RP-HPLC

An accurate and precise stability-indicating RP-HPLC method has been developed and validated for the analysis of Erlotinib HCl in tablet dosage forms. The method was developed on a Qualisil Gold C18 (250×4.6mmi.d, 5μm particle size) analytical column using methanol: water in the ratio of 58:42(v/v) as the mobile phase. The instrumental settings were flow rate 1mL/min, column temperature 23oC and the detection wavelength of 246nm using a Photo Diode Array (PDA) detector. The method is linear over a range of 5-50μg/ml. The LOD and LOQ values were found to be 0.81μg/mL and 2.466μg/mL respectively. Erlotinib was exposed to acidic, alkaline, oxidative, photolytic, thermal stress conditions and these stressed samples were analyzed by the proposed method. The drug product was completely separated from the degradants which indicates that the method is specific. Key-words: ErlotinibHCl, RP-HPLC, Stress degradation studies.

A new reverse phase high performance liquid chromatography (RP-HPLC) method for the quantitative determination of Sitagliptin in human plasma was developed and validated as per US-FDA guidelines. The drug was spiked in the plasma and extracted with mobile phase by precipitation method. The extracted analyte was injected into an INTERSIL C18 column (150 mm × 4.6 mm, 5μm), maintained at ambient temperature and effluent was monitored at 267 nm. The mobile phase consisting of acetonitrile: methanol: buffer (2:3:5 v/v). The pH of the mobile phase was adjusted to 4.0 by using O-phosphoric acid. The flow rate was maintained at 1.0 mL/min. The developed method shows high specificity for sitagliptin. Calibration curve was plotted with a range from 25-125µg/mL (r2>0.9994). The lower limit of quantification (LLOQ) was found to be 25μg/mL. The method was validated for parameters like accuracy, precision, recovery, linearity, range, stability and sensitivity. This RP-HPLC method is suitable for determining the concentration of sitagliptin in human plasma and it was applied to routine analysis for determination of the Sitagliptin from dosage form during pharmacokinetic study.

The proposed method is a simple, accurate, precise, specific and rapid method for the simultaneous estimation of dextromethorphan hydrobromide (DXM) and chlorpheniramine maleate (CPM) in bulk and syrup formulation. Stationary phase consist of Eclipse-XDB C18 column(150×4.6mm, 5μm) and mobile phase with gradient mode consisting of phosphate buffer (adjusted to pH 3.0 with o-phosphoric acid): acetonitrile (80:20 v/v) was used. The flow rate was set at 1.0 ml/min and UV detection was carried out at 272 nm. The retention time of DXM and CPM were 9.05 min and 7.53 min respectively. The % recovery of DXM and CPM was found to be 99.58 ±1.33 and 98.24 ±1.97 respectively. DXM and CPM drugs were found to be linear over the concentration range of 2-50 µg/ml and 0.8 - 20 µg/ml respectively. The proposed method can be useful in the quality control of DXM and CPM in bulk drug and drug products. 

A simple reverse phase liquid chromatographic method has been developed and subsequently validated for simultaneous determination of Ofloxacin and Ornidazole in infusion dosage form. The separation was carried out using a mobile phase containing methanol and buffer (equal proportion of 0.01M orthophosphoric acid and 0.01M sodium phosphate monobasic dihydrate) with pH 4.00 adjusted by 20% of triethylamine in the ratio of 60:40 v/v. The column used was HiQ Sil C18 (150 mm x 4.6 mm i.d, 5 µ) with flow rate of 1 mL / min using UV detection at 300 nm. The described method was linear over a concentration range of 1.25-10 µg/mL (r2>0.9991) for Ofloxacin and 3.12-25 µg/mL (r2>0.9992) for Ornidazole. Separation was achieved within 5 min. The mean % recovery was found to be 99.94% for Ofloxacin and 100.27 % for Ornidazole. The limit of detection (LOD) for Ofloxacin and Ornidazole were found to be 0.146 and 0.25 µg/mL respectively. Whereas the limit of quantification (LOQ) for Ofloxacin and Ornidazole was 0.44 and 0.77 µg/mL respectively. The results of the study showed that the proposed RP-HPLC method is simple, rapid, precise, accurate and cost effective which is useful for the routine determination of Ofloxacin and Ornidazole in bulk drug and in its infusion.

  An accurate, precise and reproducible Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method was developed and validated for the estimation of Metformin Hydrochloride, Glimepiride and Atorvastatin in Pharmaceutical dosage forms. In this method Qualisil gold C18 column (250mmx4.6mm I.D., 5µm particle size) with mobile phase containing 0.1% TFA in Water (pH adjusted to 2.92 with ammonia) and Methanol in the ratio of 28: 72v/v was used. The flow rate was 1ml/min. and the detection wavelength was 235nm1-2. The linearity was observed in the range of 50 - 225µg/ml, 0.2 - 0.9µg/ml and 1 – 4.5 µg/ml for Metformin Hydrochloride, Glimepiride and Atorvastatin with correlation coefficient of 0.9992, 0.9992, and 0.9997 respectively. Retention times were 3.1min, 7.8min, and 10.1min for Metformin Hydrochloride, Atorvastatin and Glimepiride respectively. The proposed method was validated for Linearity, accuracy, precision and Robustness. The proposed method was validated as per ICH guidelines and can be applied for routine quality control analysis of pharmaceutical dosage forms used for Multidrug therapy containing Metformin Hydrochloride, Glimepiride and Atorvastatin. Key words: RP-HPLC, OPA, Multidrug therapy, ICH, Validation.

