High-performance Liquid Chromatography Coupled With Mass Spectrometry Research Articles

Anti-breast cancer potential of Anonidium mannii (Oliv.) Engl. & Diels barks ethanolic extract: UPLC-ESI-QTOF-MS detection of anticancer alkaloids

Ethnopharmacological relevanceBreast cancer is a serious threat in low-income as well as developed countries. To face this, many herbal preparations are prescribed by traditional healers in Cameroon, among which is Anonidium mannii commonly called “wild soursop”. AimThis study was undertaken to assess the anti-tumor effect of A. mannii ethanolic extract on cancer cell growth and against DMBA-induced mammary tumors in rats. Materials and methodsThe well characterized MTT bioassay was used to assess the cytotoxic potential of A. mannii ethanolic extract in liver (HepG2), prostate (DU145 & PC3) and breast (MCF-7) cancer cell lines. Considering the fact that breast cells were the most sensitive to the extract, a 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast tumor rat model was used to assess the possible anticancer effect of A. mannii extract. Indeed, rats were treated with either tamoxifen (3.3 mg/kg BW) or A. mannii extract (16.5, 50 and 150 mg/kg BW) or vehicle (2% ethanol) for 20 weeks. Tumor incidence, tumor mass and volume, oxidative stress status in tumor as well as tumor histoarchitecture were evaluated. ResultsA 24 h incubation of tested cells with the A. mannii extract significantly slowed cell growth in a concentration-dependent manner with an interesting effect in breast cells (IC50 ~61.5 μg/mL). As compared to the DMBA rats, those treated with A. mannii extract (50 and 150 mg/kg) showed reduced breast tumor incidence (28%), tumor burden (95.34% at 50 mg/kg and 99.14% at 150 mg/kg) and tumor volume (~92%). A. mannii extract counteracted the high proliferation of terminal mammary ducts induced by DMBA, mainly at 50 mg/kg. Furthermore, the extract decreased MDA and nitrite levels but increased SOD activity in the mammary gland. High Performance Liquid Chromatography coupled with Mass Spectrometry (HPLC-MS) analysis detected potential anticancer and antioxidant alkaloids in A. manni extract, which are close to those found in Annona muricata. ConclusionThese results provide evidence on the in vitro and in vivo anticancer effects of A. mannii, and therefore support its use in traditional medicine system to fight against cancer.

Composition and profiling of essential oil, volatile and crude extract constituents of Micromeria inodora growing in western Algeria

The chemical constituents of the aerial parts of Micromeria inodora (Desf.) Benth. (Lamiaceae) collected in western Algeria, an aromatic shrub traditionally consumed as an herbal remedy, have been characterized. Secondary metabolites were extracted using sequential extractions with solvents at different polarity, and they were determined using an integrated nuclear magnetic resonance (NMR) and high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) approach. The composition of the essential oil obtained by hydrodistillation was characterized by gas chromatography-mass spectrometry (GC–MS), and the aroma profile was analysed by head-space solid phase micro extraction (HS-SPME) coupled with GC–MS. NMR and LC–MSn analyses revealed the presence of aglyconic and glycosylated flavonoids, phenylpropanoid derivatives and triterpenoid acids, mainly in the methanol, dichloromethane and n-hexane extracts. The analysis of the essential oil allowed to identify 66 components, mainly oxygenated mono- and sesquiterpenes (38.2 % and 32.0 %, respectively) and sesquiterpene hydrocarbons (10.8 %). The aroma profile revealed by HS-SPME-GC–MS was characterized by 45 volatile compounds, among which oxygenated monoterpenes appeared to be the most abundant (65.8 %), followed by sesquiterpene hydrocarbons (16.4 %) and monoterpene hydrocarbons (10.6 %). Overall, these data indicate that M. inodora could be a potential source of antioxidants and bioactive compounds, and they represent a starting point for further research on this plant species.

