validation.

The present study was designed to develop simple accurate, precise, reproducible and validating of a stability indicating reverse phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous estimation of azithromycin, ornidazole and fluconazole in bulk and its pharmaceutical dosage forms. Chromatographic separation of the three drugs was performed on a Phenomenex C18 column (250X4.6mm 5μm) as stationary phase with a mobile phase comprising of 20mM potassium dihydrogen phosphate : Acetonitrile (pH 4.8) in the ratio 30:70% v/v at a flow rate of 1ml/min and peak monitored at 254nm using PDA detector. The retention time of azithromycin, fluconazole, ornidazole and procaine hydrochloride (internal standard) was Rt1-2.8, Rt2-5.0, Rt3-6.3 and Rt-3.8minutes respectively. The linearity of azithromycin, fluconazole and ornidazole were in the range of 20-100μg/ml, 3-15μg/ml and 15-75μg/ml with an internal standard, procaine hydrochloride 5µg/ml respectively. The accuracy of the method was found to be 98-102% and %RSD was found to be less than 2% indicating high degree of accuracy and precision of the proposed HPLC method. The limit of detection for azithromycin, fluconazole and ornidazole was found to be 0.34, 2.80 and 0.76μg/ml respectively whereas, the limit of quantification was found to be 1.05, 8.6 and 2.31μg/ml respectively. Forced degradation studies were conducted to know the stability of the drug samples under various stress conditions like acid, base, peroxide and photolytic degradation according to ICH guidelines. Results are validated statistically as per ICH guidelines.

The main aim of current study was to develop U V visible spectrophotometric method for the quantitative determination of Flucloxacillin and  Ceftriaxone in bulk and  pharmaceutical formulations. Flucloxacillin  is a  broad spectrum beta -lactam antibiotic, belonging to the isoxazolyl family of penicillin’s. Ceftriaxone is a semi synthetic, broad-spectrum and third generation  cephalosporin antibiotic for intravenous or intramuscular administration. The solvents used in this method is chloroform ,double distilled water (1:1) in presence of phosphate buffer of pH 7.4. Linearity test solutions for the assay method were prepared in the range of 2-12 μg/ mL and  4-16 μg/mL respectively for determination of flucloxacillin and ceftriaxone which obeyed Beer’s law and found to linear. From the experimental results reveal that absorption maximum of 270 nm for flucloxacillin and 290 nm for ceftriaxone were found. Validation of the developed method for two drugs was carried out as per ICH requirements.

A stability indicating RP-HPLC method was developed for the simultaneous determination of Spironolactone (SRL) and Hydroflumethiazide (HFM) in pharmaceutical dosage form. Inertsil ODS - C18 (250 mm x 4.6 mm, 5 µm) column and mobile phase of methanol: acetonitrile : phosphate buffer in the ratio of 55:40:05 v/v at a flow rate of 1.0 mL/min was used for separation of the components. The components were detected at a wavelength of 221nm using UV detector. The Spironolactone and Hydroflumethiazide were separated at retention time 4.67 and 6.74 min respectively. The developed method was validated in terms of precision, accuracy, linearity, specificity, limit of detection, limit of quantitation. The range of linearity was found to be 5-30 µg/mL for Hydroflumethiazide and 5-30 µg/mL for Spironolactone. The proposed method was applied to study the stability of the drugs under different degradation conditions such as acid, alkali, peroxide, thermal and photo light. The developed method was found to be simple, sensitive and rapid and hence, It can be adopted in any laboratory for quality control analysis.

The present work describes a new simple, sensitive and precise reverse phase high performance liquid chromatographic method (RP-HPLC) for the simultaneous estimation of Losartan potassium (LP) and Perindopril erbumine (PE) in bulk and in pharmaceutical dosage forms. Chromatographic separation was performed on KNAUER High Performance Liquid Chromatographic System with C18 column of Make: Thermo Hypersil – ODS of dimensions 250 x 4.6mmwith a mobile phase comprising of 0.01M potassium phosphate buffer (pH 3.5): Acetonitrile: Methanol in the ratio of 5:55:40v/v. the pH of buffer was adjusted with ortho phosphoric acid. The flow rate was 1.0 ml/min with detection with detection at 210nm.As per International Conference on Harmonization (ICH) guidelines the method was validated for linearity, precision, limit of quantitation, limit if detection and robustness. Linearity of LP was found to be in the range of 350µg/ml-650µg/ml and 28µg/ml-52µg/ml for PE. The correlation coefficient for LP and PE was found to be 0.997 and 0.998 respectively. The mean recoveries obtained for LP and PE was found to be 100.222% and 99.844% respectively. The developed analytical method was found to be accurate, linear, specific, and precise which is evident from the statistical data.

