Anti-Helicobacter pylori Activity of Six Major Compounds Isolated from Rumex acetosa.

Gastric problems are often caused by the well-known Helicobacter pylori (H. pylori) bacterium. One of the biggest obstacles to the treatment of H. pylori infections is increasing the antibiotic resistance. During our search for naturally derived anti-H. pylori compounds, six major compounds were isolated from the methylene chloride (CH2Cl2) and ethyl acetate (EtOAc) fractions of Rumex acetosa that showed anti-H. pylori activity. Three anthraquinones and three anthraquinone glucosides were identified as the major chemical constituents of the CH2Cl2 and EtOAc fractions, respectively. The chemical structures were identified to be emodin (1), chrysophanol (2), physcion (3), emodin-8-O-ß-d-glucoside (4), chrysophanol-8-O-ß-d-glucoside (5), and physcion-8-O-ß-d-glucoside (6) by UV, 1H NMR, 13C NMR, and mass spectrometry. Anti-H. pylori activity, including the minimum inhibitory concentration (MIC) value of each compound, was evaluated against two H. pylori strains. All isolates exhibited anti-H. pylori activity with different potencies, with an MIC value ranging between 3.13 and 25 µM. However, some variations were found between the two strains. While compound 5 displayed the most potent antibacterial activity with an MIC50 value of 8.60 µM and an MIC90 value of 15.7 µM against H. pylori strain 51, compound 1 exhibited the most potent inhibitory activity against H. pylori strain 43504. The two compounds also showed moderate urease inhibitory activity, with compound 1 demonstrating activity higher than that of compound 5. Furthermore, a molecular docking study revealed the high binding ability of compounds 1 and 5 to the active site of H. pylori urease. The present study suggests that the six anthraquinones isolated from R. acetosa with the whole parts of this plant may be natural candidates for the treatment of H. pylori infection. Further studies are required to determine the exact mechanism of action and to evaluate safety issues in the human body.

6-Oxofurostane and (iso)Spirostane Types of Saponins in Smilax sieboldii: UHPLC-QToF-MS/MS and GNPS-Molecular Networking Approach for the Rapid Dereplication and Biodistribution of Specialized Metabolites.

Identifying novel phytochemical secondary metabolites following classical pharmacognostic investigations is tedious and often involves repetitive chromatographic efforts. During the past decade, Ultra-High Performance Liquid Chromatography-Quadrupole Time of Flight-Tandem Mass Spectrometry (UHPLC-QToF-MS/MS), in combination with molecular networking, has been successfully demonstrated for the rapid dereplication of novel natural products in complex mixtures. As a logical application of such innovative tools in botanical research, more than 40 unique 3-oxy-, 3, 6-dioxy-, and 3, 6, 27-trioxy-steroidal saponins were identified in aerial parts and rhizomes of botanically verified Smilax sieboldii. Tandem mass diagnostic fragmentation patterns of aglycones, diosgenin, sarsasapogenin/tigogenin, or laxogenin were critical to establishing the unique nodes belonging to six groups of nineteen unknown steroidal saponins identified in S. sieboldii. Mass fragmentation analysis resulted in the identification of 6-hydroxy sapogenins, believed to be key precursors in the biogenesis of characteristic smilaxins and sieboldins, along with other saponins identified within S. sieboldii. These analytes' relative biodistribution and characteristic molecular networking profiles were established by analyzing the leaf, stem, and root/rhizome of S. sieboldii. Deducing such profiles is anticipated to aid the overall product integrity of botanical dietary supplements while avoiding tedious pharmacognostic investigations and helping identify exogenous components within the finished products.

Daphne jejudoensis Attenuates LPS-Induced Inflammation by Inhibiting TNF-α, IL-1ß, IL-6, iNOS, and COX-2 Expression in Periodontal Ligament Cells.

