Search

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.)

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.)

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.

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.)

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.)

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.

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.)

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.

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.

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.

A simple and green sodium dodecyl sulfate-synergistic microwave-assisted extraction method was developed to extract and determine the iridoids, phenylpropanoids, and lignans in Eucommiae Cortex followed by ultra-high-performance liquid chromatography with photodiode array detection. The biodegradable solution (sodium dodecyl sulfate) was used as a promising alternative to organic solvents. The response surface methodology provided the optimum extraction conditions (2 mg/mL sodium dodecyl sulfate, 1100 W microwave power, and 6 min extraction time). The recoveries of three types of components ranged from 95.0 to 105% (RSDs < 5%). The intra- and inter-day precision and accuracy were less than 3.40% and within the range of 97.1-105%, respectively. Compared with other extraction methods, this newly established method was more efficient and environmental friendly. The results demonstrated that sodium dodecyl sulfate-synergistic microwave-assisted extraction followed by ultra-high-performance liquid chromatography with photodiode array method was applicable for the simultaneous extraction and determination of these three types of compounds for quality evaluation of Eucommiae Cortex., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This study was to develop a reliable and simple high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to detect paeoniflorin, geniposide, saikosaponin b2, liquiritin, paeonol and atractylenolide Ⅲ in beagle plasma and to study pharmacokinetic of paeoniflorin and geniposide after single-dose administration of Danzhi Xiaoyao Pill (DZXY). Chromatographic separation was performed using an Agilent C18 column, and multiple reaction monitoring (MRM) mode was used. A gradient elution procedure was used with solvent A (acetonitrile) and solvent B (0.1 % formic acid-water) as mobile phases. The elution procedure was as follows: 85 % B-30 % B (0-7 min) and 30 % B-30 % B (7.1-8 min). The flow rate was 0.3 mL/min. The column temperature was 40 ℃, and the injection volume was 10 μL. The main analytical parameters of paeoniflorin, geniposide, saikosaponin b2, liquiritin, paeonol and atractylenolide Ⅲ were m/z 525→449, m/z 433→224, m/z 780→617, m/z 417→254, m/z 167→43 and m/z 249→231, respectively. Ethyl acetate was used to extract the analytes in the plasma. Standard calibration curves of six analytes showed satisfactory linearity (r 2 ≥0.99 2) within the determined ranges. The lower limits of quantification were 0.5 ng/mL for paeoniflorin and liquiritin, 2.5 ng/mL for geniposide and saikosaponin b2 and 1.0 ng/mL for atractylenolide Ⅲ and paeonol, respectively. The intra-day and inter-day precision (RSD %) were all below 6.94 %, and the intra-day and inter-day accuracy (RE %) were within ± 6.10 %. The recovery and ME of six analytes were 85.99 %-98.10 % and 95.78%-108.06%, respectively. Additionally, the method we established in this experiment can be successfully used to study the pharmacokinetics of paeoniflorin and geniposide in beagle plasma., Competing Interests: Declaration of Competing Interest All authors declare no conflict of interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The quality evolution of 14 extra-virgin olive oils (EVOOs), with different initial polyphenol and oleic acid (64.6-77.7%) levels, was determined during real-time storage in dark conditions at room temperature for 22 months. EVOOs with low (<20-200 mg/kg), medium (450-700 mg/kg), and high polyphenolic levels (750-1400 mg/kg) were used. We found high correlations among peroxide values, K 232 , K 270 , and storage time. Oleuropein derivatives decreased by 98%, 89%, and 85% in EVOOs with low, medium, and high polyphenolic levels, respectively, with the highest depletion occurring in those with the lowest initial concentrations. Besides having higher α-tocopherol protection from oxidative phenomena, EVOOs with the highest phenolic fractions showed lower head space accumulations of off-flavour volatile substances and thus demonstrated the best retention of sensory and health benefits. We propose that the shelf life of EVOOs can be determined from their initial levels of oleuropein derivatives., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Preparations from the flowers or herb of the white dead nettle ( Lamium album L.) are recommended for the treatment of upper respiratory tract disorders or as a topical medication for mild inflammation of the throat, mouth, and skin. Taking into consideration the significance of L. album in traditional medicines across Europe, as well as the lack of studies describing the quantities of their most abundant constituents, we aimed to design a high-performance liquid chromatography coupled with diode-array detection (HPLC-DAD) method for potential standardization procedures of extracts from flowers of L. album . The HPLC-DAD method was developed and validated for quantification of iridoids (lamalbid), phenolic acids/depsides (chlorogenic acid), phenylpropanoids (verbascoside), and flavonoids (rutin; quercetin malonylhexoside; tiliroside) in aqueous and ethanolic-aqueous extracts of Lamii albi flos . The method was specific, accurate, and precise. Lamalbid was the most abundant compound both in aqueous (39.09 ± 1.02 m/g dry weight) and ethanolic-aqueous (26.66 ± 0.64 m/g dry weight) extracts. The quantities of selected compounds, except for chlorogenic acid and tiliroside, were higher in the aqueous extract than in the ethanolic-aqueous one. In conclusion, the method developed allowed for quantitation of compounds from different classes. In particular, chlorogenic acid and verbascoside have been proposed as reference compounds for routine quantitative control of Lamii albi flos .

