Total Phenolic Assay Research Articles

A-064 Improved Insulin production in Advanced glycation end-products (AGEs)-induced RIN-m5F pancreatic β-cells by using a Flavonoid-Rich Medicinal Plant, Kaempferia parviflora treatment

Abstract Background Advanced glycation end-products (AGEs) are glycated proteins that react with reducing sugar which causes the damages of function and cellular signaling of insulin production in insulinoma cells. AGEs also contribute to hyperglycemia-induced insulin resistance in type 2 diabetes. AGEs gradually form during the chronic high blood glucose condition, which is a major risk factor and harmful to β-cells function and survival, and could alter the insulin signaling and downregulate the insulin-responsive transcription factors including PDX-1, Glut-2, GCK, and Pre-INS. Kaempferia parviflora, also known as black ginger and flavonoid-rich medicinal plant, has beneficial properties for type 2 diabetes patients as a counteracts the detrimental effects of AGEs and improves the insulin production in pancreatic β-cells. Thus, we hypothesize that treated β-cells with K. parviflora, which is the most effective flavonoid-containing plant might protect against the damage of AGEs-induced insulin impairment, and these effects are also responsible for improved insulin synthesis and the essential transcription factors in pancreatic β-cells. Methods Rat pancreatic β-cell line RIN-m5F (ATCC CRL-2058) were maintained and cultured in RPMI 1640 medium containing 10% (v/v) FBS, 1% penicillin-streptomycin at 37◦C in a humidified atmosphere containing 5% CO2. RIN-m5F cells were pre-treated with the K. parviflora at concentrations 0, 25, and 50 µg/ml for 48 hours, and afterward, exposure to 250 µg/ml AGEs treatment. To quantify the bioactive compound of K. parviflora, total phenolic and flavonoid content assays were performed and the transcriptional changes of insulin-related genes (PDX-1, Glut-2, GCK, and Pre-INS) were determined by using real-time qRT-PCR. Results K. parviflora has shown a high phenolic content and flavonoid content was 50.43 ± 1.74, 26.66 ± 1.18 mg GAE/g Dry weight of sample, respectively. For the transcriptional changes of insulin-related genes, the levels of gene expression in the regulators of insulin secreting pathway (PDX-1, Glut-2, GCK, and Pre-INS) were significantly downregulated after exposure to AGEs-treated, while the pre-treatment of K. parviflora was showed statistically difference. PDX-1, Glut-2, GCK, and Pre-INS expressions were significantly upregulated after pre-treated with 50 µg/ml K. parviflora. However, under the AGEs-treated alone condition, the transcriptional levels were significantly decreased as compared to the control (untreated). Taken together, these results suggest K. parviflora could be a potential natural medicine to ameliorate the damage of insulin production in AGEs-induced β-cell damage and highlight promising alternatives for therapeutic avenue against diabetes. Conclusions Treated RIN-m5F pancreatic β-cells with K. parviflora, a flavonoid-rich medicinal plant, could upregulate the insulin-responsive transcription factors and improve the insulin production in damaging AGEs-induced pancreatic β-cells. Our findings provide a novel strategy related metabolic disease and insights into the regulation of insulin synthesis by using K. parviflora treatment as a therapeutic agent against type 2 diabetes.

Ultrasonic-Assisted Water-Rich Natural Deep Eutectic Solvents for Sustainable Polyphenol Extraction from Seaweed: A Case Study on Cultivated Saccharina latissima.

This case study introduces a green, 1 h single-step method using water-rich natural deep eutectic solvent (WRNADES) for ultrasound-assisted extraction (UAE) of polyphenols fromSaccharina latissima, a commercially cultivated brown seaweed. The extraction efficiency was evaluated using a selective quantitative NMR method (s-qNMR) and the traditional nonselective colorimetric total phenolic content assay (TPC). Initial 6 h extractions in traditional solvents (methanol, ethanol, acetone, and ethyl acetate) showed a 40-60% increase in polyphenolic yields in 50% aqueous solutions measured by the TPC method. Six different water-rich (50%) NADES (WRNADES) combinations were tested (choline chloride/betaine with lactic acid, citric acid, and 1,3-butanediol), with betaine and 1,3-butanediol (1:1) proving most effective. Parameters for the WRNADES were optimized using Box-Behnken design response surface methodology, resulting in a 1:20 w/w biomass to solvent ratio and a 1 h extraction time at 50 °C. The WRNADES extraction process was refined into a scalable, single-step procedure and compared with traditional solvent extractions (6 h, 50% aqueous methanol and acetone). A final XAD-7 polyphenol recovery step was included in all extractions. The optimized WRNADES extraction yielded 15.97 mg GAE/g of the dry weight recovered polyphenolic extract (s-qNMR), exceeding the 6 h 50% aqueous methanol (12.4 mg GAE/g) and acetone (11.4 mg GAE/g) extractions. Thus, the UAE-WRNADES method presented in this case study provides a cost-effective, sustainable, and eco-friendly alternative for the extraction of phenolic compounds from seaweed. It promotes the development of environmentally friendly production processes within the seaweed biorefinery.

