Integrated GC-MS and UPLC-ESI-QTOF-MS based untargeted metabolomics analysis of in vitro raised tissues of Digitalis purpurea L.

Digitalis purpurea L. is one of the important plant species of Nilgiris, Kashmir and Darjeeling regions of India, belonging to the family Plantaginaceae, with well-known pharmacological applications. In the present investigation, an in vitro culture technique of indirect shoot organogenesis of D. purpurea is being explored; the biochemical attributes, the antioxidant activities and the metabolomic analyses were made by utilizing untargeted Gas Chromatography-Mass Spectrometry (GC-MS) and Ultra Performance Liquid Chromatography coupled with electronspray ionization/quadrupole-time-of-flight-mass spectrometry (UPLC-ESI-QTOF-MS) approaches. Initially, the leaf explants were used for callus induction and proliferation and maximum callusing frequency (94.44%) and fresh biomass (4.9 g) were obtained on MS, fortified with 8.8 µM BAP (6-benzyl amino purine) + 0.9 µM 2,4-D (2,4-dichlorophenoxyacetic acid), subsequently shoot formation (indirect organogenesis) was noted on the same MS medium with a shoot induction frequency of 83.33%. Later on, the biochemical and antioxidant potential of in vivo-, in vitro grown leaf and leaf derived callus were assessed. Significantly higher total phenol, flavonoid, DPPH (2,2-diphenyl-1-picrylhydrazyl), POD (peroxidase) and SOD (superoxide dismutase) activities were noticed in in vitro grown callus and leaf tissues compared with field grown leaf. The GC-MS analysis of each methanolic extract (in vivo-, in vitro derived leaf and leaf derived callus) displayed the presence of more than 75 bioactive compounds viz loliolide, stigmasterin, alpha-tocopherol, squalene, palmitic acid, linoleic acid, beta-amyrin, campesterol etc. possessing immense therapeutic importance. The UPLC-MS based metabolite fingerprinting of each methanolic extracts were conducted in both positive and negative ionization mode. The obtained results revealed variation in phytochemical composition in field - and laboratory grown tissues, indicating the impact of in vitro culture conditions on plant tissues. The detected phytocompounds belongs to various classes such as flavonoids, steroids, terpenoids, carbohydrates, tannins, lignans etc. The medicinally important metabolites identified were 20, 22-dihydrodigoxigenin, digoxigenin monodigitoxoside, apigenin, luteolin, kaempferide, rosmarinic acid, nepitrin and others. The results of the present study suggest that in vitro culture of D. purpurea could successfully be utilized for the novel drug discovery by producing such important phytocompounds of commercial interest in shorter duration without harming the plants' natural population.

RP-UHPLC-ESI-QTOF-MS analyses and evaluation of the antioxidant, antimicrobial, anti-diabetic and anti-Alzheimer activities of Linum tenue Desf.

The polyphenolic compounds of the n-butanol fraction of Linum tenue Desf. (BFLTe) were characterised by RP-UHPLC-ESI-QTOF-MS analyses with the main presence of 6,8-di-C-glucosyl naringenin (11.7%), vicenin 2-isomer 2 (8.18%), luteolin-7,3'-di-O-ß-D-glucoside (7.18%), isovitexin (5.98%), luteolin-7-O-ß-D-glucoside (5.713%), myricitrin (4.41%), luteolin-4'-O-ß-D-glucoside (4.04%), chlorogenic acid (28.68%), 3-(2,6-dihydroxyphenyl)-4-hydroxy-6-methyl-3H-2-benzofuran-1-one (8.17%) and p-coumaric acid (4.0%.). The antioxidant capacity was evaluated using three complementary methods (DPPH, ABTS and Reducing power). Additionally, the antimicrobial activity was tested against eight bacterial strains and the fungi Candida albicans whereas the antidiabetic activity was performed against α-amylase. The anti-Alzheimer activity was tested by inhibiting the butyrylcholinesterase (BChE). The BFLTe showed, for the first-time, a good antioxidant potential in DPPH (IC50:68.83 ± 2.74 µg/mL), ABTS (IC50:48.73 ± 1.07 µg/mL) and Reducing power assays (A0.50:99.98 ± 1.18 µg/mL) and a moderate antimicrobial activity with 250 and 500 µg/mL MICs values. Moreover, the fraction exhibited an excellent inhibition of the BChE (IC50:33.00 ± 0.85 µg/mL) and α-amylase (IC50:1093.13 ± 12.93 µg/mL).