The crude drug ysypó hû ( Adenocalymma marginatum DC., Bignoniaceae) is used traditionally by the Guarani of Eastern Paraguayan as a male sexual enhancer. The aim of the present study was to identify the main constituents of the crude drug and to evaluate the in vitro inhibitory activity towards the enzyme phosphodiesterase-5 (PDE-5). The main compounds were isolated by counter-current chromatography (CCC). The metabolites were identified by spectroscopic and spectrometric means. The chemical profiling of the extracts was assessed by high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS/MS). The crude extract and main isolated compounds were tested for their PDE-5 inhibitory activity using commercial kits. The iridoid theviridoside and 4-hydroxy-1-methylproline were isolated as the main constituent of the crude drug. Four chlortheviridoside hexoside derivatives were detected for the first time as natural products. Chemical profiling by HPLC-MS/MS led to the tentative identification of nine iridoids, six phenolics, and five amino acids. The crude extracts and main compounds were inactive towards PDE-5 at concentrations up to 500 µg/mL. Iridoids and amino acid derivatives were the main compounds occurring in the Paraguayan crude drug. The potential of ysypó hû as a male sexual enhancer cannot be discarded, since other mechanisms may be involved.

This paper discussed the process parameters optimization of partial least-square (PLS) modeling according to quality by design (QbD) concept. D-optimal design and online near-infrared (NIR) sensor were proposed to analysis the Geniposide in Gardenia jasminoides Ellis using above process parameters to achieve robustness PLS model. Four critical model parameters (CMPs) were identified to construct a D-optimal design, which included the selection of sample set, spectra pre-processing, latent variables and variable selection methods. NIR sensor dataset was obtained under a pilot scale system. The D-optimal design optimization strategy resulted in a robust PLS model with the optimal parameters, 1/2 samples for calibration sets through Baseline spectra pre-processing with SiPLS-selecting variables under 8 factors. The critical evaluation attributes (CEAs) of PLS model were recommended as follows: the RMSEC and R cal 2 of the calibration set were 0.005901 and 0.9983. The RMSEP and R pre 2 of the validation set were 0.02002 and 0.9845. The multivariate detection limit (MDL) was 1.143 × 10 -3 . Therefore, design space of CMPs which affected CEAs of PLS model was established. The result demonstrated that the proposed method was beneficial for the robustness of PLS model, which also showed a significant guideline for the design and development of PLS model., (Copyright © 2019. Published by Elsevier B.V.)

In recent years, a large number of biological properties and an important role in the organoleptic characteristics of olive oil have been attributed to phenolic secoiridoids, such as oleacein, oleocanthal, oleuropein aglycone and ligstroside aglycone. Consequently, quantifying them is of great interest for the olive oil sector. Currently, there is no consensus in which analytical method must be use to accurately determine these compounds in olive oil, mainly owing to the lack of reference standards for calibration. In this work, analytical standards of phenolic secoiridoids have been used to develop a quantitative and rapid analytical method by UHPLC-MS/MS, in which sample extraction is not carried out. Simple dilutions of the sample with dry tetrahydrofuran and dry acetonitrile were performed before analysing them. It is worth noting that under these conditions the generation of artefacts such as acetals and hemiacetals of the aldehydic forms is highly reduced. The detection and quantification was performed with a Xevo TQS tandem quadrupole mass spectrometer. The method was validated at four concentration levels and finally applied to six samples of extra virgin olive oil.

