Ethyl Acetate Research Articles

Validation of a simple bioanalytical method for the determination of melatonin concentration in human serum using high-performance liquid chromatography with a fluorescent detector

Analysis of melatonin in biological samples is a challenge for the analyst both because of its low concentration and because of the numerous interfering substances. Therefore, chromatography, as a powerful analytical technique used for the separation, detection, and quantification of analytes, can be the method of choice for determining melatonin in biological samples for clinical use and pharmacokinetic research. The method was developed and validated according to the internal documents in accordance with Good Laboratory Practice. The concentrations of melatonin in human serum were determined using a high-performance liquid chromatography method with a fluorescence detector and mirtazapine as an internal standard. Preparation of serum samples was performed by liquid–liquid extraction with ethyl acetate: n-hexane (1:1 v/v). High linearity was exhibited over a concentration range of 25–10,000 pg/mL of melatonin, with a correlation coefficient higher than 0.999 during the course of method validation. Repeatability and intermediate precision show a relative standard deviation lower than 6.6 %, and accuracies were from 90.7 % to 110.3 %. The lower limit of quantitation was 30 pg/mL, and the limit of detection was 7.92 pg/mL. The in-house method is simple, fast, reproducible, and applicable for the determination of melatonin in human serum.

A multidimensional study for design functional foods: Chemical profiling, antioxidant potential, enzyme inhibition, and cytotoxic effects of Alkanna tubulosa extracts

This study focuses on the extraction of plant species Alkanna tubulosa growing in Turkey, using solvents of different polarities (ethyl acetate, ethanol, ethanol/water, and water) and determining their chemical composition and biological activity. The extracts were subjected to qualitative LC-MS/MS analyses, which resulted in the identification of even 50 different secondary metabolites in some extracts. The existence of total phenols and flavonoids was determined. of total phenols and flavonoids was determined. The antioxidant capacity was evaluated by various in vitro assays, as well as the enzyme inhibitory capacity against cholinesterases (AChE and BChE), tyrosinase, α-amylase, and α-glucosidase, as well as human isoforms of carbonic anhydrase. In addition, the cytotoxic activity of the extracts was also examined. The results showed that the ethanol/water extract showed the strongest antioxidant activity (225.55 mg TE/g extract in ABTS and 381.14 mg TE/g extract in CUPRAC), probably due to the high total phenol (105.17 mg GAE/g extract) and flavonoids (20.12 mg RE/g extract) content, while the ethanol extract showed the strongest enzyme inhibitory potential. These chemical and biological findings provide valuable insights for potential future products based on the A. tubulosa extracts which could be used in various industries.

Sub-chronic Toxicity Study of Heracleumlasiopetalum Extract Towards Healthy Sprague Dawley Rats

Background: Heracleum species are commonly used as spices, flavorings, and food additives. Members of the genus Heracleum offer many medicinal benefits but may also pose adverse effects on human health. Objectives: To prepare a crude leaf extract of Heracleumlasiopetalum and assess its toxicity profile towards healthy rats. Methods: The H. lasiopetalum leaf extract was prepared using pure methanol and ethyl acetate (1:10) at room temperature over a period of 72 hours. After filtration, the crude extract was obtained using a rotary evaporator at 40 - 45°C. Subsequently, various doses of the H. lasiopetalum extract were administered orally to healthy Sprague Dawley rats at three doses (300, 600, and 900 mg/kg body weight) for four weeks to test for toxicity. Blood samples were examined for hematologic and biochemical changes, while the liver, kidneys, and heart were examined for histopathological changes. Results: The toxicity study revealed no mortality at low and medium doses, as well as no clinical toxicity indicators. Additionally, there were no significant alterations observed in the haematological, biochemical, and histopathological profiles of the treated animals throughout the 28-day experiment. However, at high doses, the mortality rate was significantly elevated, accompanied by notable histopathological changes. Conclusions: Continuous administration of high doses of H. lasiopetaum may induce potential toxic effects in the treated animals.