Metabolic profiling of Vitex Pubescens Vahl bark via UPLC-ESI-QTOF/MS/MS analysis and evaluation of its antioxidant and acetylcholinesterase inhibitory activities.

BACKGROUND: Alzheimer's disease is a neurodegenerative age-related disease that primarily affects the elderly population leading to progressive memory impairments and neural deficits. It is counted as a major cause of geriatric dependency and disability. The pathogenesis of Alzheimer's disease incidence is complex and involves various hypotheses, including the cholinergic hypothesis, deposition of ß-amyloid plaques, neuroinflammation, oxidative stress, and apoptosis. Conventional treatments such as donepezil aim to delay the symptoms but do not affect the progression of the disease and may cause serious side effects like hepatoxicity. The use of natural candidates for Alzheimer's disease treatment has drawn the attention of many researchers as it offers a multitargeted approach. METHODS: This current study investigates the metabolic profiles of total defatted methanolic extract of Vitex pubescens bark and its polar fractions, viz. ethyl acetate and n-butanol, using ultra-performance liquid chromatography-electrospray ionization-quadrupole time-of-flight tandem mass spectrometry(UPLC-ESI-QTOF/MS/MS) technique as well as evaluate the antioxidant using free radical scavenging assays, viz. DPPH and ABTS assays and in-vitro acetylcholinesterase inhibitory activities using Ellman's microplate assay. RESULTS: Metabolic profiling revealed a total of 71, 43, and 55 metabolites tentatively identified in the defatted methanolic extract, ethyl acetate, and n-butanol fractions, respectively. Phenolic acids were the most abundant class, viz. benzoic acids, and acyl quinic acid derivatives followed by flavonoids exemplified mainly by luteolin-C-glycosides and apigenin-C-glycosides. Quantification of the total phenolic and flavonoid contents in the total defatted methanolic extract confirmed its enrichment with phenolics and flavonoids equivalent to 138.61 ± 9.39 µg gallic acid/mg extract and 119.63 ± 4.62 µg rutin/mg extract, respectively. Moreover, the total defatted methanolic extract exhibited promising antioxidant activity confirmed through DPPH and ABTS assays with a 50% inhibitory concentration (IC50) value equivalent to 52.79 ± 2.16 µg/mL and 10.02 ± µg/mL, respectively. The inhibitory activity of acetylcholine esterase (AchE) was assessed using in-vitro Ellman's colorimetric assay, the total defatted methanolic extract, ethyl acetate, and n-butanol fractions exhibited IC50 values of 52.9, 15.1 and 108.8 µg/mL that they proved the significant inhibition of AchE activity. CONCLUSION: The results obtained herein unraveled the potential use of the total methanolic extract of Vitex pubescens bark and its polar fractions as natural candidates for controlling Alzheimer's disease progression.

Characterization and identification of major flavonoids of bamboo leaf extract by HPLC/ESI-QTOF-MS/MS.

Bamboo leaf extract (BLE) is a pale brown powder extracted from bamboo leaves, and it is listed in the Chinese Standard GB-2760 as a legal and safe food additive. The present study aims to identify and characterize the major flavonoids in BLE. The identification of major flavonoids was carried out using ultra performance liquid chromatography combined with electrospray ionization quadruple time-of-flight tandem mass spectrometry (HPLC/ESI-QTOF-MS/MS). A total of 31 flavonoid compounds were identified and tentatively characterized base on reference standards and MS dissociation mechanisms. HPLC/ESI-QTOF-MS can serve as an important analytical platform to identification structure of bamboo leaf flavonoids (BLF).

Green banana (Musa ssp.) mixed pulp and peel flour: A new ingredient with interesting bioactive, nutritional, and technological properties for food applications.

This study aimed to characterize and evaluate the in vitro bioactive properties of green banana pulp (GBPF), peel (GBPeF), and mixed pulp/peel flours M1 (90/10) and M2 (80/20). Lipid concentration was higher in GBPeF (7.53%), as were the levels of free and bound phenolics (577 and 653.1 mg GAE/100 g, respectively), whereas the resistant starch content was higher in GBPF (44.11%). Incorporating up to 20% GBPeF into the mixed flour had a minor effect on the starch pasting properties of GBPF. GBPeF featured rutin and trans-ferulic acid as the predominant free and bound phenolic compounds, respectively. GBPF presented different major free phenolics, though it had similar bound phenolics to GBPeF. Both M1 and M2 demonstrated a reduction in intracellular reactive oxygen species (ROS) generation. Consequently, this study validates the potential of green banana mixed flour, containing up to 20% GBPeF, for developing healthy foods and reducing post-harvest losses.