Duranta erecta L.is a valuable herb used in traditional system of medicine in India and China for the treatment of malaria and abscesses. The presence of phenylethanoid (e.g., acteoside as major bioactive) in addition to iridoid and flavonoids glycosides are mainly responsible for reported pharmacological properties. Although D. erecta is widely used in many Asian countries, the quality assurance of raw herb and its derived products are still in question, due to the lack of validated protocol. Current Indian guidelines for quality assurance of phytopharmaceutical product demands accurate determination of minimum four chemicals. Moreover, there is no chromatographic method to simultaneously analyse acteoside, isoacteoside, durantoside I, quercetin, and methylapigenin-7-O-β-D-glucopyranuronate in D. erecta. The present study establishes a systematic approach for the quality evaluation of D. erecta with efficient and validated UPLC-PDA method that simultaneously analyses the five bioactive marker chemicals in D. erecta. A fast, sensitive, and reliable ultraperformance liquid chromatography-photodiode array (UPLC-PDA) method was developed for the quantification five marker compounds-acteoside, isoacteoside, durantoside-I, quercetin, and methylapigenin-7-O-β-D-glucopyranuronate in D. erecta. The optimized UPLC conditions on Reverse phase column Phenomenex Luna® (C 18 , 2.5μm, 2.0×100mm)) and binary gradient elution resulted in best separation for all five targeted phytoconstituents. Validation characteristics viz. linearity (r 2 >0.999), accuracy (%RSD<2.0), precision (%RSD, 1.62-2.59), recovery (99-101%), and sensitivity (LOD- 0.27-0.40μg mL -1 ; LOQ- 0.90-1.35μgmL -1 ) were satisfying the ICH criteria. Uncertainty in the measurement was also estimated using method validation data and other sources to fulfil the technical requirement of ISO 17025:2017. It is the first validated method that provides the simultaneous and accurate analysis of five bioactive phytoconstituents of D. erecta in short time with defined traceability and accuracy profile. Further, it delivers a holistic quality analysis of raw medicinal herb and its preparation., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A simple gradient high-performance liquid chromatography with diode array detection (HPLC-DAD) method was used to simultaneously to analyze characteristics of six indicator compounds in the traditional Chinese medicine (TCM) formulation Wen-Qing-Yin (WQY). Separate optimization was performed using a Cosmosil C18 column gradient method with 0.1% formic acid in both mobile phases of aqueous and acetonitrile (ACN), at a flow rate, detection wavelength, and sample volume of 1.8 mL/min, 268 nm, and 10 μL, respectively. The linear regression of six active compounds berberine (BER), baicalin (BAI), ferulic acid (FER), geniposide (GEN), hydorxymethoxylfurfural (HMF), and paeoniflorin (PAE) was produced at the concentration range of 10-2000 μg/mL. The method validation revealed an acceptable precision (intra- and inter-day precision < 3.39% and 4.11%, respectively) and recovery (85.60-110.45% and 86.58-110.90%), a recovery range of 86.61-109.42%, and sensitivity (limit of detection [LOD] and limit of quantification [LOQ] values were in the range of 0.03-3.13, and 0.08-9.38 μg/mL, respectively) while the calibration curves were linear with a correlation coefficient (R 2 ) ranging from 0.9966 to 0.9989. The qualitative and quantitative analyses were performed by direct comparison of the peaks of the WCY extract to retention times of reference standards. Additionally, principal component analysis (PCA) successfully discriminated four purchased commercial samples of all six indicator constituents, and the present results indicate their comprehensive potential usefulness for qualitative and quantitative analyses of the WQY decoction and its commercial products., (Copyright © 2019. Published by Elsevier Taiwan LLC.)

Geniposide (GE) is an iridoid glycoside compound with anti-inflammatory effect. The potential of sphingosine 1-phosphate (S1P) as a plasma marker in human diseases was suggested recently in the literature, which demonstrated that, in patients with inflammatory diseases, plasma S1P was elevated. It follows that the obstructive coronary artery disease can be predicted with serum S1P. Therefore, S1P can also be potentially used as a pharmacodynamic marker to study adjuvant arthritis (AA) rats. In the current study, a UHPLC-MS/MS method combined with the microdialysis sampling technique (using FTY720 phosphate as an internal standard) was adopted and validated to measure S1P levels in the hemodialysis fluid and joint cavity dialysates of AA rats after oral administration of GE. A S1P concentration-time curve in the dialysate was established in this study. It was demonstrated that GE exerted an anti-inflammatory effect by reducing AA-induced elevated S1P levels. It is showed that changes in S1P concentrations over time can be used to monitor the pharmacodynamic effects of GE in treating AA rats in pharmacodynamic studies., (© 2019 John Wiley & Sons, Ltd.)

