean autofluorescence lifetime of GE cross-linked EP also decreased. The biochemical signature of GE cross-linker and shift in collagen bands were detected and quantified using shifted excitation Raman difference spectroscopy as an innovative approach for tackling artifacts with high fluorescence backgrounds. AFM images indicated a higher and increasing Young's modulus correlated with cross-linking, as well as collagen structural changes in GE cross-linked EP, qualitatively explaining the observed decrease in the second-harmonic signal. In conclusion, we obtained detailed information about the biochemical, structural, and biomechanical effects of GE cross-linked EP using a unique combination of optical and force microscopy techniques in a nondestructive and label-free manner.

The stems of Nauclea officinalis have been utilized as a crude drug in China, so other parts of the plant are abandoned, resulting in a waste of traditional Chinese medicine resources. To determine the distribution and content of the alkaloids, phenolic acids and iridoid in different organs (stem, branch, leaf and bark) of this plant, a reliable method has been established using LC-MS/MS. Nine constituents, namely strictosamide, vincosamide, chlorogenic acid, sweroside, naucleamide B, protocatechuic acid, pumiloside, vanillic acid and cryptochlorogenic acid, were simultaneously determined in 6 min. Meanwhile, the antipyretic, anti-inflammatory and analgesic activities were evaluated for comparative analysis of the pharmacological activity of different parts of N. officinalis. The results showed that the content of active components in other organs of N. officinalis was higher than that in stems, and the pharmacological effects of branches and leaves were also better. The established approach could be helpful for the quality control of N. officinalis, and also provide necessary information for the rational utilization of resources., (© 2021 John Wiley & Sons, Ltd.)

Zhi-zi-Hou-po decoction (ZZHPD) has been used to treat depression in the clinic for thousand years in China. However, the pharmacodynamic substance of ZZHPD is still not totally clear due to its complex components. The objective of this study was to identify the effectual combination ingredients (ECIs) of ZZHPD, which could represent the antidepressant effect of the original formula. Firstly, differential plasma absorbed components with different variable importance in projection (VIP) value in chronic unpredictable mild stress (CUMS)-induced depression and control rat were revealed by untargeted metabolomics-driven strategy based on HPLC-ESI-TOF/MS, XCMS online and SIMCA-p software. Secondly, network topology scores (NTS) of plasma absorbed components were exposed by protein-protein interaction (PPI) network analysis based on components-related genes and depression-related genes, which were performed by network pharmacology tools. Finally, the ECIs were screened by considered of VIP value and NTS. Then the bioactivity was evaluated by cell viability and expression of glial fibrillary acid protein (GFAP), tumor necrosis factor α (TNFα) and interleukin 1β (IL-1β) of a lipopolysaccharide-induced astrocyte depression model. An effective combination composed with 12 components was filtrated as ECIs of ZZHPD, exposed to which the cell viability effect, the expression of GFAP and IL-1β in astrocytes were essentially equivalent with original ZZHPD (p > 0.05), and that both characteristic constituents and trace compounds of ZZHPD might exert synergistic actions through multi-targets. The result of this study provided useful information for the clinical application and modern development of ZZHPD, and the proposed strategy could be regard as an alternative solution for effective combination screening of herbal medicines., (Copyright © 2021 Elsevier B.V. All rights reserved.)

This study aimed to characterize the antioxidant potential, bioactive and volatile compounds of the stones from fruits of Cornus mas. Both fresh and roasted stones show a high antioxidant potential (166.48-509.74 μmol TE/g dw stones), which significantly depends on the cultivars. The roasted stones preserved 43.6% (DPPH; 'Raciborski') to 97.2% (FRAP; 'Alesha') of the antioxidant activity of the non-roasted stones. In the stones, two iridoids and ellagic acid were determined. During roasting, loganic acid remained stable, whereas cornuside was completely degraded. The analyses showed a 30-fold increase in the concentration of ellagic acid and in the formation of two of its derivatives. The major aroma compound of the roasted stones was furfural, but we also identified 18 pyrazine derivatives. This study is the first attempt to valorize Cornelian cherry stones via roasting. The roasted stones can be a coffee substitute, or aromatic and bioactive additions to cereal coffees., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The aim of this study was to conduct a histochemical analysis to localize lipids, terpenes, essential oil, and iridoids in the trichomes of the L. album subsp. album corolla. Morphometric examinations of individual trichome types were performed. Light and scanning electron microscopy techniques were used to show the micromorphology and localization of lipophilic compounds and iridoids in secretory trichomes with the use of histochemical tests. Additionally, the content of essential oil and its components were determined using gas chromatography-mass spectrometry (GC-MS). Qualitative analyses of triterpenes carried out using high-performance thin-layer chromatography (HPTLC) coupled with densitometric detection, and the iridoid content expressed as aucubin was examined with spectrophotometric techniques. We showed the presence of iridoids and different lipophilic compounds in papillae and glandular and non-glandular trichomes. On average, the flowers of L. album subsp. album yielded 0.04 mL/kg of essential oil, which was dominated by aldehydes, sesquiterpenes, and alkanes. The extract of the L. album subsp. album corolla contained 1.5 × 10 -3 ± 4.3 × 10 -4 mg/mL of iridoid aucubin and three triterpenes: oleanolic acid, β-amyrin, and β-amyrin acetate. Aucubin and β-amyrin acetate were detected for the first time. We suggest the use of L. album subsp. album flowers as supplements in human nutrition.