Unveiling the therapeutic potential of Canavalia rosea leaves: Exploring antioxidant, anti-inflammatory, anti-arthritic, and cytotoxic activities through biological and molecular docking evaluation with DFT analysis

BackgroundCanavalia rosea is a common tropical seacoast flowering plant from the family of Fabaceae which is reported as bay bean and coastal jack bean; has a wide range of therapeutic and nutraceutical properties. AimThe present research aims to explore some pharmacological insights of the methanol extract C. rosea of leaves (MECR) and its chloroform fraction (CFCR) and n-hexane fraction (NFCR) through in-vitro and in-silico approaches. MethodsDifferent fractions of C. rosea were subjected to ferric reduction assay and total phenolic and flavonoid content assay to explore their antioxidant potential. The anti-inflammatory activity was assessed on the hypotonic-induced human erythrocyte-lysis model, while the protein denaturation method was applied for screening anti-arthritis properties of plant extract; and the cytotoxic activity was evaluated by brine shrimp lethality bioassay. In the in-silico study, molecular docking, pass prediction and ADME/T, analysis was used to investigate anti-arthritic, anti-inflammatory, antioxidant and cytotoxic potency of five selected compounds of C. rosea. Finally, the quantum chemical density functional test analysis was applied to investigate the chemical and physical properties of those compounds. ResultsAll the soluble organic extracts of C. rosea demonstrated moderate toxicity with strong antioxidants potential, in which MECR manifested the peak level of scavenging activity (2.49) on ferric reduction assay. MECR, CFCR & NFCR significantly protected lysis of human erythrocyte membrane induced by hypotonic solution, whereas MECR and CFCR were exhibited partially equal inhibitory activity. The in-vitro anti-arthritis assay, MECR, NFCR and CFCR showed strongly significant (p˂0.001) inhibitory potency at 500 μg/mL & 1000 μg/mL concentrations. The molecular docking simulations revealed strong binding of all compounds to specific receptors, with Rutin showing the highest biological reactivity followed by Daucosterol, β-Sitosterol, Stigmasterol, and Guanidine. All the compounds showed favorable reactivity patterns, with O and H atoms poised for nucleophilic and electrophilic attacks in chemical density functional calculations. ConclusionThe recent investigation suggests that C. rosea could have the potential source of antioxidant, anti-inflammatory, anti-arthritis and cytotoxic activity, prompting further investigation into their mechanisms.

Phytochemical Investigations and In-vitro Antioxidant Analysis of Passiflora incarnata L.

The present study aims to investigate the presence of the phytochemical and in-vitro antioxidant study of hydroalcoholic (HA) and ethyl acetate (EA) preparations of Passiflora incarnata L. leaves. The hydroalcoholic (HA) and ethyl acetate (EA) extracts showed strong antioxidant activity when examined in a laboratory setting using DPPH, H2O2, NO, reducing power, total phenol, and TOC assays. Also compounds like saponin, alkaloids, flavonoids, and phenolics were all identified through phytochemical testing. In addition to expanding our understanding of the pharmacological characteristics of P. incarnata L., the results of this study have the potential to change the way medicinal botanicals are viewed forever. Nature’s treasure trove, concealed within the leaves of P. incarnata L., offers a pharmacological narrative that transcends traditional boundaries, suggesting new dimensions for antioxidant-based therapeutic strategies for the development of novel botanical interventions in preventive healthcare and disease management.

