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Gopal Rao M.,Sri Ramakrishna Institute of Paramedical science
International Journal of Pharma and Bio Sciences | Year: 2015

The objective of this study was to develop a simple, specific, precise and accurate reverse phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous estimation of Gliclazide (GLZ) and Sitagliptin Phosphate Monohydrate (SPM) in bulk and pharmaceutical dosage form. In the present work, SHIMADZU HPLC with UV detector LC 10AT VP with analytical column Phenomenex Luna (C18) A 100RP Column, 250mm x 4.6mm x 5μm, an injection volume of 20μl was injected and eluted with mobile phase Water: Acetonitrile (40:60) pumped at a flow rate of 1.0ml/min. Gliclazide (GLZ) and Sitagliptin Phosphate Monohydrate (SPM) were eluted at 3.268 and 2.260 min. The detection was carried out at a wavelength of 253nm. The method has shown good linearity in the concentration range of 5-25μg/ml for Gliclazide and 20- 100μg/ml for Sitagliptin phosphate monohydrate with a correlation coefficient of (r2) 0.9997 and 0.9940. The percentage assay values were found to be 100.01 and 99.3 for Gliclazide (GLZ) and Sitagliptin Phosphate Monohydrate (SPM). Limit of detection and limit of quantification were found to be 0.4364μg/ml and 1.3232 μg/ml for Gliclazide, 0.6 -μg/ml and 1.9 μg/ml for Sitagliptin Phosphate Monohydrate respectively. The method was validated for system suitability, linearity, accuracy, precision, robustness and ruggedness of the sample solution.


Asokkumar K.,Sri Ramakrishna Institute of Paramedical science | Sen S.,Sri Ramakrishna Institute of Paramedical science | Sen S.,Assam University | Umamaheswari M.,Sri Ramakrishna Institute of Paramedical science | And 2 more authors.
Pharmacological Reports | Year: 2014

Background Antioxidant supplements with existing drugs may confer better therapeutic efficacy in oxidative stress related diseases. The purpose of the present work was to characterize the interaction and investigate the protective effect of H2 blocker famotidine and gallic acid in combination against experimentally induced peptic ulcer. Methods Preventive effect of gallic acid and famotidine in different combinations was investigated against aspirin plus pyloric ligation induced ulcer in rat. Ulcer index, gastric juice volume, pH, other biochemical parameters of gastric juice and antioxidant activity using stomach tissue were estimated. Results Pretreatment with gallic acid and famotidine in combinations for 7 days, protected the gastric mucosa significantly (p < 0.05, 0.01), which was evidenced by decrease in ulcer index, gastric juice volume, free and total acidity, total protein, pepsin and DNA content, and increase in pH, carbohydrates concentration in gastric juice. Combination treatment increases levels of superoxide dismutase, catalase, reduced glutathione, glutathione reductase and glucose-6-phosphate dehydrogenase, and decreases lipid peroxidation, myloperoxidase in stomach tissue. Along with higher dose combination, lower dose combinations like gallic acid (50 mg/kg) plus famotidine (10 mg/kg) also offered better antiulcer activity than their individual effect. Histopathological studies confirmed their antiulcer activity. Conclusion Combination treatments confer synergistic protective effect against peptic ulcer in rats, which was related to the gastroprotective, antisecratory and antioxidant activity of combination treatment. Results proved that use of gallic acid with existing antiulcer drug will be more useful in the prevention/management of peptic ulcer. © 2014 Institute of Pharmacology, Polish Academy of Sciences.


Sonia G.,Sri Ramakrishna Institute of Paramedical science | Ravi T.K.,Sri Ramakrishna Institute of Paramedical science
Medicinal Chemistry Research | Year: 2013

Pyrrolidine carboxamides have been recognized as potent direct enoyl-acyl carrier protein reductase inhibitors. In our study, the search for new pyrrolidine carboxamides incorporated with various heterocyclic moieties, possessing antitubercular activities is been done. A series of oxadiazolyl pyrrolidine carboxamides (2a-e) were synthesized by reacting pyrrolidine carboxylic acid and oxadiazole amines using HBTU as amide forming agent. The purity of the synthesized compounds was confirmed by physical characterization like melting point and thin layer chromatography. The functional group identification was done based on spectral characterization utilizing, FT-IR, 1H NMR, 13C NMR, mass spectral data and elemental analysis. Invitro antitubercular screening was performed by alamar blue assay method on mycobacterium tuberculosis H37Rv. © 2012 Springer Science+Business Media New York.