Evaluation of Anti-angiogenic Activity of Iraqi Mesembryanthemum cordifolium using rat aorta ring assay

New blood vessels are produced during angiogenesis, which is necessary in both normal and abnormal physiological and pathologic states. The use of non-toxic plant extracts that inhibit angiogenesis might lead to more focus on the use of natural sources as a pivotal therapeutic agent in the treatment of many diseases and malignancies. Mesembryanthemum cordifolium is belongs to the Aizoaceae family and commonly known as the Heartleaf Ice Plant or Tiny Sun Rose. The plant was cultivated in the city of al Diwaniyah. No previous research has been conducted on this plant and its antiangiogenic properties. Hence, this research aimed to assess the antiangiogenic capacity of the Iraqi M. cordifolium. For this study, male albino rats aged 12 to 14 weeks were used. With a Soxhlet apparatus, the defatted plant components were extracted in 85 % methanol for 12 h. Subsequently, each fraction was fractionated using petroleum ether, chloroform and ethyl acetate and each fraction underwent phytochemical analysis. Rat aortic rings were infused with 100 ?g/mL of each plant component and placed in a growth medium. In addition, the dose-response relationship was assessed for the most active fraction at 6 different concentrations ranging from 6.25 ?g/mL to 100 ?g/mL. Compared to the negative control (1 % DMSO), the results demonstrated that fractions of M. cordifolium inhibit the growth of blood vessels in a dose-dependent manner; chloroform demonstrates the most potent effect. The results indicate that the chloroform fraction derived from M. cordifolium has a good ability to limit blood vessel development. This makes it a viable option for future investigation as an angiogenesis inhibitor.

Unlocking the functional potential of Nitraria schoberi extracts using chemical fingerprinting, biological efficiencies, in silico and network pharmacological approaches

The objective of this investigation was to assess the cytotoxic, antioxidant, and enzyme inhibitory properties of the aerial portions of Nitraria schoberi (family Nitrariaceae). Extracts were prepared by sequential maceration in hexane, dichloromethane (DCM), ethyl acetate and methanol (MeOH) and water extract was prepared by infusion. DCM and MeOH extracts accumulated the highest amount of metabolites where the former was characterized by the presence of octadecyl ferulate (16.96 mg/100 mg), methoxyluteolin (8.48 mg/100 mg) and isorhamnetin (5.35 mg/100 mg) and the alkaloids perlolyrine (5.03 mg/100 mg), (+)-5a-hydroxylemannine (2.43 mg/100 mg) and matrine (2.40 mg/100 mg). Results of antioxidant activity showed that the MeOH extract exerted the highest anti-2,2-diphenyl-1-picrylhydrazyl (DPPH) (28.13 mg Trolox equivalent (TE)/g) and Fe+++ (54.38 mg TE/g) reducing capacity. All extracts exerted significant acetylcholinesterase (3.71–4.37 mg galantamine equivalent (GALAE)/g) and tyrosinase (86.40–109.63 mg kojic acid equivalent (KAE)/g) inhibition activity. The best α-amylase and α-glucosidase inhibition was exhibited by the DCM (0.81 mmol ACAE/g) and MeOH (1.34 mmol ACAE/g) extracts respectively. The highest cytotoxic effect was exerted against the breast adenocarcinoma (MDA-MB-231) (IC50: 26.71 μg/mL) and cervix adenocarcinoma (HELA) (IC50: 29.10 μg/mL) cell lines by the MeOH and DCM extracts respectively. In addition, we performed molecular docking analysis to understand the interaction between chemical components and tested enzymes and cellular targets for cancer. Chrysoeriol-3-O-rutinoside bound strongly to AChE. Furthermore, a good interaction between vascular endothelial growth factor receptor 2 (VEGFR2) and isovitexin-2″-O-arabinoside was observed. We employed network pharmacology to understand the role of N. schoberi's compounds in neurological disorder like Alzheimer and multiple sclerosis. Therefore, the results of this study indicate that N. schoberi may serve as a viable starting point for the future creation of phytopharmaceuticals that target particular diseases associated with oxidative stress, such as diabetes and certain neurological disorders.