Unveiling the Therapeutic Potentials of Water Hyacinth (Eichhornia crassipes (Mart.) Solms) Flower against Oxidative Stress, Inflammation and Depressive Disorders: GC-MS/MS, In Vitro, In Vivo and In Silico Approaches.

Water hyacinth (Eichhornia crassipes (Mart.) Solms) is a highly invasive aquatic weed native to the Amazonia basin, known for its rapid propagation, adaptability and utilization in traditional medicine. The study aims to unveil the therapeutic potential of water hyacinth flowers methanolic extract (EC-CME) and its four kupchan fractions (EC-PESF, EC-DCMSF, EC-EASF, EC-ASF) through diversified chemical-pharmacological approaches. GC-MS/MS of EC-CME uncovered a rich tapestry of 72 phytochemical components. In vitro DPPH scavenging assay and total phenolic content determination assay deciphered promising antioxidant assays with remarkably low IC50 values of 0.353 and 0.485 µg/mL, respectively for EC-ESF and EC-ASF. Besides, different in vivo tests, including tail emersion, acetic acid-induced writhing, and thiopental-induced sleeping test of EC-CME, yielded a remarkable 8.61 ± 0.29 minutes of tail immersion time compared to the control's 2.05 ± 0.11 minutes at the highest dose (600 mg/kg). The best % inhibition of writhing was recorded as 47.96% accrued in 400 mg/kg dose, indicating robust pain-relieving properties. The onset and duration of sleep are significantly ameliorated for EC-CME, unveiling its antidepressant potential. Besides, molecular docking studies along with ADME/T analysis also validated the wet lab findings as well as their safety, efficacy and drug-likeliness profile.

Chemical Characterization of Pruning Wood Extracts from Six Japanese Plum (Prunus salicina Lindl.) Cultivars and Their Antitumor Activity.

The Japanese plum tree (Prunus salicina Lindl.) is mainly cultivated in temperate areas of China and some European countries. Certain amounts of wood (from pruning works) are generated every year from this crop of worldwide commercial significance. The main objective of this work was to value this agricultural woody residue, for which the chemical composition of pruning wood extracts from six Japanese plum cultivars was investigated, and the antiproliferative activity of extracts and pure phenolics present in those extracts was measured. For the chemical characterization, total phenolic content and DPPH radical-scavenging assays and HPLC‒DAD/ESI‒MS analyses were performed, with the procyanidin (-)-ent-epicatechin-(2α→O→7,4α→8)-epicatechin (5) and the propelargonidin (+)-epiafzelechin-(2β→O→7,4β→8)-epicatechin (7) being the major components of the wood extracts. Some quantitative differences were found among plum cultivars, and the content of proanthocyanidins ranged from 1.50 (cv. 'Fortune') to 4.44 (cv. 'Showtime') mg/g of dry wood. Regarding the antitumoral activity, eight wood extracts and four phenolic compounds were evaluated in MCF-7 cells after 48 h of induction, showing the wood extract from cv. 'Songold' and (‒)-annphenone (3), the best antiproliferative activity (IC50: 424 μg/mL and 405 μg/mL, respectively).

Utilization of Indian gooseberry (Phyllanthus emblica L.) as foods

This study aimed to investigate the consumption of amla as a food by conducting a thorough literature analysis and a field survey with unstructured, in-depth interviews. Amla or Indian gooseberry (Phyllanthus emblica L.) is rich in phytochemicals, and their pharmaceutical potentials have been extensively reported. However, their utilization as foods is not well documented. Thai recipes use whole or minced fresh amla fruits as an ingredient in some spicy dishes, such as chili pastes, soups, and salads. Like other Asian countries, amla could be processed into products such as juice, preserves, pickles, and dried amla. Ten amla dishes and products were selected for evaluation of their ascorbic acid contents and antioxidant properties (total phenolic compounds, total flavonoids, DPPH, and FRAP assays). High-heat processing resulted in a marginal reduction of ascorbic acid in amla dishes and products. Processing methods also affected antioxidant activities, and they varied depending on processing conditions and product types. The processing of amla into juice slightly decreased antioxidant activities. Thai foods that used amla as an ingredient exhibited less antioxidant activity than those made of fresh amla. The antioxidant activities of pickled and preserved amlas were substantially diminished due to their high salt and sugar content. On the contrary, dried amla demonstrated enhanced antioxidant activities as a result of its reduced moisture content and the presence of concentrated phytochemicals. Given its substantial phytonutrient content and lack of utilization, the results obtained from this research contribute to the promotion of amla as a valuable food ingredient.

