Chromatographic Studies Research Articles

Physicochemical, Phytochemical, NMR, FTIR and LC-MS Analysis of Extract of Ripe Fruits of Carissa carandus L.

Background: Carissa carandus L., commonly known as Karonda, is renowned for its distinctive tangy flavor, making it a valuable ingredient in culinary applications. Additionally, its extensive array of phytoconstituents renders it a significant component of traditional Indian herbal medicine, where it has been used to address various health conditions. Despite its availability, palatability, and diverse phytochemical profile, Karonda remains under utilized, and lacks substantial scientific validation. Further research is needed to substantiate its therapeutic potential and elevate its status in modern scientific and medical contexts. Methods: An ethanolic extract was prepared from the ripe fruits using a hot continuous extraction method. Phytochemical screening of the extract identified the presence of tannins, flavonoids, phenolic compounds, saponins, steroids, and carbohydrates. Subsequent analysis using spectrometric and spectro chromatographic techniques, including NMR, FTIR, and LC-MS, were conducted, supplemented by chromatographic studies, to further elucidate the extract's chemical profile and validate the presence of these bioactive compounds. Results: The analysis of the ripe fruit extract identified several functional groups, including O-H, N-H, C=O, C-N, C-H, and –COOH, based on IR absorption bands observed in the high wave region at 3850 cm−1 and 1728 cm−1. Active compounds were further characterized by comparing these findings with standard reference charts. Proton environments and their electronic states were examined using 1H-NMR spectroscopy. The 1H-NMR spectrum revealed signals at specific δ ppm values corresponding to R-CH3, R-CH2-R, RO-C-H, and F-C-H groups, as detailed in the accompanying table. Conclusion: These results provide a comprehensive chemical fingerprint of CA fruits offering scientific validation of its chemical nature and supporting its potential therapeutic applications.

Phyotochemical Analysis of Bridelia scandens Willd a folk lore herbal drug a Review

Analytical study deals with the modern pharmaceutical analysis of the leaves of the drug. Here, organoleptic characters, physico-chemical parameters, UV spectrophotometric study, chromatographic study which were carried out for the standardization of both leaf powder and leaf oil samples are documented. The leaf powder sample of the plant yielded moisture, 10.1`%, ash, 6.65%, water soluble extractive 16.5% and methanol soluble extractive 22.3%. Whereas the leaf oil sample showed specific gravity 0.9650; refractive index 1.473; acid value 1.17; saponification value 241.4; ester value 240.23 and unsaponifiable matter 0.55%. UV spectrophotometric study of the leaf powder sample showed three absorptions peaks: the absorption being maximum at 218 nm followed by 272 nm and 666 nm. The spectra of the unsaponifiable matter of the leaf oil sample also showed three absorption peaks at 209 nm, 235 nm and 287 nm. Comparison of these two spectra reveals that their pattern is different. The chromatogram of both the samples obtained in day light showed 8 spots in the leaf powder sample and no spots in the other sample. On viewing under short wave UV radiation, the leaf powder sample revealed 3 spots and the other sample 2 spots. Among the spots the spot at Rf 0.66 was present in both the samples. After exposing to Iodine vapour almost all the spots viewed in day light were revealed in the first sample and 5 spots were found in the second sample. On spraying with vanillin sulphuric acid 6 spots were revealed in leaf powder sample and 5 spots in second sample. Among these 2 spots at Rf 0.46 and 0.66 were common in both the samples.

Development of a liquid chromatography method for the analysis of oil extract of artemisia cina