Amarogentin (AG) is one of the bitter secoiridoid glycosides, which exerts various pharmacological activities as a bitter stomachic. Recently, there is an increasing demand for AG-containing plants in Japan due to their use as folk medicines and food additives; hence, it is crucial to develop analytical techniques that are specific for AG. In this study, a new magnetic particles-based enzyme immunoassay (MPs-EIA) using a specific monoclonal antibody against AG (MAb 1E9) for the rapid determination of AG in plants of the family Gentianaceae was described. AG directly immobilized onto magnetic particles (MPs) was used as a competitor for free AG against MAb 1E9, thereby increasing the surface area of the solid phase and decreasing the immunoreaction time. In addition, the blocking step required in case of the conventional enzyme-linked immunosorbent assay could be avoided in the proposed MPs-EIA, which enables an even more rapid performance for the immunoassay. In the developed MPs-EIA, AG exhibited linearity in the range of 15.6-500 ng mL -1 , with a limit of detection of 8.58 ng mL -1 . Validation analysis revealed that MPs-EIA is a sufficiently sensitive and rapid for the quantitative analysis of AG in plant samples. To the best of our knowledge, this is the first MPs-EIA that has been applied to plant samples., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A new high performance liquid chromatography (HPLC) method has been established for quantitative and qualitative analysis of three tetracyclic iridoids: ML-2-3 (1), molucidin (2), and ML-F52 (3), which are responsible for anti-trypanosomal and anti-leishmanial activities of Morinda lucida Bentham leaves. Separation of 1-3 from dried 80% aqueous (aq.) ethanol extract was achieved on a reversed-phase cholester column packed with cholesteryl-bonded silica using an acetonitrile-0.1% aq. formic acid mobile phase system. Ultraviolet-visible (UV-VIS) spectroscopy was employed for detection of compounds, and their contents were determined by measuring absorbance at 254 nm. Depending on the above system, several factors potentially affecting the concentration of tetracyclic iridoids were evaluated resulting in several variation on plant organs, seasonality, variation between individual trees, and branch positions within the trees. Moreover, we developed a simple, quick, and effective method for tetracyclic iridoid isolation from M. lucida leaves that consisted of extraction by sonication into 80% aq. ethanol, basic hydrolysis, acid neutralization, liquid-liquid extraction into an organic solvent, and reverse phase open column chromatography. Employing this method, we have succeeded to obtain 1 as a colorless crystal yielding of 0.23%, which was 28 times higher than that of previous isolation method. Setting up methodology in this paper may be important for future in vitro and in vivo studies of tetracyclic iridoids and moreover for their applications in new drug design and development., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Inter- and intrapopulation variability in six natural populations of the rare species Gentiana pneumonanthe was examined based on morphological and chemical data. Population size and linear morphometric parameters differed significantly among populations, but without a clear connection to habitat conditions, i. e. water supply and light availability. Leaf shape varied from ovate to lanceolate in all populations, and one population was distinctive in having the largest number of leaves of transitional shape. HPLC analyses of six secondary metabolites were performed separately for belowground parts, and aboveground vegetative and reproductive parts of individual plants (6 populations ×7 individuals ×3 plant parts, n=126) in order to examine differences at the population and individual levels. Three secoiridoids (swertiamarin (SWM), sweroside (SWZ), and gentiopicrin (GP)), one xanthone (mangiferin (MGF)), and two flavones (isoorientin (IO) and isovitexin (IV)) were detected and quantified in the analyzed samples: sweroside dominated in the aboveground reproductive part, mangiferin in the aboveground vegetative part, and gentiopicrin in the belowground part. At the population level, differences in contents of the analyzed chemicals among populations were significant only for a few metabolites. At the individual level, a pronounced organ-dependent distribution of secondary metabolites was revealed. The results of this study contribute to a better understanding of natural variability within populations of the rare and threatened G. pneumonanthe, and provide data on the contents and within-plant distribution of secondary metabolites, which are important as pharmacologically active compounds and may be useful for further biotechnological procedures regarding this species., (© 2019 Wiley-VHCA AG, Zurich, Switzerland.)

Cornus officinalis-Rehmannia glutinosa herb couple is widely used herb medicine in clinical practice to treat chronic kidney disease (CKD). However, the in vivo integrated metabolism of its main bioactive components in CKD rats remains unknown. In this study, UPLC-Q-TOF/MS technique combined with Metabolynx ™ software, was developed and successfully applied for analysis of metabolic profiles of the bioactive components of the herb couple in normal and CKD rat biological samples. Main parent components of the herb couple extract such as loganin, morroniside and catalpol were absorbed into the blood circulation of the normal and CKD rats. Another parent component acteoside was almost completely degraded. Seventeen metabolites involved in the in vivo metabolism processes were tentatively identified. These metabolites indicated that loganin was mainly metabolized to the demethylated product, and morroniside was firstly deglycosylated to the aglycone and the latter was subsequently demethylated and acetylated. Additionally, hydrogenation and deglycosylation were the principal metabolic reactions of catalpol; while O-glucuronide and O-sulphate conjugates were observed as major metabolites for methylated caffeic acid and hydroxytyrosol released from acteoside. Compared with the normal group, the CKD rat showed lower conversion capability. Few kinds and minor amounts of the metabolites appeared in the CKD rat samples. While considerable amounts of the parent compounds were detected in the CKD plasma. This will help maintain a high blood drug concentration which might be beneficial for the treatment of CKD. The proposed method could develop an integrated template approach to analyze screening and identification of the bioactive components in plasma, urine and feces after oral administration of herb medicines. Additionally, this investigation might provide helpful chemical information for further pharmacology and active mechanism research on herb medicines., (Copyright © 2018. Published by Elsevier B.V.)

Iridoid blue-based pigments (IBBP) extract of Genipa americana L. represents a natural alternative additive for food applications and also exerts desirable biological effects on human health. In this study the iridoids present in the extract were identified, the influence of pH and temperature on color difference (ΔE) of IBBP was evaluated using a central composite design (CCD) and finally the antioxidant capacity was monitored before and after its in vitro digestion. Ten glucoside iridoids were detected and the main compounds identified were genipin, genipin 1-β-gentiobioside and geniposide. It was also observed an increase of 17-18% of antioxidant capacity after the in vitro digestion, respectively. Among the conditions tested, the color of extract was more stable at 12-20 °C and low pH (3.0-4.0), suggesting that it is compatible for coloring acidic foods. Finally, the in vitro digestion also increased the antioxidant capacity (ORAC assay) by 39%., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: Swertia nervosa (Wall. ex G. Don) C. B. Clarke, a promising traditional herbal medicine for the treatment of liver disorders, is endangered due to its extensive collection and unsustainable harvesting practices., Objective: The aim of this study is to discuss the diversity of metabolites (loganic acid, sweroside, swertiamarin, and gentiopicroside) at different growth stages and organs of Swertia nervosa using the ultra-high-performance LC (UPLC)/UV coupled with chemometric method., Methods: UPLC data, UV data, and data fusion were treated separately to find more useful information by partial least-squares discriminant analysis (PLS-DA). Hierarchical cluster analysis (HCA), an unsupervised method, was then employed for validating the results from PLS-DA., Results: Three strategies displayed different chemical information associated with the sample discrimination. UV information mainly contributed to the classification of different organs; UPLC information was prominently responsible for both organs and growth periods; the data fusion did not perform with apparent superiority compared with single data analysis, although it provided useful information to differentiate leaves that could not be recognized by UPLC. The quantification result showed that the content of swertiamarin was the highest compared with the other three metabolites, especially in leaves at the rooted stage (19.57 ± 5.34 mg/g). Therefore, we speculated that interactive transformations occurred among these four metabolites, facilitated by root formation., Conclusions: This work will contribute to exploitation of bioactive compounds of S. nervosa, as well as its large-scale propagation., Highlights: The roots formation may influence the distribution and accumulation of metabolites.