The fruits of Cornus mas and Cornus officinalis have been known and appreciated in folk medicine for years and have a high biological value, which is mainly connected with their polyphenols and iridoids content. However, hybrids of C. mas × C. officinalis have not been investigated. The aim of this study was to evaluate the iridoids, anthocyanins, and flavonols content, and antioxidant capacity of Cornus mas , Cornus officinalis , and C. mas × C. officinalis . Iridoids and flavonoids were quantified by the High-Performance Liquid Chromatography (HPLC) method. Antioxidant capacity (AC) was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH • ), 2,2'-azino-bis (3-ethyl benzothiazoline-6-sulfonic acid (ABTS •+ ), and ferric reducing antioxidant power (FRAP) tests. Total phenolic content (TPC) was evaluated using the Folin-Ciocalteu reagent. Among the C. mas cultivars and C. officinalis genotypes, there was considerable variation in the content of iridoids, flavonoids, and AC. Interspecific hybrids C. mas × C. officinalis contained more iridoids than C. mas and more anthocyanins than C. officinalis and additionally had higher AC and TPC than C. officinalis and most C. mas . AC, TPC, and the presence of iridoids, anthocyanins, and flavonols in hybrids C. mas × C. officinalis are reported for the first time. The Cornus species deserve special attention due to their highly biologically active substances, as well as useful medicinal properties.

The effect of micronization of granulometrically fractionated olive pomace (OP) on the bioaccessibility of polyphenols and the antioxidant capacity was investigated during sequential in vitro static digestion. Crude OP was fractionated in a 2-mm sieve (F1: > 2 mm; F2: < 2 mm) and then micronized (300 r min -1 , 5 h) generating F1AG (17.8 μm) and F2AG (15.6 μm). Micronization increased the release of hydroxytyrosol, oleuropein, caffeic acid, and decarboxymethyl oleuropein aglycone (3,4-DHPEA-EDA) in the salivary and gastric phase, beyond luteolin in the gastric phase. Micronization also increased the intestinal bioaccessibility of hydroxytyrosol, 3,4-DHPEA-EDA, oleuropein, luteolin, and apigenin; it was more effective for F2AG than F1AG. Micronized samples increased antioxidant capacity in the gastric phase. F2AG exhibited the highest antioxidant capacity in the insoluble intestinal fraction. Thus, micronization can be further exploited to improve the nutraceutical properties of OP by increasing the bioaccessibility and antioxidant capacity of phenolic compounds., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Oil mills could benefit by preparing their own aqueous extracts from olive leaves. Accordingly, the present study aimed to measure the bioactive compounds richness of such extracts, especially oleuropein. A water-based microwave extraction procedure was developed and a selective and precise high-performance liquid chromatography with diode array detection (HPLC-DAD) method was validated for the determination of oleuropein and others bioactive compounds from olive leaves., Results: The water solubility of oleuropein was determined to be 9.5 g L -1 . The extraction procedure was optimized in terms of power, olive leaf weight/water volume ratio and time of extraction, and the results revealed that 2 mg mL -1 and a microwave irradiation at 800 W for 30 s resulted in the greatest efficiency. Oleuropein was determined by the new validation method, which showed good linearity (r 2 = 0.996), precision (% relative standard deviation < 10%), recovery (118.6%), and limits of detection (17.48 mg L -1 ) and quantification (21.54 mg L -1 ). Good correlation (r 2 = 0.979) was obtained between oleuropein of the olive leaf extracts determined by HPLC-DAD and by UV-visible spectrophotometry., Conclusion: A simple extraction method was developed and validated to obtain aqueous extract from olive leaves by microwave extraction, determining for the first time oleuropein water solubility. Validation of the method showed that oleuropein in olive leaves could be quantified when it is at least 1% of dry weight by means of HPLC-DAD. UV-visible spectrophotometry can be useful in oil mills because it enables the content of oleuropein and other bioactive compounds content to be determined in situ in such leaf aqueous extracts. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Olive (Olea europaea) is an important crop in Europe, with high cultural, economic and nutritional significance. Olive oil flavor and quality depend on phenolic secoiridoids, but the biosynthetic pathway of these iridoids remains largely uncharacterized. We discovered two bifunctional cytochrome P450 enzymes, catalyzing the rare oxidative C-C bond cleavage of 7-epi-loganin to produce oleoside methyl ester (OeOMES) and secoxyloganin (OeSXS), both through a ketologanin intermediary. Although these enzymes are homologous to the previously reported Catharanthus roseus secologanin synthase (CrSLS), the substrate and product profiles differ. Biochemical assays provided mechanistic insights into the two-step OeOMES and CrSLS reactions. Model-guided mutations of OeOMES changed the product profile in a predictable manner, revealing insights into the molecular basis for this change in product specificity. Our results suggest that, in contrast to published hypotheses, in planta production of secoxy-iridoids is secologanin-independent. Notably, sequence data of cultivated and wild olives point to a relation between domestication and OeOMES expression. Thus, the discovery of this key biosynthetic gene suggests a link between domestication and secondary metabolism, and could potentially be used as a genetic marker to guide next-generation breeding programs., (© 2020 The Authors. New Phytologist © 2020 New Phytologist Foundation.)

