High-performance Liquid Chromatography Coupled With Mass Spectrometry Research Articles

Mechanism of unactivated peroxymonosulfate-induced degradation of amino-G acid: Kinetics and transformation pathway

Amino-G acid (7-amino-1,3-naphthalenedisulfonic acid), a pollutant produced during textile preparation and in dyeing industries, is discarded in wastewater and causes ecological problems. In this study, amino-G acid (50.0 μM) could be completely removed by unactivated peroxymonosulfate (PMS) (1.0 mM) within 240 min, and the reaction kinetics exhibited strong pH dependence. A species-specific parallel reaction describes this pH dependence well, and the species-specific reaction rate constants of [amino-G acid]2− and [amino-G acid]− with HSO5− were calculated to be 0.0826 ± 0.0019 and 000122 ± 0.0027 M−1 s−1. The reactivity of the six naphthalene sulfonic acids and unactivated PMS varied according to the structure, and the amino group in naphthalene sulfonic acid, a typical electron-donating group, was more vulnerable to attack by unactivated PMS, which was further confirmed by the density functional theory results. The high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS) spectral analysis results indicated that naphthalene sulfonic acid was first oxidized to hydroxylamine products and then further oxidized to nitroso compounds, which exhibited more ecological toxicity. However, as the reaction processing, the toxicity of the product gradually decreased. Our results provide new insights into the in-situ oxidation of unactivated PMS.

Investigating the anti-inflammatory potential of Elaeagnus an-gustifolia L. through enzyme inhibition assays

Elaeagnaceae family, which is common in the steppe regions of Asia, Europe, and North America, contains about 45 species, including three genus: Hippophae, Shepherdia, and Elaeagnus. There are two species in the genus Hippophae rhamnoides and Elaeagnus angustifolia. The species of the Elaeagnaceae family is widely used in public and traditional medicine due to its widespread prevalence in the world and in Turkey. Ethnobotanical field studies have revealed that the fruits of E. angustifolia L. are used in Anatolia as an anti-inflammatory, antipyretic, urolithiatic, antidiarrheal, and anti-inflammatory agent in eye infections. Furthermore, the fruit of this plant is consumed as an appetizer and a source of nutrition. The study aims to evaluate the in vitro anti-inflammatory activity of extracts in different polarities from the leaves, branches, and fruits of E. angustifolia and to establish the content of polyphenolic compounds (flavones and polyphenolic carboxylic acids) in the ethyl acetate extract branches using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). In this context, the anti-inflammatory effects of n-hexane, ethyl acetate, and methanol extracts prepared from the plant have been evaluated through in vitro cyclooxygenase (COX-1 and COX-2) enzyme inhibition, inhibition of protein denaturation and proteinase enzyme, and anti-lipoxygenase activity tests. The results showed the highest activity of the ethyl acetate extract from branches. The HPLC-MS analysis revealed that the ethyl acetate extract prepared from the branches at a concentration of 100 mg/g contains p-coumaric acid (123.91 mg%), ferulic acid (62.07 mg%), kaempferol (43.11 mg%), sinapic acid (9.82 mg%), and quercetol (12.46 mg%). It was concluded that the data obtained in this context supported the use of the plant in folk medicine.

Antiparasitic Activity of Isolated Fractions from Parthenium incanum Kunth against the Hemoflagellate Protozoan Trypanosoma cruzi.