Extra virgin olive oils (EVOOs) containing more than 5 mg/20 g tyrosol, hydroxytyrosol, and their secoiridoids can be recognized by health claims related to the protection of blood lipids from oxidative stress. Therefore, a reliable, accurate, and standardized analytical procedure is needed to determine these markers of EVOO quality. In order to overcome the limitations of current methods, a detailed investigation of sample preparation and chromatographic conditions was performed by UHPLC-UV-HRMS. The use of a C18 fused-core column and nonacidified gradient elution provided single, sharp peaks for oleocanthal and oleacein, allowing their reliable quantitation in UV profiles. Positive- and negative-UHPLC-HRMS/MS characterization of methanolic extracts revealed the presence of dimethyl acetal, methyl hemiacetal, and monohydrate derivatives of all secoiridoids. These artifacts were formed in aqueous methanol, which is usually employed to extract and analyze EVOO phenols, making the HPLC profiles more complex and the measurements less accurate and reproducible. Acetonitrile proved to be a suitable solvent to avoid the formation of secoiridoid dimethyl acetals and methyl hemiacetals and to efficiently extract EVOO bioactive phenols. Finally, the phenolic contents of Italian EVOO samples were determined by UHPLC-UV analysis of acetonitrile extracts before (direct method) and after acid hydrolysis (indirect method). The results indicated that the use of tyrosol and hydroxytyrosol as reference standards allowed more accurate quantitative data to be obtained. Direct and indirect methods provided comparable levels of EVOO phenols, highlighting the usefulness of acid hydrolysis in routine analyses. The improved procedure defines the most reliable conditions to provide an analytical method with suitable accuracy and repeatability in the analysis of healthy and functional EVOO phenols.

A mass spectrometry-guided elution-extrusion countercurrent chromatography protocol was developed to separate chemical components from Cornus officinalis Sieb. et Zucc. In this study, ion current extraction, a mass spectrometry-based data postacquisition method, was utilized to boost the separation power and scope of countercurrent chromatography technique. As a peak repicking and denoising tool, ion current extraction was carried out to process the liquid chromatography with mass spectrometry and the countercurrent chromatography with mass spectrometry data. So the target components were reacquired in the created extracted ion current patterns with enhanced responses and diminished background noise, which facilitated the distribution constant determination (by liquid chromatography with extracted ion current) and the targets fractionation (by countercurrent chromatography with extracted ion current). Under the guidance of the extracted ion currents of the target components and with the support of elution-extrusion mode in the countercurrent chromatography separation, six pairs of minor iridoid isomers were obtained in shortened experimental duration. Besides, a reciprocal shifted symmetry plot was established to represent the elution-extrusion countercurrent chromatography chromatogram. The results demonstrated the capability of the ion current extraction-guided elution-extrusion countercurrent chromatography protocol in discovery, analysis, and fractionation of low-concentration and structurally similar chemicals from a complicated sample., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Olive leaves are rich in polyphenolic compounds that are known to have antioxidant, antimicrobial, and anti-inflammatory activities. Therefore, olive leaf extract (OLE) is considered as a natural supplement. In this study we evaluated the antibacterial and the anti-inflammatory effect of OLE and its individual phenolic components in vitro. Polymorphonuclear cells (PMNCs) were isolated from the whole blood using Histopaque solution and cultured in RPMI-enriched medium. Tumor necrosis factor α (TNFα) level was determined by ELISA after 24 h of lipopolysaccharide stimulation. The antibacterial activity of OLE was determined by well diffusion assay. We found a significant decrease in TNFα secretion level in PMNCs culture treated with OLE. Oleuropein is the only OLE component that has shown anti-inflammatory effects at a concentration of 20 μg/mL. Furthermore, OLE exhibited antibacterial activity against some gram positive bacterial strains; however, gram negative bacterial strains were resistant to OLE. Downregulation of TNFα secretion in PMNCs culture in response to OLE treatment indicates that this polyphenol-rich extract has an anti-inflammatory effect, and oleuropein is the major OLE component responsible for this effect. The antibacterial activity of OLE is limited to gram positive bacteria.

