Methanol Solvent System Research Articles

Anti-Diabetic Activity And Anti-Oxidant Activity Of Kalanchoe Pinnata And Their Characterization Studies.

Species of Kalanchoe have been used as a remedy for various infectious diseases and cancer treatment. This study evaluates the anti-diabetic properties of the plant, Kalanchoe pinnata. Phytochemical components, anti-oxidant assays were also carried out. Phytochemical analysis showed the presence of alkaloids, flavonoids, steroids, phenolic compounds. The free radical scavenging activity was determined in which the ethanol extract showed the maximum inhibitory effect when compared with aqueous extract .Thin layer chromatography was done to determine the compound in a solvent system of methanol, acetic acid, formic acid and water in the ratio 3:1:1:0.5 . Characterization of the leaf extract was done by UV-visible spectroscopy which showed the peak between 300-350nm. FTIR analysis showed the presence of the various functional group present in the plant. Aqueous and ethanol extract was subjected to α-amylase inhibitory effect and α-glucosidase inhibitory effect to find the antidiabetic potential activity of the plant extract.

Superstructures of copper nanoclusters as NIR TADF emitters: solvent-dependent optical and morphological modulation.

Herein, we report 2-mercaptopyridine-templated copper nanoclusters (CuNCs) which display near infra-red (NIR) emission, both in the solid and colloidal states. Interestingly, the NIR emission can be modulated to orange emission by preparing the CuNCs in a mixed solvent system of chloroform and methanol instead of water. The drastic change in the photo-physical properties of the CuNCs when prepared in two different solvent systems is accompanied by a unique morphological tuning. Further studies reveal that the strong NIR emission is the result of thermally activated delayed fluorescence (TADF) which is confirmed by the long excited state lifetime (∼4 μs at room temperature), time resolved emission spectroscopy (TRES) measurements, temperature-dependent photoluminescence studies, temperature-dependent lifetime studies, and excitation-transmittance dependent TRES intensity measurements. The CuNCs exhibit an exceptionally small singlet-triplet energy gap of 58.2 meV, indicating a highly efficient TADF in the system. Moreover, the solvent-dependent morphological tuning of the nanocluster superstructures rendering a drastic change in the photo-physical signatures is the consequence of different ΔE(S1-T1) values for the CuNCs in different solvent environments. Further findings corroborate that the electronic structure of the surface ligands can also help us to tune the ΔE(S1-T1) energy gap for these nanoclusters.

Solid-liquid equilibrium, molecular simulation and thermodynamic properties of sulfaguanidine in pure and binary solvents from 288.15 K to 343.15 K

The solubility of sulfaguanidine(SG) in water, methanol, ethanol, n-propanol, isopropanol, n-butanol and binary solvent systems (water + methanol/n-propanol) was determined by the synthetic method at local pressure values in the temperature range of 288.15 K to 343.15 K. The experimental data were compiled and plotted, and it can be seen that the solubility of SG in the six pure solvents and two binary solvent systems in the selected temperature range increased monotonically with increasing temperature. The solubility of SG in pure solvents was in the order of methanol > ethanol > n-propanol ≈ isopropanol > n-butanol > water. The solubility of SG in the mixture of water + methanol increased with the composition of the benign solvent, while the solubility in the mixture of n-propanol + water increased first and then decreased with the increase of the benign solvent. In addition, the Van’t Hoff model, Apelblat model, λh model and Wilson model were cited to correlate the solubility data of SG in pure and binary solvents, and additionally the binary models Apelblat-Jouyban-Acre (A-JA) model and Van't Hoff-Jouyban-Acre model were used to fit the solubility of SG in two binary solvents. The RAD and RMSD values show that the chosen models both show good correlation. The solvent effect was analyzed using the KAT-LSER model and the solubility of SG in the solvents used was mainly influenced by non-polarity and viscosity. After that, the inter-molecular forces of SG were explored using Hirshfeld surface analysis and a two-dimensional fingerprint was plotted. Finally, the thermodynamic properties of SG in all solvent systems, including Gibbs free energy, entropy and enthalpy, were calculated using Van’t Hoff equation and the experimentally measured solubility data.