Introduction. The problem of development and implementation of own drugs in Kazakhstan today is acute and urgent. The solution to this problem is possible through the use of own resources – domestic medicinal plant raw materials. A promising medicinal raw material is Artemisia cina – an endemic medicinal plant of the of the Turkestan region of Kazakhstan.Aim. Designing a liquid chromatography methodology to analyze the oil extract obtained from Artemisia cina.Materials and methods. Standard sample (SS) of santonin, Artemisia cina’s oil extract. The substance was identified and quantitatively determined using an Agilent Technologies 1200 liquid chromatograph (USA) equipped with ChemStation software.Results and discussion. The main active substance of the oil extract was analyzed using high-performance liquid chromatography (HPLC) method. Choosing the separation conditions for the column involves determining the best composition of the mobile phase and the rate of elution. There were used a reversed-phase system with sorbent C18 and a mobile phase consisting of acetonitrile and a phosphate buffer with a pH of 6.8. Chromatographic studies of the substance were carried out in a gradient mode at an analytical wavelength of 237 nm. The retention time of the standard sample (SS) of santonin matched the retention time (tR) of santonin isolated from the oil extract of Artemisia cina and amounted to 14.3 minutes.Conclusion. A liquid chromatography method has been established for the identification and quantitative determination of the oil extract, focusing on the main active substance, santonin.

Targeted isolation of extracellular vesicles from cell culture supernatant using immuno-affinity chromatography

Extracellular vesicles (EVs) have emerged as promising therapeutics with broad clinical applications as diagnostic biomarkers and therapeutic drug delivery systems. Yet, these biopharmaceuticals pose a challenge in terms of manufacturing due to their complexity and heterogeneity. Despite advancements in the field, current purification technologies lack scalability and/or selectivity. Affinity chromatography (AC) − coupling unmatched specificity and scalability − could be used to simplify purification processing and generate clinical-grade EVs with higher titers and purity.In the present work, we report the implementation of an immuno-AC resin to capture and purify EVs directly from clarified cellular feedstocks. Firstly, to guide and support marker selection, vesicle phenotype characterization was conducted using single particle interferometric reflectance image sensing (SP-IRIS) coupled with immunofluorescence. CD81 was the marker which shown to be more present and more likely to have the other markers (CD63 and CD9). Thus, anti-CD81 VHH ligand was generated and evaluated towards recombinant CD81 protein and CD81 bearing EV particles using surface plasmon resonance (SPR). Different chromatographic studies with Anti-CD81 ligand immobilized onto agarose beads resin were conducted to optimize the process parameters (residence time, dynamic binding capacity and impurity clearance). At residence time of 2 min, on average 40 % of pure triple tetraspanin-positive EV fraction was recovered. The enrichment in EV particles herein obtained, based on scale-up calculations, it would be possible to produce 1 × 1013 EVs from a 1L cell culture, while meeting impurity requirements in a single-step purification process (impurity removal over 2 log reduction value). A single-step purification process is possible, enabling the successful isolation of homogeneous EVs population, counting with a final HCP titer of 60 ng/mL and 9 ng/mL of dsDNA impurities. EV’s morphological integrity and internalization ability were also demonstrated, showcasing elution’s efficiency under mild conditions.Overall, this work contributes to the development of a novel, highly specific, AC technology using a camelid-derived affinity ligand which, bridging the scalability requirements demanded of large-scale production, could potentiate the advent of EV-based therapies.

NOVEL METHOD FOR DETERMINATION OF POMALIDOMIDE IN HUMAN PLASMA BY ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY MASS SPECTROSCOPY / MASS SPECTROSCOPY

The current investigation studied and verified an accurate Ultra performance liquid chromatography mass spectroscopy/ mass spectroscopy technique for the high-throughput detection of pomalidomide in human plasma with celecoxib as an internal standard. Chromatographic studies were carried out on Hypersil BDS (50 mm x 4.1 mm, 5m) with an isocratic mobile phase of acetonitrile: ammonium formate (80:20, V/V), at a flow rate of 0.5 mL min-1 and a total run time of 3 min. A triple quadruple Tandem Mass spectroscopy with electrospray ionization in positive mode was utilized to identify the analyte. Using multiple reaction monitoring modes and precursor-to-product ion transitions of 274.43 m/z for pomalidomide and 382.12 m/z for standard, respectively, pomalidomide and internal standard were measured. The developed technique was verified as per the regulatory standards for bioanalytical procedures validation. Other validation results also match.

Electrokinetics of Nitrite to Ammonia Conversion in the Neutral Medium Over A Platinum Surface.

