validation

Fluindione is an oral anticoagulant drug. A simple and rapid and was validated stability indicating HPLC method for Fluindione was successfully developed and was validated. This method is based on HPLC separation followed by UV detection at 285 nm. HPLC method was developed on a Symmetry thermo C18 (4.6 x 250 mm) column with a mobile phase consisting of 10mM di-sodium hydrogen phosphate buffer pH 3.5: methanol 15:85 v/v, pumped at 1.0 ml min-1 flow rate. The pH of buffer was adjusted to 3.5 with ortho phosphoric acid. The column was maintained at ambient temperature and 20μl of solutions were injected. The eluted compound was detected by using PDA detector. Fluindione was eluted at 2.4 ± 0.2 min. Stress degradation study shows that sample degraded with acid and base hydrolysis, under oxidation, thermal and photolytic stress conditions. The method was validated in accordance with requirement of ICH guidelines.

A simple and sensitive high performance thin layer chromatography (HPTLC) method has been developed for the quantitative estimation of Donepezil HCL in its bulk and tablet dose tablet formulation (5 mg). Donepezil HCL was chromatographed on silica gel 60 F254 TLC plate using  methanol : toluene (2:8, v/v) as mobile phase. Donepezil HCL showed Rf value 0.54 + 0.008 and scanned at 245 nm using a camag TLC scanner 3. The method was validated in terms of linearity (100 – 800 ng/spot), precision (system precision = 0.0123 and method precision = 0.0084), accuracy (100.3 ± 0.76) and specificity. The limit of detection and limit of quantification for Donepezil HCL were found to be 1.72 ng/spot and 2.07 ng/spot, respectively. The developed method was successfully used for the assay of Donepezil HCL tablet formulation. This method also contain forced degradation studies for standard and tablet. The method was found to be simple, sensitive, specific, accurate and precise and can be used for the routine quality control testing of Donepezil HCL in tablet dosage form.

A simple, specific, accurate and precise reverse phase high performance liquid chromatographic (RP-HPLC) method was developed with high sensitivity for determination of Valsartan and Hydrochlorothiazide drugs in combined dosage forms. The separation was achieved on Zorbax CN (25 cm x 4.6 mm, 5 µm) at flow rate of 1.8 ml/min with 70: 30 mixture of phosphate buffer: acetonitrile (pH 6) as the mobile phase. The quantification was achieved with PDA detector at 265 nm. The injection volume was 20 μl. The retention times of Valsartan and Hydrochlorothiazide were 6.04 min and 2.27 min, respectively. The method was validated for linearity, precision, specificity, robustness and recovery according to the ICH guidelines. The linearity of response for Valsartan and Hydrochlorothiazide were assessed by analysis of five independent levels of calibration curve in range of 20-100 µg/ml and 2-10 µg/ml respectively. The recovery data was between 98.77-99.73% and 98.75-100.56% for Valsartan and Hydrochlorothiazide respectively. The limit of detection and quantification for Valsartan were 0.98 and 2.97μg/ml respectively and for Hydrochlorothiazide were 0.18 and 0.57μg/ml, respectively. The method was found to be simple and highly sensitive and can be useful in the routine quality control of Valsartan and Hydrochlorothiazide in bulk manufacturing and pharmaceutical dosage forms.

A novel reverse phase Ultra performance liquid chromatographic technique was developed for the determination of balofloxacin in bulk and pharmaceutical dosage forms. The method was developed using waters Acquity BEH 50mm, 2.1mm, 2μm, C 18 column with mobile phase containing a gradient mixture of 0.1% phosphoric acid and acetonitrile. Detection was carried out at wavelength 295 nm. The retention time of balofloxacin was 0.89 min. The method showed good linearity in the range 0.5, 1, 1.5,2,3 µg/ml with correlation coefficient for balofloxacin. The proposed method has been validated as per ICH guidelines and successfully applied to the estimation of balofloxacin in their tablet dosage form.

A simple, accurate, rapid and precise isocratic High performance liquid chromatographic (HPLC) method was developed and validated for the quantification of betaine in Achyranthes aspera. The method employs Reversed Phase Phenomenex (US) C18 Column 250 x 4.6mm I.D, 5µ Particle Size system consisted of prominence LC-20AT gradient pumps, Prominence SIL- 20A Auto sampler and a Prominence SPD-M20A Diode Array Detector (SHIMADZU), Japan and flow rate of 1.0 ml/min with a load of 20μl. Water and acetonitrile was used as mobile phase in the composition of 10:90. The detection was carried out at 242 nm. Linearity range was 0.32-0.48 mg/ml. Retention time of betaine was found to 8.42 min. The recovery studies 98-103%. This method was validated for accuracy, precision, linearity and Robustness as per ICH guidelines. The method was found to be specific, selective, and robust.