Grewia flava twig extracts: phytochemical, antioxidant, and antimicrobial evaluations

BackgroundGrewia flava infusions are consumed to assist with kidney problems and stomach ailments. However, there are no scientific data on the phytochemical profile or biological properties to validate its folklore use. Thus, the study aimed to assess the phytochemical profile, antioxidant, and antimicrobial activities of Grewia flava twig extracts.MethodologyThe antioxidant activities of the extracts were assayed using 1,1-diphenyl-2-picrylhydrazyl radical scavenging, reducing power, metal chelation, and total phenolic and flavonoid content assays. The agar well diffusion and microdilution methods were used for crude extracts and fractions (from 80% methanol extract) for antimicrobial screening against P. aeruginosa, S. aureus, E. coli, B. subtilis, A. niger, and R. oryzae.ResultsThe 80% methanol twig extract (250.00 ± 2 GAE/g) exhibited a high concentration of phenolic content, followed by the distilled water extract (192.00 ± 2 mg GAE/g) and the hexane extract (43.10 ± 0.2 mg GAE/g). Fraction 14 of the methanol twig extract exhibited MIC values of 0.21–0.31 mg/mL against all test microorganisms. The root and twig extracts exhibited significant antioxidant and antimicrobial activities, which were attributed to the extracts of bioactive phytochemical compounds such as alkaloids, flavonoids, saponins, steroids, glycosides, anthraquinones, and tannins that were detected in the extracts. Also, the root and twig non-polar extracts were subjected to gas chromatography–mass spectrometry analysis, which identified several bioactive compounds like betulin, β-amyrin, palmitic acid, lupenone, and phytol, highlighting the potential of the plant species as a botanical drug.ConclusionsThe study supports the traditional use of plant roots and twigs for treating various ailments, indicating their medicinal value. The twigs can be used in place of the roots to guarantee Grewia flava harvesting that is sustainable. However, a comparison of the quantities of the active compound in the twigs and roots using LC–MS is crucial.

New potentiometric sensor for total phenolic assay of plant extracts

Abstract In this study, a sensitive, fast, and inexpensive new potentiometric determination method based on the Folin–Ciocalteu reagent (FCR) is proposed to determine the total phenolic content in plant extracts. For this purpose, a potentiometric-based all-solid-state-contact polyvinyl chloride membrane sensor was developed. Gallic acid compound was used as the active component (ionophore) in the developed sensor and the potentiometric behavior of the sensor was characterized. The detection limit of the selective sensor to the FCR was determined as 0.022 mol L−1 and showed a linear potential change in the concentration range of 0.0039 to 0.5 mol L−1 and a fast response time of 40 to 45 s. The developed sensor was also applied to the plant extracts and the obtained measurement results were found to be in agreement with the spectrometer results in the literature.

Improvement of the antioxidant potential: impact of drying and extraction techniques on polyphenols in Arbutus unedo L. leaf aqueous extract

The search for alternatives to chemical pharmaceutical products remains an urgent and crucial step for humanity. In this regard, the use of plants presents an ideal approach for such biological studies. Arbutus unedo L., which belongs to the family Ericaceae, is one of the species that holds significant importance in traditional pharmaceutical uses. In Algeria, the use of this species in traditional treatment is generally rare. Therefore, this study aims to highlight its medicinal importance and the proper method for extracting its bioactive elements. To achieve this, the biological activity of leaves including total phenolic content (TPC), DPPH assay, and total flavonoid content (TFC), were assessed under four different drying temperatures (shade-drying, sun-drying, 40°C and 60C°) and two extraction methods after maceration (centrifugation, filtration). The results have indicated that these processes have affected the final accumulation of polyphenols and antioxidant activity in the leaf extracts. Additionally, it has been highlighted that the centrifugation method extracts a higher biochemical amount, especially after drying at 40°C. Furthermore, Arbutus unedo L. leaves should be considered a promising source of natural compounds to be used as ingredients in various fields.