Periodontitis is a common disease involving inflammation and tissue destruction in the periodontal region. Although uncontrolled long-term inflammation in the gingiva may lead to loss of the periodontal ligament, treatments or preventive solutions for periodontitis are scarce. The aim of this study is to find anti-inflammatory material from a natural source that can be used to treat or protect against periodontitis. Daphne species (Thymelaeaceae) are important and popular components of traditional Chinese medicine and are used as anti-inflammatory agents. Daphne jejudoensis is an endemic plant that grows on Jeju Island and was identified as a new species in 2013. In this study, for the first time, we investigated the anti-inflammatory effect of D. jejudoensis leaf extract (DJLE) on human periodontal ligament cells. The gene expression levels of pro-inflammatory cytokines (interleukin-1ß and 6 and tumor necrosis factor-α) and inflammation-inducible enzymes (inducible nitric oxide synthase and cyclooxygenase-2) were reduced after DJLE treatment with/without lipopolysaccharide stimulation. The findings of this study indicate that D. jejudoensis possesses anti-inflammatory activities, suggesting that DJLE may be a potential preventive and therapeutic agent for periodontitis.

A Comprehensive Workflow for the Analysis of Bio-Macromolecular Supplements: Case Study of 20 Whey Protein Products.

The presence of bio-macromolecules as major ingredients is a primary factor in marketing many biologically derived macromolecular supplements. Workflows for analyzing these supplements for quality assurance, adulteration, and other supply-chain difficulties must include a qualitative assessment of small-molecule and macromolecular components; however, no such integrated protocol has been reported for these bio-macromolecular supplements. Twenty whey protein supplements were analyzed using an integrated workflow to identify protein content, protein adulteration, inorganic elemental content, and macromolecular and small-molecule profiles. Orthogonal analytical methods were employed, including NMR profiling, LC-DAD-QToF analysis of small-molecule components, ICP-MS analysis of inorganic elements, determination of total protein content by a Bradford assay, SDS-PAGE protein profiling, and bottom-up shotgun proteomic analysis using LC-MS-MS. All 20 supplements showed a reduced protein content compared to the claimed content but no evidence of adulteration with protein from an unclaimed source. Many supplements included unlabeled small-molecule additives (but nontoxic) and significant deviations in metal content, highlighting the importance of both macromolecular and small-molecule analysis in the comprehensive profiling of macromolecular supplements. An orthogonal, integrated workflow allowed the detection of crucial product characteristics that would have remained unidentified using traditional workflows involving either analysis of small-molecule nutritional supplements or protein analysis.

Comparative analysis of five Salvia species using LC-DAD-QToF.

Several Salvia species, commonly known as sage plants, are an integral part of various culinary and folklore preparations for the perceived wide range of effects from organoleptic to psychological. As a result, many of these species are an integral part of botanical drug applications, highlighting the need for accurate identification and quality control for consumer's safety. Five closely related Salvia species (S. officinalis, S. miltiorrhiza, S. divinorum, S. mellifera, and S. apiana) within a same botanical family were analyzed and differentiated using LC-QToF. Accurate mass measurement (<5 ppm) of protonated and deprotonated molecules together with resulting fragments and product ions allowed unequivocal or tentative identification of more than 180 compounds either by comparison with reference standards or literature data. The leaf part were identified based on various phenolic acids, flavonoids as well as di- and tri-terpenoids. Polyphenolics, viz., salvianolic A/B and rosmarinic acids in S. officinalis, lipophilic diterpenoids, viz., tanshinones in S. miltiorrhiza, abietatriene diterpenes and triterpenoids (ursane-/olean-type) in S. mellifera, and S. apiana were identified as characteristic, significant components. In comparison, salvinorins and divinorins representing a class of neoclerodane diterpenoids were detected only in S. divinorum. The presented methodology can successfully be applied to qualitatively assess sage-based ingredients in various finished products and formulations.

Protective Effect of Carotenoid Extract from Orange-Fleshed Sweet Potato on Gastric Ulcer in Mice by Inhibition of NO, IL-6 and PGE2 Production.

Ipomoea batatas (L.) Lam., Convolvulaceae is widely distributed in Asian areas from tropical to warm-temperature regions. Their tubers are known for their antioxidant, anti-bacterial, anti-diabetic, wound healing, anti-inflammatory, and anti-ulcer activities. The preventive and therapeutic effects of orange-fleshed sweet potato on gastric ulcers have not been investigated. In this study, the carotenoid extract (CE) of orange-fleshed sweet potato was found to protect against gastric ulcers induced by HCl/ethanol in mice. The anti-inflammatory and antioxidant activities of the carotenoid pigment extract were also evaluated as possible evidence of their protective effects. Administration of CE reduced gastric ulcers. Oral administration of CE (100 mg/kg) protected against gastric ulcers by 78.1%, similar to the positive control, sucralfate (77.5%). CE showed potent reducing power and decreased nitric oxide production in a mouse macrophage cell line, RAW 264.7, in a concentration-dependent manner. The production of the inflammatory cytokine interleukin-6 and prostaglandin E2 was also reduced by CE in a dose-dependent manner. The high carotenoid content of orange-fleshed sweet potato could play a role in its protective effect against gastric ulcers. This result suggests the possibility of developing functional products using this nutrient-fortified material.