Stable Fe3O4 submicrospheres with SiO2 coating for heterogeneous Fenton-like reaction at alkaline condition

In the traditional Fenton process, the efficient generation of hydroxyl radical (HO) strongly relies on an acidic circumstance and the iron ions would precipitate and form large amounts of hazardous iron-containing sludge at alkaline pH. To realize stable heterogeneous Fenton-like catalytic degradation at alkaline condition, Fe3O4 submicrospheres with SiO2 coating were successfully synthesized by using water glass as the silica sources via a facile ultrasound assisted method. The as-obtained Fe3O4@SiO2 spheres were further used as catalysts for the Fenton-like degradation of tetracycline hydrochloride (TC). The Fe3O4@SiO2 submicrospheres exhibited superior catalytic activity in higher pH environment (pH value = 11), and the degradation efficiency toward TC was ca. 80% after ten successive runs. The kinetics for the catalytic degradation of TC were agreed well with the second-order kinetic model. The reaction rate constant (k) over the Fe3O4@SiO2 submicrospheres at a pH value of 11 was 7.69 times greater than that at a pH value of 3. Reactive species scavenging experiments revealed that HO and superoxide radical (O2− / HO2−) played a dominant role during the Fenton-like degradation of TC at pH 3 and pH 11, respectively. Possible Fenton-like degradation pathways of TC were proposed through the identification of intermediates using the high performance liquid chromatography coupled with mass spectrometry (HPLC-MS), which involved cleavage of methyl groups, N-dimethyl group, and hydroxy groups, ring-opening reaction, etc. The degradation efficiency of TC was close to 91.5% and total organic carbon (TOC) in solution was eliminated by about 41.4% at the optimized conditions. In a word, with the unique acidic surface properties and abundant Si-OH bonds, the Fe3O4@SiO2 submicrospheres exhibited well dispersion, good catalytic activity, strong alkali resistance and excellent recyclability in an ultrasonic-Fenton-like system.

Isolation of Microalgae from Mediterranean Seawater and Production of Lipids in the Cultivated Species.

Isolation and identification of novel microalgae strains with high lipid productivity is one of the most important research topics to have emerged recently. However, practical production processes will probably require the use of local strains adapted to commanding climatic conditions. The present manuscript describes the isolation of 96 microalgae strains from seawater located in Bay M’diq, Morocco. Four strains were identified using the 18S rDNA and morphological identification through microscopic examination. The biomass and lipid productivity were compared and showed good results for Nannochloris sp. (15.93 mg/L/day). The lipid content in the four species, namely Nannochloropsis gaditana, Nannochloris sp., Phaedactylum tricornutum and Tetraselmis suecica, was carried out by high performance liquid chromatography coupled to mass spectrometry (HPLC-MS ) highlighting the identification of up to 77 compounds.

Liquid chromatography and mass spectrometry for the analysis of acylglycerols in art and archeology.

Lipid characterization in art and archeology, together with the study of lipid degradation processes, is an important research area in heritage science. Lipid-based materials have been used as food since ancient times, but also employed as illuminants and as ingredients in cosmetic, ritual, and pharmaceutical preparations. Both animal and plant lipids have also been processed to produce materials used in art and crafts, such as paint binders, varnishes, waterproofing agents, and coatings. Identifying the origin of the lipid materials is challenging when they are found in association with artistic historical objects. This is due to the inherent complex composition of lipids, their widespread occurrence, and the chemical alterations induced by ageing. The most common approach for lipid characterization in heritage objects entails profiling fatty acids by gas chromatography/mass spectrometry after saponification or transesterification. New developments in high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) for the characterization of acylglycerols, together with more efficient sample treatments, have fostered the introduction of liquid chromatography for characterizing the lipid profile in heritage objects. This review reports the latest developments and applications of HPLC-MS for the characterization of lipid materials in the field of heritage science. We describe the various approaches for sample pretreatment and highlight the advantages and limitations of HPLC-MS in the analysis of paint and archeological samples. © 2020 John Wiley & Sons Ltd.