Solubility determination and thermodynamic model analysis of nefopam hydrochloride in eight plain solvents and three binary solvents from 273.15 K to 323.15 K

Nefopam hydrochloride (NPH) is a non-narcotic analgesic with antipyretic, analgesic, and muscle relaxant effects. Solubility is an important molecular property that can inform the crystallization and purification of drugs. In the present study, NPH solubility was measured in eight plain solvents and three binary solvents at 273.15 K-323.15 K. Results showed that in the plain solvents, NPH had the greatest solubility in methanol and the least in acetone. The solubility in the solvent system of methanol + acetone and ethanol + acetone becomes greater with the increase of the molar fraction of the positive solvent, while it becomes greatest in the system of water + acetone when the molar fraction of the positive solvent is 0.8 and at molar fractions>0.8 the solubility begins to decrease, which may be owing to the existence of the phenomenon of co-solubility. NPH solubility in plain solvents and binary solvents is positively correlated with temperature. By XRD and TG-DSC, the crystal structure of NPH in solvent did not change and the weight loss and melting temperatures were 500.49 K and 518.34 K respectively, which were much higher than the experimental temperatures. Selection of Hansen solubility parameter to predict the solubility effect of NPH in different solvents. The analysis was carried out using five thermodynamic models, ranging from Modified Apelblat model, Buchowski-Ksiazaczak λh model, CNIBS/R-K model, Jouyban-Acree model and SUN model, to fit the solubility data, out of which Modified Apelblat model and CNIBS/R-K model had more accurate prediction results.

Phytochemical Investigation and Thin Layer Chromatography of Methanol Extract of Psoralea corylifolia and Emblica officinalis Leaves

The current study focuses on phytochemical screening of Psoralea corylifolia and Emblica officinalis leaf extracts in methanol solvent. The antibacterial compounds found in both leaf extracts of Psoralea corylifolia and Emblica officinalis plants were investigated using phytochemical studies. The extracts contained flavonoids, terpenoids, tannins, alkaloids, saponins, and phenolic chemicals, according to preliminary phytochemical screening. The solvent systems of toluene: ethyl acetate: methanol (24:5:2) and chloroform: ethyl acetate: acetic acid (50:50:1) yielded the most phytochemicals from methanolic extracts of Psoralea corylifolia and Emblica officinalis plants, respectively. On TLC plates, these chemicals were separated, resulting in the discovery of different spots in both leaf extracts. The Rf values of methanol extract of P. corylifolia run under toluene: ethyl acetate: methanol (24:5:2) solvent system was 0.12, 0.19, 0.30, 0.41, 0.53, 0.65, 0.77, 0.84, 0.89, and 0.92, respectively, while Rf values of methanol leaf extract of E. officinalis run under chloroform: ethyl acetate: acetic acid (50:50:1)solvent system was 0.23, 0.31, 0.41, 0.64, 0.76 and 0.88 respectively. The results of the investigation will be used to confirm the proper identification of antibacterial fractions from P. corylifolia and E. officinalis crude plant extracts by GC-MS. The optimum solvent for extracting antibacterial components from P. corylifolia and E. officinalis leaves was methanol.

Quantitative Estimation of Quercetin of Some Selected Edible Plants of West Bengal by High-performance Thin-layer Chromatography Densitometry Method

ABSTRACT Introduction: Quercetin is a polyphenol, widely found in most edible fruits and vegetables. This plant pigment provides cardiovascular, anti-inflammatory, antidiabetic, asthma, and many more health benefits. Objective: The aim of this study is to quantify the quercetin by high-performance thin-layer chromatography (HPTLC) method from the methanolic leaf extracts of three edible plants of Bengal, viz., Nyctanthes arbor-tristis L., Oxalis corniculata L., and Polygonum plebeium R.Br. Materials and Methods: HPTLC was carried out on aluminum-backed plate precoated with silica gel 60 F254, using the solvent system of toluene, ethyl acetate, formic acid, and methanol (5.5:3:1:0.5 v/v/v/v). The method validation was done according to the International Conference on Harmonization guidelines in terms of linearity, limit of detection, limit of quantification, accuracy, precision, and specificity. Results: The desired compound was well separated using the solvent system toluene, ethyl acetate, formic acid, and methanol (5.5:3:1:0.5 v/v/v/v) which gave a sharp and well-defined band with Rf 0.60 for quercetin. The calibration curves showed a good linear correlation coefficient (R 2 = 0.9774). The linearity range was found to be 20–100 (μg/spot). Among the selected plants, the highest amount of quercetin was found in the leaf extracts of O. corniculata L. (220.035 ± 0.06 mg/100 g) and the lowest for N. arbor-tristis L. (31.253 ± 0.99 mg/100 g). Conclusion: The proposed HPTLC method for quercetin estimation was found to be simple, cost-effective, precise, and accurate and can be used for the quality control of plant material. O. corniculata L. was found to be a potent source of quercetin as it possesses a decent amount of quercetin.