Chemical Profiling and Characterization of Anthraquinones from Two Bulbine Species and Dietary Supplements Using Liquid Chromatography-High Resolution Mass Spectrometry.

BACKGROUND: Bulbine natalensis Baker and Bulbine frutescens (L.) Willd., belonging to the family Asphodelaceae, are widely distributed in South Africa and traditionally used as an aphrodisiac and skin remedies. OBJECTIVE: The aim of this study is to develop an analytical method for chemical profiling and identification of components in Bulbine species, which would be useful for herbal identification and understanding of the biological activity of B. natalensis in terms of safety and benefits to human health. METHOD: The anthraquinone-type compounds were structurally characterized from the extracts of dried stem and roots of Bulbine species and dietary supplements using liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QToF) with negative and positive ion electrospray. The calculated accurate masses of the protonated and deprotonated molecules and fragment ions were used for identification of the components from two Bulbine species. RESULTS: A total of 55 anthraquinone-type compounds, including 11 standard compounds, were identified in the crude extracts of two Bulbine species. Two Bulbine species and dietary supplements were clustered into different groups and possible chemical markers were identified. CONCLUSIONS: The developed analytical method provided a fast and economic method for quality assessment of Bulbine species in dietary supplements based on anthraquinone-type compounds. HIGHLIGHTS: This study reports holistic chemical profiling of Bulbine species using LC-QToF. The developed analytical method enabled non-targeted analysis of components in B. natalensis and B. frutescens, and is recommended for commercial and regulatory purposes.

Quantitative determination and characterization of polyphenols from Cissus quadrangularis L. and dietary supplements using UHPLC-PDA-MS, LC-QToF and HPTLC.

Stem and leaf of Cissus quadrangularis L. (Vitaceae), indigenous to Asia and Africa, were used for medicinal and dietary purposes with limited information about the plant's phytochemistry. Stem and leaf samples were assessed for the simultaneous determination of polyphenolic compounds (catechin, epicatechin, quercetin-3-O-ß-glucopyranoside, kaempferol-3-O-ß-glucoside, quercetin-3-O-ß-rhamnoside, leachianol F, amurensin A, pallidol, resveratrol, and quadrangularin A), using UHPLC-PDA-MS. The validation data showed that the method is precise, specific, accurate, and linear over the range of 0.5-100 µg/mL. Reversed-phase ultra-high performance liquid chromatography (UHPLC) fingerprints of the crude methanolic stem and leaf extracts of C. quadrangularis were obtained at different wavelengths based on their λmax. Polyphenolics were characterized using both UHPLC-PDA-MS and LC-QToF analysis. From liquid chromatography quadrupole time of flight-electrospray ionization mass spectrometry (LC-QToF) spectra, over 40 components were structurally correlated, and confirmation was based on the fragmentation characteristics and also from the information available in the literature. In addition to the LC-QToF method, a simple, fast HPTLC method was developed as a visual aid for the rapid qualitative analytical tool to help establish the quality assessment of botanical raw materials and dietary supplements claiming to contain Cissus.

Dibenzocyclooctadiene Lignans in Plant Parts and Fermented Beverages of Schisandra chinensis.

The fruit of Schisandra chinensis, Omija, is a well-known traditional medicine used as an anti-tussive and anti-diarrhea agent, with various biological activities derived from the dibenzocyclooctadiene-type lignans. A high-pressure liquid chromatography-diode array detector (HPLC-DAD) method was used to determine seven lignans (schisandrol A and B, tigloylgomisin H, angeloylgomisin H, schisandrin A, B, and C) in the different plant parts and beverages of the fruit of S. chinensis grown in Korea. The contents of these lignans in the plant parts descended in the following order: seeds, flowers, leaves, pulp, and stems. The total lignan content in Omija beverages fermented with white sugar for 12 months increased by 2.6-fold. Omija was fermented for 12 months with white sugar, brown sugar, and oligosaccharide/white sugar (1:1, w/w). The total lignan content in Omija fermented with oligosaccharide/white sugar was approximately 1.2- and 1.7-fold higher than those fermented with white sugar and brown sugar, respectively. A drink prepared by immersion of the fruit in alcohol had a higher total lignan content than these fermented beverages. This is the first report documenting the quantitative changes in dibenzocyclooctadiene-type lignans over a fermentation period and the effects of the fermentable sugars on this eco-friendly fermentation process.