INTESTINAL PARASITISM IN WORKING HORSES AND ASSOCIATED ZOONOTIC RISKS IN LOWLANDS OF NEPAL

The presence of intestinal parasites influences equines' well-being and working performance. However, the scenario of parasitism in working horses in the lowlands of Nepal is yet to be explored. The present study aimed to reveal the prevalence and diversity of intestinal parasites (protozoa and helminths) and to list the zoonotic species in working horses in the lowlands of Nepal. Fresh fecal samples (N=102) from horses were collected at two locations (Chitwan and Birgunj) in the lowlands of Terai and were transferred to the research laboratory. Coproscopy was carried out via direct wet mount, formalin ethyl acetate (FEA) sedimentation, saturated salt flotation, and acid-fast staining techniques. Coproscopy revealed an overall prevalence rate of 90.2% (92/102) with 15 known diverse species of parasites (Protozoa: 5 and Helminths: 10) and an unknown coccidian, out of which eight possess zoonotic potential. The prevalence and diversity of intestinal parasites were higher in adult than in young animals (90.7%; 15 spp. vs. 88.9%; 11 spp.) The overall prevalence of helminths was double that of protozoa (89.2% vs. 43.1%). Furthermore, polyparasitism was much more prevalent than monoparasitism (85.3% vs 4.9%). Co-infection with two parasite species (37%) was higher in young horses. In comparison, triplet infection (34%) was higher in adults, and a maximum concurrency of up to six species of parasites at a time was recorded. Following it, the differences in the prevalence rate of parasites based on the predictor of risks, like sex, grazing, domestication type, nature of the floor, and medication practices, were statistically significant. Working horses in the lowlands of Terai harbored a significant variety of intestinal parasites with important prevalence. Since eight of the reported parasitic species were zoonotic, infected horses pose a zoonotic risk to the owners. Therefore, timely deworming, pasture management, and reduction in working pressure are highly recommended.

Research on triamterene equilibrium solubility in mono-solvents and aqueous solutions according to solvent effect, models, thermodynamic analysis and quantum chemical calculation

The equilibrium solubilities of triamterene in pure solvents {isopropanol, methanol, n-propanol, water, 1,2-dichloroethane, 2-methoxyethanol, ethyl acetate, acetonitrile, N-methylformamide (NMF), n-butanol, isobutanol and dimethylsulfoxide (DMSO)}, as well as solvent mixtures of methanol/DMSO + water, were determined by dint of the saturation shake-flask technique. Triamterene solubility (105x) rose as the determination temperature rose and fell in the order at 298.15 K: 2-methoxyethanol (453.9) > DMSO (281.3) > NMF (142.6) > methanol (8.424) > n-propanol (1.432) > n-butanol (1.152) > ethyl acetate (0.9441) > isobutanol (0.8738) > acetonitrile (0.7991) > isopropanol (0.7419) > 1,2-dichloroethane (0.5232) > water (0.1795). No crystal transition or solvation in the trial process was shown via X -ray power diffraction patterns. The solvent effects, in which the interactions of solvent–solvent and solvent–solute were considered, were studied via linear solvation energy relationship. Using equations such as Wilson, Apelblat, λh, and NRTL for mono-solvents and Jouyban-Acree as well as modified van't Hoff-Jouyban-Acree for solvent mixtures, the magnitudes of equilibrium solubility were associated. The greatest relative average deviation of 5.787 × 10−2 and the maximum deviation value of root mean square of 219.1 × 10−4 were computed for neat solvents. According to the solubility information in pure solvents, the Wilson equation was used to compute the mixing solution parameters. At 298.15 K, the inverse Kirkwood-Buff integral was utilized here to examine the preferred solvation of triamterene. In blends of DMSO/methanol + water with middle and rich DMSO/methanol compositions, the parameters of preferred solvation for DMSO/methanol were positive, indicating the preferential solvation of triamterene by DMSO/methanol. In addition, the electrostatic acidity-basicity properties were demonstrated using the lowest negative electrostatic potential in company with the minimum m local ionization energy of molecular surface. The N in ring of the triamterene is the preferential target for electrophilic assault; and –NH2 of the triamterene, for nucleophilic assault. The weak triamterene-solvent interactions were graphically displayed by the use of the independent gradient approach on the basis of Hirshfeld partition analysis.