Polycrystalline Pt electrode was employed to selectively convert nitrite ions ( ) into useful nitrogenous compound through electrochemical reduction reaction in neutral medium. According to adsorptive stripping analysis, the reduction process produced nitric oxide (NO) on the surface of Pt electrode. The spectroscopic test and gas chromatographic studies discovered the presence of ammonia (NH3) in the electrolyzed solution, suggesting the transformation of adsorbed NO into NH3 during the reverse scan. Scan rate dependent investigation was performed to elucidate kinetic information relating to this reaction on Pt surface. From Ep vs scan rate (υ) and jp vs υ (logarithmic plot), it was found that the conversion of ion into NO is an irreversible reaction which relies on the diffusion of ions to electrode surface. The Tafel analysis unveiled that the first electron transfer sets the overall reaction rate, having formal reduction potential, E0'=-0.46 V and standard heterogeneous rate constant, k0= cm s-1. Reductive transfer coefficient (α) is another kinetics parameter, which was found to be approximate 0.77 from the difference between Ep and Ep/2 of the voltammograms obtained over scan rate range 0.005 V s-1 to 0.250 V s-1, indicating a stepwise process. According to temperature-dependent voltammograms, the nitrite reduction reaction on Pt had a calculated activation energy of about 19.8 kJ mol-1 and a pre-exponential factor of about 8.39×103 mA cm-2.

Synthesis of phosphinic structural analogue of Met-Glu-His-Phe

The synthesis of a phosphinic structural analogue of the tetrapeptide Met-Glu-γ-His-Phe by adding the dipeptide component His-Phe to the adamantyl ester of the phosphinic pseudo-Met-[P]-Glu-peptide in the form of cyclic glutamate anhydride is proposed. The conditions for the interaction of phosphinic pseudo-Met-[P]-Glu-anhydride with His-Phe in free form to form phosphinic Met-[P]-Glu-γ-His-Phe tetrapeptide have been found. A chromatographic mass-spectrometry study, including MS2, and NMR of the phosphinic tetrapeptide on 1H, 13C, 31P nuclei was carried out using the methods of two-dimensional 1H–1H COSY, 1H–13C HSQC and 1H–13C HMBC NMR spectroscopy.

Development of a Method for Optimization of the Process of Extraction of Values of Numerical Characteristics of Oil Feedstock Components from Graphical Data of Chromatographic Analysis

A variant of the algorithm has been developed to perform the procedure of automated recovery of numerical values of graphically represented chromatograph signal function, studying the component composition of heavy oil feedstock samples. The problem, which the developed method aims to solve, consists in the poor adaptation of chromatographs to the oil industry: oil is a natural raw material, which is not chemically pure, therefore not all numerical characteristics of the components contained in the investigated sample are fixed within the chromatographic study. In the current configuration of the method, the values from the chromatogram are recorded manually. The developed method takes as input data the images of oil chromatograms obtained in the laboratory, presented in the original black and white colour scheme. The output data of the method is an array of numerical values of coordinates reconstructed with a step of one pixel. The size of the error in the reconstruction of the values by the method is much smaller than the threshold set by the petrochemical laboratory. In addition to automating the indicated task, the array of obtained coordinate values was vectorized in order to use the vector as input data in the Transformer model to solve the problem of predicting the redistribution of hydrocarbon components of heavy oil under the influence of catalysts. As a result of the change in input data representation, the time required to obtain a prediction and the training time were reduced by a multiple, while the value of the average prediction error decreased.

Synthesis of Hetaryl Substituted Pyrazolo[3,4-b]Quinolinone Systems by Multicomponent Cyclocondensation Reaction

In this study, the synthesis of pyrazolo[3,4-b]quinolinone compounds was carried out via one-pot three-component reactions. These reactions proceed as domino processes, making them easier to occur than the conventional multistep organic reactions. By employing this method, new organic molecules can be synthesized in a single step, using minimal time and number of trials. In the first phase of this two-step study, heteroaromatic carbaldehydes (quinoline-8-carboxaldehyde and quinoline-4-carboxaldehyde) were prepared to be used as substrates in subsequent reactions by oxidation of 8-methylquinoline and 4-methylquinoline with selenium dioxide, a mild oxidant. In the second step, heteroaromatic carbaldehyde reacted with aminopyrazole and dimedone in anhydrous ethanol by one-pot multicomponent condensation method to synthesize six new compounds with heteryl-substituted pyrazolo[3,4-b]quinolinone ring system. The crude products were obtained in excellent yields and further purified by crystallization. The structures of the compounds, which were found to be completely pure as a result of chromatographic studies, were elucidated by spectroscopic methods and elemental analysis.