Modulation of CYP3A4 and CYP2C9 activity by Bulbine natalensis and its constituents: An assessment of HDI risk of B. natalensis containing supplements.

BACKGROUND: Bulbine natalensis is an African-folk medicinal plant used as a dietary supplement for enhancing sexual function and muscle strength in males by presumably boosting testosterone levels, but no scientific information is available about the possible herb-drug interaction (HDI) risk when bulbine-containing supplements are concomitantly taken with prescription drugs. PURPOSE: This study was aimed to investigate the HDI potential of B. natalensis in terms of the pregnane X receptor (PXR)-mediated induction of major drug-metabolizing cytochrome P450 enzyme isoforms (i.e., CYP3A4 and CYP2C9) as well as inhibition of their catalytic activity. RESULTS: We found that a methanolic extract of B. natalensis activated PXR (EC50 6.2 ± 0.6 µg/ml) in HepG2 cells resulting in increased mRNA expression of CYP3A4 (2.40 ± 0.01 fold) and CYP2C9 (3.37 ± 0.3 fold) at 30 µg/ml which was reflected in increased activites of the two enzymes. Among the constituents of B. natalensis, knipholone was the most potent PXR activator (EC50 0.3 ± 0.1 µM) followed by bulbine-knipholone (EC50 2.0 ± 0.5 µM), and 6'-methylknipholone (EC50 4.0 ± 0.5 µM). Knipholone was also the most effective in increasing the expression of CYP3A4 (8.47 ± 2.5 fold) and CYP2C9 (2.64 ± 0.3 fold) at 10 µM. Docking studies further confirmed the unique structural features associated with knipholones for their superior inductive potentials in the activation of PXR compared to other anthraquinones. In a CYP inhibition assay, the methanolic extract as well as the anthraquinones strongly inhibited the catalytic activity of CYP2C9 while, inhibition of CYP3A4 was weak. CONCLUSIONS: These results suggest that consumption of B. natalensis may pose a potential risk for HDI if taken with conventional medications that are substrates of CYP3A4 and CYP2C9 and may contribute to unanticipated adverse reactions or therapeutic failures. Further studies are warranted to validate these findings and establish their clinical relevancy.

Quantification and Characterization of Phenolic Compounds from Northern Indian Propolis Extracts and Dietary Supplements.

BACKGROUND: Propolis is a resinous substance produced by bees. Propolis extracts have been used for anti-inflammatory and antimicrobial activities. The use of propolis dietary supplements has been increasing in the United States and the rest of the world. OBJECTIVE: A simple, economic, and valid analytical method is needed for quality assessment of dietary supplements and extracts claiming to contain propolis. METHODS: A ultra-high performance liquid chromatography (UHPLC) quadropole time-of-flight-MS method was used to characterize the chemical composition of northern Indian propolis. Fourteen major phenolic compounds were quantified using a UHPLC-DAD method. An HPTLC method was used to develop chemical fingerprinting profiles for propolis extracts and dietary supplements. The seven propolis extracts and 14 dietary supplements purchased in the U.S. were analyzed using the UHPLC-DAD-QToF method. RESULTS: Fifty-seven compounds belonging to phenolic, coumarin, fatty acid, and terpene classes were identified in propolis extracts. Based on quantification results, the content of 14 phenolic compounds in propolis extracts varied from 19-32% in dietary supplements, a significant variation to the recommended daily intake (0.2-94 mg/day). CONCLUSIONS/HIGHLIGHTS: The developed analytical methods can be used for quality assessment of propolis extracts and dietary supplements.

Chemical profiling and characterization of phenolic acids, flavonoids, terpene glycosides from Vangueria agrestis using ultra-high-performance liquid chromatography/ion mobility quadrupole time-of-flight mass spectrometry and metabolomics approach.