Copper sulphate immobilized on P(AN-NIPAM-MBAM) terpolymer as a highly efficient catalyst for the selective reduction of nitro-arenes

A new copper sulphate anchored to an acrylonitrile-N-isopropylacrylamide-bisacrylamide based P(AN-NIPAM-MBAM) terpolymer support has been employed as a catalyst for the hydrogenation of varied substituted nitro-arenes affording corresponding amines with excellent yield. Application of hydrogen source from NaBH4 in combination with NaOH under water and/or methanol solvent system at 50-55°C has been investigated. The terpolymer and catalyst have been characterized by ICP-OES, FT-IR, TGA, SEM and EDAX analysis methods. Different parameters supporting chemo-selective hydrogenation such as concentration of NaBH4, TP-CuSO4, assisting base and solvents have been investigated. Controllable vigorous H2 gas evolution, low-temperature requirement, feasibility under air atmosphere, low catalyst loading, use of non-toxic reagents and solvents, high isolated yield, easy work-up procedure, purification without involving tedious column or flash chromatographic separation techniques, makes this protocol highly significant from the implementation point of view.

Natural vitamins as food antimicrobials in stem and thorn extracts of Hippophae species studied by HPLC-ESI-MS

Plants have always been and still on, a source of lifelines i.e., food and bioactive drugs for humans. Natural plant extracts are counted as natural antimicrobial agents and free radical scavengers and thus increasingly become important food additives. They can be added to food products in many forms viz., powder, paste, or liquid. Further food must be soft to eat and digest with safe and free from toxins. The plant extracts are safe and soft candidates as food preservatives over toxic and carcinogenic synthetic compounds due to their natural origin. The renewability of these plant extracts further compliments their use in the food industry to develop products for human well-being. In the present study, the metabolic composition of Hippophae tibetana, Hippophae rhamnoides, and Hippophae salicifolia stem and thorn extracts was studied. The extracts were tested in methanol, water, and hydroalcoholic solvent systems with High-performance liquid chromatography (HPLC). Ascorbic acid is found to be the highest percent content followed by tocopherol and coumaric acid in Hippophae species stem and thorn extracts. The presence of coumaric acid, vitamins C and E in these extracts soars the possibility of their essential use in the food and drug industry.

Solubility and solvation thermodynamics of l-tryptophan in aqueous methanol solvent mixtures

This article presents mole fraction solubilities of l-tryptophan in binary aqueous methanol solvent systems and applied them to estimate thermodynamical parameters like transfer Gibbs free energetics and entropies of solutions. The estimated thermodynamic energies were linked with interaction parameters like dipole–dipole interactions, enthalpy due to transfer of l-tryptophan from water to binary water–methanol solvent mixtures. The transfer Gibbs free energetics for chemical interactions and entropies for solvent–solvent interaction were calculated by mathematical relations. The chemical transfer energetics was applied to explain the probable stability of l-tryptophan in the mixed solvents. Many associated thermophysical parameters were also calculated at standard temperature (298.15 K).

Regioselective 2,7-dimethyl-9-phenyl xanthen-9-yl protection and deprotection of nucleosides under mild conditions

A simple and efficient protocol is developed for regioselective protection of primary hydroxyl group of nucleosides using 2,7-dimethylpixyliumtrifluoroacetate, prepared in-situ from 2,7-dimethyl-9-phenylxanthen-9-ol (DMPx-OH) and trifluoroacetic anhydride (TFAA). Furthermore deprotection of DMPx group is accomplished with TFAA in shorter reaction times with excellent yields in DCM:Methanol solvent system. Both procedures are successfully implemented on gram scale for synthesis of natural as well as other modified nucleosides. Various functional groups such as benzoyl, benzyl, tetrahydropyranyl, TBDPS, and formamidine groups are intact during protection with DMPx-group.