The aim of the study was to evaluate the effect of the different methods of preparation of the blue honeysuckle (Lonicera caerulea L. var. Kamtschatica) cv. 'Wojtek' and their influence on the iridoid content of fruit juices and fresh and freeze-dried pomaces. Compounds were identified by UPLC-ESI-qTOF-MS/MS in negative ion mode. Five iridoids - loganic acid, 7-epi-loganic acid 7-O-pentoside, loganin, sweroside, and secologanin - were determined in blue honeysuckle fruits and products. The results revealed no differences in iridoid content between the fresh pomace from skin and flesh, or between the juice from skin and flesh. A higher content of iridoids was obtained in juice from crushed than from whole berries, and in freeze-dried pomace from whole than from crushed berries. Freeze-dried pomace from blue honeysuckle can be an excellent source of bioactive iridoids., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Herbal medicines are widely used as therapeutic products in many countries. Fructus Corni, a traditional herb medicine, has been clinically used to cure chronic nephropathy for thousands of years. It could be converted by gut microflora in vivo to shape its pharmacological profiles. Thus, metabolic profiles of major active constituents in Fructus Corni extracts by gut microflora from rats in healthy and nephropathy state were firstly investigated in vitro by ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) in this study. According to the features of protonated ions, five metabolites (M1, M2, M3, M5 and M6) were found and preliminarily authenticated. Intestinal bacteria were capable of converting N0 (loganin) to its aglycone M1 (loganetin). The latter was further hydrogenated to the corresponding M2 (hydrogenated loganetin) and subsequently to M3 (hydrogenated and demethylated loganetin) by demethylation; While M5 (demethylated morronisid aglycone) and M6 (dehydroxylated morronisid aglycone) were identified as the two metabolites of N4 (morronisid) through demethylation and dehydroxylation. Gut microflora from healthy and nephropathy rats could degrade loganin and morronisid to the above metabolites. However, healthy rat intestinal bacteria showed more powerful degradation and much more amounts of M1 and M6 were obtained in their samples. Additionally, this work demonstrated that UPLC-Q-TOF/MS approach connected with MetaboLynx™ analysis software was rapid and reliable for screening and authentication of natural product metabolites., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The search for compounds with functional properties from natural sources has grown in recent years as people have developed healthier habits. Therefore, the aim of this study was to evaluate the extraction of bioactive compounds from various parts of unripe genipap fruit (Genipa americana L.) by using pressurized ethanol to verify which part of the fruit provides the greatest recovery of the iridoids genipin and geniposide. Two process variables were studied: temperature (50 and 80°C) and pressure (2, 12 and 20 bar). The whole fruit and the peel, mesocarp, endocarp, endocarp+seeds and seeds of the fruit were studied. The endocarp presented with the highest recovery of genipin (48.6±0.6mg/g raw material) and the extraction from the mesocarp allowed a greater recovery of geniposide (59±1mg/g raw material). The highest values of total phenolic content were obtained with mesocarp extracts. The endocarp and mesocarp extracts presented the highest antioxidant activity as measured by FRAP and DPPH. These results are promising and support the use of unripe genipap fruit as a source of iridoids and natural antioxidants., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A novel methodology is presented for the enhanced electrochemical detection of oleuropein in complex plant matrices by Graphene Oxide Pencil Grahite Electrode (GOPGE) in combination with a buffer modified with a Natural Deep Eutectic Solvent, containing 10% (v/v) of Lactic acid, Glucose and H 2 O (LGH). The electrochemical behavior of oleuropein in the modified-working buffer was examined using differential pulse voltammetry. The combination of both modifications, NADES modified buffer and nanomaterial modified electrode, LGH-GOPGE, resulted on a signal enhancement of 5.3 times higher than the bare electrode with unmodified buffer. A calibration curve of oleuropein was performed between 0.10 to 37 μM and a good linearity was obtained with a correlation coefficient of 0.989. Detection and quantification limits of the method were obtained as 30 and 102 nM, respectively. In addition, precision studies indicated that the voltammetric method was sufficiently repeatable, %RSD 0.01 and 3.16 (n = 5) for potential and intensity, respectively. Finally, the proposed electrochemical sensor was successfully applied to the determination of oleuropein in an olive leaf extract prepared by ultrasound-assisted extraction. The results obtained with the proposed electrochemical sensor were compared with Capillary Zone Electrophoresis analysis with satisfactory results., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Lider-7-tang, a medicine used for the treatment of respiratory diseases especially pneumonia and fever in Mongolian Traditional Medicine, was selected for this phytochemical and pharmacological study. The objectives of the study were to determine total biological active substances and analyze the effects of Lider-7-tang treatment in rats with acute lung injury (ALI). Quantitative determination of the total active constituents (phenolic, flavonoid, iridoid and alkaloid) of the methanol extract of Lider-7-tang was performed using Folin-Ciocalteu reagent, aluminum chloride reagent, Trim-Hill reagent, and Bromocresol green reagent, respectively. A total of fifty 8-10-week-old male Wistar rats (200-240 g) were randomized into three groups: control group, lipopolysaccharide (LPS) group (7.5 mg/kg) and LPS+Lider-7 group (90 mg/kg Lider-7-tang before LPS administration). The total content of alkaloids was 0.2±0.043%, total phenols 7.8±0.67%, flavonoids 3.12±0.206%, and iridoids 0.308±0.0095%. This study also evaluated the effects of Lider-7 on levels of inflammatory mediators by observing histopathological features associated with LPS-induced ALI. The rats pretreated with Lider-7 had significantly lower levels of IL-6 (at 3 and 6 h), and TNF-α (at 3, 6, 9, and 12 h). The current study showed that Lider-7 exerted a preventive effect against LPS-induced ALI, which appeared to be mediated by inhibiting the release of pro-inflammatory cytokines.