Vangueria agrestis is a shrub indigenous to tropical Africa, belonging to family Rubiaceae and is traditionally used as a decoction for treatment of fever, pain, and malaria. This study was undertaken to investigate the chemical constituents based on precursor exact mass and fragment ion information. The chemical profiling and structural characteristics of chemical constituents from methanolic extracts of dried aerial parts and roots of V. agrestis and dietary supplements were analyzed using ultra-high-performance liquid chromatography/ion mobility quadrupole time-of-flight mass spectrometry coupled with UNIFI platform and multivariate analysis in both negative and positive ion modes. A non-targeted ultra-high-performance liquid chromatography-mass spectrometry analysis was carried out to profile the chemical constituents of crude extracts of V. agrestis, and 73 compounds, including reference compounds, were identified. The fragments of flavonoids, monoterpene, and triterpene glycosides revealed the characteristic cleavage of glycosidic linkages, and the fragmentation pattern provided the identity of the sugars. This analytical method provides a quick method for quality assessment of dietary supplements. Finally, a chemometrics approach with multivariate statistical tools was used to visualize the differences between root and aerial parts of plant samples and to find the potential chemical markers that differentiate among these parts of V. agrestis samples and dietary supplements.

The scoop on brain health dietary supplement products containing huperzine A.

Context: Public health concerns are emerging surrounding huperzine A commonly found in dietary supplements. We sought to determine the actual content of products claiming to contain huperzine A and whether the ingredients on the supplement facts labels matched the analyses.Methods: We identified and analyzed 22 dietary supplement products listing huperzine A on product labels. We found these products were listed in Natural Medicines and Dietary Supplement Databases and being queried by Military Service Members for enhanced mental focus, alertness and energy. Analyses were conducted by using Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry.Results: Sixteen (73%) products had at least one ingredient claimed on the supplement facts label not detected through analysis. Compounds not reported on the label were detected in 16 (73%) products analyzed. Nine products (41%) listed ingredients not meeting the regulations for being a dietary supplement ingredient according to the FDA. Ingredients of most concern detected include stimulants: demelverine, 1,5-dimethylhexylamine, 1,3-dimethylhexylamine, N-phenethyl dimethylamine, halostachine, higenamine, noopept, ß-PEA, vinpocetine, sulbutiamine; and hordenine, currently on the FDA advisory list. Quantitative analysis showed the presence of huperzine A in the range from detected under the limits of quantification (DUL) to 267.1 µg/serving. Only two supplements showed huperzine A content within 10% of the declared amount.Conclusions: In a study of dietary supplements claiming to contain huperzine A, we found products that had at least one ingredient claimed on the supplement facts label not detected through analysis. Moreover, some ingredients not on the label could be dangerous and likely do not meet the definition of a dietary supplement ingredient according to the FDA. Quantitative analysis of huperzine A showed the amount detected was not in line with what appeared on the product label. Consumers should be aware of deceptive label claims and warned not to purchase products containing potentially dangerous ingredients.

Analysis of prenylflavonoids from aerial parts of Epimedium grandiflorum and dietary supplements using HPTLC, UHPLC-PDA and UHPLC-QToF along with chemometric tools to differentiate Epimedium species.

An UHPLC method was developed for the determination of 15 prenylflavonoids from aerial parts of Epimedium grandiflorum and related species (Berberidaceae). The separation was achieved using a reverse phased column and water/acetonitrile gradient as a mobile phase at a temperature of 40°C. The developed analytical method was validated for linearity, limits of detection (LOD) and limits of quantification (LOQ), stability and repeatability. The LOD and LOQ were found to be in the range from 0.1-0.5 µg/mL and 0.3-1 µg/mL, respectively. The wavelength used for quantification with the photodiode array detector was 269 nm. The total content of 15 prenylflavonoids was 9.1-20.6 mg/g for E. grandiflorum (except for sample #2899 and #20862), 5.6-35.4 mg/g for E. brevicornu and 10.8-30.5 mg/g for E. sagittatum. Twenty dietary supplements contained in the range from 0.1 to 81.7 mg/day. The developed method is simple, rapid and especially suitable for quality assessment of E. grandiflorum and dietary supplements containing E. grandiflorum. Liquid chromatography quadrupole time-of-flight-mass spectrometry (LC-QToF) is described for the identification and confirmation of compounds in plant samples and dietary supplements. This technique is also used for chemical profiling of Epimedium samples. This method involved the use of protonated ions in the positive ion mode and deprotonated ions in the negative ion mode with extracted ion chromatogram (EIC). Chemometric analytical tools for visualizing the plant and commercial samples quality were used for discriminating between Epimedium species and dietary supplements with regards to the relative content or presence of components. A HPTLC method was also developed for the fast chemical fingerprint analysis of Epimedium species.