Development of bioactive films loaded with extract and polysaccharide of Pinus brutia bark.

Society's interest in natural and clean products in many areas, such as food and cosmetics, has increased considerably. It has led to the development of new techniques in the packaging of products so that the wastes from the preferred products can be recycled. In this context, Pinus brutia bark was preferred within the scope of the study to transform natural wastes into functional components and use them as packaging material. P. brutia bark (PBB) samples were collected from Bursa, Turkey. PBB samples were ultrasonically extracted using various solvents (acetone, butanol, ethanol, ethyl acetate, hexane, methanol, petroleum ether, and water) and a solvent-acidic hydrolysis system. The phenolic content profile of PBB samples was determined using high-performance liquid chromatography with diode-array detection, and total flavonoid content, antioxidant capacity, and total phenolic content were determined. Chitosan-polyvinyl alcohol (CS-PVA) films loaded with polysaccharides and containing methanolic extract were developed. The physical, chemical, and mechanical properties of the films were characterized. It is known that the thickness of the films determines the mechanical properties required to maintain the integrity of the packaging during storage and transport. From the results of the study, it was concluded that the elongation at break value was higher in CS-PVA-PBB-M films (111.08%±10.46%), Young's modulus (31.74±21.37N/mm2), and tensile strength (3.01±0.50N/mm2) values were higher in CS-PVA films. In this case, it was concluded that adding proanthocyanidin to edible films gives flexibility to the films.

Evaluation of TPC, TFC, and Antioxidant Activity of Extracts of Piper longum L.

Evaluation of TPC, TFC, and Antioxidant Activity of Extracts of Piper longum L.

Magnetic field assisted centrifugal acceleration thin-layer chromatography: An approach to investigate the magnetic field effects on separation parameters including retention factor difference, selectivity factor, and resolution

The effect of internal and external magnetic fields on the separation of antifungal drugs by centrifugal acceleration thin-layer chromatography was reported for the first time. External and internal magnetic fields were applied using neodymium magnets and CoFe2O4@SiO2 ferromagnetic nanoparticles. Separation of ketoconazole and clotrimazole was performed using a mobile phase consisting of n-hexane, ethyl acetate, ethanol, and ammonia (2.0:2.0:0.5:0.2, v/v). The influence of the magnetic field on the entire chromatographic system led to changes in the properties of the stationary and mobile phases and the analytes affecting the retention factor, shape, and width of the separated rings. The extent of this impact depended on the structure of the analyte and the type and intensity of the magnetic field. In the presence of the external magnetic field, there were more significant changes in the chromatographic parameters of the drugs, especially the width of the separated rings, and ketoconazole was more affected than clotrimazole. The changes are conceivably due to the effect of the magnetic field on the analyte distribution between the stationary and mobile phases, which is also caused by the possibility of the magnetic field affecting the viscosity, surface tension, and surface free energy between the stationary and mobile phases.

In-vitro and molecular docking studies of plant secondary metabolites isolated from Hypericum oblongifolium as antibacterial agents and lipoxygenase (5-LOX) inhibitors