A cocultivation system of Astragalus membranaceus hairy root cultures (AMHRCs) and immobilized food-grade fungi was established for the enhanced production of calycosin (CA) and formononetin (FO). The highest accumulations of CA (730.88 ± 63.72 μg/g DW) and FO (1119.42 ± 95.85 μg/g DW) were achieved in 34 day-old AMHRCs cocultured with immobilized A. niger (IAN) for 54 h, which were 7.72- and 18.78-fold higher than CA and FO in nontreated control, respectively. IAN deglycosylation could promote the formation of CA and FO by conversion of their glycoside precursors. IAN elicitation could intensify the generation of endogenous signal molecules involved in plant defense response, which contributed to the significantly up-regulated expression of genes in CA and FO biosynthetic pathway. Overall, the coupled culture of IAN and AMHRCs offered a promising and effective in vitro approach to enhance the production of two health-promoting isoflavone aglycones for possible nutraceutical and pharmaceutical uses.

This study was aimed to investigate antioxidation and α-glucosidase inhibition of the bioactive compounds in three cultivars of blueberry ('Northland' (NL) from the hybrid blueberry (V. corymbosum L.×V. angustifolium Aiton), 'Britewell' (BW), and 'Gardenblue' (GB) from the rabbiteye blueberry (Vaccinium ashei Reade)). A total of eighteen anthocyanins were identified and quantified in Anthocyanins (ACNs), among which four acylated anthocyanins were exclusively found in Northland. The blueberry anthocyanin extracts (BAEs) were further measured the antioxidant activity and α-glucosidase inhibition effect. Northland BAEs exhibited significantly superior antioxidant activity compared with BAEs of other cultivars, and the antioxidant activity was correlated with the content of anthocyanins. However, α-glucosidase inhibition test showed that Britewell BAEs had the strongest inhibitory effect. BAEs were further separated into anthocyanin fraction (AF) and copigment fraction (CF). Fifteen phenolic acids and four iridoids were identified in CF. In terms of α-glucosidase inhibition effects, the CF from three cultivars could inhibit α-glucosidase in a dose-dependent manner, while the AF did not show significant inhibitory effects. The blueberry exhibits excellent antioxidant activity and α-glucosidase inhibition., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The aim of this study was to investigate the changes in the phenolic profile in different organs (buds, flowers, fruits and leaves) from Chemlali olive variety cultivated in the Center of Tunisia according to development stage. Phenolics recovery was carried out using aqueous methanol from freeze-dried powdered tissues. The extracts were then analyzed by using high performance liquid chromatography coupled to time of flight and ion trap mass spectrometry detectors. Qualitatively, secoiridoids, flavonoids, simple phenols, cinnamic acid derivatives and lignans were identified in the analyzed extracts. Quantitatively, floral buds showed the highest phenolic contents compared to the other tissues under study. The highest content of secoiridoids was observed for unripe fruit extract, whereas the highest content of flavonoids was registered for floral buds., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The analysis of the n-butanol extract of Linaria tingitana (BELT) by HPLC-DAD-ESI/MS allowed the identification of four iridoids structurally confirmed by NMR and ESI-MS/MS extensive experiments data. The subjection of BELT to the anti-inflammatory activity showed that it exhibited a concentration dependent stabilisation of HRBC membrane, inhibition of protein denaturation and nitric oxide scavenging effect in the in vitro process. These results were confirmed in the in vivo experiments which showed that BELT was found to be most pronounced at 200 mg/kg after carrageenan injection which significantly reduced the swelling in both early and late phases of carrageenan-evoked oedema, as well as a significant reduce at the accumulation of infiltrating cells, inhibition of the myeloperoxidase activity and suppressed the lipid peroxidation. These results were supported by the histological analysis which revealed the reduction of oedema and cells infiltration, this might be influenced by the synergistic action of the above isolated compounds.