Antioxidant Guided Fractionation of Blackberry Polyphenols Show Synergistic Role of Catechins and Ellagitannins

In the present study, blackberry extract was prepared using a previously optimized solid-liquid extraction method in 70% aqueous acetone aimed at the recovery of its principal phenolics. Subsequently, 0.5 g of freeze-dried extract was subjected to flash chromatography fractionation, which was conducted on a C18 column using a binary solvent system of water and methanol at 10 mL/min. The total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) activities of the obtained 42 flash fractions were determined, and a strong positive correlation (r ≥ 0.986) was exhibited among them. Furthermore, the graph of the antioxidant indices of the flash fractions resembled the flash chromatogram, suggesting a good correlation among the compounds within the chromatographic peaks and the antioxidant indices. LC-MS/MS identified as many 28 phenolics, including cinnamtannin A2 reported for the first time in blackberries. This study further established the role of dominant anthocyanins (cyanidin-3-O-glucoside and cyanidin-3-O-rutinoside), but uniquely those of ellagitannins and catechins on the antioxidant capacity of blackberries.

RP-HPTLC fingerprinting of secondary metabolites from Nephrolepis exaltata and Cycasrevoluta

This study aims to develop a quantitative fingerprinting between two evolutionary close plant species Nephrolepis exaltata (L.) Schott and Cycas revoluta Thunb. The crude plant extracts were prepared in 1:1:1, v/v/v: Ethyl Acetate: n-Hexane: Methanol (AR) solvent system through cold extraction for 24 Hrs. The plant extracts were diluted (10 mg/mL) with a similar solvent system spotted (10 μL) on silica gel 60 F254 thin-layer chromatography plates. Five mobile phases (i) tetrahydrofuran (THF): toluene: formic acid: water (16:8:2:1) (ii) toluene: ethyl acetate: diethylamine (7:2:1) (iii) toluene: ethyl acetate: formic acid (7:3:0.1) (iv) n-hexane: ethyl acetate (7.2:2.9) (v) toluene: methanol (9:1) were used. The results were scanned at 254 nm, 366 nm, and in visible white light. This study gave minimum compact spots between Rf 0.06 to 0.051 corresponding to terpenoids, simultaneously for flavonoids spots were found between Rf 0.004 to 0.910. Total eight phenolic bands were found in Cycas revoluta among which five was found in leaf extract with Max RF of 0.033, 0.093, 0.273, 0.901 and 0.964 whereas three bands were found in stem extract with Max Rf of 0.007, 0.897 and 0.954. The presence of fifteen different types of flavonoids was determined by the flavonoids profile at Rf 0.004, 0.027, 0.047, 0.073, 0.134, 0.230, 0.303, 0.369, 0.369, 0.427, 0.514, 0.631, 0.703, 0.757, 0.846 and 0.910 for Nephrolepis exaltata. HPTLC fingerprint has shown several peaks with different Rf for different mobile phases. The fingerprinting would be helpful in the identification, and authentication of these species ecologically and therapeutically.

Unravelling the regulation mechanism of nanoflower shaped Na3V2(PO4)3 in methanol–water system for high performance sodium ion batteries

Researchers are accustomed to utilizing traditional solid-state and sol–gel routes to synthesize Na3V2(PO4)3 (NVP). No regulation mechanism is proposed to illustrate the growth process of NVP in solvothermal environment. In current work, uniform spherical particles with positive charge are synthesized by molecular self-assembly of nanosphere micelles using structural guiding agent polyethylene glycol (PEG) and surfactant Cetyltrimethylammonium bromide (CTAB). Nanoflower-shaped NVP sample is successfully prepared in the 100 % methyl alcohol medium, derived from the plate V2O5 (negative charge) combining with the nuclear nanospheres (positive charge) with the induction of electrostatic attraction. Based on the multi-dimensional tests and molecular dynamic calculation, the protective mechanism of polar solution for NVP system is proposed for the first time, that is, methanol solvent system can better protect the morphological features of NVP than water environment to maintain the original special nanoflower morphology. Due to the distinctive and stabilized nanoflower construction, the optimized NVP60 sample can provide more extra active sites for the efficient de-intercalation of Na+ and possess a large contacting area with electrolyte, resulting in the facilitated kinetic characteristics and superior electrochemical performance. The optimized NVP60 cathode possesses superior sodium storage property in both half cell and full cell. This work proposes a synergetic strategy combining theory and experiment to design and synthesize the modified NVP-based cathode materials from the molecular bonding aspect, further promoting the development of high performance cathode for SIBs.