Multi-Residue Analysis of Thyreostats in Animal Muscle Tissues by Hydrophilic Interaction Liquid Chromatography Tandem Mass Spectrometry: A Thorough Chromatographic Study

Τhyreostats (TSs) are veterinary drugs used in livestock farming for fattening. Their administration is banned in the European Union since 1981, and their monitoring for food quality and safety control requires sensitive and confirmatory methods. The present study describes the development and validation of a hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC-MS/MS) method for the simultaneous determination of 2-thiouracil (TU), 6-methyl-2-thiouracil (MTU), 6-propyl-2-thiouracil (PTU), 6-phenyl-2-thiouracil (PhTU), tapazole (TAP), and 2-mercaptobenzimidazole (MBI) in bovine muscle tissues. Investigation of the retention mechanism of the six analytes on the selected amide-based stationary phase showed that hydrophilic partition was the dominant interaction. The sample preparation included extraction with ACN/H2O (80/20), followed by dispersive solid-phase extraction (d-SPE) with C18 sorbent and hexane partitioning. The method was validated according to European guidelines using internal standards, including isotopically labelled ones. The method’s LODs ranged between 2.8 ng g−1 (6-phenyl-2-thiouracil) and 4.1 ng g−1 (2-thiouracil). Application of the proposed method to 48 bovine tissue samples showed non-detectable results.

Preparation, Characterization and Sustained-release Study of Encapsulated Cinnamon Essential Oil Microcapsules

Aims: The aim of this study is to study preparation, characterization and sustained release of Encapsulated Cinnamon Essential Oil Microcapsules. Background: In this work, encapsulated cinnamon essential oil (CEO) microcapsules were prepared, characterized, and analyzed for their sustained-release properties. CEO microcapsules were encapsulated from an alginate polymer using homogenization and extrusion, and the encapsulation mechanism used was ionic gelation. The potent antibacterial properties of natural cinnamon oil extracts and their shelf-life activity can be reduced or eliminated as a result of deterioration caused by light, heat, and oxygen exposure during production. High-speed homogenization was utilized for the encapsulation, which encloses and protects the volatile compounds from degradation. Objective: The objective of this study is to synthesize sustained-release encapsulated cinnamon essential oil (CEO) microcapsules. Method: The preparation of encapsulated cinnamon oil was achieved through homogenization. The extrusion method was employed to obtain microcapsules encapsulating liquid active ingredients (AI) with alginate polymer to induce ionic gelation. Results: SEM and Optical images reveal that all microcapsules maintain their spherical shape with clearly defined membranes. XPS analysis indicates the presence of oxygen (O), carbon (C), and sodium (Na) on the surface, suggesting the presence of an alginate-based ionic gelation. Chromatographic studies demonstrate a high encapsulation efficiency of 99%. The average microcapsule size is 261.5 nm for the fresh sample and 278 nm after 3.5 months. The zeta potential is -29.8 mV for the fresh sample and -28.2 mV after 3.5 months. Notably, there is no evidence of microcapsule agglomeration during the 3.5-month storage period as observed in the study. TGA data reveals that only 7.5% of the adsorbed water and essential oil mixture is lost at 40°C over 4 hours, in contrast to 11.7% for the adsorbed water material, indicating a sustained release of the encapsulated CEO from the microcapsules. Conclusion: The microcapsules exhibited an impressive encapsulation efficiency of 99%, demonstrating stability over the 3.5-month investigation period and showcasing sustained-release properties.

Funerary vs. domestic vessels from the Hallstatt period. A study on ceramic vases from the Milejowice settlement and the Domasław cemetery

Clay vessels have a wide variety of functions in social activities in the Hallstatt period. In addition to food storage and processing, they were used for ritual purposes and as funerary vessels. The paper presents the results of archaeological and chromatographic studies of 31 vases from two different Hallstatt culture sites in lower Silesia (Poland). The investigations included vessels fragments from the Domasław cemetery and from the Milejowice settlement. The chromatographic analyses focused on fatty acids and biomarkers and made it possible to identify the most likely sources of substances they came into contact with during use. The c-means and hierarchical cluster analyses showed that grave vessels differed from settlement ceramics. Thus, conclusions on the diverse vessel functions could be made.