This study focused on isolating, identifying, and evaluating the trypanocidal potential against the hemoflagellate protozoan Trypanosoma cruzi of compounds from Parthenium incanum (Mariola), a plant used in traditional Mexican medicine to treat stomach and liver disorders. P. incanum has a wide distribution in Mexico. This study found that methanolic extracts of P. incanum, obtained by static maceration and successive reflux, had promising results. The fractions were compared using thin-layer chromatography (TLC) and those that showed similarities were mixed. A bioguided assay was performed with Staphylococcus aureus ATCC 25923, using agar diffusion and bioautography techniques to determine the preliminary biological activity. The fractions with antimicrobial activity were purified using a preparative thin-layer chromatography (PTLC) plate, obtaining the bioactive bandages that were subjected to a trypanocidal evaluation against the Ninoa strain of T. cruzi in its epimastigote stage. This revealed an IC50 of up to 45 ± 2.5 µg/mL, in contrast to the values obtained from the crude extracts of less than 100 µg/mL. The TLC, Fourier-transform infrared spectroscopy (FT-IR), and high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) techniques were used to identify the compounds, demonstrating the presence of sesquiterpene lactones, parthenin, and coronopolin. We concluded that these compounds have the potential to inhibit T. cruzi growth.

Does Erodium trifolium (Cav.) Guitt exhibit medicinal properties? Response elements from phytochemical profiling, enzyme-inhibiting, and antioxidant and antimicrobial activities

Abstract Geraniaceae are typically used as diuretic, anti-diarrhoeal, stomachic, and anti-hemorrhagic drugs. This study examined the phytochemicals and bioactivities in methanolic extract (ME) and petroleum ether extract (PEE) of the Erodium trifolium aerial part. Inductively coupled plasma mass spectrometry was used to assess the mineral profiles, high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and high-performance liquid chromatography with diode-array detection (HPLC-DAD) were used to assess the phenolic content of ME, and gas chromatography-mass spectrometry (GC-MS) was used to assess the fatty acid and volatile composition of the PEEs. In addition, the bioactivities of extracts were evaluated by using the 1,1-diphenyl-2-picrylhydrazyl, ABTS, and cupric reducing antioxidant capacity assays, including enzyme inhibition against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, α-glucosidase activities, and antibacterial properties. HPLC-MS identified eight compounds in ME: rutin, catechin, and caffeine were the major phenolic compounds detected. HPLC-DAD analysis showed that 11 compounds detected among epicatechin, catechin, chlorogenic acid, and gallic acid were predominant. GC-MS analysis revealed 30 fatty acids in PEE, with palmitic acid and oleic acid being predominant. The mineral content showed that Fe was the abundant microelement, and Ca and K were the abundant macroelements. The ME exhibited the highest activity compared to the PEE in all tests regarding antioxidant and antibacterial activities. Furthermore, all the extracts showed moderate inhibition against AChE, BChE, α-amylase, and α-glucosidase. In conclusion, E. trifolium may be employed to separate novel bioactive metabolites with potential pharmaceutical activities.

Identification of Benzodiazepine Use Based on Dried Blood Stains Analysis.

Biological matrices are typically used in forensic toxicological or pharmacological analysis: mainly blood, vitreous humor or urine. However, there are many cases in which crimes are a consequence of drug intoxication or drug abuse and they are not closed because over the months or years the samples become altered or decomposed. A dried blood stains test (DBS-MS) has recently been proposed to be used in drug toxicology when blood is found at a crime scene. This test could help an investigator to reveal what a person had consumed before the perpetration of the crime. In order to check the possibilities of this test, we analyzed several dried blood stains located on a cotton fabric. Therefore, the aim of this study was to determine if the analysis of a dried blood spot located on a cotton fabric could be an alternate source of obtaining toxicological results, particularly regarding benzodiazepines. We splashed blood stains on cotton fabric with different concentrations of the following benzodiazepines: alprazolam, bromazepam, clonazepam, diazepam and lorazepam, which were dried for 96 h and subsequently quantified by high-performance liquid chromatography coupled mass spectrometry (HPLC-MS). Our results show that it is possible to identify several benzodiazepines contained in a cotton fabric blood stain; consequently, this method may add another sample option to the toxicological analysis of biological vestiges found at a crime scene.