Natural Products are bases for modern pharmaceuticals as they are precursors for many synthetic drugs. People also use natural products directly in the form of herbal medicines due to their less harmful effects. This research project aimed to investigate the new pharmacological potential of natural products found in Hypericum oblongifolium. Five compounds were identified from chloroform and ethyl acetate soluble fractions of H. oblongifolium. These compounds were identified as 21-β-hydroxyursolic acid (1), β-sitosterol (2), 7,4′-dihydroxy-5,3′-dimethoxyisoflavone (3), α-D-glucopyranosyl-6′-O-hexadecanoate (4), Quercetin-3′-O-ß-D-glucopyranoside (5). When these compounds were tested for antibacterial potential compound 5 showed maximum inhibition against gram-positive Bacillus subtilis and was highly active against Escherichia coli gram-negative bacterium. Minimum inhibition was shown by compounds 2 and 3 against Shigella flexneri among gram-negative bacteria. Compound 5 also exhibited the least binding energy in the molecular modeling of these compounds against Escherichia coli proteins GlcN-6-P synthase (PDB ID: 1MOQ). Compound 5 also showed good inhibition of lipoxygenase (5-LOX) enzyme with an IC50 value of 30.1 ± 0.20 as it exhibited maximum hydrogen bondings and minimum energy when docked with LOX-5 (PDB ID 3V99) using a patch dock.

Development of Simultaneous HPTLC Method and Validation for the Quality Assessment of Ayurvedic Formulation-Ayush Kvatha Churna by Using Marker Compound Rosmarinic Acid, Trans-Cinnamaldehyde and Piperine.

Ayurveda emphasizes the propagation of nature in maintaining health. In the present scenario, we have seen the faith of people in herbal drugs during the Covid 19 outbreak. The raises in the number of peoples have been using herbal drugs to boost immunity against infectious diseases shows the popularity of this ancient system of medicine. The standardization of Ayush Kvatha Churna (AKC), work set out to establish a straightforward, accurate and sensitive HPTLC method for the identification and quantification of marker compounds. The Rosmarinic acid, trans-Cinnamaldehyde and Piperine were used for the estimation of markers in Ayush Kvatha Churna by using HPTLC with a solvent system, consisting of Toluene: Ethyl acetate: Ethyl alcohol: Formic acid (5.6:2.4:2: 0.3v/v/v/v). The Rf value 0.33 for Rosmarinic Acid, 0.69 for Piperine and 0.77 for trans-Cinnamaldehyde was observed and it is exactly complying with the corresponding bands in Ayush Kvatha Churna. The technique has been effectively verified and validated, enabling it to be used for the standardization or quantitative analysis of Rosmarinic acid, trans-Cinnamaldehyde and piperine in Ayush Kvatha Churna.

Marine seaweed endophytic fungi-derived active metabolites promote reactive oxygen species-induced cell cycle arrest and apoptosis in human breast cancer cells.

Endophytic fungi have an abundant sources rich source of rich bioactive molecules with pivotal pharmacological properties. Several studies have found that endophytic fungi-derived bioactive secondary metabolites have antiproliferative, anti-oxidant, and anti-inflammatory properties, but the molecular mechanism by which they induce cell cycle arrest and apoptosis pathways is unknown. This study aimed to determine the molecular mechanism underlying the anticancer property of the endophytic fungi derived active secondary metabolites on human breast cancer cells. In this study, we identified four endophytic fungi from marine seaweeds and partially screened its phytochemical properties by Chromatography-Mass Spectrometry (GC-MS) analysis. Moreover, the molecular mechanism underlying the anticancer property of these active secondary metabolites (FA, FB, FC and FE) on human breast cancer cells were examined on MCF-7 cells by TT assay, Apoptotic assay by Acridine orang/Ethidium Bromide (Dual Staining), DNA Fragmentation by DAPI Staining, reactive oxygen species (ROS) determination by DCFH-DA assay, Cell cycle analysis was conducted Flow cytometry and the apoptotic signalling pathway was evaluated by westernblot analysis. Doxorubicin was used as a positive control drug for this experiment. The GC-MS analysis of ethyl acetate extract of endophytic fungi from the marine macro-algae revealed the different functional groups and bioactive secondary metabolites. From the library, we observed the FC (76%), FB (75%), FA (73%) and FE (71%) have high level of antioxidant activity which was assessed by DPPH scavenging assay. Further, we evaluated the cytotoxic potentials of these secondary metabolites on human breast cancer MCF-7 cells for 24h and the IC50 value were calculated (FA:28.62 ± 0.3µg/ml, FB:49.81 ± 2.5µg/ml, FC:139.42 ± µg/ml and FE:22.47 ± 0.5µg/ul) along with positive control Doxorubicin 15.64 ± 0.8µg/ml respectively by MTT assay. The molecular mechanism by which the four active compound induced apoptosis via reactive oxygen species (ROS) and cell cycle arrest in MCF-7 cells was determined H2DCFDA staining, DAPI staining, Acridine orange and ethidium bromide (AO/EtBr) dual staining, flowcytometry analysis with PI staining and apoptotic key regulatory proteins expression levels measured by westernblot analysis. Our findings, revealed the anticancer potential of endophytic fungi from marine seaweed as a valuable source of bioactive compounds with anticancer properties and underscore the significance of exploring marine-derived endophytic fungi as a promising avenue for the development of novel anticancer agents. Further investigations are necessary to isolate and characterize specific bioactive compounds responsible for these effects and to validate their therapeutic potential in preclinical and clinical settings.