An accurate, precise and economic reversed phase high performance liquid chromatographic (RP-HPLC) method was developed and validated for the estimation of lamivudine, zidovudine and nevirapine in pharmaceutical dosage forms. In this method Qualisil BDS C8 column (250mmx4.6mm i.d., 5µm particle size) with mobile phase containing water and acetonitrile in the ratio of 70: 30 v/v with pH adjusted to 5 with ortho phosphoric acid (OPA). The flow rate was 1mL/min and the detection wavelength was 250nm. The linearity was observed in the range of 1-15µg/mL for lamivudine, 3-24 µg/mL for zidovudine and 2.5-20 µg/mL for nevirapine. Retention times were 3.1min, 4.4min, and 7.0min for lamivudine, zidovudine and nevirapine respectively. The proposed method was validated as per ICH guidelines for linearity, accuracy, precision and robustness and can be applied for routine quality control analysis of pharmaceutical dosage forms used for multidrug therapy containing lamivudine, zidovudine and nevirapine.

  An isocratic reverse phase liquid chromatography (RP-HPLC) method has been developed and subsequently validated for the determination of Dasatinib in Bulk and its pharmaceutical formulation. Separation was achieved with a Cosmicsil BDS C18 ((Make: Nomura chemicals (Japan); 150 x 4.6mm I.D; particle size 5 μm)) Column and Triethlyamine buffer (pH adjusted to 6.5 ± 0.05 with diluted orthophosphoric acid): Maethanol and Acetonitrile (50:50) v/v as eluent at flow rate 1.0 mL/min and the Column temperature was 35°C. UV detection was performed at 315 nm and sample temperature was maintained at 5°C. The method is simple, rapid, and selective. The described method of Dasatinib is linear over a range of 3.821 μg/mL to 57.314 μg/mL. The method precision for the determination of assay was below 2.0% RSD. The percentage recoveries of active pharmaceutical ingredient (API) from dosage forms ranged from 98.5 to 99.8 %. The method enables accurate, precise, and rapid analysis of Dasatinib. It can be conveniently adopted for routine quality control analysis of Bulk and pharmaceutical formulations.

  The present work deals with development and validation for simultaneous determination of antihistaminic drugs in pharmaceutical formulations. A rapid, precise and specific high performance liquid chromatography (RP-HPLC) method was developed for Levocetirizine dihydrochloride and Phenylephrine. Chromatographic separations was achieved on Waters Younglin system C-18 (5μm, 250×4.6 mm) HPLC column within a short runtime of 10 min. HPLC system having isocratic mode, with mobile phase containing methanol : water (pH 3) (70:30% v/v) and flow rate maintained at 1.0 mL/min was used. Effluents were monitored at 230 nm. Retention time of Levocetirizine dihydrochloride and Phenylephrine were found to be 2.6 and 4.6 min respectively. Linearity was studied in the concentration range of 2 to 12 μg/mL and 12 to 72 μg/ mL for Levocetirizine dihydrochloride and Phenylephrine respectively, with a correlation coefficient of 0.998 and 0.999 respectively. The proposed method was validated according to the ICH guidelines with respect to specificity, linearity, accuracy, precision and robustness.

Present work describes a selective, precise and accurate Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for simultaneous estimation of Escitalopram Oxalate (ESC) and Etizolam (ETI) on Kromasil 100 C18, 5µ(150×4.6 mm) Column using Acetonitrile:0.005 M Hexane Sulfonic Acid pH 3.0 (adjusted with o-phosphoric acid) (40:60 v/v) as mobile phase, at a flow rate of 1.0 ml/min and the detection wavelength was 254 nm. The retention time for ESC and ETI was found to be 3.66 and 8.07 min, respectively. The ion-pairing reagent improved the retention of polar ESC on Reverse-phase column. The method was validated for linearity, precision, accuracy, LOD, LOQ robustness, solution stability and specificity. The method was linear in the concentration range of 20-160µg/ml for ESC and 2-16 µg/ml for ETI with a correlation coefficient of 0.9994 and 0.9993 for respective drugs. The percent recovery was found in the range of 98.14-101.72% and 98.83-101.12 % for ESC and ETI, respectively. The specificity of method was established based on peak purity data. The proposed method was successfully applied for quantitative determination of escitalopram oxalate and Etizolam in combined tablet dosage form for routine analysis.

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