A Simple, selective, accurate, precise and linear RP-HPLC method was developed and subsequently validated for estimation of cinacalcet in bulk & tablet dosage form. Gradient elution at a flow rate of 0.8ml/min was used for separation of drugs in reversed-phase mode using Waters HPLC 22695 model on a INERTSIL ODS C18 column (150 x 4.6 mm; 5µ) at a ambient temperature. Mobile phase consisted of water: methanol: acetonitrile (20:60:20). The UV detection wavelength was 235nm 20 µl was injected. The retention time for cinacalcet was 3.7min. The percentage RSD for precision and accuracy of the method was found to be less than 2%. The % recovery was within the range between 99.73% and 99.85%. The method was validated as per the ICH guidelines. Key words: cinacalcet, RP-HPLC, validation  

Iloperidone is a novel antipsychotic drug widely used to treat schizophrenia. Objective of this investigation was to develop a validated stability indicating high performance thin layer chromatographic method for the quantification of iloperidone in bulk and pharmaceutical dosage form. Aluminium backed TLC plates precoated with silica gel 60F-254 was employed as the stationary phase and n-propanol: chloroform (5:5 v/v) as the mobile phase. Densitometric analysis was performed at 275 nm in the reflectance mode. Compact spots of iloperidone with Rf value 0.36 were observed. Validation of the method as per ICH guidelines produced satisfactory results of linearity (r2>0.999), limit of detection (5.349 ng/spot), limit of quantification (16.2099 ng/spot), precision (< 2%), and accuracy (99.66 ±0.341 to 100.34±0.292%). Degradation products were found to be well separated from the pure drug with significantly different Rf values suggesting a stability indicating analysis method for the estimation of iloperidone in pharmaceutical preparations and as bulk drug. The proposed method is selective, sensitive, precise, and accurate. It is also simple, economic and time saving as compared to reported HPLC methods. 

  A rapid and sensitive UV-Visible spectroscopic method was developed for the estimation of cefuroxime in pure and its Pharmaceutical formulations. The method was based on the measurement of absorbance of Cefuroxime active moiety of Cefuroxime tablet at 277 nm using methanol as solvent. The absorbance was found to increase linearly with increase in concentration of Cefuroxime which was corroborated by correlation coefficient values. The standard solution of Cefuroxime obeyed Beer’s law over the concentration range of 9.20–27.60 µg/mL. The method is linear (from 9.20-27.60 µg/mL) with an R2 of 0.999, accurate (% recovery 100.56%) and precise (% RSD 0.316%). The method is specific and robust for Cefuroxime. Key words: Cefuroxime, Spectrophotometric method, validation, accuracy

A novel, precise and selective high performance liquid chromatographic method was developed for the estimation of paliperidone using paracetamol as the internal standard. Separation was achieved on a LiChrospher® RP-18 HPLC column (5 μ particle size and 25 cm × 4.6 mm internal diameter) using 10 mM ammonium acetate: methanol in the ratio of 10:90 (v/v) as the mobile phase, at flow rate of 0.7 ml/min and the eluate was monitored at 277 nm. The method was validated in compliance with ICH guidelines. The correlation coefficient of the calibration graph was 0.99946±0.00037 over the concentration range 1 to 5 µg mL-1. The limit of detection and limit of quantification were 0.569 µg mL-1 and 1 µg mL-1, respectively. Overall percentage recovery of paliperidone ranged between 98.92±0.595 to 100.30±0.693. Relative standard deviations for intra- and inter-day precision studies were

  A simple, specific, accurate and precise reverse phase high performance liquid chromatographic method was developed for simultaneous estimation of diazepam and propranolol hydrochloride in pharmaceutical tablet formulation. The separation was achieved on Phenomenex C18 column (250 mm i.d., 4.6 mm, 5 µm particle size) using methanol: acetonitrile: water (50 : 25 : 25, v/v/v, pH adjusted to 2.8 ± 0.05 with ortho- phosphoric acid) as the mobile phase at a flow rate of 1.0 ml min-1. The quantification was achieved with PDA detector at 235 nm. The injection volume was 20 µl. The retention times of diazepam and propranolol hydrochloride were 5.38 ± 0.29 min and 3.80 ± 0.15 min, respectively. The method was validated for linearity, precision, specificity, robustness and recovery according to the ICH guidelines. The linearity was obtained in the concentration range of 0.1-5.0 µg/ml for both drugs with mean recovery of 100.3 ± 0.47 and 100.2 ± 0.78 % for diazepam and propranolol hydrochloride, respectively. The limit of detection and quantification for diazepam were 0.015 and 0.050 µg/ml, respectively and for propranolol hydrochloride were 0.014 and 0.045 µg/ml, respectively. The method was found to be simple and highly sensitive and can be useful in the routine quality control of diazepam and propranolol hydrochloride in bulk manufacturing and pharmaceutical dosage forms.   Key words: Diazepam, propranolol hydrochloride, RP-HPLC, validation, simultaneous, tablet