Amutha Gnana Arasi M.A.S.,Sri Ramakrishna Institute of Paramedical science | Gopal Rao M.,Sri Ramakrishna Institute of Paramedical science | Bagyalakshmi J.,Sri Ramakrishna Institute of Paramedical science
International Journal of Biological Macromolecules | Year: 2016

This study deals with the optimization of microwave assisted extraction of polysaccharide from Psidium guajava L. fruit using Response surface methodology. To evaluate the effect of three independent variables, Water to plant material ratio, microwave power used for extraction and Irradiation time, central composite design has been employed. The yield is considered as dependent variable. The design model estimated the optimum yield of 6.81677% at 200 W microwave power level, 3:1 water to plant material ratio and 20 min of irradiation time. Three factors three levels Central composite design coupled with RSM was used to model the extraction process. ANOVA was performed to find the significance of the model. The polysaccharide extracted using microwave assisted extraction process was analyzed using FTIR Spectroscopy. © 2016 Elsevier B.V.


Varghese S.J.,Sri Ramakrishna Institute of Paramedical science | Ravi T.K.,Sri Ramakrishna Institute of Paramedical science
Journal of AOAC International | Year: 2010

This paper describes validated HPLC and HPTLC methods for the simultaneous determination of rosuvastatin (ROS) and ezetimibe (EZE) in a combined tablet dosage form. The isocratic RP-HPLC analysis was performed on a Chromolith C18 column (100 × 6 mm id) using 0.1% (v/v) orthophosphoric acid solution (pH 3.5)-acetonitrile (63 + 37, v/v) mobile phase at a flow rate of 1 mL/min at ambient temperature. Quantification was carried out using a photodiode array UV detector at 245 nm over the concentration range of 0.5-10 mg/mL for ROS and EZE. The HPTLC separation was carried out on an aluminum-backed sheet of silica gel 60F254 layers using n-butyl acetate-chloroform-glacial acetic acid (1 + 8 + 1, v/v/v) mobile phase. Quantification was achieved with UV densitometry at 245 nm over a concentration range of 0.1-0.9 μg/spot for ROS and EZE. The analytical methods were validated according to International Conference on Harmonization guidelines. Low RSD values indicated good precision. Both methods were successfully applied for the analysis of the drugs in laboratory-prepared mixtures and commercial tablets. No chromatographic interference from the tablet excipients was found. These methods are simple, precise, and sensitive, and are applicable for simultaneous determination of ROS and EZE in pure powder and tablets.


Varghese S.J.,Sri Ramakrishna Institute of Paramedical science | Ravi T.K.,Sri Ramakrishna Institute of Paramedical science
Journal of Liquid Chromatography and Related Technologies | Year: 2011

Simple, accurate, precise, sensitive, and validated HPLC and HPTLC-densitometric methods were developed for simultaneous determination of amlodipine (AML), valsartan (VAL), and hydrochlorothiazide (HYD) in combined tablet dosage form. Method A, the gradient RP-HPLC analysis was performed on a Phenomenex Luna C18 (4.60 mm x 150 mm, 5 μ particle size) column, using a mobile phase consisting of 10 mM ammonium acetate buffer (pH 6.7) and methanol in solvent gradient elution for 20 min at a flow rate of 1 mL min-1. Quantification was carried out using a photodiode array UV detector at 238 nm. The employment of a diode array detector allowed selectivity confirmation by peak purity evaluation. Method B, the HPTLC analysis was carried out on an aluminum-backed sheet of silica gel 60F254 layers using chloroform: glacial acetic acid:n-butyl acetate (8:4:2, v/v/v) as the mobile phase. Quantification was achieved with UV densitometry at 320 nm. The analytical methods were validated according to International Conference on Harmonization (ICH) guidelines. No chromatographic interference from tablet excipients was found and hence these methods are applicable for simultaneous determination of AML, VAL, and HYD in pharmaceutical formulations and biological fluids. Copyright © Taylor & Francis Group, LLC.


Varghese S.J.,Sri Ramakrishna Institute of Paramedical science | Ravi T.K.,Sri Ramakrishna Institute of Paramedical science
Journal of AOAC International | Year: 2013

A simple, rapid, and sensitive LC/electrospray ionization (ESI)-MS method was developed and validated for the simultaneous determination of rosuvastatin (ROS) and ezetimibe (EZE) in human plasma. Following liquid-liquid extraction, the analytes and an internal standard, atorvastatin (ATO), were separated using an isocratic mobile phase comprising 0.1% (v/v) formic acid-methanol (20 + 80, v/v) on an RP-C18 column. Detection was performed on a mass spectrometer by selected ion monitoring using their respective [M-H]- ions, m/z 480 for ROS, m/z 408 for EZE, and m/z 557 for ATO. For both analytes, the method was linear in the range of 0.1 to 10 ng/mL. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 4 min made it possible to determine many plasma samples/day. The validated LC/ESI-MS method can be used to study pharmacokinetics, bioavailability, and bioequivalence of combined dosage forms of ROS and EZE. © 2013 Publishing Technology.