Identification of in vitro phase I metabolites of benzotriazole UV stabilizer UV-327 using HPLC coupled with mass spectrometry

The benzotriazole UV stabilizer (BUVS) 2-(5-chloro-benzotriazol-2-yl)-4,6-di-(tert-butyl)phenol (UV-327) is used in various plastic products to protect them against harmful UV radiation. Meanwhile, there are concerns about potential adverse health effects on humans, as residues of UV-327 and other BUVSs have already been detected in various environmental matrices. However, information on the metabolism of UV-327 is not yet available. Therefore, in vitro experiments with human liver microsomes (HLMs) were performed in order to identify phase I metabolites to be used as specific biomarkers of exposure in biomonitoring studies. The samples were analyzed by HPLC coupled with mass spectrometry (HPLC/MS). Potential metabolites, which were formed by hydroxylation and further oxidation to carboxylic acid, were tentatively identified. Special metabolite structures were suspected and custom-synthesized as reference substances for verification. In total, seven phase I metabolites, which may be suitable biomarkers for the assessment of exposure to UV-327, have been identified and quantified. The results of the present study provide initial insights into the metabolic pathway of UV-327, which is essential for further research on its human metabolism.

Box-Behnken experimental design for a green extraction method of phenolic compounds from olive leaves

Olive leaves are important olive by-products due to their high content of phenolic compounds and elenolic acids, which possess antioxidant, antimicrobial, anti‐atherogenic and anti‐inflammatory properties, among others. Thus, pressurized liquid extraction was used to obtain a high recovery of these compounds from olive leaves. A Box-Behnken design was performed to optimize the PLE conditions of temperature (50−200 °C), % ethanol-water (0–100 %) and extraction time (5−20 min) in order to obtain the highest content of simple phenols, secoiridois, flavonoids, elenolic acids, total compounds and extraction yield from olive leaves. Olive leaf extracts were analyzed by high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). The highest content of total compounds was 158.7 ± 0.4 mg g-1 d.w obtained at 105 °C, 100 % ethanol and 5 min, whereas the highest extraction yield was 43 ± 3% obtained at 198 °C, 100 % ethanol and 5 min. Therefore, multi-response analysis was carried out using desirability function to optimize the PLE conditions for both total compounds and yield. These optimal conditions were 138 °C, 100 % EtOH and 5 min obtaining 144 mg g-1 d.w. of total compounds and 42.2 % of yield. The optimal temperature of 138 °C has shown a great phenolic recovery and 100 % ethanol has been shown to be a safe solvent to use in the food industry. In addition, short extraction times (5 min) mean lower energy consumption and lower costs. Therefore, these PLE optimum conditions could be implemented on an industrial scale.

Bioremediation of di-(2-ethylhexyl) phthalate contaminated red soil by Gordonia terrae RL-JC02: Characterization, metabolic pathway and kinetics

Di-(2-ethylhexyl) phthalate (DEHP) is the most widely used plasticizer and a representative endocrine disrupting chemical. The toxicological effects of DEHP on environmental and human health have been widely investigated. In this study, the DEHP-degrading bacterial strain RL-JC02 was isolated from red soil with long-term usage of plastic mulch, and it was identified as Gordonia terrae by 16S rRNA gene analysis coupled with physiological and biochemical characterization. The biodegrading capacity of different phthalic acid esters and related intermediates was investigated as well as the performance of strain RL-JC02 under different environmental conditions, such as temperature, pH, salinity and DEHP concentration. Specifically, strain RL-JC02 showed good tolerance to low pH, with 86.6% of DEHP degraded under the initial pH of 5.0 within 72 h. The metabolic pathway of DEHP was examined by metabolic intermediate identification via a high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) analysis in which DEHP was hydrolyzed into phthalic acid (PA) and 2-ethylhexanol (2-EH) via mono (2-ethylhexyl) phthalate (MEHP). PA and 2-EH were further utilized through the protocatechuic acid metabolic pathway and β-oxidation via protocatechuic acid and 2-ethylhexanoic acid, respectively. The application potential of strain RL-JC02 was confirmed through the bioremediation of artificial DEHP-contaminated red soil showing 91.8% DEHP degradation by strain RL-JC02 within 30 d. The kinetics analysis of DEHP degradation by strain RL-JC02 in soil demonstrated that the process followed the modified Gompertz model. Meanwhile, the cell concentration monitoring of strain RL-JC02 in soil with absolute quantification polymerase chain reaction (qPCR) suggested that strain RL-JC02 survived well during bioremediation. This study provides sufficient evidence of a robust degrader for the bioremediation of PAE-contaminated red soil.