Exploring antioxidative, cytotoxic and neuropharmacological insights into Bixa orellana leaves: Experimental and in silico approaches

Background StudyThe aim of this research was to examine possible antioxidant, cytotoxic and neurological activity of methanol and n-hexane extracts of Bixa orellana leaves. Additionally, we aimed to identify potential lead compounds through in-silico analysis. MethodsIn-vitro antioxidative properties were investigated through different assays, including: total phenolic content assay (TPC), total flavonoid content assay (TFC), DPPH free radical scavenging assay and reducing power assay. Also, the cytotoxic effect of the samples was assessed using the brine shrimp lethality test. In addition, anxiolytic, locomotor, and CNS depressant activities were assessed utilizing various established methods. Moreover, reported compounds were used in the in silico study to explore the best-fit phytoconstituents against gamma-aminobutyric acid (GABAA) receptor. ResultsMBOL displayed substantial antioxidative activities in various established assays compared to NBOL. In brine shrimp lethality bioassay, both MBOL and NBOL revealed cytotoxic activity in a concentration-dependent approach. Again, in Elevated Plus Maze test, 200 and 400 mg/kg of NBOL and MBOL demonstrated significant anxiolytic activities evident from time spent in open arms. In addition, maximum number of head dipping was demonstrated by MBOL at 400 mg/kg (53.90 ± 1.16) in Hole Board test. NBOL and MBOL at both doses significantly diminished the magnitude of movements from the 2nd to 5th observation periods in Open Field test. Furthermore, in Hole Cross test, MBOL remarkably dwindled the locomotor activity at 120 min and 180 min (3.60 ± 0.40 and 2.40 ± 0.51) at 400 mg/kg. Finally, in silico analysis revealed 13 compounds as promising leads with strong binding affinity to GABAA receptor along with good pharmacokinetics and toxicity profiles. ConclusionTherefore, the present study's findings advocate the traditional usage of this plant and recommend both MBOL and NBOL as as a potential source of therapeutic candidate for the management of neurological disorders.

Neuroprotective Potential of Thunbergia laurifolia Lindl Leaf Extracts Against Beta Amyloid-induced Neurotoxicity: An in vitroModel of Alzheimer’s Disease

Background: Beta-amyloid peptide (Aβ) induces oxidative stress, contributing to Alzheimer’s disease (AD) initiation and progression. This study aims to explore Thunbergia laurifolia ( T. laurifolia) leaf extract’s protective effects on Aβ25–35-induced oxidative stress and cell injury in SH-SY5Y cells and investigate underlying mechanisms. Materials and Methods: SH-SY5Y cells were treated with T. laurifolia leaf extract in the presence or absence of Aβ25–35. After 24 h, neuroprotective effects were assessed using cell viability and lactate dehydrogenase (LDH) assays. Caspase-3/7 activity, intracellular reactive oxygen species (ROS) levels, catalase (CAT), and superoxide dismutase (SOD) activities were measured to examine mechanisms. Total flavonoid and phenolic content assays were performed. Results: The findings showed that exposure to Aβ25–35 led to a notable rise in oxidative stress in SH-SY5Y cells, as evidenced by increased levels of ROS. Additionally, Aβ25–35 treatment increased caspase-3/7 activity and LDH release and decreased cell viability. However, T. laurifolia extract effectively suppressed ROS production, attenuated caspase-3/7 activity, and concentration-dependently reduced Aβ25–35-induced neurotoxicity. LDH release decreased, and cell viability increased. SOD and CAT activities also increased after T. laurifolia treatment. The extract had total phenolic and flavonoid contents of 178.5 ± 6.86 and 32.51 ± 1.26 mg/g, respectively. Conclusion: T. laurifolia extract demonstrated neuroprotective effects against Aβ25–35-induced injury in SH-SY5Y cells. These effects were attributed to reduced oxidative stress, elevated SOD and CAT activity, and suppressed caspase-3/7 activity. T. laurifolia extract shows potential as an alternative or therapeutic approach to AD mediated by Aβ. Nevertheless, further research is needed to elucidate the mechanism by which T. laurifolia ameliorates neuronal cell death induced by Aβ25–35.