In the last two decades, phenolic compounds occurring in olive oils known as secoiridoids have attracted a great interest for their bioactivity. Four major olive oil secoiridoids, i.e., oleuropein and ligstroside aglycones, oleacin and oleocanthal, were previously characterized in our laboratory using reversed-phase liquid chromatography with electrospray ionization-Fourier transform-mass spectrometry (RPLC-ESI-FTMS). The same analytical approach, followed by multivariate statistical analysis (i.e., Principal Component Analysis), was applied here to a set of 60 Italian extra-virgin olive oils (EVOO). The aim was to assess the secoiridoid contents as a function of olive cultivars, place of cultivation (i.e., different Italian regions) and olive oil processing, in particular two- vs. three-phase horizontal centrifugation. As expected, higher secoiridoid contents were generally found in olive oils produced by two-phase horizontal centrifugation. Moreover, some region/cultivar-related trends were evidenced, as oleuropein and ligstroside aglycones prevailed in olive oils produced in Apulia (Southern Italy), whereas the contents of oleacin and oleocanthal were relatively higher in EVOO produced in Central Italy (Tuscany, Lazio and Umbria). A lower content of all the four secoiridoids was generally found in EVOO produced in Sicily (Southern Italy) due to the intrinsic low abundance of these bioactive compounds in cultivars typical of that region.