Network pharmacology and metabolomics study on the intervention of traditional Chinese medicine Huanglian Decoction in rats with type 2 diabetes mellitus

Ethnopharmacological relevanceType 2 diabetes mellitus (T2DM) is currently one of the most prominent and global chronic conditions. Huanglian Decoction (HLD) is a traditional Chinese medicine (TCM) preparation that has been used to treat T2DM for thousands of years in China. However, its mechanism of action at the metabolic level is still unclear.The purpose of this work is to study the mechanism of HLD in treating T2DM based on metabolomics and network pharmacology. Materials and methodsIn this study, metabolomics combined with network pharmacology was used to elucidate the therapeutic mechanism of HLD in T2DM. Serum samples were collected from rats with T2DM, induced by a high-sugar and high-fat diet combined with streptozotocin (STZ), to measure the levels of biochemical markers. Urinary metabolomics-based analysis using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was conducted to evaluate the differential metabolites from multiple metabolic pathways. ResultsAfter treatment with HLD for 4 weeks, biochemical indicators, including fasting blood glucose (FBG), blood lipid, fasting insulin (FINS), insulin sensitivity index (ISI), and homeostasis model assessment of insulin resistance (HOMA-IR), were significantly improved. Metabolomics results revealed that HLD regulated the biomarkers, such as cytosine, L-carnitine, betaine, phenylalanine, glucose, citrate, phenylpyruvate, and hippuric acid in glyoxylate and dicarboxylate metabolism, phenylalanine metabolism, and tricarboxylic acid (TCA) cycle. The combination of network pharmacology, metabolomics, western blot, and PCR showed that HLD can treat T2DM by enhancing the gene and protein expression levels of glucose transporter 4 (GLUT4), insulin receptor (INSR), and mitogen-activated protein kinase 1 (MAPK1) to interfere with glyoxylate and dicarboxylate metabolism. ConclusionsThe study based on metabolomics and network pharmacology indicated that HLD can improve T2DM through multiple targets and pathways, and it may be a useful alternative therapy for the treatment of T2DM.

Assessment of substrate biodegradability improvement in anaerobic Co-digestion using a chemometrics-based metabolomic approach

Anaerobic co-digestion (AcoD) can increase methane production of anaerobic digesters in plants treating wastewater sludge by improving the nutrient balance needed for the microorganisms to grow in the digesters, resulting in a faster process stabilization. Substrate mixture proportions are usually optimized in terms of biogas production, while the metabolic biodegradability of the whole mixture is neglected in this optimisation. In this aim, we developed a strategy to assess AcoD using metabolomics data. This strategy was explored in two different systems. Specifically, we investigated the co-digestion of wastewater sludge with different proportions of either grass or fish waste using untargeted High Performance Liquid Chromatography coupled to Mass Spectrometry (HPLC-MS) metabolomics and chemometrics methods. The analysis of these data revealed that adding grass waste did not improve the metabolic biodegradability of wastewater sludge. Conversely, a synergistic effect in the metabolic biodegradability was observed when fish waste was used, this effect being the highest for 25% of fish waste. In conclusion, metabolomics can be regarded as a promising tool both for characterizing the biochemical processes occurring during anaerobic digestion, and for providing a better understanding of the anaerobic digestion processes.

Protective effect assessment of Moringa oleifera against cadmium-induced toxicity in HCT116 and HEK293 cell lines.