MICROMERIA MYRTIFOLIA BOISS. & HOHEN’İN BİYOLOJİK AKTİVİTELERİNİN BELİRLENMESİ

Objective: Lamiaceae family has a wide variety of well-known and lesser-known plants with strong medicinal qualities. The genus Micromeria Benth. is a member of this family consisting of herbaceous plants with a variety of significant biological, phytochemical, and ethnobotanical uses. In this study, the biological activities of methanol and ethanol extracts of Micromeria myrtifolia were evaluated. Material and Method: To demonstrate the antioxidant activity DPPH radical scavenging activity and total phenolic content assays were done. The effects of the extracts on acetylcholinesterase (AChE) and monoamine oxidase-A were then assessed. Result and Discussion: Methanol extract showed the highest DPPH scavenging activity, at the dose of 10 mg/ml with a value of 96.55%. For the highest concentration that can be applicable, AChE inhibitions for the methanol and ethanol extracts were 25% and 27%, respectively. On the other hand, the inhibitory effects of the ethanol and methanol extracts of the plant on MAO-A were determined; for the ethanol extract IC50 value was found as 32.5876 ± 0.89 g/ml, and for the methanol extract it was found as 34.6544 ± 0.76 g/ml. It can be told that M. myrtifolia can act as a potential antioxidant. With further research and investigation, it is thought that Micromeria myrtifolia could be used as a natural source for the treatment of various neurological diseases.

Phytochemical Screening and Quantitative Determination of Antioxidant Properties of Watermelon Rinds and Seeds

In this present age of food insecurity, it is essential to source for agricultural waste that can be nutritionally beneficial to humans and animals. Watermelon rinds and seeds are regarded as agricultural wastes which need to be utilized for human and animal nutrition. This research work is aimed at examining the extractive values, qualitative and quantitative determination of phytochemicals and antioxidants properties in watermelon rinds and seeds and their solvent extracts using six different solvents. The rinds and seeds of watermelon fruits were obtained, rinsed, cut into smaller pieces, air-dried, ground into powdery sample, and sieved with 40 mm mesh size. 20 g each of sample was extracted using 200 mL of six different solvents (acetone, chloroform, ethyl acetate, ethanol, methanol and water) for 72 h. Each solvent extract was screened for twelve phytochemicals (flavonoid, phenol, reducing sugar, tannin, saponin, alkaloid, volatile oil, quinone, cardiac glycoside terpenoids, steroids and phlobatannin). The extractive values of watermelon rinds ranged from 1.51±0.26% to 13.56±0.20% while that of watermelon seeds ranged from 3.54±0.16% to 10.17±0.19% in all the six solvents used. Acetone and methanol had the higher extractable phytochemicals (41.67%) in watermelon rinds while methanol, ethyl acetate and ethanol (58.33 – 66.67%) had the highest extractable phytochemicals in watermelon seeds. Water and methanol extracts of watermelon rinds had higher total phenol, DPPH and Iron (Fe2+) chelation assay than raw sample of watermelon rinds. Water and methanol extracts of watermelon rinds had higher iron (Fe2+) chelation assay and ferric reducing antioxidant power (FRAP) than raw sample of watermelon seeds. Methanol extract of watermelon seeds had higher total flavonoid, total phenol and DPPH than raw sample of watermelon seeds.

Simultaneously quantification of eight marker compounds by HPLC, and HPTLC analysis for the marker-based shelf-life kinetic study for the standardization of polyherbal drug AYUSH SG-5, medicinal properties and computational studies