LC-ESI-QTOF-MS/MS characterization of phenolic compounds in Australian native passion fruits and their potential antioxidant activities.

Passion fruits, renowned globally for their polyphenolic content and associated health benefits, have enjoyed growing attention from consumers and producers alike. While global cultivar development progresses, Australia has pioneered several native cultivars tailored for its distinct planting conditions. Despite their cultivation, comprehensive studies on the phenolic profiles and antioxidant capacities of these Australian-native passion fruits are notably lacking. This study aims to investigate and compare the polyphenolic content present in the by-products, which are peel (L), and consumable portions, which are the pulp and seeds (P), of four indigenous cultivars: 'Misty Gem' (MG), 'Flamengo' (FG), 'Sweetheart' (SW), and 'Panama' (SH). Employing LC-ESI-QTOF-MS/MS for profiling, a comprehensive list of polyphenols was curated. Additionally, various antioxidant assays-DPPH, FRAP, ABTS, RPA, FICA, and •OH-RSA-were performed to evaluate their antioxidant potential. A total of 61 polyphenols were identified, categorized into phenolic acid (19), flavonoids (33), and other phenolic substances (9). In the antioxidant assays, the SHP sample exhibited the highest •OH--RSA activity at 98.64 ± 1.45 mg AAE/g, while the FGL sample demonstrated prominent DPPH, FRAP, and ABTS activities with values of 32.47 ± 1.92 mg TE/g, 62.50 ± 3.70 mg TE/g, and 57.84 ± 1.22 mg AAE/g, respectively. Additionally, TPC and several antioxidant assays had a significant positive correlation, including DPPH, FRAP, and ABTS. The Australian-native passion fruits revealed distinct polyphenolic profiles and diverse antioxidant capacities, establishing a foundation for deeper health benefit analyses. This study accentuates the significance of understanding region-specific cultivars and their potential nutraceutical applications.

Argentinian Rose Petals as a Source of Antioxidant and Antimicrobial Compounds.

The total phenolic, flavonoid, and anthocyanin contents were evaluated in 11 cultivars of Argentinian roses of different colors. HPLC-ESI-QTOF/MS was used to identify the components where ellagic and quinic acids, quercetin, and kaempferol glycosylated derivatives were found. The phenolic contents ranged from 78.8 ± 3.2 to 203.4 ± 3.1 mg GAE/g dw, the flavonoid content ranged from 19.1 ± 3.8 to 125.9 ± 6.5 mg QE/g dw, and the anthocyanin content ranged from less than 0.01 to 5.8 ± 0.1 mg CE/g dw. The dark red cultivars exhibited the greatest levels of the analyzed compounds and of the antioxidant activities, even higher than those of certain plants known for their high phenolic contents and antioxidant activity. Moreover, the addition of these extracts decreased the population of L. innocua and P. aeruginosa to undetectable levels 24 h after inoculation. Rose petal extracts, mainly those with a dark red color, can be used as natural additives in food, feed, and cosmetics, as they contain a high proportion of bioactive compounds with antioxidant and antimicrobial effects.

Optimization of Ultrasonic Extraction Parameters for the Recovery of Phenolic Compounds in Brown Seaweed: Comparison with Conventional Techniques.

Seaweed, in particular, brown seaweed, has gained research interest in the past few years due to its distinctive phenolic profile that has a multitude of bioactive properties. In order to obtain the maximum extraction efficiency of brown seaweed phenolic compounds, Response Surface Methodology was utilized to optimize the ultrasound-assisted extraction (UAE) conditions such as the amplitude, time, solvent:solid ratio, and NaOH concentration. Under optimal conditions, UAE had a higher extraction efficiency of free and bound phenolic compounds compared to conventional extraction (stirred 16 h at 4 °C). This led to higher antioxidant activity in the seaweed extract obtained under UAE conditions. The profiling of phenolic compounds using LC-ESI-QTOF-MS/MS identified a total of 25 phenolics with more phenolics extracted from the free phenolic extraction compared to the bound phenolic extracts. Among them, peonidin 3-O-diglucodise-5-O-glucoside and hesperidin 5,7-O-diglucuronide are unique compounds that were identified in P. comosa, E. radiata and D. potatorum, which are not reported in plants. Overall, our findings provided optimal phenolic extraction from brown seaweed for research into employing brown seaweed as a functional food.