The cadmium (Cd) is considered one of the widespread toxic metals in the aquatic and terrestrial environments, which is due to its long half-life, non-degradable characteristic, and toxicity. Aqueous extract of freeze-dried Moringa oleifera (Moringaceae family) leaves was examined for protective effect and antioxidant power against Cd toxicity. The results revealed that Moringa aqueous extract (MAE) has contents of total polyphenols and flavonoids about 30.14mg GAE/g and 18.35mg QE/g respectively. Furthermore, phenolic compounds in leaves of Moringa were studied using a high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). Results showed that the largest number of phenolic compounds determined in leaves of Moringa belongs to flavonoids. Moreover, biological properties were determined by radical scavenging capacity (DPPH) and ferric-reducing power (FRAP). Cytoprotective effect and antioxidant power of Moringa extract were assessed using the mitochondrial activity testing method (MTT test), malondialdehyde (MDA), and reactive oxygen species (ROS) production. Results indicate that Moringa aqueous extract have a significant (i) proliferative, (ii) antioxidant, and (iii) cytoprotective effect on HCT116 and HEK293 cells against metal toxicity.

Phenolic Profile, Toxicity, Enzyme Inhibition, In Silico Studies, and Antioxidant Properties of Cakile maritima Scop. (Brassicaceae) from Southern Portugal.

Cakile maritima Scop. (sea rocket) is an edible halophyte plant with several ethnomedicinal uses. This work reports the chemical profile and bioactivities of food grade extracts from sea rocket organs. Toxicity was determined on mammalian cells, and phenolic profiling and the quantitation of the main metabolites were made by high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS). Enzymatic inhibition was determined towards acetyl- and butyrylcholinesterase (AChE, BuChE), α-glucosidase, α-amylase, and tyrosinase. Docking studies were performed to tyrosinase, on the major metabolites, and samples were tested for antioxidant properties. Extracts were not toxic, were constituted mainly by flavonoids, and some compounds (roseoside and oleuropein) are here described for the first time in the species. The aerial organs’ ethanol extract had relevant activity towards 2,2-diphenyl-1-picrylhydrazyl [DPPH, half maximal inhibitory concentration (IC50) = 0.59 mg/mL], and ferric-reducing activity power (FRAP, IC50 = 0.99 mg/mL). All samples were more active towards AChE than on BuChE. The ethanol fruits’ extract inhibited α-glucosidase [2.19 mmol of equivalent of acarbose (ACAE)/g]. Samples were active against tyrosinase, especially the aerial organs’ ethanol extracts [25.9 mg of equivalent of kojic acid (KAE)/g]. Quercetin and kaempferol glycosides fit well into the enzymatic pocket of tyrosinase. Our results suggest sea rocket as a candidate to be further explored as a source of bioactive products.

Detection Method, Distribution, and Risk Assessment of Pharmaceuticals and Personal Care Products in the Yellow Sea and the East China Sea

At present, research findings on pharmaceuticals and personal care products (PPCPs) in coastal areas are still unclear, and there is a need to develop a method to detect more PPCPs simultaneously in seawater. In this study, nine compounds of non-steroidal anti-inflammatory drugs, antibiotics, lipid regulators. and stimulants were selected as analytes. Solid phase extraction (SPE) was used to extract the compounds, which were then analyzed by high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS). The optimum experimental conditions, such as the filler, eluent, pH, flow rate, and the reduction of matrix effect were optimized during the SPE. The results showed that the best extraction column was CNW HLB, the best eluent was methanol:acetonitrile (1:1, volume ratio), the best eluent volume was 6 mL, the best pH was 7, the best flow rate was 5 mL·min-1, the amount of EDTA-Na2 added was 1 g, and the best concentration multiple was 500. The linear regression equations of all PPCPs had good linearity. Correlation coefficients were>0.999, recovery rates were between 82%-106%, relative standard deviations were between 1.6%-14%, and detection limits were between 0.01-2 ng·L-1, thus satisfying the requirement of trace analysis in seawater. Distribution characteristics and sources of PPCPs were studied in the Yellow Sea and the East China Sea during summer 2018. All nine PPCPs were detected and the main pollutants were NAP, IBU, GEM, CAF, and ASA. High concentrations of PPCPs were generally detected in the nearshore area and displayed conspicuous decreasing tendencies from the inshore towards the offshore. The concentrations of PPCPs in the Yellow Sea were higher than of those in the East China Sea, and this was related to there being more sources of pollutions and poor water exchange capacity in the Yellow Sea. Principal component analysis showed that the main source of PPCPs was terrestrial input. The environmental risk assessment of PPCPs indicated that risk quotients (RQs) of IBU and NAP (0.1-1) posed a medium risk to the aquatic environment, while others posed low risk to organisms.