A method for simultaneously and accurately quantify seven quinolones in matrix reference materials by HPLC-MS

Reference materials (RM) are commonly employed in analytical chemistry for tasks such as instrument calibration, validation of analytical methods and quality control. Determining the values of target drugs in the matrix is a key step in the development of matrix reference materials. However, there is almost no sharing of the certification methodologies for the development of RM. Hence, a detailed method was established for the value determination of seven quinolones in minced fish matrix, which was the candidate materials for preparing matrix reference materials. Target analysis of quinolone drugs in complex matrix was performed by “quick, easy, cheap, effective, rugged and safe” (QuEChERS) extraction and high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). The accuracy, precision, and sensitivity of this method has been thoroughly validated, which provided a robust analytical tool for the accurate quantification of quinolones in quality control samples and actual samples with a detection limit of 0.5–0.6 μg/kg and a quantification limit of 1 μg/kg. The actual samples prepared by three different spiking methods was determined, the outstanding performance show that this method contributing significantly to the development of matrix reference materials in aquatic products, which can be used as a template for the determination of other quinolone compounds in minced fish matrix. This work can not only provide reference for the development of matrix reference materials, but also promote the development of metrology.

Cobalt-catalysed bicarbonate-activated peroxide as a promising system for the advanced oxidation of epirubicin in wastewaters

Epirubicin (EPI) is a common cytostatic drug often found as a pollutant in hospital wastewater. While many studies have focused on the degradation kinetics of its epimer, doxorubicin (DOX), none have reported the kinetics of EPI degradation. Herein, we report a new cost-efficient, environmentally friendly system for the advanced degradation of EPI consisting of cobalt-catalysed bicarbonate-activated peroxide. We propose a new kinetic model for EPI degradation that shows the hydroperoxyl radical is the most important reactive oxygen species (ROS) involved in the degradation process. Seven degradation products are identified using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). The effect of surfactants, commonly found in wastewaters, is also investigated and the presence of the cationic surfactant hexadecylpyridinium chloride (CPC) in the reaction medium leads to a 6.1-fold increase in the degradation rate. Modelling the reaction kinetics using the Piszkiewicz model unveils the rate constant for the micellar degradation of EPI to be 4.66×10‐3s‐1. The ecotoxicity study of the degradation products shows a dramatic decrease in both acute and chronic toxicity. Although the level of mineralisation of the products is less than 10%, this method is still suitable as a pre-treatment of wastewaters. The same system is successfully used for the degradation of DOX, mitoxantrone, and quinizarin. In the case of DOX, a rate constant of 3.74×10‐2s‐1 is obtained, the highest reported so far.

Structural and Biochemical Characterization of a New Phage-Encoded Muramidase, KTN6 Gp46.

Endolysins are phage-encoded lytic enzymes that degrade bacterial peptidoglycan at the end of phage lytic cycles to release new phage particles. These enzymes are being explored as an alternative to small-molecule antibiotics. The crystal structure of KTN6 Gp46 was determined and compared with a ColabFold model. Cleavage specificity was examined using a peptidoglycan digest and reversed-phase high-performance liquid chromatography coupled to mass spectrometry (HPLC/MS). The structure of KTN6 Gp46 could be determined at 1.4 Å resolution, and key differences in loops of the putative peptidoglycan binding domain were identified in comparison with its closest known homologue, the endolysin of phage SPN1S. Reversed-phase HPLC/MS analysis of the reaction products following peptidoglycan digestion confirmed the muramidase activity of Gp46, consistent with structural predictions. These insights into the structure and function of endolysins further expand the toolbox for endolysin engineering and explore their potential in enzyme-based antibacterial design strategies.