The herbal or alternative medicines having their own significances in each country. The major problem behind the non-acceptance of the alternative medicines is the poor standardization and quality control. Maximum times, it’s impossible to quantify the complex matrix of polyherbal medicines. The pharmacopeial standards of these medicines are limited to physicochemical and pharmacognostic analysis only, which are insufficient for the proper quality control of ingredients used for the preparation of polyherbal formulations. Multi-markers-based analysis of alternative medicines and their polyherbal formulations by using the sophisticated analytical techniques like HPLC, and LC-MS is the demand of current scenario. In our recent report we have reported pharmaceutical standard operating procedure for a coded polyherbal formulation (CPF i.e. AYUSH-SG-5) which was prepared for the rheumatoid arthritis. The marker-based quantification and identification of major metabolites and shelf-life (marker-based) studies were not reported in that study. In this report we have quantified eight markers (6-Gingerol, 8-Gingerol, Guggulsterone-E, Guggulsterone-Z, Resveratrol, Salicin, Colchicine, and Caffeic acid) simultaneously by HPLC chromatography. To identify the phytochemicals in polyherbal formula (CPF), LC-QTOF-MS/MS analysis was performed, where 32 main phytochemicals were identified out of 80 peaks. In herbal industry, HPTLC is the main tool for drug quantification, so we have reported a HPTLC method for the real time shelf-life study. In HPTLC study, we have used three major markers (6-Gingerol, Guggulsterone-E, Guggulsterone-Z) that are reproducible in hydroalcoholic extract and not in aqueous and methanolic extracts of CPF. The kinetic study-based shelf-life of CPF formulation was performed, the observed shelf-life of CPF was more than 41 months. In addition, evaluation of antioxidant activity like total phenolic (TPC), total flavonoid (TFC) and DPPH assays were performed, and results exhibited that hydroalcoholic extract having more values than aqueous and methanolic extracts of CPF. The phytochemicals observed in LC-QTOF-MS/MS studies were tested for in silico anti-inflammatory activity and anti-arthritic activities, which need to be explored after the detailed pharmacological and biological studies. Overall, analytical studies result of present study could use to standardize the CPF as well as the all relevant polyherbal formulations where these raw drugs have been used.

Phytochemical analysis and biological activities of various parts of Bulbine natalensis (Baker): A comparative study

Introduction: Bulbine natalensis is a succulent plant native to South Africa, used to treat various skin conditions. The purpose of this investigation was to analyse the phytochemical content, as well as antioxidant and antibacterial activities of various parts of B. natalensis. Methods: Phytochemical screening tests were performed to investigate the presence of ten compounds from B. natalensis methanol extract. Total phenolic, flavonoid, tannin, and proanthocyanidin content assays were followed to determine their concentrations in B. natalensis. The 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2 O2 ), and iron chelating assays were used to assess the antioxidant activity. To determine the antibacterial properties of B. natalensis against Staphylococcus aureus and Escherichia coli, the agar-well diffusion method was adopted. Results: The phytochemical screening revealed more compounds present in the leaves. The underground stems yielded higher concentrations of total phenolics (1909.2 ± 4.8 mg gallic acid equivalent [GAE]/g), total flavonoids (259.7 ± 27.2 mg quercetin equivalent [QE]/g), and total proanthocyanidins (858.3 ± 1.7 mg catechin equivalent [CE]/g) in comparison to the leaves and roots. The roots showed stronger DPPH (0.36 ± 0.02 mg/mL) and H2 O2 (0.24 ± 0.04 mg/mL) scavenging activities. Only the underground stems and roots showed inhibition against S. aureus with the values of 15.33 ± 0.67 mm and 15.67 ± 0.33 mm, respectively, whereas the leaves displayed the highest inhibition against E. coli (18.33 ± 0.88 mm). Conclusion: The methanolic extracts of B. natalensis leaves, underground stems, and roots possess significant phytochemical content, in conjunction with antioxidant and antibacterial activities.

Vitro bioactivities and simulated gastrointestinal digestion studies on Guras-based traditional fermented drinks collected from Singalila ridge of the Himalayas

Guras or Rhododendron wine and its distilled form (Raksi) are popular traditional and therapeutic drinks served in the Rhododendron growing regions of the Himalayas, mainly in the northern and northeastern part of India, Nepal and Indo-Nepal Singalila ridge. In this research, lali Guras (Rhododendron arboreum Sm.) based traditional beverages such as unfermented decoction (as control), fermented Guras wine and distilled spirit Guras ko Raksi were collected from Singalila ridge of the Himalayas for analysis. In vitro experiments such as physicochemical and qualitative and quantitative biochemical tests; antioxidant assays (DPPH and iodometric assay); antibacterial assay; and simulated gastrointestinal digestion were carried out. Phytochemically rich unfermented decoction exhibited the best results in total phenolic content assay (82.18±1.13 mg/100 mL gallic acid equivalent) and fatty acid quantification (1.86±0.08%) test while the wine was found to be the most potential sample in all bioactivity tests. The wine was found to contain high amounts of glycoside and fermented distilled liquor Guras Raksi showed high presence of glycerol. Both the fermented samples exhibited high rate of digestion in simulated human digestion system compared to the phenolic rich unfermented decoction. This research validated the therapeutic potential of these Guras-based beverages in high-altitude conditions like Singalila by focusing on their biochemistry.