Encapsulation of β-alanine model amino-acid in zirconium(IV) metal organic frameworks: Defect engineering to improve host guest interactions

Metal-Organic Frameworks (MOFs) are porous coordination networks assembled through metal complexes with organic linkers. Due to their chemical versatility, these materials are being investigated for various applications including gas storage and separation, biomedicine and catalysis. The aim of this work is the encapsulation of the model β-alanine amino-acid in the nanostructured zirconium-based MOF (UiO-66) which contains the ligand H2BDC (1,4-benzenedicaboxylic acid). Additionally, ligand functionalization (by using H2doBDC (2,5-dihydroxy-1,4-benzenedicarboxylic acid) and defect engineering have been carried out to produce UiO-66 derivatives, in order to modify the host-guest interactions, and hence study their influence on the β-alanine loading capacity and release kinetics. The as-obtained materials have been characterized by X-ray diffraction (XRD), X-ray thermo diffraction (TDX), infrared (IR) spectroscopy, thermogravimetric analysis-differential scanning calorimetry (TG-DSC) and elemental analysis (EA). Morphology of nanoscale MOFs has been explored by transition electron microscopy (TEM). Adsorption isotherms have been constructed, and the concentration of β-alanine in the post-adsorption solution (supernatant) has been quantified by high performance liquid chromatography coupled with mass spectroscopy (HPLC-MS) and EA. Adsorption capacity values indicate that the presence of hydroxyl groups at the organic linker H2doBDC enhances the host-guess affinity between the framework and the adsorbate β-alanine. The influence of defect engineering, on the adsorption however, is not that obvious. On the other hand, desorption experiments show similar behaviour for H2doBDC-based derivatives. An adsorption mechanism has been proposed consisting of a combination of host-guest interaction at low concentrations, and covalent anchoring/ligand displacement by β-alanine at the inorganic clusters.

Acylation of o-Cresol with Pyromellitic Dianhydride

The acylation of o-cresol with pyromellitic dianhydride (PMDA) has been experimentally studied. The optimum conditions of the acylation reaction of o-cresol with PMDA have been determined. The synthesis has been performed in o-cresol melt at 120°C for 30 h at a reactant molar ratio of PMDA : o-cresol : SnCl4 = 1.0 : 4.1 : 2.0. The yield of 3,3,7,7-tetrakis(3-methyl-4-oxyphenyl)pyromellitide is 62.0% of the theoretical value. The structure of the target synthesis product has been proven by instrumental methods, namely, elemental analysis; thin layer chromatography (TLC); UV, IR, and 1H NMR spectroscopy; DSC; and high-performance liquid chromatography coupled with mass spectrometry (HPLC/MS). The revealed properties of 3,3,7,7-tetrakis(3-methyl-4-oxyphenyl)pyromellitide allow it to be recommended as a tracer for oil reservoir evaluation.

Gastroprotective effects of Erythrina speciosa (Fabaceae) leaves cultivated in Egypt against ethanol-induced gastric ulcer in rats.

