Validation

A simple, specific, accurate and precise reverse phase high performance liquid chromatographic method was developed and validated for the estimation of Venlafaxine hydrochloride in pure and Pharmaceutical dosage form. Kromasil C18 column having 150 mm x 4.6 mm internal diameter, 5 µm particle sizes in isocratic mode with mobile phase containing mixture of methanol and water in the ratio of 65:35 v/v was used. The flow rate was 1.0 ml/min and effluents were monitored at 225 nm. The retention time for Venlafaxine was 2.424 min. The method was validated for linearity, accuracy, precision, specificity, limit of detection, limit of quantification and robustness. Limit of detection and limit of quantification were found 2.97µg/ml and 9.92 µg/ml respectively and recovery of Venlafaxine from tablet formulation was found 100.4 %. The proposed method was successfully applied for the quantitative determination of Venlafaxine in tablet dosage form.

A new, simple, specific, accurate and precise RP-HPLC method was developed for determination of Baclofen. In the present study, stress testing of Baclofen was carried out according to ICH guidelines Q1A (R2). Baclofen was subjected to stress conditions of hydrolysis, oxidation, photolysis and neutral decomposition. Extensive degradation was found to occur in acidic, condition. Mild degradation was observed in basic and at thermal conditions. Successful separation of drug from degradation products formed under stress conditions was achieved on a Hypersil BDS C18 column (250 mm × 4.6 mm, 5.0 μ particle size) using acetonitrile: acetate buffer (pH 3.7 ± 0.05) (50:50 v/v), at a flow rate of 1.0 mL/min and column was maintained at 40˚C. Quantification and linearity was achieved at 272 nm over the concentration range of 5 - 100 μg/mL for Baclofen. The method was validated for specificity, linearity, accuracy, precision, LOD, LOQ and robustness.

Sensitive spectrophotometric method is described for the determination of tetracycline HCl in bulk and in pharmaceutical formulations. The method is based on the coupling of tetracycline HCl with diazotized 8-hydroxy quinoline reagent in basic medium at room temperature to form a reddish to brown mono azo dye soluble in water with maximum absorption at 395 nm. The reaction was followed up spectrophotometrically by measuring the increase in absorbance at 395 nm. The analytical performance of the method, in terms of accuracy and precision, was statistically validated; the results were satisfactory. The calibration graph is linear in the concentration range 1-20 μg ml-1 ,with 0.005 μg ml-1 detection limit and 0.324 μg ml-1 , 0.980 μg ml-1  Limit of detection(LOD) and Limit of quantification(LOQ) respectively. The method has been successfully applied to the determination of the studied drugs in commercial pharmaceutical formulations.

A simple, specific, accurate, precise and sensitive RP- HPLC method has been developed for the rapid estimation of Ganciclovir in bulk and its formulations. The chromatographic separation was carried on Grace smart RP-18 column (250 x 4.6 mm, 5 μm), using Methanol: Citrate buffer (0.05M) at PH-5.2 with KOH 70:30 (v/v) as mobile phase, at a flow rate of 1.0 ml/min.  The detection was carried out at 254 nm and drug eluted with a retention time of 2.982 min. Beer’s law was obeyed in the concentration range of 10-60μg/ml with correlation coefficient 0.999. The method has been validated according to ICH guidelines for specificity, linearity, accuracy, precision, robustness, ruggedness, LOD and LOQ. The method was found to be specific, accurate, and precise, robust, rugged and sensitive. The developed method was good linearity, novel, rapid for the estimation of Ganciclovir in bulk and capsule dosage form. Thus it can be employed for the routine analysis.

An accurate, simple, rapid, precise and economical RP- HPLC method has been developed for the rapid estimation of ImatinibMesylate in bulk and pharmaceutical formulation. The separation was achieved on Phenomenex C18 G column ( 250 x 4.6 mm i.d, 5 μm), using Methanol : 1-octanesulphonic acid (0.05M) at PH-8 with KOH  70:30 (v/v) as mobile phase, at a flow rate of 1.0 ml/min. Detection was carried out at 269 nm and drug eluted with a retention time of 5.548 min. Beer’s law was obeyed in the concentration range of 2-12μg/ml with correlation coefficient 0.999. The method had been validated according to ICH guide lines for specificity, linearity, accuracy, precision, robustness, ruggedness, LOD and LOQ. The method was found to be specific, accurate, and precise, robust, rugged and sensitive. The proposed method was convenient for quantitative routine analysis and quality control of ImatinibMesylate in bulk and pharmaceutical dosage form. Key words: Imatinib Mesylate , RP-HPLC, Validation, 1-Octanesulphonic acid.