Elucidating Chiral Resolution of Aromatic Amino Acids Using Glycopeptide Selectors: A Combined Molecular Docking and Chromatographic Study.

An asymmetric synthesis is a favorable approach for obtaining enantiomerically pure substances, but racemic resolution remains an efficient strategy. This study aims to elucidate the chiral resolution of aromatic amino acids and their elution order using glycopeptides as chiral selectors through molecular docking analysis. Chiral separation experiments were conducted using Vancomycin as a chiral additive in the mobile phase (CMPA) at various concentrations, coupled with an achiral amino column as the stationary phase. The Autodock Vina 1.1.2 software was employed to perform molecular docking simulations between each enantiomer (ligand) and Vancomycin (receptor) to evaluate binding affinities, demonstrate enantiomeric resolution feasibility, and elucidate chiral recognition mechanisms. Utilizing Vancomycin as CMPA at a concentration of 1.5 mM enabled the separation of tryptophan enantiomers with a resolution of 3.98 and tyrosine enantiomers with a resolution of 2.97. However, a poor chiral resolution was observed for phenylalanine and phenylglycine. Molecular docking analysis was employed to elucidate the lack of separation and elution order for tryptophan and tyrosine enantiomers. By calculating the binding energy, docking results were found to be in good agreement with experimental findings, providing insights into the underlying mechanisms governing chiral recognition in this system and the interaction sites. This comprehensive approach clarifies the complex relationship between chiral discrimination and molecular architecture, offering valuable information for creating and improving chiral separation protocols.

Nitrosamine formation driven by electrochemical chlorination of urine-containing source waters: Effects of operational conditions

We investigated the formation of nitrosamines from urine during electrochemical chlorination (EC) using dimensionally stable anodes. Short-term electrolysis (< 1 h) of urine at 25 mA cm−2 generated seven nitrosamines (0.1–7.4 µg L−1), where N-nitrosodimethylamine, N-nitrosomethylethylamine, and N-nitrosodiethylamine were predominant with concentrations ranging from 1.2 to 7.4 µg L−1. Mechanistic studies showed that the formation kinetics of nitrosamines was influenced by urine aging and composition, with fresh urine generating the highest levels (0.9–5.8 µg L−1) compared with aged, centrifuged, or filtered urine (0.2–4.1 µg L−1). Concurrently, studies on urine pretreatment through filtration and centrifugation underscored the significance of nitrogenous metabolites (such as protein-like products and urinary amino acids) and particle-associated humic fractions in nitrosamine formation during EC of urine. This finding was confirmed through chromatographic and spectroscopic studies utilizing LCOCD, Raman spectra, and 3DEEM fluorescence spectra. Parametric studies demonstrated that the ultimate [nitrosamines] increased at a pH range of 4.5–6.2, and with increasing [bromide], [ammonium], and current density. Conversely, sulfate and carbonate ions inhibited nitrosamine formation. Moreover, the implications of EC in urine-containing source waters were evaluated. The results indicate that regardless of the urine source (individual volunteers, septic tank, swimming pool, untreated municipal wastewater), high levels of nitrosamines (0.1–17.6 µg L−1) were generated, surpassing the potable reuse guideline of 10 ng L−1. Overall, this study provides insights to elucidate the mechanisms underlying nitrosamine formation and optimize the operating conditions. Such insights facilitate suppressing the generation of nitrosamine byproducts during electrochemical treatment of urine-containing wastewater.

