IMPURITY PROFILING OF FIRST LINE ANTI-TB DRUG-TERIZIDONE USING CHROMATOGRAPHIC AND RELATED TECHNIQUES

Objective: The objective of the present study was to investigate the stability of TRZ against different stressors and to prepare impurity profile for potential impurities and degradation products (DPs) formed under stress degradation of TRZ bulk drug and formulation. Methods: Three analytical methods were developed; the stability-indicating method that was developed using HPLC instrument with 0.01M ammonium acetate buffer (pH 4.0 using glacial acetic acid (GAA)) and acetonitrile in gradient program. The second method was a UPLC/ESI-MS method using 0.1 % Formic acid in Milli Q water (pH= 2.70) and 0.1%Formic acid in Milli Q water: Acetonitrile (10:90) in gradient program for identification of TRZ and DPs while the third, preparative HPLC method was used for isolation of impurities using (A) 0.05% ammonia (NH3) in water and (B) Acetonitrile+20% mobile phase A in gradient sequence. Gradient sequences are described in the main text. Results: The analytical method for stability study was developed and validated using ICH (Q2) R1 guidelines. The result of stability study by stress degradation showed that TRZ was susceptible to degradation in acid (7 DPs), alkaline, neutral (9 DPs) and oxidative conditions (10 DPs); major DPs were identified (where it was possible) and the chemical structure was elucidated by combining the data of ESI/MS, NMR and/or Tandem MS. The Impurity profiling was completed by reporting all the DPs, either major or minor for TRZ bulk drug and formulation. Conclusion: The complete Impurity profiling for TRZ is reported for the first time in literature. The study data would be add-on for formulation storage condition and further development

STABILITY INDICATING HPTLC METHOD FOR DETERMINATION OF CLEVIDIPINE BUTYRATE IN SYNTHETIC MIXTURE

Objective: To develop and validate stability indicating HPTLC method for determination of clevidipine butyrate in synthetic mixture.Methods: The present study deals with development and validation of stability indicating HPTLC method for estimation of clevidipine butryate. Chromatographic separation was performed on aluminum plate pre coated with Silica Gel 60 F254 using toluene: ethyl acetate (8:2) as mobile phase. TLC scanner was set at wavelength of 370 nm.Results: Retention factor Rf of clevidipine was found to be 0.49. The method was validated as per ICH guidelines. Calibration curve was in the range of 1000-6000ng/band. The correlation coefficient was found to be 0.999. The precision expressed by RSD was less than 2%. The accuracy of method was confirmed by recovery studies using standard addition method and recovery was found to be 99.03-99.57%. The drug was subjected to ICH prescribed hydrolytic, oxidative, photolytic and thermal stress conditions. Clevidipine and its degradation products were well resolved under experimental conditions. The method was validated according to ICH guidelines. The drug showed significant degradation in alkaline and acidic condition and slight degradation in oxidative condition. The drug was stable in thermal condition.Conclusion: A new, Simple, Accurate, Precise, Sensitive and economic stability indicating HPTLC method has been developed and validated for the determination of clevidipine and can be employed for stability indicating analysis

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING METHOD RP-HPLC METHOD OF ACOTIAMIDE

Objective: The objective of present work was to develop and validate simple, precise, accurate and specific stability indicating method for determination of acotiamide in presence of its degradation products.Methods: An isocratic RP-HPLC method has been developed using C-8 Thermo Hypersil BDS Column (250 x 4.6 mm i.d., 5µparticle size) with the mobile phase composition of acetonitrile: 0.1 % triethylamine in 0.2% formic acid (30: 70) at column oven temperature of 40 °C. The flow rate was 1.0 ml min-1 and effluent was detected at 282 nm. The method was validated in terms of linearity, accuracy, precision, LOD (Limit of Detection), LOQ (Limit of Quantification) and robustness as per ICH guidelines.Results: The method was found to be linear in the range of 10-60µg/ml. Limit of detection and limit of quantification was found to 0.36µg/ml and 1.10 µg/ml.% Recovery was found to be in the range of 99.45%-99.75%and precision less than 2%. The developed method was successfully applied for estimation of Acotiamide in marketed tablet formulation and percentage assay was found to be 100.45%. Acotiamide was subjected to stress degradation under acid, base, neutral hydrolysis, oxidation, dry heat, photolysis conditions. Significant degradation was observed in acid and base degradation.Conclusion: The developed RP-HPLC method was simple, rapid, accurate, precise and stability indicating for the estimation of Acotiamide in bulk and tablet dosage form