Comparison of phenolic composition in Australian-grown date fruit (Phoenix dactylifera L.) seeds from different varieties and ripening stages.

In this research, different seeds of Australian-grown date palm (Phoenix dactylifera L.) were studied to evaluate the antioxidant potential and analyze their phenolic constituents. Phenolic compounds were extracted from seeds of various Australian-grown date varieties at different ripening stages. Eight varieties of date seeds (Zahidi, Medjool, Deglet nour, Thoory, Halawi, Barhee, Khadrawy, and Bau Strami) at three ripening stages (Kimri, Khalal, and Tamar) were investigated in this study. Date seeds at Khalal (9.87-16.93 mg GAE/g) and Tamar (9.20-27.87 mg GAE/g) stages showed higher total phenolic content than those at Kimri stage (1.81-5.99 mg GAE/g). For antioxidant assays like DPPH, FRAP, ABTS, RAP, FICA, and TAC, date seeds at Khalal and Tamar stages also showed higher antioxidant potential than Kimri stage. However, date seeds at Kimri stage (55.24-63.26 mg TE/g) expressed higher radical scavenging activity than Khalal (13.58-51.88 mg TE/g) and Tamar (11.06-50.92 mg TE/g) stages. Phenolic compounds were characterized using LC-ESI-QTOF-MS/MS, revealing the presence of 37 different phenolic compounds, including 8 phenolic acids, 18 flavonoids, and 11 other phenolic compounds. Further, phenolic compounds were quantified using LC-DAD, revealing that Zahidi variety of date seeds exhibited the highest content during the Kimri stage. In contrast, during the Khalal and Tamar stages, Deglet nour and Medjool date seeds displayed higher concentrations of phenolic compounds. The results indicated an increase in phenolic content in date seeds after the Kimri stage, with significant variations observed among different date varieties.

Identification and characterization of the in-vivo metabolites of the novel soluble epoxide hydrolase inhibitor EC5026 using liquid chromatography quadrupole time of flight mass spectrometry.

EC5026 is a novel soluble epoxide hydrolase inhibitor being developed clinically to treat neuropathic pain and inflammation. In the current study, we employed the LC-ESI-Q-TOF-MS/MS technique to identify four in-vivo phase-I metabolites of EC5026 in rat model, out of which three were found to be novel. The identified metabolites include aliphatic hydroxylation, di-hydroxylation, terminal desaturation, and carboxylation. No phase-II metabolites were found. The pharmacokinetic profile of identified metabolites was established after a single oral dose of EC5026 to Wistar rats. The Tmax of the drug and metabolites were found to be in the range of 1-2 hours and 4-12 hours, respectively. The major metabolites M1 and M2 were found to have more than 2-fold (263.87% AUC) and equivalent exposure (96.33% AUC) compared to the parent drug, respectively. Further, the docking study revealed that the mono-hydroxylated and terminally desaturated metabolites possess better binding affinity than the parent drug. Therefore, these metabolites may hold sEH inhibition potential and can be followed through future research.

Characterization, antioxidant and antitumor activities of phenolic compounds from Amomum villosum Lour.

Amomum villosum Lour. (A. villosum), known as Sharen in China, is widely used for culinary and medicinal purposes due to containing a diverse set of bioactive compounds. In this study, the optimum ethanol extraction process was optimized and the composition and biological activities (antioxidant and antitumor) of five different fractions (dichloromethane, petroleum ether, ethyl acetate, n-butanol and H2O) extracted from the ethanol extract of A. villosum were investigated. The results showed that the optimal extraction conditions were extraction temperature 80°C, extraction time 120 min, ethanol concentration 40% and solid-liquid ratio 1:25 g/mL. Moreover, 35 bioactive compounds were successfully identified by UPLC-ESI-QTOF-MS/MS from five factions for the first time, including 12 phenolic acids and derivatives, 2 organic acids, 12 flavonoids and derivatives, 2 oxylipins and 7 proanthocyanidins. Among them, ethyl acetate fraction (Fr-EtOAc) exhibited the highest content of total phenolic (374.01 mg GAE/g DW) and flavonoid (93.11 mg RE/g DW), where vanillic acid, catechin, epicatechin and protocatechuic acid were the predominant phenolic compounds that accounting for 81.65% of the quantified bioactive compounds. In addition, Fr-EtOAc demonstrated excellent total antioxidant activity (IC50 of DPPH and ABTS assays were 0.23, 0.08 mg/mL, respectively, and FRAP assay was 322.91 mg VCE/100 g DW) and antitumor activity (1,000 µg/mL, 79.04% inhibition rate). The results could provide guidance for the industrial production and application of A. villosum.