Assessing Matrix Solid-Phase Dispersion Extraction Strategies for Determining Bisphenols and Phthalates in Gilthead Sea Bream Samples

Microplastics (MPs) and nanoplastics (NPs) are widely spread in the environment, generating significant concern due to their potential impact on environmental health. Marine species usually ingest plastic fragments, mistaking them for food. Many toxic compounds, such as plastic additives that are not chemically bound to the plastic matrix, can be released from MPs and NPs and reach humans via the food chain. This paper highlights the development and validation of a straightforward solid–liquid extraction clean-up procedure in combination with a matrix solid-phase dispersion method using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) detection, enabling facile, precise, and reliable identification and quantitation of a total of six bisphenols and phthalates in gilthead sea breams. Under the optimized conditions, the developed method showed good linearity (R2 > 0.993) for all target compounds. The recoveries obtained were between 70 and 92%. The relative standard deviations (RSDs) for reproducibility (inter-day) and repeatability (intra-day) were less than 9% and 10%, respectively. The limit of detection (LOD) and limit of quantification (LOQ) for the target compounds ranged from 0.11 to 0.68 µg/kg and from 0.37 to 2.28 µg/kg, respectively. A new, efficient extraction methodology for the determination of BPA, BPS, BPF, DBP, DEP, and DHEP in gilthead seabream has been optimized and validated.

TRACING THE COORDINATED CONTROL OF NAD+ HOMEOSTASIS USING C. ELEGANS

Abstract Nicotinamide adenine dinucleotide (NAD+) is a vital coenzyme involved in redox reactions and cellular signaling. Its levels decline with age in humans, mice, and worms, leading to metabolic disturbances associated with age-related diseases. Interestingly, our studies in aged mice indicate that the rate of NAD+ synthesis remains unchanged, ruling out a lack of precursors as the cause of NAD+ decline. Instead, we found that tissues with lower NAD+ concentrations in aged mice exhibit higher fractional turnover, suggesting increased consumption as the primary driver of NAD+ decline during aging. This led us to hypothesize that the loss of NAD+ metabolic robustness and resiliency with age is linked to the hyper-activation and competition of NAD+-consuming enzymes. To investigate the physiological impact of NAD+ consuming enzymes on metabolic robustness, we used genetic tools and mass spectrometry in C. elegans. Surprisingly, we identified novel reproductive and aging phenotypes in mutant animals lacking specific NAD+ consuming enzymes, including Poly ADP-Ribose Polymerase (parp-1 and parp-2), TANKyrase (tank-1), and Toll and Interleukin 1 Receptor (tir-1). These mutants displayed significant reductions in fecundity and extensions in longevity. Additionally, employing High-Performance Liquid Chromatography coupled with Mass Spectrometry (HP-LC-MS), we examined the NAD+ metabolome across different developmental stages in our mutant strains, revealing notable metabolic disruptions associated with the loss of NAD+ consumers. In summary, our findings establish a novel connection between the hyper-activation of NAD+ degradation and the onset of reproductive aging. This research provides valuable insights into the role of NAD+ metabolism and its impact on aging-related processes.

Modified Biejia Jianwan decoction restrains PD-L1-mediated immune evasion through the HIF-1α/STAT3/NF-κB signaling pathway