Analysis of complexation between new bidentate bis-NHC ligand and some metal cations at different temperature

In this research, the determination and complexation process between 3,3'-(2,2'-(4-methyl-phenylenesulfonamido)bis(ethane-2,1-diyl))bis(1-benzyl-3H-benzo[d]imidazol-1-ium)dibromide with Ni2+, Zn2+, Pd2+, Ag+, and Hg2+ cations in the binary mixture of methanol (MeOH) and water (H2O) at different temperatures (15, 25, 35 and 45ºC) were studied using a conductometric method. The results show that the stoichiometry of the complexes in all binary mixed solvents for Ni2+, Zn2+, and Pd2+ were 1:1 (M:L), while in other cases 1:2 (M:L) and 2:1(M:L). The stability constants (log ) of complex formation have been determined by fitting molar conductivity curves using a computer program (GENPLOT). The obtained data shows that in the pure methanol solvent system, the stability order is Ni2+< Pd2+<Zn2+<Hg2+<Ag+ and the complexation process seems more stable in pure methanol in most cases. The thermodynamic parameters were determined conductometrically. The complexes in all cases were found to be enthalpy destabilized but entropy stabilized. The experimental data was tested by using an artificial neural network (ANN) program and was in good agreement with the estimated data.

Spectrodensitometric and ultra-performance liquid chromatographic quantification of dapagliflozin and saxagliptin in their dosage form and human plasma


 
 
 
 Purpose: To simultaneously quantify dapagliflozin (DAPA) and saxagliptin (SAX) in a pharmaceutical product and human plasma.
 Methods: Separation and quantification of DAPA and SAX were performed on pre-coated TLC plates in TLC-densitometric method using a solvent system of chloroform, ethyl acetate and methanol at a volume ratio of 8:1:1 as the mobile phase. The developed spots were scanned at 225 and 210 nm in absorbance mode. Moreover, the studied drugs were concurrently determined in human plasma using ultra-performance liquid chromatography (UPLC). The separation process was carried out in WatersTM Acquity C18 BEH column using a solvent system of 0.02 M KH2PO4 buffer, pH 4; MeOH and acetonitrile (2:1:1, v:v:v) isocratically at a flow speed of 0.5 mL/min. The absorbance of each eluent was read at 220 nm.
 Results: Concurrent evaluation of DAPA and SAX was carried without separation using TLC-densitometric method, and it was successful in determination of DAPA and SAX in concentration ranges of 10 – 70 μg/band and 5 – 60 μg/band, respectively. In addition, retardation factor (Rf) values for SAX and DAPA were 0.17 and 0.31, respectively. Furthermore, the studied drugs were concurrently determined in human plasma using UPLC, which was sensitive enough to quantify DAPA and SAX in concentration ranges of 100 – 1000 and 20 – 200 ng/mL, respectively.
 Conclusion: These methods can be utilized for sensitive monitoring of DAPA and SAX in pharmacokinetic and bioequivalence studies.
 