DEVELOPMENT OF HPLC AND CHEMOMETRIC ASSISTED SPECTROPHOTOMETRIC METHODS FOR THE SIMULTANEOUS DETERMINATION OF FIVE ACTIVE INGREDIENTS IN COUGH AND COLD TABLETS AND THEIR APPLICATION TO DISSOLUTION STUDY

Objective: Chemometric assisted spectrophotometry, and HPLC methods have been developed for the simultaneous determination of phenylephrine hydrochloride (PEPH), paracetamol (PCM), guaifenesin (GNF), chlorpheniramine maleate (CPM) and bromhexine hydrochloride (BRM).Methods: The chromatographic separation was carried out on a Phenomenex RPC18 column. An isocratic elution was carried out with the mobile phase comprising methanol, acetonitrile and 10 mM phosphate buffer (pH 3) in the ratio of 27.5:22.5:50 respectively at a wavelength of 218 nm. Two chemometric methods i.e. principal component regression (PCR) method and partial least squares (PLS) method were also developed to quantify each drug in the mixture using the information included in the UV absorption spectra of appropriate solutions in the range 210–320 nm with the intervals of 2 nm at 51 wavelengths.Results: The three methods were successfully applied to a tablet formulation and the results were compared statistically by applying ANOVA, which showed no significant difference among the three methods. The methods were applied to the dissolution study of the five components in tablet formulation and the percentage release of all the five components was found to be greater than 85% within 45 min by all the three methods.Conclusion: Thus, the proposed methods, i.e., PLS, PCR and HPLC, were found to be suitable and can be successfully used for the determination of the PEPH, PCM, GNF, CPM and BRM in pharmaceutical tablet formulation as well as for dissolution study.Â

RP-HPLC and chemometric assisted UV-spectrophotometric methods for simultaneous in vitro analysis of atrovastatin calcium, ezetimibe and fenofibrate in their pharmaceutical formulation

One RP-HPLC and two chemometric assisted UV spectrophotometric methods were developed and validated for the simultaneous in vitro analysis of Atrovastatin calcium (ATV), Ezetimibe (EZET) and Fenofibrate (FEN) in their pharmaceutical preparation and ternary mixtures. The chromatographic separation was achieved on a reversed-phase, Hypersil BDS C8 column (250X4.6 mm i.d, 5 particle size) with a mobile phase consisting of methanol and 0.05M phosphate buffer (pH-6.3 adjusted with sodium hydroxide) in the ratio of (85:15)% v/v. The total run time was 7 min. Quantitation was achieved with UV detection at 248nm based on peak area. Linearity was observed over concentration range of 5 - 12μg mL-1 for ATV and EZET and 80 - 192 μg mL-1 for FEN. The two chemometric methods applied were inverse least square (ILS) and classical least square (CLS). These approaches were successfully applied to quantify each drug in their mixture using the information included in the UV absorption spectra of appropriate solutions in the wavelength range 220-310nm with the intervals of 5nm (Δλ = 5nm) at 19 wavelength points. For the chemometric calibration, 18 ternary solutions were prepared as training set and 10 ternary solutions were prepared as validation set. The developed methods were successfully applied for laboratory prepared mixtures as well as commercial tablet formulation for ATV, EZET and FEN concentration. The results obtained for pharmaceutical formulation by ILS and CLS methods were compared with isocratic HPLC method and a good agreement was found

Centrality evolution of the charged-particle pseudorapidity density over a broad pseudorapidity range in Pb-Pb collisions at root s(NN)=2.76TeV