Ethnopharmacological relevanceModified Biejia Jianwan (M-BJJW), a Traditional Chinese Medicine (TCM) decoction, has exhibited great potential in treating hepatocellular carcinoma (HCC). However, its underlying functional mechanism still remains unknown. Aim of the studyThe study aimed to explore the anti-hepatocarcinogenic effects of M-BJJW, specifically its influence on PD-L1-mediated immune evasion in hypoxic conditions, and elucidate the related molecular mechanisms in HCC. Materials and methodsTo investigate the therapeutic efficacy and mechanisms underlying M-BJJW's effects on HCC, we employed a diethylnitrosamine (DEN)-induced rat model maintained for 120 days. Following model establishment, flow cytometry was utilized to assess the distribution of immune cell populations in peripheral blood, spleens, and tumor tissues after M-BJJW administration. Simultaneously, enzyme-linked immunosorbent assays (ELISA) were conducted to analyze cytokine profiles in serum samples. Immunohistochemistry was employed to determine the expression levels of crucial proteins within tumor tissues. Furthermore, HCC cells exposed to CoCl2 underwent Western blot analysis to validate the expression levels of HIF-1α, PD-L1, STAT3, and nuclear factor kappa B (NF-κB) p65. The modulatory effects of STAT3 and NF-κB p65 were investigated using specific inhibitors and activators in wild-type cell lines. High-performance liquid chromatography coupled with mass spectrometry (HPLC/MS) was utilized to identify the chemical constituents present in M-BJJW-medicated serum. The immunomodulatory properties and the anti-tumor activities of M-BJJW were evaluated by co-culturing with peripheral blood mononuclear cells (PBMC) and the CCK-8 assay. Additionally, we assessed M-BJJW's impact on hypoxia-induced alterations in HCC cell lines using immunofluorescence and Western blot assessments. ResultsM-BJJW exhibited substantial therapeutic advantages by effectively alleviating pathological deterioration within the HCC microenvironment. In the DEN-induced rat model, M-BJJW administration notably reduced tumor growth. Flow cytometry analyses revealed an increased proportion of Cytotoxic T lymphocytes (CTLs) accompanied by a simultaneous decrease in regulatory T cells (Tregs). ELISA data supported a marked decrease in pro-inflammatory cytokines, including interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor α (TNF-α). Immunohistochemistry confirmed the suppressive effect of M-BJJW on the expression of HIF-1α and PD-L1. Notably, western blotting unveiled the role of HIF-1α in regulating PD-L1 expression via the STAT3 and NF-κB signaling pathways in HCC cell lines, which was validated using activators and inhibitors of STAT3 and NF-κB. The CCK-8 assay and co-culture techniques demonstrated the anti-tumor activity of M-BJJW. Immunofluorescence and western blotting further confirmed that M-BJJW-containing serum dose-dependently inhibited HIF-1α, PD-L1, p-STAT3, and p-p65 in hypoxic HCC cell lines. ConclusionsM-BJJW demonstrates significant therapeutic potential against HCC by influencing the hypoxic microenvironment, thereby regulating the immunosuppressive milieu. Specifically, M-BJJW modulates the HIF-1α/STAT3/NF-κB signaling pathway, leading to reduced PD-L1 expression and an elevated ratio of cytotoxic T lymphocytes (CTLs), while concurrently decreasing T regulatory cells (Tregs) and immunosuppressive factors. These synergistic effects aid in countering PD-L1-mediated immune evasion, presenting compelling pharmacological evidence supporting the clinical application of M-BJJW as a therapeutic approach for HCC.

Microwave-assisted extraction of β-Sitosterol: A by-product from Agave angustifolia Haw bagasse

β-sitosterol (BSS) and β-sitosterol glucoside (BSSG) were extracted from Agave angustifolia Haw bagasse using microwave-assisted extraction (MAE). The quantification and characterization of BSS and BSSG were performed through high-performance thin-layer chromatography (HPTLC), Fourier-transform infrared spectroscopy (FT-IR), high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS), and gas chromatography-mass spectrometry (GC-MS). With an extraction time of 9 seconds using MAE, a higher amount of BSS (103.6 mg of β-sitosterol per gram of dry weight of extract) was obtained compared to BSSG (61.6 mg of β-sitosterol glucoside per gram of dry weight of extract). MAE emerges as a promising method for the efficient recovery of β-sitosterol (BSS) and β-sitosterol glucoside (BSSG) from A. angustifolia bagasse. This enriches scientific knowledge regarding phytochemical extraction and holds great potential for various industries, such as pharmaceuticals and nutraceuticals. Additionally, by valorizing the by-products of the agave industry, this research contributes to the sustainable utilization of resources, aligning with the principles of green and circular economy practices.