Members of the genus Erythrina have been traditionally used in the treatment of various ailments such as inflammation and gastrointestinal disorders. Erythrina speciosa (Fabaceae) is a spiny, deciduous shrub or small tree native to Southern America in Brazil. It is cultivated in Africa and Asia. The traditional usage of E. speciosa indicated its antibacterial, analgesic, and anti-inflammatory activities. Evaluation of the phytochemical constituents, gastroprotective effects and possible mechanism of action of the ethyl acetate fraction obtained from the methanol extract of E. speciosa leaves (ESLE). Chemical characterization of ESLE was done using high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). The gastroprotective activity of ESLE was evaluated using ethanol-induced gastric-ulcer model in rats. Rats were pre-treated with ESLE 25, 50 and 100 mg/kg 1 h before the administration of absolute ethanol. Histological analysis, mucin content, and total acidity were evaluated. The possible mechanism of action of ESLE was studied through the examination of oxidative stress and inflammatory markers, PGE2, and NF-κB, iNOS, COX-2, and HSP-70 immunoexpression. In vitro, anti-Helicobacter pylori activity of ESLE was also studied using micro-well dilution method. Fourteen compounds were tentatively identified including alkaloids, flavonoids, and saponins. ESLE exerted a powerful gastroprotective effect. The pre-treatment with ESLE at different doses resulted in a significant reduction in gastric lesions and significant elevation in the mucin production. These effects could be partially mediated by the potent anti-inflammatory activity of ESLE as evidenced by the significant reduction in the immunoexpression of NF-κB, COX-2, iNOS and the reduction in the pro-inflammatory marker, TNF-α. ESLE counteracted the ethanol-induced oxidative stress by increasing the levels of depleted GSH and catalase as well as significantly attenuating the ethanol-induced lipid peroxidation tissue levels. In addition, ESLE exhibited in vitro antibacterial activity against H. pylori. The chemical constituents of ESLE strongly support its potent gastroprotective effect suggesting its future potential application in the management of gastric ulcer by eliminating its symptoms and causes including H. pylori.

Qualitative Fingerprint Analysis and Multidirectional Assessment of Different Crude Extracts and Essential Oil from Wild Artemisia santonicum L.

Artemisia species are used as folk medicines in several countries. This work was aimed to shed more light on the effect of methanol, water, ethyl acetate extracts, and essential oil (EO) of A. santonicum on selected enzymes (cholinesterase, tyrosinase α-amylase, and α-glucosidase) as well of their antioxidant and pharmacological effects. The chemical profile of the essential oil was determined using gas chromatography coupled to mass spectrometry (GC-MS) analysis, while the extracts were chemically characterized by high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). Forty-nine constituents were identified and camphor (36.6%), 1,8-cineole (10.2%), α-thujone (10.1%), borneol (4.5%), and β-thujone (3.6%) were the major components. Overall, 45, 74, and 67 components were identified from the ethyl acetate, methanol, and water extracts, respectively. The EO and extracts showed significant antioxidant properties, in a cell-free model; particularly, methanol and water extracts revealed promising sources of antioxidant compounds. Additionally, we evaluated protective effects of EO and extracts in isolated rat colon tissue challenged with lipopolysaccharide (LPS), as an ex vivo model of colon inflammation, and human colon cancer HCT116 cell line. Particularly, we observed that, among all tested samples, A. santonicum ethyl acetate displayed the best pharmacological profile, being able to blunt LPS-induced levels of all tested biomarkers of inflammation and oxidative stress, including colon nitrites, lactate dehydrogenase, prostaglandin E2, and serotonin. Additionally, this extract was also able to reduce HCT116 cell viability, thus suggesting potential antiproliferative effects against colon cancer cells. Based on our results, A. santonicum has great potential for developing novel functional agents including pharmaceuticals, cosmeceuticals, and nutraceuticals.

TiO2 and N-TiO2-photocatalytic degradation of salicylic acid in water: characterization of transformation products by mass spectrometry.