Phenols are responsible for the only health claim of virgin olive oil (VOO) recognized by the European Commission EU 432/2012 and the European Food Safety Authority. In this research, we studied the decrease in the phenolic content of 160 extra VOOs (EVOOs) after 12 months storage in darkness at 20 °C. Phenolic concentration was decreased 42.0 ± 24.3% after this period and this reduction strongly depended on the initial phenolic profile. Hence, EVOOs with predominance in oleacein and oleocanthal experienced a larger decrease in phenolic content than oils enriched in other phenols. Complementarily, hydroxytyrosol and oleocanthalic acid increased significantly in aged EVOOs, which allowed their discrimination from recently produced EVOOs. These changes are explained by degradation of main secoiridoids during storage due to their antioxidant properties. Hydroxytyrosol and oleocanthalic acid can be considered markers of olive oil ageing, although they can also provide information about quality or stability., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In the present research, seven different cornelian cherry (Cornus mas L.) cultivars and selections were examined. In vitro and in silico methods were applied for determining and correlating phytochemical constituents and biological potential. Loganic acid, cornuside, cyanidin3-galactoside, and pelargonidin 3-galactoside were determined as the most dominant compounds, presenting ≥90% of the all detected iridoid and phenolic constituents in the extracts. Cornelian cherry fruits were characterized by high antioxidant capacity and antiproliferative activity on human colon cancer cells (HT29). It was observed the strong inhibitory potential of α-amylase, α-glucosidase, and dipeptidyl peptidase III (DPP III) enzyme activities. Principal component analysis (PCA) was used as a very helpful tool to discriminate the constituents with the highest contribution to tested bioactivities and to highlight the most potent genotypes. PCA, together with binding energies measurements and docking analysis, pointed out pelargonidin 3-robinobioside as the strongest inhibitor of α-glucosidase., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Amarogentin (AG), a biologically active secoiridoid glycoside, is responsible for the efficacy of Gentianaceae based medications. Thus, qualitative and quantitative analyses of AG are of significance for batch to batch quality control purposes. By conjugating colloidal gold nanoparticles with the AG-specific monoclonal antibody, MAb 1E9, we were able to develop a single-step competitive immunochromatographic assay (ICA) for simple quantification of the AG content in plant samples. With a limit of detection of 250 ng/mL, the analytical results were obtained after immersing the ICA test strip in the detection mixture for 15 min. This new ICA is superior to conventional ICAs as it is considerably faster due to the speed with which the test strips can be produced and the omission of the time-consuming preparation phase that was previously required to make the fiber pad. Moreover, our ICA only needs a small amount of analyte (20 µL).The reliability of the reported test strip was confirmed by comparing its semi-quantitative results with those obtained via an indirect competitive enzyme-linked immunosorbent assay (icELISA). The positive correlation between these methods (R 2  = 0.984) indicated that this new ICA could be applied for the semi-quantitative analysis of the AG content in plant samples.

Background: Nowadays, the pharmacological effects of Plantaginis semen was getting more and more attention because of the great effect of treating diuresis, hypertension, hyperlipidemia, and hyperglycemia. According to the Chinese Pharmacopoeia, Plantaginis semen is the seed of Plantago asiatica L. or P. depressa Willd. This was verified by examining chemical composition differences in a preliminary experiment, predicting their differences in pharmacology., Purpose: In this study, we aimed to compared the the differences in main components and anti-obesity effects of Plantago asiatica L. seed extract (PASE) and P. depressa Willd. seed extract (PDSE)., Study Design and Methods: The ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) analysis was used to characterize and compare the differences chemical constituents of PASE and PDSE. The difference therapeutic effects between PASE and PDSE on obesity and associated metabolic disorders was investigated by high-fat (HF) diet induced mice model., Results: The fingerprint of Plantaginis semen were established by screening and identified 15 main components, including iridoids, phenethanol glycosides, flavonoids, guanidines, and fatty acids. Pentahydroxy flavanone was observed only in PDSE but not in PASE. The quantitative analysis results indicated that the main bioactive components in PASE were geniposidic acid and acteoside; their concentrations were three times higher in PASE than in PDSE. In anti-obesity effects, the result show the levels of fasting blood glucose were improved in both PASE and PDSE when compared with the HF group, while the PASE is show a significant effect then the PDSE group and improved the glucose tolerance but not in PDSE. The results also displayed that the Plantaginis semen did not modify food intake or body weight but decreased abdominal white/brown adipocyte size, serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-c), hepatic TG and TC, fecal TG and TC concentrations when compared with the HF group. Among these indicators, serum TG, liver TG, fecal TC and TG levels were significantly improved in PASE compared with PDSE. The results indicated that PASE treatment more effectively improved lipid and glucose metabolism in HF diet-induced obese mice than did PDSE., Conclusion: As Plantaginis semen sources, P. asiatica L. seeds demonstrated more bioactive components and favorable metabolic disorder treatment outcomes than did P. depressa Willd. seeds., (Copyright © 2020. Published by Elsevier GmbH.)

Chinese materia medica processing is a distinguished and unique pharmaceutical technique in traditional Chinese medicine (TCM), which has played an important role in reducing side effects, increasing medical potencies, altering the properties and even changing the curative effects of raw herbs. The efficacy improvement in medicinal plants is mainly caused by changes in the key substances through an optimized processing procedure. Thus, the use of a rapid method for determining suitable chemical markers between raw and processed TCM is critical in order to elucidate how the bioactive compounds influence the clinical effects. In this study, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry combined with MS/MS-based molecular networking (MN) and a multivariate statistical analysis method is proposed for the first time. This combination was used to identify the complex chemical composition and clarify the changed constituents between raw and processed Cistanche tubulosa (C. tubulosa). The chemical analysis results demonstrated that a total of 85 compounds were identified in the crude and processed C. tubulosa. Moreover, 34 compounds were detected as chemical markers. This systematic research into chemical constituents and chemical markers of crude and processed C. tubulosa lays a solid foundation for further study of the quality control of C. tubulosa. Moreover, the study provides a new and valuable technical strategy for analyzing chemical components and identifying potential chemical markers for the processing of herbal medicines.Graphical abstract.