Synthesis, Spectral, Chromatographic and Antimicrobial Studies of Transition Metal Complexes of Chromium (III) and Cobalt (II) Ions with Alprazolam Drug

Synthesis, Spectral, Chromatographic and Antimicrobial Studies of Transition Metal Complexes of Chromium (III) and Cobalt (II) Ions with Alprazolam Drug

Bivalent affinity binding-inspired PPARγ immobilization with selective conformation and improved ligand-binding activity

Functional protein immobilization forms the basis for bio-detections. A series of one-point, site-specific immobilization methods have been developed, however, it still remains as a challenge how to avoid the proteins to move in all directions as well as conveniently regenerate the bio-devices. Herein, we have developed a bivalent affinity binding-inspired method for PPARγ immobilization using DNA aptamer and nickel-nitrilotriacetic acid (Ni2+-NTA) chelation. The specific DNA aptamer (Apt 2) was selected by an on-column systematic evolution of ligands by exponential enrichment (SELEX) method with affinity of (1.57 ± 0.15) × 105 M−1, determined by isothermal titration calorimetry (ITC). Apt 2 and nickel-nitrilotriacetic acid (Ni2+-NTA) were modified on macroporous silica gels via L-α-allylglycine as a linker. They respectively interacted with PPARγ and 6×His tag via bivalent affinity binding for the receptor immobilization. After comprehensive surface characterization, PPARγ was proved to be successful immobilized. Chromatographic studies revealed that the immobilized PPARγ has conformation selectivity, which discriminated agonist and antagonist of the receptor. Ligand-binding parameters (affinity and rate constant) of four agonists (rosiglitazone, pioglitazone, troglitazone, and magnolol) with PPARγ were determined. Troglitazone showed the lowest dissociation rate constant. The binding affinities (3.28 × 107, 1.91 × 106, 2.25 × 107, and 2.43 × 107 M−1) were highly consistent with the data obtained using purified receptor in solution (2.16 × 107, 4.52 × 106, 1.20 × 107, and 1.56 × 107 M−1), offering reliable bio-detection method for PPARγ and its ligands. Due to the biocompatibility of nuclear receptor with DNA, it is conceivable that the bivalent affinity-based method will be a general method for the immobilization of other nuclear receptors, which may provide selective conformation and improved ligand-binding activity for the receptors.

A comprehensive strategy based on ultra-high performance liquid chromatography with diode array detector fingerprinting and multi-component ultra-performance liquid chromatography with tandem mass spectrometry technology for quality control of Jiawei Huoxiang Zhengqi Pill.

Jiawei Huoxiang Zhengqi Pill (JHZP) is a commonly used Chinese patent medicine for the clinical treatment of headache, dizziness, chest tightness as well as abdominal distension, and pain caused by wind-cold flu. In this study, a comprehensive strategy combining ultra-high performance liquid chromatography with diode array detector (UHPLC-DAD) fingerprinting and multi-component quantitative analysis was established and validated for quality evaluation of JHZP. A total of 49 characteristic common peaks were selected in a chromatographic fingerprinting study to assess the similarity of 15 batches of JHZP. Furthermore, 109 compounds were identified or preliminarily identified from JHZP by coupling with an advanced hybrid linear ion trap-Orbitrap mass spectrometer. For quantification, the optimized ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method was employed for the simultaneous determination of 13 target compounds within 12min. The sensitivity, precision, reproducibility, and accuracy of the method were satisfactory. This validated UPLC-MS/MS method was successfully applied to analyzing 15 batches of JHZP. The proposed comprehensive strategy combining UHPLC-DAD fingerprinting and multi-component UPLC-MS/MS analysis proved to be highly efficient, accurate, and reliable for the quality evaluation of JHZP, which can be considered as a reference for the overall quality evaluation of other Chinese herbal formulations.

Analytical method development and validation for the evaluation of related substances in Apalutamide by RP-HPLC

Apalutamide is an anti-cancer agent used for the management of prostate cancer. A new stability indicating RP-HPLC method (Gradient mode) has been developed for the estimation of Apalutamide and its related substances using Waters Alliance system (Model no. 2996 and 2695) with Inertsil ODS-3 (250 × 4.6 mm, 5μm) column (PDA detector) was used for the present study. A mixture of Ammonium phosphate buffer solution and Acetonitrile (30: 70, v/v) was used as the mobile phase for the chromatographic study (Flow rate: 1.0 mL/min; Detection wavelength: 243 nm). Stress degradation studies were performed and the method was validated as per ICH guidelines. Keywords: Apalutamide, RP-HPLC, Related substances, Impurities, Stability indicating, Validation, ICH guidelines.