Influence of Post-Harvest Processing on Functional Properties of Coffee (Coffea arabica L.).

Coffee is one of the most popular beverages worldwide, valued for its sensory properties as well as for its psychoactive effects that are associated with caffeine content. Nevertheless, coffee also contains antioxidant substances. Therefore, it can be considered a functional beverage. The aim of this study is to evaluate the influence of four selected post-harvest coffee fruit treatments (natural, full washed, washed-extended fermentation, and anaerobic) on the antioxidant and psychoactive properties of Arabica coffee. Additionally, the impact of coffee processing on the selected quality parameters was checked. For this purpose, results for caffeine content, total phenolic content (TPC), DPPH assay, pH, titratable acidity, and water content were determined. The results show that natural and anaerobic processing allow the highest caffeine concentration to be retained. The selection of the processing method does not have a significant influence on the TPC or antiradical activity of coffee. The identified differences concerning water content and pH along with lack of significant discrepancies in titratable acidity may have an influence on the sensory profile of coffee.

The effect of soaking of white and red varieties of Sorghum bicolor flour on its antioxidant, antidiabetic and physicochemical properties

Sorghum is rich in beneficial phytochemicals such as phenolic acids, anthocyanins, phytosterols, tannins, and policosanols. This study aimed to study comprehensively the antioxidant and antidiabetic activities of red and white sorghum compared to soaked sorghum. The treatment process of soaked sorghum was carried out by soaking white and red sorghum grains for 2 h at 50 ℃ followed by drying in the sun for 8 h. All the sorghum grains were processed into flour. The antioxidant assay by DPPH showed that red sorgum flour (RSF) had the highest antioxidant activity with IC50 of 98.85 ppm, followed by soaked red sorgum flour (RSF2), white sorghum flour (WSF) and soaked white sorghum flour (WSF2). Meanwhile, for the antidiabetic assay, WSF and RSF had similar alpha-glucosidase inhibitory activity as of 97.43% and 97.42%, respectively at concentrations of 1000 ppm. Total phenolic assay using Folin-Ciocalteau reagent showed that RSF2 had the highest content as of 5031.6 mg GAE/g. The physico-chemical properties of sorghum flour were also studied by proximate analysis, SEM (Scanning electron microscopy), FTIR (Fourier transform infrared spectroscopy), XRD (X-ray diffraction), and XRF (X-ray fluorescence spectroscopy).The result showed that sorghum flour had good potential properties as flour substitution in functional food products since its good antioxidants and antidiabetic properties.

Wild-Grown Romanian Helleborus purpurascens Approach to Novel Chitosan Phyto-Nanocarriers-Metabolite Profile and Antioxidant Properties.

The current nanomedicinal approach combines medicinal plants and nanotechnology to create new scaffolds with enhanced bioavailability, biodistribution and controlled release. In an innovative approach to herb encapsulation in nanosized chitosan matrices, wild-grown Romanian Helleborus purpurascens was used to prepare two new chitosan nanocarriers. The first carrier preparation involved the nanoencapsulation of hellebore in chitosan. The second carrier emerged from two distinct stages: hellebore-AgNPs phyto-carrier system succeeded by nanoencapsulation in chitosan. The morphostructural characteristics and thermal behavior of these newly prepared nanocarriers were examined using FT-IR, XRD, DLS, SEM, EDS and thermogravimetric analyses. In addition, the encapsulation yield, encapsulation efficiency and encapsulation contents were investigated. The antioxidant activity was estimated using four in vitro, noncompetitive methods: total phenolic assay; 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay; phosphomolybdate (i.e., total antioxidant capacity); and iron(III)-phenanthroline antioxidant assay. Moreover, this study reports the first low-molecular-weight metabolite profile of wild-grown Romanian Helleborus purpurascens Waldst. & Kit. A total of one hundred and five secondary metabolites were identified in the mass spectra (MS)-positive mode from fourteen secondary metabolite categories (alkaloids, butenolides, bufadienolides, phytoecdysteroids, amino acids and peptides, terpenoids, fatty acids, flavonoids, phenolic acids, sterols, glycosides, carbohydrates, nucleosides and miscellaneous). The collective results suggest the potential application is a promising new antioxidant vehicle candidate in tumor therapeutic strategy.