Acetylcholine-Binding Protein Affinity Profiling of Neurotoxins in Snake Venoms with Parallel Toxin Identification

Snakebite is considered a concerning issue and a neglected tropical disease. Three-finger toxins (3FTxs) in snake venoms primarily cause neurotoxic effects since they have high affinity for nicotinic acetylcholine receptors (nAChRs). Their small molecular size makes 3FTxs weakly immunogenic and therefore not appropriately targeted by current antivenoms. This study aims at presenting and applying an analytical method for investigating the therapeutic potential of the acetylcholine-binding protein (AChBP), an efficient nAChR mimic that can capture 3FTxs, for alternative treatment of elapid snakebites. In this analytical methodology, snake venom toxins were separated and characterised using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and high-throughput venomics. By subsequent nanofractionation analytics, binding profiling of toxins to the AChBP was achieved with a post-column plate reader-based fluorescence-enhancement ligand displacement bioassay. The integrated method was established and applied to profiling venoms of six elapid snakes (Naja mossambica, Ophiophagus hannah, Dendroaspis polylepis, Naja kaouthia, Naja haje and Bungarus multicinctus). The methodology demonstrated that the AChBP is able to effectively bind long-chain 3FTxs with relatively high affinity, but has low or no binding affinity towards short-chain 3FTxs, and as such provides an efficient analytical platform to investigate binding affinity of 3FTxs to the AChBP and mutants thereof and to rapidly identify bound toxins.

Preparation of nickel-doped iron-based bimetallic organic framework carbonized derivatives for heterogeneous catalytic hydrogen peroxide degradation of 4-NP

A nickel-doped iron-bimetallic organic framework carbon-derived material (Fe-Ni-MOF@C) was successfully constructed by microwave-pyrolysis and used as a new non-homogeneous Fenton-like catalyst for the catalytic degradation of p-nitrophenol (4-NP) by hydrogen peroxide (H2O2). Several characterization results such as SEM and XRD show that Fe-Ni-MOF@C is successfully synthesized and has porous, strong stability and magnetic properties. Fe-Ni-MOF@C materials exhibit excellent catalytic performance for the degradation over a wide temperature (15–45 °C) and pH (3.00–9.00). Complete removal of 4-NP (30 mg/L) was achieved in 30 min at 25 °C and pH = 3. The material properties can be restored by calcination. According to the radical burst experiments, electron paramagnetic resonance spectroscopy (EPR) combined with the characterization results, the radicals acting in the Fe-Ni-MOF@C/H2O2 system are mainly •OH and, to a lesser extent, •O2−. The synergistic effect between Fe and Ni species accelerates electron transfer and radical generation, which is the reason for the material's high catalytic performance. High-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was used to analyze the main degradation products and pathways. This study demonstrates that bimetallic organic framework carbon-derived materials with synergistic effects have great application prospects as non-homogeneous Fenton-like catalysts in the field of H2O2 oxidation of organic pollutants.

The Comparison of Serum Exosome Protein Profile in Diagnosis of NSCLC Patients.

A thorough study of the exosomal proteomic cargo may enable the identification of proteins that play an important role in cancer development. The aim of this study was to compare the protein profiles of the serum exosomes derived from non-small lung cancer (NSCLC) patients and healthy volunteers (control) using the high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS) method to identify potentially new diagnostic and/or prognostic protein biomarkers. Proteins exclusively identified in NSCLC and control groups were analyzed using several bioinformatic tools and platforms (FunRich, Vesiclepedia, STRING, and TIMER2.0) to find key protein hubs involved in NSCLC progression and the acquisition of metastatic potential. This analysis revealed 150 NSCLC proteins, which are significantly involved in osmoregulation, cell-cell adhesion, cell motility, and differentiation. Among them, 3 proteins: Interleukin-34 (IL-34), HLA class II histocompatibility antigen, DM alpha chain (HLA-DMA), and HLA class II histocompatibility antigen, DO beta chain (HLA-DOB) were shown to be significantly involved in the cancer-associated fibroblasts (CAFs) and tumor-associated macrophages (TAMs) infiltration processes. Additionally, detected proteins were analyzed according to the presence of lymph node metastasis, showing that differences in frequency of detection of protein FAM166B, killer cell immunoglobulin-like receptor 2DL1, and olfactory receptor 52R1 correlate with the N feature according to the TNM Classification of Malignant Tumors. These results prove their involvement in NSCLC lymph node spread and metastasis. However, this study requires further investigation.