Recovery of organic acids from lactic acid-enriched fermentation broth via salting-out assisted solvent extraction

The organic acids (OAs) market has been expanding due to their versatility and wide range of uses in several industrial applications, thus prompting a transition from chemical to biological approaches to generate such high-value products. Nevertheless, downstream processes to separate OAs from the fermentation broth represent, to date, the main cost in biological OAs manufacturing. With the goal of limiting this issue, the present study investigated OAs separation from a lactic acid (LA)-rich fermentation broth through physical, i.e., using ethyl acetate (EA), and physico-chemical, i.e., using tributyl phosphate (TBP), solvent extraction. In addition, the salting-out effect of different salts on OAs recovery was investigated. Among the investigated conditions, a combination of pure TBP and 40% ammonium sulfate ensured the highest LA extraction efficiency of 65%. Besides, mixing TBP with EA (50:50) enabled to maintain the same LA extraction efficiency while reducing the process costs, limiting the toxicity of the chemicals involved, and obtaining an extracted OAs mix with lower co-metabolites, i.e. volatile fatty acids, compared to using pure TBP. Overall, solvent extraction assisted by salting-out agents were shown to be a promising method to separate OAs from a fermentation broth in liquid form, especially for fermentation processes operated at low pH.

Production and Physicochemical Characterization of the Gel Obtained in the Fermentation Process of Blue Corn Flour (Zea mays L.) with Colletotrichum gloeosporioides.

Food gels are viscoelastic substances used in various gelled products manufactured around the world. Polysaccharides are the most common food gelling agents. The aim of this work was the production and characterization of a gel produced in a blue corn flour fermentation process, where different proportions were used of blue corn (Zea mays L.) flour and Czapek Dox culture medium (90 mL of culture medium with 10 g of blue corn flour, 80 mL of culture medium with 20 g of blue corn flour, and 70 mL of culture medium with 30 g of blue corn flour) and were fermented for three different durations (20, 25, and 30 days) with the Colletotrichum gloeosporioides fungus. A characterization of the gel was carried out studying the rheological properties, proximal analysis, toxicological analysis, microscopic structure, and molecular characterization, in addition to a solubility test with three different organic solvents (ethanol, hexane, and ethyl acetate, in addition to water). The results obtained showed in the rheological analysis that the gel could have resistance to high temperatures and a reversible behavior. The gel is soluble in polar solvents (ethanol and water). The main chemical components of the gel are carbohydrates, especially polysaccharides, and it was confirmed by FT-IR spectroscopy that the gel may be composed of pectin.