Development and Validation of a UHPLC-PDA-MS Method for the Quantitative Analysis of Anthraquinones in Bulbine natalensis Extracts and Dietary Supplements.

A UHPLC-photodiode array-MS method was developed and validated for the quantification of one chromone and six anthraquinone type of compounds from Bulbine natalensis plant samples and dietary supplements. Metabolites 1:  -  7: were identified based on their retention times and electrospray ionization-MS spectra compared with a mix of previously isolated compounds. The quantification of 1:  -  7: was based on photodiode array detection. The optimized separation was achieved using a CORTECS C18 column with a gradient of water/acetonitrile as the mobile phase. Seven compounds were separated within 15 minutes with detection limits of 50 pg on the column. The analytical method was validated for linearity, repeatability, accuracy, limits of detection, and limits of quantification. The relative standard deviations for intra- and inter-day experiments were less than 5% and the recovery efficiency was 98 - 101%. Nine dietary supplements labeled as containing B. natalensis were examined. Anthraquinone-type compounds were detected in only five out of nine dietary supplements, with the total amount ranging from 11.3 to 90.4 mg per daily dose. The analytical method is simple, economic, rapid, and can be applied for quality assessment of B. natalensis and dietary supplements. Electrospray ionization-MS was used for the identification of these compounds in plant samples and dietary products.

Anthraquinone-Based Specialized Metabolites from Rhizomes of Bulbine natalensis.

The rhizomes of Bulbine natalensis furnished six previously unreported anthraquinone derivatives (1-6), together with eight known specialized metabolites. Their structures were determined by interpretation of 1D and 2D NMR and HRESIMS data. The absolute configurations of compounds 1-6 were determined by specific rotation and circular dichroism experiments. The isolated compounds were evaluated for antimicrobial activities, and compound 1 was found to be a moderate inhibitor (IC50 0.02 µM) against methicillin-resistant Staphylococcus aureus (MRSA).

Liquid chromatography-quadrupole time of flight mass spectrometric method for targeted analysis of 111 nitrogen-based compounds in weight loss and ergogenic supplements.

The use of supplements for weight loss and in sports as pre-workout (ergogenic) products is widespread. Many of these supplements were found to contain active components, which were not claimed on the products labels. A validated liquid chromatography high-resolution mass spectrometry quadrupole time-of-flight (LC-QToF-MS) method was developed for the simultaneous analysis of 111 amine-based compounds belonging to ergogenics, anorectics and other active components including phenethylamines (amphetamines, ephedrines), sibutramine or yohimbine. This method involves the detection of [M+H]+ ions and the separation was achieved using a C18 column, water/acetonitrile gradient as the mobile phase. The method was validated for linearity, repeatability, accuracy, stability, system suitability, limits of quantification (LOQ) and limits of detection (LOD). The limits of detection were in the range from 0.001-0.5 µg/mL. The validated method was applied to the analysis of twenty-seven weight loss and ergogenic dietary supplements. Two-thirds of the supplements contained compounds that were not listed on the product's label. These include several phenethylamines (PEA) such as demelverine, hordenine, N, N-dimethyl-phenethylamine, synephrine, N-methyl-ß-phenethylamine, and methylsynephrine. In addition, the PEA mimics such as dimethylamylamine, dimethylbutylamine other stimulants including fursultiamine, evodiamine, phenibut and theophylline were also observed. One or more of the ingredients listed on the labels were not detected in forty-four percent of the products analyzed. Positive identification was based on retention time, accurate mass and fragment ions in comparison with the respective reference standards. Development of such methods is anticipated to be of aid to regulatory agencies for the identification of undeclared exogenous components that are found in many dietary supplement products.

A new anti-Helicobacter pylori juglone from Reynoutria japonica.