Fructus Corni has been used for nourishing liver and kidney in clinical practice for many years. However, the in vivo integrated metabolism profile of its bioactive components remains unknown. In the present study, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry coupled with the MetaboLynxTM software was established for screening and identification of the metabolites of the loganin and the morroniside in normal and chronic kidney disease (CKD) rat plasma, urine and feces after oral administration of Fructus Corni extract. The results showed that 11 metabolites including taurine and glucuronide conjugates, deglycosylated, dehydrated, (de)hydroxylated, (de)methylated, acetylated products were tentatively detected in normal rat samples while only eight metabolites were obtained in the model samples. Two parent compounds were both absorbed into the blood circulation of the normal and CKD rats. However, the compounds in the CKD rat showed lower metabolic capability. Few kinds and minor amounts of the metabolites were appeared in the CKD rat plasma, urine and feces. While considerable amounts of the parent compounds were detected in the CKD plasma. This helped maintain a high blood drug concentration which might be beneficial for the treatment of CKD. These results will be helpful for the further investigation of the pharmacokinetic study of Fructus Corni in vivo., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Cultivated blueberry (Vaccinium corymbosum, Vaccinium angustifolium, Vaccinium darrowii, and Vaccinium virgatum) is an economically important fruit crop native to North America and a member of the Ericaceae family. Several species in the Ericaceae family including cranberry, lignonberry, bilberry, and neotropical blueberry species have been shown to produce iridoids, a class of pharmacologically important compounds present in over 15 plant families demonstrated to have a wide range of biological activities in humans including anti-cancer, anti-bacterial, and anti-inflammatory. While the antioxidant capacity of cultivated blueberry has been well studied, surveys of iridoid production in blueberry have been restricted to fruit of a very limited number of accessions of V. corymbosum, V. angustifolium and V. virgatum; none of these analyses have detected iridoids. To provide a broader survey of iridoid biosynthesis in cultivated blueberry, we constructed a panel of 84 accessions representing a wide range of cultivated market classes, as well as wild blueberry species, and surveyed these for the presence of iridoids. We identified the iridoid glycoside monotropein in fruits and leaves of all 13 wild Vaccinium species, yet only five of the 71 cultivars. Monotropein positive cultivars all had recent introgressions from wild species, suggesting that iridoid production can be targeted through breeding efforts that incorporate wild germplasm. A series of diverse developmental tissues was also surveyed in the diversity panel, demonstrating a wide range in iridoid content across tissues. Taken together, this data provides the foundation to dissect the molecular and genetic basis of iridoid production in blueberry.

Kigelia africana is a tree native to Africa, with a local employment in numerous fields, ranging from traditional medicine to cosmetics and religious rituals. Parts of the plant generally used are stem bark, fruits, roots and leaves. The fruits, which have a singular 'sausage' shape, are widely exploited by local folk, in particular for applications/products involving genito-urinary apparatus of both human genders. The scope of this work was to make a consistent chemical investigation on this plant species, in order to clarify and increase the information at present available in literature. To this aim, ethanolic extracts of K. africana fruits were analysed by high-performance liquid chromatography with photodiode array (HPLC-PDA) and electrospray-mass spectrometry (HPLC-ESI-MS) detection, revealing the presence of polyphenols and iridoids. The two detection systems used along with standard co-injection and comparison with previous reports, led to the identification and quantification of six phenolic compounds and three iridoids.

Olive leaves represent a quantitatively significant by-product of agroindustry. They are rich in phenols, mainly oleuropein, which can be hydrolyzed into several bioactive compounds, including hydroxytyrosol. In this study, water extract from olive leaves 'Biancolilla' was analyzed for polyphenol profile, DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity and protective effect on differentiated Caco-2 cells. The efficacy of two enzymatic treatments in promoting the release of bioactive phenols was investigated: a) enzymatic extract from Wickerhamomyces anomalus, characterized by β-glucosidase and esterase activities; b) commercial β-glucosidase. Composition and bioactivity of the resulting extracts were compared. The results showed that the yeast-treated extract presented hydroxytyrosol content and DPPH radical scavenging activity comparable to those obtained using commercial β-glucosidase; however, it was showed the additional presence of hydroxycinnamic acids. In experiments on Caco-2 cells, the leaf extracts promoted the recovery of cell membrane barrier at different minimum effective concentrations. The high specificity of W. anomalus enzymatic extract may represent an effective tool for the release of bioactive phenols from olive by-products.

Valeriana jatamansi Jones is an important medicinal plant and its quality is closely related to its region of origin. In the current study, we utilized a flexible and powerful strategy for comprehensive evaluation of the quality diversity for 15 regions in China. The method was based on a hybrid linear ion trap-Orbitrap mass spectrometry platform. For structure characterization, fragmentation patterns were detected by analyzing a series of standard compounds using data dependent multistage mass spectrometry acquisition. A fragment ion database for valepotriates was established, and the acquired data were high throughput filtered by fragment ion search for compound identification. For quantitative purposes, we normalized the mass spectrometry data of 15 samples using SIEVE 2.0 and the differences in composition were analyzed using principal component analysis combined with hierarchical clustering analysis. The results identified a total of 92 compounds from Valeriana jatamansi Jones. Samples from Dali, Kunming, and Baoshan have better qualities and concentrations of the main active constituents. To verify our strategy, we compared the valtrate, acevaltrate, and baldrinal contents using high-performance liquid chromatography with diode array detector. We developed and validated a comprehensive qualitative and quantitative analytical method to achieve quality control of Valeriana jatamansi Jones., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)