A comprehensive evaluation of different responses of supplementary light qualities on physiological and biochemical mechanisms of ‘Kyoho’ grape

Grape is an economically important fruit crop worldwide, for which light qualities play a diverse role in vineyard growth and development. However, the regulatory mechanisms between different light qualities and the quality of plant growth at the morphophysiological and pomological levels remain unknown. Therefore, in the current study, we studied plant physiological, biochemical and genetical analysis to investigate the response of all seven light qualities, including red (R), blue (B), white (W), 1-red, 1 blue (1R1B), 2-red, 1-blue (2R1B), 1-red, 2-blue (1R2B), and natural (N, was used as control group) to evaluate ‘Kyoho'grape (Vitis labrusca L. × V. vinifera) morphophysiological, pomological traits at light responsive transcriptional pathway, which revealed the possible networking metabolism controlling fruit quality and quantity of grape plant. Our results showed that photosynthetic indicators such as leaf stomata, number of stomata, and longitudinal and transverse diameter of stomata improved under red and blue light qualities as compared to white light. Leaf gas exchange indicators showed that net photosynthetic rate, transpiration rate, intercellular CO2 concentration rate, total conductance to CO2, stomatal conductance, conductance to water vapor, maximal quantum yield of PS (II), actual photochemical quantum efficiency, electron transfer chain, and non-photochemical quantum efficiency improve under blue and red light. Photosynthetic pigments showed that chlorophyll a and b, carotenoid, and total pigment contents improved under blue and red light. Nutrient elements showed that potassium, phosphorous, magnesium, calcium, iron, and zinc contents improved under blue and red light. Morphological and organic acid indicators such as the appearance of fruit shape, firmness, and titratable acidity improved under blue and light. However, there was a notable increment in the total soluble sugar under blue and red light as compared to natural light. The composition of sugar including (sucrose, fructose, glucose) and organic acids (tartaric acid, citric acid, and malic acid) improved under blue and red light (fruits) and showed notable reductions for tartaric acid under blue and red light (leaves) as compared to natural light. Aromatic compounds showed that esters, aldehydes, acetaldehyde, 2-hexenal, ethyl acetate, n-propyl acetate, butanoic acid, ethyl ester, 1-hexanol, 2-ethyl, 2-propanone, and 1‑hydroxyl, improved under blue and red light. Further, changes in enhanced expression of light-responsive genes including VvCRYI, VvCRY2, VvPHYA, VvPHYB, VvCOP1, VvHY5, VvHYH, and VvPIF4 upregulated under blue and red light, while VvPIF3.1 down-regulated as compared to natural light. Also, chlorophyll synthesis-related genes including VvSGR, VvPAO, and VvCHLH upregulated under blue and red light, while VvNYC1, VvPPH, and VvPOR showed noticeable changes under blue and red light as compared to natural light. Interestingly, most of the relative genes were highly expressed in fruits as compared to leaves. Finally, we have established correlation and principal component analysis for indicators in grapes. Current findings provide novel mechanisms by which different responses of supplementary light control the plant fruit quality of grapes.

Antiurolithiatic activity of Indian medicinal plant: Ocimum kilimandscharicum Gurke (Lamiaceae)

Urolithiasis is the most prevalent condition of the urinary system, characterized by the formation of stones inside the urinary tract. It is urgent to look for a natural urolithiasis therapy due to the serious side effects of conventional medications. Hydro-alcoholic (80% v/v) extract of the aerial parts of Ocimum kilimandscharicum (OK) and its ethyl acetate, chloroform, n-butanol, aqueous, and n-hexane fractions were subjected to in vitro antiurolithiatic screening as well as preliminary screening of phytochemicals. The in vitro antiurolithiatic activity of O. kilimandscharicum was studied using its hydroalcoholic extract (HAEOK). Calcium phosphate test using a colorimetric approach and calcium oxalate assay using a titrimetric model were used to determine the proportion of calcium oxalate crystals that dissolved. Total phenolic content (TPC) and total flavonoid content (TFC) were measured for the extract and fractions of OK. Ethyl acetate fraction (EAFOK) had a greater capacity to suppress crystal formation in both the calcium phosphate and calcium oxalate assays. The percent dissolution of calcium oxalate by HAEOK and EAFOK (31.48 ± 0.920% and 39.21 ± 0.903%) and calcium phosphate crystals by HAEOK and EAFOK (59.03 ± 0.820% and 66.62 ± 0.468%) was determined, respectively. At p < 0.05 and p < 0.01, differences between the results were regarded as significant. Cystone was employed as a standard drug. This study revealed that EAFOK showed significant antiurolithiatic activity. The antiurolithiatic activity of the extract/fraction was attributed to the steroids, triterpenoids, and flavonoid content of OK.