 
 

Development, Validation and Forced Degradation Study of Emtricitabine and Tenofovir Alafenamide in its Pharmaceutical Dosage Form Using RP-HPLC

Aims: The present research was aimed to develop and validate a reverse phase high performance liquid chromatographic (RP-HPLC) method for the quantification of Emtricitabine (EMT) and Tenofovir Alafenamide (TEN) in combination.
 Methodology: Separation was achieved under optimized chromatographic condition on an Inertsil C18, 250 x 4.6 mm, 5μm column. Various composition of mobile phase was tried. Separation of EMT and TEN was started with Methanol: Buffer and Methanol finally using solvent system of Buffer (pH 3.5) and Methanol in ratio of (30:70) and flow rate adjust at 1.0 ml/min was used as solvent system, the detection was carried out at 262nm using Shimazdu UV-visible detector. The mobile phase run time for the developed analytical method was 10 minutes.
 Results: The standard curve was found linear in the concentration range of 20-60 μg/ ml (r2- 0.9994) and 2.5-7.5 μg/ ml (r2-0.9992) for EMT and TEN respectively. The %RSD was found to be 0.80-0.95% and 0.63-1.09 for EMT and TEN respectively. Percentage (%) recoveries for EMT and TEN to be in range of 100%-100.6% and 99.32%-100.83% respectively. The limit of detection and the limit of quantification were found to be 4.80 μg/ ml and 14.7 μg/ ml respectively for EMT and 0.11 μg/ ml and 0.33μg/ ml respectively for TEN. Results of forced degradation study showed EMT degradation in acid and base medium while TEN was showed degradation in oxidative stress. The proposed developed RP-HPLC method was validated statistically and the values were found to be within the acceptable limits.
 Conclusion: In conclusion, the developed RP-HPLC method was found to be simple, specific, and rugged for simultaneous estimation of EMT and TEN. Validation results of method was found within the acceptable limits. Hence it can be used for analysis of EMT and TEN.

Development and Validation of UV Spectrophotometric Method for Estimation of Tea Tree Oil in Bulk and Cosmeceutical Creams

Abstract: Background: Tea tree oil, also known as Melaleuca alternifolia essential oil is widely used in skin care cosmeceuticals for its antibacterial, antifungal, analgesic and anti-inflammatory effect. The complex and variable composition of the oil poses many challenges to the analytical chemist and a recent survey of analytical methods indicated only a few simple and validated methods for estimation of the oil. Aim: A quality by design approach has therefore been adopted to develop a simple and novel UV spectrophotometric method for estimation of tea tree oil in bulk and cosmeceutical creams. Materials and Methods: UV spectrophotometric method was developed for estimation of Tea tree oil using dichloromethane- methanol (54 % v/v) solvent system. The method was then validated under optimized conditions for accuracy, precision and ruggedness, limit of detection and limit of quantification as per ICH: Q2 (R1) guidelines. Results: A characteristic spectral peak was observed at 267 nm and linearity was established over a concentration range of 10-160 mcg/ml with R2 value of 0.9961. Accuracy based on recovery studies, 100.5-113.6 %, precision and ruggedness based on % RSD values less than 2 %, LOD, 0.1277mcg/ml and LOQ and 4.195 mcg/ml indicated that the method is sensitive enough to be used for routine estimation. Conclusion: The method is robust and has been used to determine the essential oil content of two cosmeceutical creams. Key words: Tea tree oil, UV spectrophotometry, Box Behnken design, Optimization, Routine determination, Cosmeceutical cream.

A simple protocol for the isolation of amentoflavone from two species of Biophytum DC. (Oxalidaceae) and evaluation of its antiproliferative potential