Madeswaran A.,Sri Ramakrishna Institute of Paramedical science | Asokkumar K.,Sri Ramakrishna Institute of Paramedical science
Oriental Pharmacy and Experimental Medicine | Year: 2015

Angiotensin-converting enzyme (ACE) is known to be key factor for hypertension. Alkaloids are the principal constituents that may play a crucial role in the management of hypertension. The current objective of the study is to identify inhibitory affinity potential of the certain commercially available alkaloids, against crystal structure of human angiotensin-converting enzyme (4APH) using AutoDock 4.2. In this perspective, alkaloids like quinidine, quinine, tubocurarine, vinblastine, vincamine, vincristine and yohimbine were selected. Captopril, a known ACE inhibitor was used as the standard. All the selected compounds were analysed for Lipinski’s rule of five. In the docking studies, conformational site analysis and docking parameters like binding energy, inhibition constant and intermolecular energy were determined using AutoDock 4.2. The selected compounds and the standard showed the following amino acid residues are responsible for the inhibitory affinity potential ASP 1, ARG 2, VAL 3, TYR 4, ARG 522 along with hydrogen bonding interactions. The selected compounds exhibited the binding energy ranging between −9.56 and −5.82 kcal/mol when compared with that of the standard (−5.38 kcal/mol). Inhibition constant (97.64 nM to 53.96 μM) of the alkaloids also coincide with the binding energy. The compounds exhibited the intermolecular energies ranging between −11.40 and −9.14 kcal/mol. The current molecular simulation study predicted the ACE inhibitory activity of the selected compounds. Hence, these compounds can be further evaluated to develop potential chemical molecules for the prevention and management of hypertension. © 2015, Institute of Korean Medicine, Kyung Hee University and Springer Science+Business Media Dordrecht.


Umamaheswari M.,Sri Ramakrishna Institute of Paramedical science | Madeswaran A.,Sri Ramakrishna Institute of Paramedical science | Asokkumar K.,Sri Ramakrishna Institute of Paramedical science
Iranian Journal of Pharmaceutical Research | Year: 2013

Allopurinol, the xanthine oxidase inhibitor, is the only drug available for the treatment of gout. We examined the xanthine oxidase inhibitory activity of some commercially available favonoids such asepigallocatechin, acacatechin, myricetin, naringenin, daidzein and glycitein by virtual screening and in-vitro studies. The interacting residues within the complex model and their contact types were identifed. The virtual screening analysis were carried out using AutoDock 4.2 and in-vitro xanthine oxidase inhibitory activity was carried out using xanthine as the substrate. In addition, enzyme kinetics was performed using LineweaverBurkplot analysis. Allopurinol, a known xanthine oxidase inhibitor was used as the standard. The docking energy ofglycitein was found to be -8.49 kcal/mol which was less than that of the standard (-4.47 kcal/ mol). All the selected favonoids were found to exhibit lower binding energy (-8.08 to -6.03 kcal/ mol) than allopurinol. The docking results confrm that favonoids showed greater inhibition of xanthine oxidase due to their active binding sites and lesser binding energies compared to allopurinol. This may be attributed to the presence of benzopyran ring in the favonoids. In the xanthine oxidase assay, IC50 value of glycitein was found to be 12±0.86 μg/mL, whereas that of allopurinol was 24±0.28 μg/mL. All the remaining compounds exhibited IC50 values ranging between 22±0.64 to 62±1.18 μg/mL. In the enzyme kinetic studies, favonoids showed competitive type of enzyme inhibition. It can be concluded that favonoids could be a promising remedy for the treatment of gout and related infammatory disorders. Further in-vivo studies are required to develop potential compounds with lesser side effects. © 2013 by School of Pharmacy.


Sonia G.,Sri Ramakrishna Institute of Paramedical science | Thachil K.K.,Sri Ramakrishna Institute of Paramedical science | Parameswaran M.K.,Sri Ramakrishna Institute of Paramedical science | Kochupappy R.T.,Sri Ramakrishna Institute of Paramedical science
Medicinal Chemistry Research | Year: 2014

2-[2-(3-Methyl-5-oxo-4,5-dihydro-1H-pyrazol-1-yl)-2-oxoethyl]-2H-1, 4-benzoxazin-3(4H)-one (3) was obtained starting from methyl-(3,4-dihydro-3-oxo- 2H-1,4-benzoxazin-2-yl) acetate (1) through the corresponding hydrazide (2). Condensation of (3) with different aromatic aldehydes under Knoevengel condensation afforded 2-{2-[3-methyl-4-(2-methylbenzylidine)-5-oxo-4,5-dihydro- 1H-pyrazol-1-yl]-2-oxoethyl}-2H-1,4-benzoxazin-3(4H)-ones (4a-k). The structures of the compounds were determined by FT-IR, 1H NMR, 13C NMR, mass spectral data, and elemental analysis. All the synthesized compounds were screened for their in vitro antimicrobial and antioxidant studies. © 2013 Springer Science+Business Media New York.

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