Two chromatographic methods for determination of cytarabine and dexamethazone; two co-administered drugs for the treatment of leukemia: green analytical perspective

Two sensitive, straightforward and repeatable chromatographic techniques were developed for the determination of Cytarabine HCl and Dexamethazone in their pure form and spiked human plasma without prior separation. The drugs are used co-administered for the treatment of Leukemia, a certain type of blood cancer. Method (A) is an isocratic chromatographic HPLC method; separation was accomplished on C18 column using the eluting mixture of 6.9 g/L Monobasic Sodium Phosphate pH 3: methanol (70:30, v/v) and detection was at 275 nm. Concentrations were in the range of 0.2–15 μg/mL for both CYT and DEX. Method (B) is a HPTLC method in which separation was attained on HPTLC F254 plates using methanol: ethyl acetate: ammonia, (7.8:2:0.2, by volume) as eluting solvents and detection was at 275 nm. Concentrations were in the range of 0.1–4 μg/band for both CYT and DEX. The parameters for system suitability testing were evaluated to determine the effectiveness of the developed chromatographic procedures in terms of performance. The recently developed techniques were applied for the determination of the drugs under investigation in spiked human plasma. Validation parameters were examined in accordance with US-FDA criteria. All results were found to be within the acceptable ranges. To evaluate the greenness characters of the proposed methods to the environment; three greenness assessment tools including eco-scale assessments (ESA), green analytical procedure index (GAPI), and Analytical Greenness calculator (AGREE) were used. Acceptable and satisfying results that demonstrated the greenness characteristics of the suggested methods were attained.

Antibacterial Activity of Ethanol, Ethyl Acetate, and n-Hexane Extracts of Bidara Laut Lignum (Strychnos lucida R.Br) against Staphylococcus aureus and Escherichia coli

Strychnos lucida is empirically believed to have efficacy in treating various diseases such as fever, malaria, toothache, and stomachache and to treat wounds. The secondary metabolite of Strychnos lucida has various polarity. The purpose of this study was to compare the inhibitory activity of Strychnos lucida extracts against E. coli and S. aureus bacteria. The test was carried out by measuring the Diameter of the Inhibitory Region (DIR) using the disc diffusion method on a concentration of 90%, 75%, 60%, 45%, 30%, and 15%. The Minimum Inhibitory Concentration (MIC) was carried out by agar dilution method on a concentration of 15%, 12.5%, 10%, 7.5%, 5%, and 2.5%. The positive control was 1% ampicillin, and the negative control was 10% DMSO. The ethyl acetate extract showed the highest DIR of 14.30 mm against S. aureus and E. coli on a concentration of 45%, ethanol extract was 8,38 mm against S. aureus and 8.71 mm against E. coli, and n-hexane had no inhibitory activity on a concentration 45%. The result of the MIC test for ethanol and ethyl acetate extract for both bacteria was found at a concentration of 15%, while n-hexane extract against Staphylococcus aureus was found at a concentration of 90% and a concentration of 75% against Escherichia coli.

Supercritical water depolymerization of black liquor, refining and comprehensive analysis of products including biopolyols

Black liquor (BL) has valuable lignin and its derived compounds, which can be used for many purposes, such as polyol synthesis. Herein, we reveal polyols presence in depolymerized black liquor (DBL) and propose a fast and scalable approach to increase the yield of this fraction. Hydrothermal treatment (HTL) of black liquor (385°C, 26 MPa) was performed in a custom designed supercritical water (SCW) pilot-plant with rapid reaction times of around 0.4 s. Ash-free, total ethyl acetate extracted bio-oil yield was found 77% w/w. In addition to the contribution of the detailed fractionation methods, this study highlights continuous operation (>1 hour), and short reaction times (∼0.4 s) of raw black liquor in SCW to produce biopolyols and aromatic bio-oil.