Jasminum is an important genus in the olive family (Oleaceae), comprising about 200 species distributed all over the world. In the current study, the methanolic extract of Jasminum grandiflorum subsp. floribundum aerial parts and its respective fractions; dichloromethane and n-butanol fractions were analyzed using ultra-performance liquid chromatography coupled to high resolution mass (UPLC-HRMS) for profiling and characterization of the plant metabolites. More than seventy metabolites were identified belonging to different classes including phenolic acids, flavonoids, secoiridoids, iridoids, lignans, fatty acids, and triterpenes. The samples were also assessed for their angiotensin-I-converting enzyme (ACE) and renin inhibitory activity along with their antioxidant potential using five complementary assays: TAC (total antioxidant capacity), DPPH (1,1-diphenyl-2-picrylhydrazyl), ABTS (2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid), FRAP (ferric reducing antioxidant power) and iron reducing power. The results revealed that the n-butanol fraction showed a potent ACE and renin inhibition as compared to Lisinopril and Aliskiren standard drugs (24.66 ± 2.41 ng/mL vs. 18.37 ± 1.21 ng/mL and 141.14 ± 5.28 ng/mL vs. 447.87 ± 3.2 ng/mL, respectively) and also a strong antioxidant activity. Interestingly, the secoiridoids, dominated metabolites detected in the n-butanol fraction, revealed the potential of them for management of the hypertension diseases. The total extract and fractions were also standardized using HPLC analysis of the major secoiridoid glycoside; oleuropein. Finally, J. grandiflorum standardized extract could be considered as a target for further studies to discover a new therapeutic anti-hypertensive drug., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Herein we report for the first time the levels of phenylpropanoids and iridoids in extracts and infusions of V. minutiflora consumed in Brazil to treat urinary and infectious disorders. An in house validation study demonstrated good accuracy and precision to determine the bioactive compounds in V. minutiflora by HPLC-DAD. Phenylpropanoids varied in the extracts (leaves 139.70 to 221.20 mg g-1, flowers 106.43 to 227.22 mg g-1, stems 42.18 to 56.48 mg g-1). Verbascoside occurred in higher concentration in extracts of leaves (87.66 - 136.16) mg g-1 and flowers (58.12 - 148.96) mg g-1 than in stems (19.24 - 24.62) mg g-1. Iridoids in extracts were as follows: leaves (46.60 - 54.79) mg g-1, flowers (55.88 - 93.87) mg g-1 and stems (40.05 to 61.74) mg g-1. High levels of iridoids (314.70 - 415.10) μg mL-1, phenylpropanoids (1996.39 - 2674.13) μg mL-1 and verbascoside (1029.38 - 1456.42 μg mL-1) in infusions support the popular consume of V. minutiflora.

Soybean is one of the most important commodities in the world, being applied in feed crops and food, pharmaceutical industries in different ways. Soy is rich in isoflavones that in aglycone forms have exhibited significant anti-obesity and anti-lipogenic effects. Obesity is a global problem as several diseases have been related to this worldwide epidemic. The aim of this work was to verify the effect of free and immobilized β-glucosidase, testing Lentikats, and sol-gel as carriers. Moreover, we wanted to examine if the different types of hydrolysis would generate extracts with distinct biological activity concerning lipid accumulation, PPAR-α regulation, and TNF-α, IL-6, and IL-10 concentrations using in vitro assays. Our results show that all formulations of β-glucosidase could hydrolyze soy isoflavones. Thus, after 24 h of incubation, daidzein content increased 2.6-, 10.8-, and 12.2-fold; and genistein content increased 11.7, 11.4, and 11.4 times with the use of free enzyme, Lentikats®, and sol-gel immobilized enzyme, respectively. Moreover, both methodologies for enzyme immobilization led to promising forms of biocatalysts for application in the production of soy extracts rich in isoflavones aglycones, which are expected to bring about health benefits. A mild lipogenic effect was observed for some concentrations of extracts, as well as a slight inhibition in PPAR-α expression, although no significant differences were noticeable in the cytokines TNF-α, IL-10, and IL-6 as compared with the control.