RP-UHPLC-ESI-QTOF-MSn analyses, antioxidant, antimicrobial, analgesic activities and toxicity of Achillea ligustica All.

Fourty five polyphenols were identified in the hydroethanol extract of Achillea ligustica All. by LC-HRMS/MS with caffeoyl-6-oleside (5.74%), eucommin A (4.03%), quercetin-3-O-ß-D-glucoside (3.13%) and cirsimaritin (2.95%) as the major compounds. A good antioxidant potential was shown in DPPH, ABTS and phenanthroline tests and the highest antioxidant activity (A0.5:36.23 ± 3.07 µg/mL), which was close to the standard α-tocopherol, was shown in Reducing power. The extract inhibited the growth of all tested microorganisms with MICs ranging between 10 and 190 µg/mL. In the acute toxicity test, no death was observed at doses of 100, 750 and 1500 mg/Kg with DL50 higher than 2000 mg/Kg. In analgesic in vivo assay, the extract showed a very important capacity to reduce pain, whether central or peripheral, with a certain dose-dependent relationship. For the three tests (tail flick, hot plate and acetic acid assay), the effective dose was 750 mg/kg.

Influence of the Encapsulating Agent on the Bioaccessibility of Phenolic Compounds from Microencapsulated Propolis Extract during In Vitro Gastrointestinal Digestion.

The aim of this work is to develop different encapsulated propolis ingredients by spray-drying and to evaluate their bioaccessibility using simulated in vitro digestion. To achieve these goals, first, microparticles of a propolis extract with inulin as the coating polymer were prepared under the optimal conditions previously determined. Then, a fraction of inulin was replaced with other encapsulating agents, namely sodium alginate, pectin, and chitosan, to obtain different ingredients with controlled release properties in the gastrointestinal tract. The analysis of the phenolic profile in the propolis extract and microparticles showed 58 compounds tentatively identified, belonging mainly to phenolic acid derivatives and flavonoids. Then, the behavior of the free extract and the formulated microparticles under gastrointestinal conditions was studied through an in vitro gastrointestinal digestion process using the INFOGEST protocol. Digestion of the free extract resulted in the degradation of most compounds, which was minimized in the encapsulated formulations. Thus, all developed microparticles could be promising strategies for improving the stability of this bioactive extract under gastrointestinal conditions, thereby enhancing its beneficial effect.

Mechanochemistry of Strecker degradation: Interaction of glyoxal with amino acids.

Mechanochemistry is rapidly evolving into a versatile and green method for chemical synthesis. Due to its unique reaction conditions, ball milling of sugars and amino acids mainly leads to the formation of Amadori products with minimum degradation. In this study, we milled glyoxal trimer dihydrate with twenty proteogenic amino acids to demonstrate the formation of Strecker degradation products. HS-GC/MS studies indicated that Strecker degradation proceeded to selectively generate Strecker aldehyde and unsubstituted pyrazine as the major volatiles. Moreover, ESI/qToF/MS studies demonstrated for the first time the formation of the proposed key Strecker degradation intermediates, such as the condensation products and their decarboxylated products, indicating the similarity of the mechanism of Strecker reaction under ball milling to that proposed under hydrothermal reaction conditions. These studies provided supporting evidence that ball milling at ambient temperatures could be used as a novel synthetic approach to prepare precursors of aroma-active volatiles through Strecker degradation.

Metabolomic profiling and quantification of polyphenols from leaves of seven Acacia species by UHPLC-QTOF-ESI-MS.