Bioassay-guided isolation of anti-inflammatory and antinociceptive metabolites among three Moroccan Juniperus leaves extract supported with in vitro enzyme inhibitory assays

Ethnopharmacological relevanceHerbs of the genus Juniperus (family Cupressaceae) have been commonly used in ancestral folk medicine known as “Al'Araar” for treatment of rheumatism, diabetes, inflammation, pain, and fever. Bioassay-guided isolation of bioactives from medicinal plants is recognized as a potential approach for the discovery of novel drug candidates. In particular, non-addictive painkillers are of special interest among herbal phytochemicals. Aim of the studyThe current study aimed to assess the safety of J. thurifera, J. phoenicea, and J. oxycedrus aqueous extracts in oral treatments; validating the traditionally reported anti-inflammatory and analgesic effects. Further phytochemical investigations, especially for the most bioactive species, may lead to isolation of bioactive metabolites responsible for such bioactivities supported with in vitro enzyme inhibition assays. Materials and methodsFirstly, the acute toxicity study was investigated following the OECD Guidelines. Then, the antinociceptive, and anti-inflammatory bioactivities were evaluated based on chemical and mechanical trauma assays and investigated their underlying mechanisms. The most active J. thurifera n-butanol fraction was subjected to chromatographic studies for isolating the major anti-inflammatory metabolites. Moreover, several enzymatic inhibition assays (e.g., 5-lipoxygenase, protease, elastase, collagenase, and tyrosinase) were assessed for the crude extracts and isolated compounds. ResultsThe results showed that acute oral administration of the extracts (300–500 mg/kg, p. o.) inhibited both mechanically and chemically triggered inflammatory edema in mice (up to 70% in case of J. thurifera) with a dose-dependent antinociceptive (tail flick) and anti-inflammatory pain (formalin assay) activities. This effect was partially mediated by naloxone inhibition of the opioid receptor (2 mg/kg, i. p.). In addition, 3-methoxy gallic acid (1), quercetin (2), kaempferol (3), and ellagic acid (4) were successfully identified being involved most likely in J. thurifera extract bioactivities. Nevertheless, quercetin was found to be the most potent against 5-LOX, tyrosinase, and protease with IC50 of 1.52 ± 0.01, 192.90 ± 6.20, and 399 ± 9.05 μM, respectively. ConclusionJ. thurifera extract with its major metabolites are prospective drug candidates for inflammatory pain supported with inhibition of inflammatory enzymes. Interestingly, antagonism of opioid and non-opioid receptors is potentially involved.

Gas chromatography-mass spectrometry characterization of the aroma of Ossetian cheeses

The aim of the study was the qualitative and quantitative determination of volatile aroma compounds and their formation pathways in brine Ossetian cheeses. Volatile components of cheeses were isolated by steam distillation and extraction with dichloromethane, with their subsequent determination and quantification by gas chromatography-mass spectrometry. The results of the analysis are presented according to the structural classes of the main chemical components and the corresponding microbial metabolic processes. Four processes were found to be the main contributors to flavor formation: lipolysis, proteolysis, glycolysis, and a number of oxidative enzymatic transformations. Lipolysis of the fatty fraction of cheeses is a source of formation of volatile carboxylic acids and their esters. Proteolysis of the casein fraction yields branched alcohols, aldehydes, and a number of aromatic and heteroaromatic compounds. Glycolysis of the carbohydrate fraction is a source of ethanol formation, which is the main cause of the dominance of ethyl esters in the ester fraction. Redox enzymatic transformations mainly determine the biosynthesis of unbranched aldehydes, ketones and lactones. A clear distinction between retail and homemade cheeses was observed, due to the different technological approaches to the cheese preparation. The structuralchemical and quantitative evolution of the volatile composition of the studied cheese samples during ripening is tentatively shown. From the authors’ point of view, the aromatic composition of the Tib cheese sort is the most consistent with the Ossetian cheese standard. This study represents the first gas chromatographic study of Ossetian cheeses and aims to create objective criteria for controlling technological processes and product quality during production and storage in the food industry.