Peer reviewe

STABILITY INDICATING HPTLC METHOD FOR DETERMINATION OF CLEVIDIPINE BUTYRATE IN SYNTHETIC MIXTURE

Objective: To develop and validate stability indicating HPTLC method for determination of clevidipine butyrate in synthetic mixture.Methods: The present study deals with development and validation of stability indicating HPTLC method for estimation of clevidipine butryate. Chromatographic separation was performed on aluminum plate pre coated with Silica Gel 60 F254 using toluene: ethyl acetate (8:2) as mobile phase. TLC scanner was set at wavelength of 370 nm.Results: Retention factor Rf of clevidipine was found to be 0.49. The method was validated as per ICH guidelines. Calibration curve was in the range of 1000-6000ng/band. The correlation coefficient was found to be 0.999. The precision expressed by RSD was less than 2%. The accuracy of method was confirmed by recovery studies using standard addition method and recovery was found to be 99.03-99.57%. The drug was subjected to ICH prescribed hydrolytic, oxidative, photolytic and thermal stress conditions. Clevidipine and its degradation products were well resolved under experimental conditions. The method was validated according to ICH guidelines. The drug showed significant degradation in alkaline and acidic condition and slight degradation in oxidative condition. The drug was stable in thermal condition.Conclusion: A new, Simple, Accurate, Precise, Sensitive and economic stability indicating HPTLC method has been developed and validated for the determination of clevidipine and can be employed for stability indicating analysis

Biological Evaluation of Polyherbal Ayurvedic Cardiotonic Preparation “Mahamrutyunjaya rasa”

Mahamrutyunjaya rasa (MHR), an Ayurvedic formulation, used as cardiotonic, contains potentially toxic compounds like aconitine, which are detoxified during preparation using traditional methods. Comparative toxicological evaluation of laboratory prepared formulation (F1) and two marketed formulations (F2 and F3) were performed based on their effects on viability of H9c2 cells and after single oral dose administration in mice. Cardioprotective effect of formulations at 25 and 50 mg/kg doses were studied in isoproterenol- (ISO-) induced myocardial infarcted rats. F1 and F2 did not affect the cell viability, while F3 decreased the cell viability in concentration and time-dependent manner. Rats administered with ISO showed significant increase in the serum levels of glutamate oxaloacetate transaminase, alkaline phosphotase, creatinine kinase isoenzymes, lactate dehydrogenase, and uric acid, while F1 and F2 treatment showed significant reduction in the same. F3 showed further increase in the serum levels of enzymes and uric acid in ISO-challenged rats. High pressure liquid chromatographic analysis of formulations showed higher concentration of aconitine in F3. Study shows that F1 and F2 possess cardioprotective property with higher safety, while formulation F3 cannot be used as cardioprotective due to its cytotoxic effects. Thus, proper quality assessment methods are required during preparation of traditional formulations

Simultaneous Resolution of Clopidogrel Bisulphate and Aspirin in their Combined Dosage Form by Ratio First Derivative Spectrophotometry

In the present work, the ratio spectra first derivative spectrophotometry is proposed for the resolution of Clopidogrel bisulphate (CLP) and Apirin (ASP) in synthetic mixtures prepared in random fashion and tablets available in 1:1 and 1:2 ratio. The calibration graph follows Beer’s law in the range of 6 to 30 μcg/ml for clopidogrel bisulphate and from 4-20 μcg/ml for aspirin. The mean recoveries & relative standard deviations were found as 99.40% & 0.442% for CLP and 100.09% and 0.905% for ASP, respectively in 1:1 ratio. Similarly the mean recoveries & relative standard deviations were found as 999.84% & 1.024% for CLP and 99.95% and 0.578% for ASP, respectively in 1:2 ratio. The graphical treatment of the overlapping spectra depends on division of the absorption spectrum of the binary mixture by a standard spectrum of one of the components and then calculating the first derivative of the ratio spectrum. These approaches were successfully applied to quantify CLP combined with ASP in tablets respectively and validated accordingly for random ratios in synthetic mixtures