A 70% ethanol extract from the root portion of Reynoutria japonica afforded one new and three known juglone derivatives, namely, 2-methoxy-6-acetyl-7-methyljuglone (1), 2-ethoxy-6-acetyl-7-methyljuglone (2), 2-methoxy-7-acetonyljuglone (3), and 3-acetyl-7-methoxy-2-methyljuglone (4) together with two phenolics (5 and 6), an anthraquinone (7), a stilbene (8) and a phthalide (9). Their structures were elucidated on the basis of comprehensive spectroscopic studies including IR, MS, and 1H, 13C, 2D NMR spectra. Compound 3 is a new compound in nature, and compounds 4-6 have been isolated for the first time from R. japonica. The isolates were evaluated for their antibacterial activity against three strains (43504, 51, and 26695) of Helicobacter pylori. The four isolated juglone derivatives (1-4) showed potent growth inhibitory activity. Among them, compounds 1-3 exhibited stronger inhibitory activity than those of the positive controls, juglone and metronidazole, for the three strains and that of another reference, clarithromycin, for the 43504 and 51 strains. Specifically, the new juglone compound 3 displayed the most potent antibacterial activity against all three strains, 43504, 51, and 26695, with MIC values of 0.06, 0.06 and 0.13 µM, respectively, and MIC50 values of 0.14, 0.11 and 0.15 µM, respectively.

Decaffeinated Green Tea Extract Does Not Elicit Hepatotoxic Effects and Modulates the Gut Microbiome in Lean B6C3F1 Mice.

The main purpose of this study was to investigate the hepatotoxic potential and effects on the gut microbiome of decaffeinated green tea extract (dGTE) in lean B6C3F1 mice. Gavaging dGTE over a range of 1X-10X mouse equivalent doses (MED) for up to two weeks did not elicit significant histomorphological, physiological, biochemical or molecular alterations in mouse livers. At the same time, administration of dGTE at MED comparable to those consumed by humans resulted in significant modulation of gut microflora, with increases in Akkermansia sp. being most pronounced. Results of this study demonstrate that administration of relevant-to-human-consumption MED of dGTE to non-fasting mice does not lead to hepatotoxicity. Furthermore, dGTE administered to lean mice, caused changes in gut microflora comparable to those observed in obese mice. This study provides further insight into the previously reported weight management properties of dGTE; however, future studies are needed to fully evaluate and understand this effect.

Quantitative determination and pharmacokinetic study of fusaricidin A in mice plasma and tissues using ultra-high performance liquid chromatography-tandem mass spectrometry.

Fusaricidins are a family of cyclic lipodepsipeptides that convey antifungal and antibacterial activity. Fusaricidin A (FA) is one of the Fusaricidins major compounds and it is showing promising activity against fungi and bacteria. In the present study, a fast and sensitive ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed for the analysis of FA in mice plasma, liver, kidney and brain tissues. The instrument was operated in positive electrospray ionization mode. Multiple reaction monitoring (MRM) mode was performed with ion pairs of m/z: 883.5→256.3, 883.5→197.2 and 883.5→72.1 for FA. The method was validated for linearity, repeatability, accuracy, stability, limits of detection (LOD) and limits of quantification (LOQ). The LOD and LOQ were 0.01 and 0.05 ng/mL for plasma and tissues, respectively. The calibration curve (10-200 ng/mL) was linear ( r2 = 0.99). Precision and accuracy values were found to be < 10% (within acceptable limit). The pharmacokinetic and tissue distribution characteristics of FA were determined in plasma, liver, kidney and brain of CD1 mice after I.V. administration of a single dose of 15 mg/kg body weight. Highest plasma concentration (Cmax) was calculated to be 4169.97 ± 50 ng/mL with a tmax of 0.08 h. The plasma clearance rate of FA was 397.6 ± 203 mL/h with a t1/2 of 2.2 ± 0.5 h and apparent volume of distribution during the terminal phase (Vz) of 979.2 ± 318 mL. The highest tissue concentration (Cmax) was found in the liver (219 ± 14 ng/mg) at a tmax of 0.08 h followed by the kidneys (38.6 ± 16 ng/mg) at tmax of 0.2 h. FA was poorly distributed to the brain with a Cmax of 0.45 ± 0.2 ng/mg and a tmax of 0.08 h. The method for quantitative analysis and pharmacokinetic data provided will support the development of various formulation approaches and therapeutic application for future clinical studies.