A novel, simple, precise, rapid, reproducible and cost effective RP-HPLC method was developed and validated for the simultaneous estimation of Nadifloxacin and Clobetasol propionate in its pharmaceutical dosage form. The chromatographic separation was carried out using C18 Shim pack XR ODS II (250 mm × 4.6 mm, 5µm) column with mobile phase comprising of Acetonitrile : Water (50:50)(%v/v). Flow rate was maintained 1.0 mL/min and quantitation was carried out using UV detection at 242nm. Retention time of Nadifloxacin and Clobetasol propionate were found to be 2.64 min and 6.19 min respectively. The method was validated by assessing different parameters such as specificity, linearity, precision, accuracy, robustness, LOD and LOQ for the developed method. The linearity range 20-240 µg/mL and 1-12 µg/mL were selected for Nadifloxacin and Clobetasol propionate respectively. The correlation coefficient (r2) for Nadifloxacin and Clobetasol propionate were found to be 0.9995 and 0.9999 respectively. The limit of detection for Nadifloxacin and Clobetasol propionate were found to be 0.029 µg/mL and 0.21 µg/mL. The limit of quantitation for Nadifloxacin and Clobetasol propionate were found to be 0.089 µg/mL and 0.64 µg/ml respectively. Percentage recovery was found to be 99% to 100.56% for Nadifloxacin and 99.37% to 99.79% for Clobetasol propionate. All the validation parameters were with-in the acceptance limit. The relative standard deviation (%RSD) was found to beless than 2% in all the assessed parameters. The developed HPLC method can successfully used for the quantitative estimation of both the drugs in its formulation.

A simple, cheap, fast and accurate Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method was developed and validated for determination of Trifluoperazine Hydrochloride and its degraded products in pharmaceutical dosage form. This method was developed by using an analytical Zodiac C18 Column (250 mmx4.6mm, 5µm) and mobile phase comprises of 70% methanol and 30% acetonitrile. The method was validated and found to be linear, selective, accurate, robust, rugged and precise. The lower Limit of Detection (LOD) and lower Limit of Quantification (LOQ) respectively were 1.50 µg/ml and 5.0 µg/ml.  The methods utilized in this study are the degradation of drug i.e. aqueous, 0.1N HCl, 0.1N NaOH, 3% H2O2, thermal, photolytic and UV light. Developed HPLC method is able to separate all degrading products from any stress condition from drug peak by resolution of more than 2. This developed method can be used for routine analysis for the estimation of Trifluoperazine Hydrochloride in bulk and in tablet dosage form in pharmaceutical industry because it is simple, cheap and accurate.

The article aims at developing simple, fast and effective dissolution method for Dabigatran etexilate mesylate capsules by RP-HPLC and validate as per ICH guidelines. The optimized RP-HPLC method for dissolution studies uses a reverse phase column, Phenomenex Luna C18 (250 X 4.6 mm;5μ), a mobile phase of triethylammonium phosphate buffer (pH 3.0):acetonitrile in the proportion of 40:60 v/v, diluent as 0.01N HCl, flow rate of 1.0ml/min, injection volume as 20µl. and a detection wavelength of 341nm using a UV detector. The optimized dissolution conditions include, 0.01N HCl as dissolution media, apparatus as USP Type 1 Basket, rpm as 100, dissolution media temperature as 37±0.5ºC, dissolution volume as 500ml, dissolution time point as 30mts, working concentration of standard and sample as 5µg/ml and a detection wavelength of 341 nm. The developed method resulted in Dabigatran etexilate exhibiting linearity in the range 1.25-10μg/ml. System precision and intra-day precision is exemplified by relative standard deviation of 1.59% and 2.21% respectively. Method was found to be rugged/inter day precise as %RSD was found to be 3.25. Percentage Mean recovery was found to be greater than 80% at all the three levels by absolute method during accuracy studies. LOD and LOQ for Dabigatran etexilate were found to be 0.05ng/ml and 5ng/ml respectively. Hence it can be concluded that effective dissolution method by RP-HPLC is developed and validated as per ICH guidelines which can be applicable in various pharmaceutical industries.

A simple, rapid, accurate, precise and economical reverse phase high performance liquid chromatographic method was developed for simultaneous quantification of two anti-hypertensive drugs Olmesartan and Chlorthalidone. The separation of both the drugs was achieved on BDS C18 250mm x 4.6 mm, 5m using a mobile phase of 10 mM orthophosphoric acid buffer and acetonitrile (45:55v/v) at a flow rate of 1.0 mL min-1 and detection was performed at 212 nm using photodiode array (PDA) detector. The drug was subjected to various ICH prescribed stress conditions including hydrolysis (neutral, acid and alkaline), oxidation, photolysis and thermal degradation. The proposed method was validated with respect to specificity, linearity, accuracy, and precision, limit of detection (LOD), limit of quantitation (LOQ), stability and robustness as per ICH guidelines. The proposed analytical method could effectively separate the drug from its degradation products employed as stability indicating studies.

The present manuscript describes sensitive, rapid, accurate, and precise spectrophotometric method for simultaneous estimation of allopurinol and aceclofenac in pharmaceutical dosage form. Simultaneous equation method used at 250nm & 273nm as λmax of allopurinol and aceclofenac respectively in 0.1N HCl. Both the drugs follow beer-lamber’s law on the concentration range 2-12µg/ml and 2-24µg/ml respectively for allopurinol & aceclofenac. The mean percentage recovery is 100.01% and 99.4% for allopurinol and aceclofenac respectively by simultaneous equation. The method has been validated as per ICH guidelines. The proposed method effectively applied to bulk drug and pharmaceutical formulation. The accuracy and reproducibility are close to 100% with acceptance range of %R.S.D. The method was successfully applied to pharmaceutical dosage form because no interference was found from excipient.