Knotwood and Branchwood Polyphenolic Extractives of Silver Fir, Spruce and Douglas Fir and Their Antioxidant, Antifungal and Antibacterial Properties.

The extractive contents of three softwood species largely used in the wood industry, namely Abies alba (Silver fir), Picea abies (spruce) and Pseudotsuga menziesii (Douglas fir), have been determined quantitatively for knots and at different points chosen along their branches, before analysis using high-performance liquid chromatography coupled with Mass Spectrometry (HPLC-MS). The results indicated that branchwood samples located in close proximity to the stem present high contents of extractives similar to those recorded for the knots. HPLC analysis showed quite similar chemical compositions, indicating that first cm of the branches could be considered as an additional source of knotwood. The antibacterial, antifungal and antioxidant activities of knot's extractives have been investigated with the dual objective of better understanding the role of high levels of secondary metabolites present in the knot and evaluating their potential for biorefinery applications. The antioxidant activity study showed that crude extracts of Douglas fir knotwood presented higher radical scavenging activity levels than the extracts of Silver fir and spruce, which presented more or less the same activities. Silver fir and spruce knotwood extracts presented higher antibacterial activity levels than the Douglas fir knotwood extracts did, while Douglas fir knotwood extracts presented more fungal growth inhibition than the spruce and fir knotwood extracts did. The structure-activity relationships indicate that radical scavenging and antifungal activities are associated with a higher relative quantity of flavonoids in the crude extracts, while higher relative quantities of lignans are associated with antibacterial activity.

HPLC-Mass spectrometry analysis of phenolics comparing traditional bilberry and blueberry liqueurs

The purpose of this study was to investigate the differences between the phenolics compositions of bilberry and blueberry liqueurs, to determine whether these are comparable. Both bilberry and blueberry liqueurs have the same name on the market, however they differ in phenolic composition and content. A total of 43 compounds were identified, 36 in the bilberry liqueur, and 21 in the blueberry liqueur. High performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was used to identify and quantify these compounds, where they were fragmented to the MS4 fragments. Anthocyanins were the major phenolics group detected in the bilberry liqueurs, but only the second most abundant phenolics group in the blueberry liqueurs. The most abundant phenolics group in the blueberry liqueurs was hydroxycinnamic acids. The lowest content of individual phenolic compounds was for blueberry liqueur made from whole fruit. This study will provide valuable data to consumers in their choice of a more expensive bilberry liqueur or a cheaper blueberry liqueur.

LC-MS guided isolation of N,β-glucopyranosyl vincosamide and other compounds from the curare ingredient Strychnos peckii

Strychnos peckii (Loganiaceae) is an important active ingredient in curare poisons in the Amazon rainforest. Although previous studies have identified this species as a promising source of monoterpene indole alkaloids (MIAs), knowledge about other natural products is still scarce. Thus, to detect and guide the isolation of unprecedented bioactive compounds from the leaves of S. peckii, an untargeted high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) analysis was performed with the leaf aqueous extract. The HPLC-MS analysis allowed the detection of eleven compounds, including the alkaloids harman-3-carboxylic acid (5) and N,β-glucopyranosyl vincosamide (6), and the flavonoids quercetin 3-O-rhamnopyranoside (9) and kaempferol 3-O-rhamnopyranoside (10), all not previously reported in the Loganiaceae family. These compounds, along with strictosidine (3), 5-carboxystrictosidine (7), and desoxycordifoline (8) were isolated through modern chromatographic techniques and determined by using NMR spectroscopy in combination with MS. Overall, the untargeted HPLC-MS analysis proved to be a simple and effective approach to guide the isolation of substances not yet identified from S. peckii.