The genus Acacia (Fabaceae) comprises >1350 species and has been used in traditional medicine as infusions and decoctions to treat wounds, sores, headaches, diarrhea, and cough. The leaf methanolic extracts of seven Acacia species growing in Egypt namely: Acacia saligna, Acacia seyal, Acacia xanthophloea, Acacia tortilis subsp. raddiana., Acacia tortilis, Acacia laeta, Acacia albida were analyzed using UPLC-QTOF-ESI-MS. A total of 37 polyphenols were identified and discussed in detail. They included phenolic acids, flavonoids, and procyanidins, among which sixteen polyphenols were identified in Acacia for the first time. Folin-ciocalteau assay and ferric reducing antioxidant power, cupric reducing antioxidant capacity, 2,20 -azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) cation radical and the scavenging capacity against 2,2-diphenyl-1- picrylhydrazyl radical were performed to investigate the total phenolic content and the antioxidant activity of the Acacia extracts, respectively. Furthermore, the absolute quantification of eighteen polyphenols common to most of the species was performed using UPLC-MS. It was evident that the differences in the chemical composition among the species accounted for the difference in antioxidant activity which was in line together with the total phenolic content.

Steroidal saponins profiling of Rohdea chinensis (Baker) N.Tanaka by high performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry.

The rhizome of Rohdea chinensis (Baker) N.Tanaka (RRc) is a famous folk medicine for the treatment of carbuncles and pharyngitis. Steroidal saponins (SSs) were considered to be the most abundant active constituents in RRc. However, to date, the in-depth study of SSs is still lacking. This study was aimed to investigate the SSs profiles of RRc extract by HPLC-ESI-QTOF-MS/MS. Analysis was performed on an Agilent poroshell 120 EC-C18 column (2.1 mm × 100 mm, i.d., 2.7 µm) with 0.1% formic acid aqueous solution and acetonitrile as the mobile phase under gradient conditions. The results showed that 32 SSs including 20 furospirostanol, 11 spirostanol and 1 pseudo-spirostanol saponins were identified, 5 of which were reported in this plant for the first time. This is the first report on the analysis of SSs in RRc. This novel analysis method may stimulate further research regarding the identification of SSs in other plant species.

Mango (Mangifera indica L.) seed kernel extract suppresses hyperglycemia by modulating pancreatic ß cell apoptosis and dysfunction and hepatic glucose metabolism in diabetic rats.

This study investigated the anti-hyperglycemic action of mango seed kernel extract (MKE) and various mechanisms involved in its actions to improve pancreatic ß cells and hepatic carbohydrate metabolism in diabetic rats. An intraperitoneal injection of 60 mg/kg of streptozotocin (STZ) followed by 30 consecutive days of treatment with MKE (250, 500, and 1000 mg/kg body weight) was used to establish a study group of diabetic rats. Using liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS) for identification, 26 chemical compounds were found in MKE and the high-performance liquid chromatography (HPLC) analysis of the MKE also revealed the existence of mangiferin, gallic acid, and quercetin. The results confirmed that in each diabetes-affected rat, MKE mitigated the heightened levels of fasting blood glucose, diabetic symptoms, glucose intolerance, total cholesterol (TC), and low-density lipoprotein-cholesterol (LDL-C). As demonstrated by a remarkable increment in serum and pancreatic insulin, the diabetic pancreatic ß cell function was potentiated by treating with MKE. The effect of MKE on diabetic pancreatic apoptosis clearly reduced the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells, which was related to diminished levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and Bax and an increase in Bcl-xL protein expression. Furthermore, diabetes-induced liver damage was clearly ameliorated along with a notable reduction in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels and abnormal liver histology. By enhancing anti-oxidant superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities, MKE alleviated diabetes-induced pancreatic and liver oxidative damage, as demonstrated by diminished levels of malondialdehyde. In minimizing the expression levels of glucose 6-phosphatase and phosphoenolpyruvate carboxykinase-1 proteins in the diabetic liver, MKE also enhanced glycogen content and hexokinase activity. Collectively, these findings indicate that by suppressing oxidative and inflammatory processes, MKE exerts a potent anti-hyperglycemic activity in diabetic rats which serve to protect pancreatic ß cell apoptosis, enhance their function, and improve hepatic glucose metabolism.

Development and Validation of LC-MS/MS Method for Determination of Cytisine in Human Serum and Saliva.