Spent coffee ground (SCG) is the remaining residue produced after extraction of coffee, and it is considered a source of unextracted bioactive compounds. For this, in the latest years, the attention has been focused to innovative reuses that can exploit the potentiality of SCG. Unfortunately, the content of bioactive compounds has not been thoroughly studied yet, and the major of publication has investigated the caffeine and chlorogenic acids levels, total polyphenol contents, and total flavonoid content. Hence, these approaches have determined only an estimation of flavonoids and polyphenols content and lack on single polyphenols investigation. Therefore, the objective of the current work was to provide a deep characterization of bioactive compounds in SCG. For this purpose, a new analytical method for the quantification of 30 molecules, including caffeine, chlorogenic acids, phenolic acids, flavonoids, and secoiridoids, has been developed using high-performance liquid chromatography tandem mass spectrometry. Moreover, several extraction procedures, that is, liquid-solid extraction assisted and not by ultrasounds, testing diverse solvents, were evaluated. Liquid-solid extraction assisted by sonication, with water/ethanol (30/70, v/v), resulted the best in terms of total bioactive compounds, and, once validated, the new analytical method was applied to five different espresso SCG samples. Data showed that caffeine (means: 1193.886 ± 57.307 mg kg -1 ) and chlorogenic acids (means of total CQAs: 1705.656 ± 88.694 mg kg -1 ) were the most abundant compounds in all SCG samples followed by phenolic acids such as caffeic, ferulic, gallic, p-coumaric, syringic, trans-cinnamic, and vanillic acid. Moreover, some flavonoids, that is, rutin, cyanidin 3-glucoside, and quercetin, occurred in almost all samples. This work provided a deepened characterization of bioactive compounds in SCG and can contribute to develop new strategies of reuses., (© 2020 John Wiley & Sons, Ltd.)

The awareness of the functional and nutraceutical properties of olives and olive oil bioactive constituents contributed to oliviculture recent increase. Olives' metabolism and nutritional quality are determined by how the olive-tree is coping to climate change-related episodes, which increasingly occur in the Mediterranean. We characterize the most relevant lipophilic and phenolic compounds of olives from Olea europaea cultivars [Cobrançosa, Cordovil de Castelo Branco and Cordovil de Serpa (C.Serpa)] exposed to drought + heat. Olives from the three cultivars presented a similar qualitative profile but differed in their relative richness. Cobrançosa olives are richer in organic acids, esters and carbohydrates, while C.Serpa olives have higher levels of phenolic compounds, particularly under control conditions. Drought + heat changed the quantitative profile of olives, in a way dependent on the cultivar, and C.Serpa olives showed the highest stress susceptibility. Climate change-related conditions stimulate the accumulation of relevant bioactive compounds in olives, contributing to increasing its nutritional value., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The huge interest in the health-related properties of foods to improve health has brought about the development of sensitive analytical methods for the characterization of natural products with functional ingredients. Greek olive leaves and drupes constitute a valuable source of biophenols with functional properties. A novel ultra-high-performance liquid chromatography-quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS) analytical method was developed to identify biophenols through target and suspect screening in Greek olive leaves and drupes of the varieties: Koroneiki, Throumbolia, Konservolia, Koutsourelia, Kalamon, Petrolia, Amigdalolia, Megaritiki, Mastoeidis, Agouromanakolia, Agrilia, Adramitiani and Kolovi. The method's performance was evaluated using the target compounds: oleuropein, tyrosol and hydroxytyrosol. The analytes demonstrated satisfactory recovery efficiency for both leaves (85.9-90.5%) and drupes (89.7-92.5%). Limits of detection (LODs) were relatively low over the range 0.038 (oleuropein)-0.046 (hydroxytyrosol) and 0.037 (oleuropein)-0.048 (hydroxytyrosol) for leaves and drupes, respectively For leaves, the precision limit ranged between 4.7 and 5.8% for intra-day and between 5.8 and 6.5% for inter-day experiments, and for drupes, it ranged between 3.8 and 5.2% for intra-day and between 5.1 and 6.2% for inter-day experiments, establishing the good precision of the method. The regression coefficient (r 2 ) was above 0.99 in all cases. Furthermore, the preparation of herbal tea from olive leaves is suggested after investigating the optimum infusion time of dried leaves in boiling water. Overall, 10 target and 36 suspect compounds were determined in leaves, while seven targets and thirty-three suspects were identified in drupes, respectively.