The biflavonoid, amentoflavone, isolated earlier from the medicinally important species, Biophytum sensitivum, is reportedly a potent bio-active compound. In the present study, extraction protocols were standardized for the isolation of amentoflavone from two other species of Biophytum viz. B. reinwardtii (Zucc.) Klotzsch and B. veldkampii Shanavas et al and subsequent characterization of the same (using various chromatographic and spectroscopic methods) followed by screening of its bioactive potential using selected in vitro cytotoxicity assays (MTT assay, Trypan Blue Exclusion Assay and Flow cytometry).The shade-dried plant parts of the two species of Biophytum were Soxhlet extracted with ethyl alcohol (70%) and used as the raw material for the isolation of amentoflavone. Column chromatography, with a gradient solvent system of chloroform, methanol and water, followed by the thin layer chromatography of the pooled fractions with a solvent combination of chloroform, acetone and formic acid was adopted. A single fraction in both species, showed a clear prominent band in TLC, [a distinct fluorescent yellow band under short UV (100 nm) and blackish brown colour under long UV (320 nm)]. The band crystallized into a yellow powdery substance. Co-TLC (Rf value- 0.6) and High Performance Liquid Chromatography (Rt- 5.3 min) was used to confirm the isolated compound as amentoflavone. Yield, physical (solubility and melting point) and chemical properties Ultraviolet- Visible spectroscopy (UV–vis), High Resolution Mass Spectroscopy (HRMS) and Nuclear Magnetic Resonance Spectroscopy (1H, 13C, DEPT) of amentoflavone were also noted for both species. The yield of amentoflavone was higher in B. reinwardtii (86.66 mg /100 g dried plant powder) than in B. veldkampii (77.50 mg/100 g). The data generated from the different spectral techniques corresponded well with the chemical configuration of amentoflavone in the database. Therefore it could be confirmed that the isolated pure compound from both species of Biophytum was amentoflavone (C30H18O10). The isolated compound, amentoflavone was tested against cancer and normal cell lines. Amentoflavone was not cytotoxic to Human Peripheral Lymphocytes (normal cell lines) in trypan blue exclusion assay. MTT assay clearly showed that the exposure of amentoflavone at different concentrations resulted in decrease of MCF- 7(cancer cell lines) cell proliferation in a dose- dependent manner. Apoptosis was found to be dose-dependent, which was further confirmed by flow cytometry. Significant cell cycle arrest occurred in S phase and G2/M phase. Thus, anticancer studies (trypan blue exclusion assay, MTT assay and flow cytometry) proved useful for the scientific validation of the anticancer activity of the isolated pure compound, amentoflavone. The present study is the first report on the isolation of amentoflavone from Biophytum reinwardtii and B. veldkampii. Since the amentoflavone content in the two species of Biophytum is high, it can be extracted in sufficient amounts and both species can be considered potential source of the compound.

Kinetics Study of Dye Decolorization by Oxidation in Aqueous and Aqueous Alcohol Medium

The kinetics of oxidation of methylene blue with potassium bromate was investigated by observing decolorization of methylene blue in aqueous, aqueous methanol and aqueous isopropanol solvent system. The rate of reaction was studied as function of concentrations of methylene blue and potassium bromate in acidic medium at temperature 303 and 313 K. The increase in concentration of potassium bromate enhances the rate of decolorization of methylene blue in acidic medium; no significant change in rate was observed with variation of methylene blue. The rate of decolorization was found to be fastest in aqueous methanol compared to aqueous isopropanol and slow in aqueous medium. From the investigation, it was observed the reaction follows pseudo-first-order kinetics. The kinetic data were used to propose the mechanism of reaction. Thermodynamic parameters, enthalpy of activation (ΔH*), entropy of activation (ΔS*) and Gibbs free energy of activation (ΔG*), were also calculated to support kinetic results.

Simultaneous identification and estimation of glycyrrhizin, glabridin, and 18β‐glycyrrhetinic acid in de‐glycyrrhized Ayurvedic lipid‐based formulation of Glycyrrhiza glabra using dual wavelength reverse phase‐high‐performance liquid chromatography

AbstractPlant‐based Ayurvedic formulations such as medicated oils, confectioneries, etc. are developed with a rationale of selecting specific compounds for targeted action and minimal side effects. It is imperative to develop an analytical method to simultaneously identify and quantify the targeted compounds for good resolution with low retention time. The present assay using reverse phase‐high‐performance liquid chromatography is optimized to resolve glycyrrhizin, glabridin, and 18β‐glycyrrhetinic acid simultaneously at retention times of 6.6, 8.1, and 10.2 min, respectively, using acidified mobile phase from Glycyrrhiza glabra utilized in Ayurvedic lipid (cow's ghee and sesame oil) based formulations. Raw material, its decoction, and residues formed during preparation steps were extracted in methanol while lipid formulations were extracted using a binary solvent system of methanol and n‐hexane. The separation was performed on Hypersil gold column maintained at 40°C using 0.2% ortho‐phosphoric acid with pH 3.5 in water and acetonitrile as binary gradient mobile phase. The compounds were detected at wavelengths 230 (glabridin) and 254 (glycyrrhizin and 18β‐glycyrrhetinic acid) nm. The method revealed de‐glycyrrhized finished products containing glabridin and 18β‐glycyrrhetinic acid having medicinal value.