Cytisine (CYT) is a quinolizidine alkaloid used for nicotine addiction treatment. Recent clinical trial data regarding cytisine confirm its high effectiveness and safety as a smoking cessation treatment. CYT's popularity is growing due to its increased availability and licensing in more countries worldwide. This increased use by smokers has also resulted in an urgent need for continued drug research, including developing appropriate analytical methods for analyzing the drug in biological samples. In this study, a simple, fast, and reliable method combining hydrophilic interaction liquid chromatography and electrospray ionization quadrupole time of flight mass spectrometry (HILIC/ESI-QTOF-MS) for the determination of CYT in human serum and saliva was developed and validated. This was undertaken after the previous pre-treatment of the sample using solid-phase extraction (SPE). A hydrophilic interaction liquid chromatography (HILIC) column with a silica stationary phase was used for chromatographic analysis. In a linear gradient, the mobile phase consisted of acetonitrile (ACN) and formate buffer at pH 4.0. The proposed method was fully validated and demonstrated its sensitivity, selectivity, precision, and accuracy. The method was successfully applied to determine CYT in serum and, for the first time, in saliva. The findings indicate that saliva could be a promising non-invasive alternative to measure the free concentration of CYT.

First Chemical Profile Analysis of Acacia Pods.

This study intended to evaluate the potential industrial applications of various Acacia species (Acacia melanoxylon, Acacia longifolia, Acacia cyclops, Acacia retinodes, Acacia pycnantha, Acacia mearnsii, and Acacia dealbata) by examining their chemical composition, antioxidant, and antimicrobial properties. Using high-resolution mass spectrometry, a comprehensive analysis successfully identified targeted compounds, including flavonoids (flavonols/flavones) and phenolic acids, such as 4-hydroxybenzoic acid, p-coumaric acid, and ellagic acid. Additionally, p-coumaric acid was specifically identified and quantified within the hydroxycinnamic aldehydes. This comprehensive characterization provides valuable insights into the chemical profiles of the studied species. Among the studied species, A. pycnantha exhibited a higher concentration of total phenolic compounds, including catechin, myricetin, quercetin, and coniferaldehyde. Furthermore, A. pycnantha displayed notable antibacterial activity against K. pneumoniae, E. coli, S. Typhimurium, and B. cereus. The identified compounds in Acacia pods and their shown antibacterial activities exhibit promising potential for future applications. Moreover, vibrational spectroscopy was a reliable method for distinguishing between species. These significant findings enhance our understanding of Acacia species and their potential for various industrial applications.

Evaluation of the antioxidant potential of brown seaweeds extracted by different solvents and characterization of their phenolic compounds by LC-ESI-QTOF-MS/MS.

Seaweeds, serving as valuable natural sources of phenolic compounds (PCs), offer various health benefits like antioxidant, anti-inflammatory properties, and potential anticancer effects. The efficient extraction of PCs from seaweed is essential to harness their further applications. This study compares the effectiveness of different solvents (ethanol, methanol, water, acetone, and ethyl acetate) for extracting PCs from four seaweed species: Ascophyllum sp., Fucus sp., Ecklonia sp., and Sargassum sp. Among them, the ethanol extract of Sargassum sp. had the highest content of total phenolics (25.33 ± 1.45 mg GAE/g) and demonstrated potent scavenging activity against the 2,2-diphenyl-1-picrylhydrazyl radical (33.65 ± 0.03 mg TE/g) and phosphomolybdate reduction (52.98 ± 0.47 mg TE/g). Ecklonia sp. had the highest content of total flavonoids (0.40 ± 0.02 mg QE/g) in its methanol extract, whereas its ethyl acetate extract contained the highest content of total condensed tannins (8.09 ± 0.12 mg CE/g). Fucus sp. demonstrated relatively strong antioxidant activity, with methanolic extracts exhibiting a scavenging ability against 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical (54.41 ± 0.24 mg TE/g) and water extracts showing ferric-reducing antioxidant power of 36.24 ± 0.06 mg TE/g. Likewise, liquid chromatography-mass spectrometry identified 61 individual PCs, including 17 phenolic acids, 32 flavonoids, and 12 other polyphenols. Ecklonia sp., particularly in the ethanol extract, exhibited the most diverse composition. These findings underscore the importance of selecting appropriate solvents based on the specific seaweed species and desired compounds, further providing valuable guidance in the pharmaceutical, nutraceutical, and cosmetic industries. PRACTICAL APPLICATION: The PCs, which are secondary metabolites present in terrestrial plants and marine organisms, have garnered considerable attention due to their potential health advantages and diverse biological effects. Using various organic/inorganic solvents during the extraction process makes it possible to selectively isolate different types of PCs from seaweed species. The distinct polarity and solubility properties of each solvent enable the extraction of specific compounds, facilitating a comprehensive assessment of the phenolic composition found in the seaweed samples and guiding industrial production.