The aim of this work is to study the byproducts formed as a result of the photocatalytic process under different conditions of light wavelength and photocatalyst doping, rendering valuable information about the fate of pollutants for water treatment applications. Salicylic acid was selected as a model emerging pollutant and powders of nitrogen-doped titanium dioxide (N-TiO2) and TiO2 were prepared by the sol-gel process, using TiO2 P-25 Degussa as benchmark. Two light sources, UVA fluorescent tubes (372 nm) and blue LEDs (462 nm), were employed for photolysis and photocatalysis experiments. Transformation products formed during the process were studied by high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS). Major differences were found in the amount and identity of the transformation products due to the different light sources, detecting similar transformation products among the studied catalysts. Under UVA light, hydroxylated and carbonylated byproducts were the first intermediates to reach maximum abundances whereas presumed ring opening products were the last ones. On the other hand, under blue LED illumination byproducts accumulated with decreased mineralization. Photocatalytic degradation pathways were proposed based on the findings.

Determination of minimum inhibitory concentration and half maximal inhibitory concentration of antibiotics and their degradation products to assess the eco-toxicological potential

Numerous studies have been reported on today’s ubiquitous presence of antibiotics in surface waters causing the emergence of multidrug-resistant bacteria. Advanced water treatment procedures aim at the complete prevention and elimination of antibiotics from entering natural water bodies. In this study, photoinduced degradation processes using UVC-irradiation were applied toward selected fluoroquinolone, tetracycline, macrolide, and sulfonamide antibiotics. Photodegradation products were elucidated using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). Rate constants and quantum efficiencies were determined for the active substances and their photo-transformation products. As a measure of ecotoxicity, minimum inhibitory and half-maximal inhibitory concentrations were determined against the Gram-negative bacterium Pseudomonas fluorescens and the Gram-positive Bacillus subtilis in a standard assay format. These values were further recorded as a function of UV exposure time. The irradiation time-dependent increase of minimum inhibitory concentration (MIC) values agreed well with the kinetic models. After 10 min of irradiation, the compound solutions ceased to inhibit bacterial cell growth, indicating removal of the pharmaceutical activity. These findings were found in agreement with quantitative structure–activity relationship analysis. The combination of microbiological activity testing, molecular structure analysis, chemical kinetic investigation, and quantitative structure–activity relationship models proved to be able to predict irradiation times and evaluate potential ecotoxicological hazard of the irradiated drugs.

Determination of corticosterone from rat hair samples by HPLC-MS method

Abstract Corticosterone is an adrenocortical steroid hormone with glucocorticoid and mineralocorticoid effects. Based on previous studies, the plasma level of corticosterone correlates with the stress exposure of rodents. Because the half-life of corticosterone in blood is short, its plasma concentration can be used as an acute stress marker. But hair is accumulating the systemic and locally produced corticosterone, therefore it can be used to study chronic stress. However, the accurate quantification of corticosterone is an analytical challenge owing to the very low amount of hormone found in a complicated biological matrix. The high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) can provide the required selectivity and sensitivity for this purpose. Currently published methods for corticosterone quantification involve complicated sample preparation and long run time. Accordingly, the aims of the study were to simplify the extraction method of the corticosterone from rat hair samples and to develop an optimized HPLC-MS method for the accurate quantification. The rat hair samples were washed with methanol, dried and cut, then extracted with methanol at room temperature for 24 hours. The lipids were precipitated with formic acid aqueous solution and eliminated by centrifugation. The corticosterone was separated from other compounds with reverse phase chromatography using acetonitrile and 0,1% aqueous solution of formic acid as mobile phase. The detection was performed in positive SIM mode measuring the 347 m/z molecular ion. A six point calibration was performed in the range of 0,5-20 ng/ml, the accuracy was tested with quality control samples at two different concentration level. The total run time is only 4,2 minutes and the lower limit of quantification (LLOQ) is 0,5 ng/ml, with 10 pg absolute sensitivity. By determining the quantity of the hormone for a well-defined hair region, based on the speed of hair growth, we can characterize the retrospective stress exposure of the animals in different conditions.