Eugenia sonderiana O. Berg leaves: Phytochemical characterization, evaluation of in vitro and in vivo antidiabetic effects, and structure-activity correlation

Several medicinal plants have drawn the attention of researchers by its phytochemical composition regarding their potential for treating chronic complications of diabetes mellitus. In this context, plants of the Myrtaceae family popularly used in Brazil for the treatment of diabetes mellitus, including Eugenia sonderiana, have shown beneficial effects due to the presence of phenolic compounds and saponins in their chemical constitution. Thus, the present work aimed to perform the phytochemical characterization of the hydroethanolic extract of E. sonderiana leaves using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS), along with in vitro and in vivo studies of antidiabetic activity. The chemical characterization revealed the presence of phenolic compounds, flavonoids, neolignans, tannins, and saponins. In addition, the extract exhibited minimum inhibitory concentrations of alpha-amylase and alpha-glycosidase higher than the acarbose in the in vitro tests. Also, the in vivo tests revealed a slight increase in body mass in diabetic rats, as well as a significant decrease in water and feed consumption provided by the extract. Regarding serum biochemical parameters, the extract showed significant activity in decreasing the levels of glucose, hepatic enzymes, and triglycerides, in addition to maintaining HDL cholesterol levels within normal ranges, protecting the cell membranes against oxidative damage. Thus, the extract of E. sonderiana leaves was considered promising pharmaceutical ingredient in the production of a phytotherapy medication.

Cannabis sativa L. roots from Northeast Brazil reduce abdominal contortions in a mouse model of primary dysmenorrhea

Ethnopharmacological relevanceAlthough the root of Cannabis sativa L. has been mentioned in some regions, such as the Vale do São Francisco, for its potential traditional medicinal use as an anti-inflammatory, anti-asthmatic, and against gastrointestinal diseases, it has received little exploration and discussion. Aim of the studyThis study aimed to perform a chemical analysis of an aqueous extract of Cannabis sativa roots (AqECsR) and evaluate its pharmacological effects against uterine disorders, in vivo and ex vivo, in rodents. Materials and methodsThe roots were provided by the Brazilian Federal Police, and the freeze-dried extract was used for the chemical analysis of the AqECsR by high performance liquid chromatography coupled with mass spectrometry (HPLC-MS). The sample was subsequently used in three doses for pharmacological assays (12.5, 25, and 50 mg/kg), which included the spasmolytic activity test and the primary dysmenorrhea test. The primary dysmenorrhea test aimed to verify the effect of AqECsR on induced abdominal contortions in female mice in vivo and to perform a morphometric analysis of the organs. Association tests at subtherapeutic doses of AqECsR with antidysmenorrheic drugs were also performed. ResultsThe data obtained by HPLC-MS suggested the presence of four substances: cannabisativine, anhydrocannabisativine, feruloyltyramine, and p-coumaroyltyramine. In the pharmacological assays, the AqECsR showed no spasmolytic effect. However, in the antidysmenorrheal activity test, AqECsR demonstrated a significant in vivo effect of reducing oxytocin-induced abdominal contortions. Morphometric analysis of the uterus showed no significant organ enlargement effect, and the association of AqECsR with subtherapeutic doses of three drugs used in antidysmenorrheal therapy (mefenamic acid, scopolamine, and nifedipine) showed an effect in reducing abdominal contortions. ConclusionsIn conclusion, AqECsR contains four chemical compounds and exhibits an antidysmenorrheic effect both alone and in association with drugs, reducing abdominal contortions in female mice without generating organ enlargement in the animals. Further studies are needed to prove the mechanism of action by which AqECsR promotes its effect on primary dysmenorrhea and to explore its associations.