Present  study  describes  the  spectrophotometric  method  development  and  subsequent  validation  of  dolutegravir  with  greater  precision  and  accuracy.  AIDS  is  the  most  dreadful  disease  in  the  society;  this  has  a  very  high  mortal  rate  among  the  countries,  as  results  loosing  many  beneficial  personalities.  There  are  many  antiviral  and  antiretroviral  drugs,  brought  forward  by  many  efficient  scientists. A  simple,  accurate,  novel,  safe,  and  precise  method  could  be  developed  for  the  estimation  of  Dolutegravir.  Spectrophotometric  measurements  were  carried  out  using  Schimadzu    double  beam(UV-1800  model)  Ultra  violet  visible  spectrophotometer  with  10mm  matched  quartz  cells  and  water  as  solvent. Linearity  for  the  method  was  found  in  the  range  of  2-14μg/ml  (r2=0.997).  Tablet  formulation  was  analyzed  and  %  assay  for  the  absorption  maxima  was  found  to  be  95.6%. The proposed method was validated as  per  ICH  guidelines.  Validated  studies  demonstrated  that  proposed  method  is  simple,  accurate,  precise,  specific,  rapid,  reliable,  and  reproducible.

A new, rapid, precise, selective and sensitive validative forced degradative UV spectroscopy method was developed for estimation of Montelukast sodium in bulk and pharmaceutical formulation in the developed method, the absorbance was measured at  272.19nm.the drugs obeyed the Beer’s law in the  concentration range of 30-150μg/mL. Accuracy studies of the method were determined by recovery studies and were found to be 99.9 for Montelukast sodium. It can be used for routine analysis of drug in bulk and in pharmaceutical formulation.

A simple, rapid reverse phase high performance liquid chromatographic method (RP- HPLC) has been developed and validated for simultaneous estimation of Aceclofenac and Pregabalin in tablet dosage form. Chromatographic separation was achieved C-18 (250 mm × 4.6 mm, 5.0μ) as stationary phase and mobile phase containing phosphate (pH adjusted to 5.0 ± 0.05 using NaOH.) Buffer: Acetonitrile (30:70 v/v) at flow rate of 1 ml/min using UV detection at 210 nm. The retention time for Aceclofenac and Pregabalin was found to be 3.177 and 5.530 min respectively. The method was validated as per International Conference on Harmonization guideline and successfully used for the quantitative analysis of commercially available tablet. The calibration curve was linear over the concentration range of 20-60μg/ml for Aceclofenac and 15-45μg/ml for Pregabalin.. Lower values of Limit of Detection (0.60μg/ml for Aceclofenac and 0.88μg/ml for Pregabalin) and Limit of Quantification (1.84μg/ml for Aceclofenac and 2.68μg/ml for Pregabalin ) indicated good sensitivity of the method. The method was validate with respect to linearity, robustness, precision and accuracy and was successfully applied for the simultaneous determination of aceclofenac and pregabalin from the combined dosage formulation. The percent amount for both the drugs were found to be within limits in the tablet dosage form for both the methods.

A simple UV-Visible spectrophotometric method is developed for the simultaneous determination of Ebastine and Phenylephrine hydrochloride in pharmaceutical dosage form using  the  first order derivative spectrophotometric method. The determination of both the drugs is based on the respective zero crossing point (ZCP) of their first order derivative spectra obtained in methanol. The first order derivative spectra were obtained using methanol as a solvent and the determinations were made at 241.0 nm (ZCP of Phenylephrine HCl) for Ebastine and 232.0 nm (ZCP of Ebastine) for Phenylephrine hydrochloride. The linearity was obtained in the concentration range of 4-24 μg/ml for both drugs and correlation coefficient (r2) were found to be 0.9994 and 0.9991 for Ebastine and Phenylephrine hydrochloride respectively. The percentage purity of drugs in combined tablet dosage form was found to be 100.02 % for Ebastine and 99.89 % for Phenylephrine hydrochloride. The % recoveries were found to be 99.88% for Ebastine and 99.24% for Phenylephrine hydrochloride.  The method was found to be simple, accurate and precise and was applicable for the simultaneous determination of Ebastine and Phenylephrine in tablet dosage form.

A simple, precise, economical and accurate difference spectroscopic method has been developed for the valacyclovir in bulk and in pharmaceutical dosage form. The proposed method is based on the principle that valacyclovir exhibits two different chemical forms that differs in the absorption spectra in acidic and basic solution. The absorptions were measured in acidic and basic solution separately against reagent blank. Valacyclovir has exhibited maximum absorbance at 252 and 262nm in acidic and basic solution respectively. Difference in absorbance between these two maxima was calculated to find out the amplitude. The amplitude plotted against concentration showed linear response in the concentration range of 0.5 – 2.5μg/mL with linear regression value 0.999. The proposed method was applied to pharmaceutical formulation and the common excipient present in the formulation does not interfere in the analysis of the drug. The method was validated as per ICH guidelines and statistical results of analysis were found to be satisfactory.

A simple, precise and economical UV Spectrophotometric method has been developed for the estimation of Tolterodine tartarate in bulk and pharmaceutical dosage form. The method is based on measurement of absorption at maximum wavelength of 283.0 nm. Linearity for detector response was observed in the concentration range of 10-50 μg/ml. The accuracy of the method was assessed by recovery studies and was found to be 99.80%. The LOD and LOQ were found to be 0.1865 and 0.5621 respectively. The developed method was validated with respect to linearity, accuracy (recovery), precision, specificity and robustness and ruggedness. The results were validated statistically as per ICH Q2 R1 guideline and were found to be satisfactory. The proposed method was successfully applied for the determination of Tolterodine tartarate in commercial pharmaceutical dosage form.