RP-HPLC

Enrofloxacin (EFX) is a third generation Fluoroquinolone with a broad spectrum antibacterial activity. Enrofloxacin hydrochloride is 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-4-oxo-1, 4-dihydroquinoline-3-carboxylic acid. A Sensitive, simple and rapid reverse phase high performance liquid chromatographic method was developed for the determination of Enrofloxacin (EFX) in tablet dosage form. The chromatographic separation was performed on a Kromasil C-18 column (250mm x 4.6 mm x 5µ) in isocratic mode using phosphate buffer pH 3:Methanol (40:60 v/v), pH adjusted to 3.0 using orthophosphoric acid as mobile phase at a flow rate of 1.0 ml/min with column temperature 30 OC. The quantification was performed at 280 nm. The method showed good linearity over the concentration range of 5-25 µg/ml with correlation coefficient r2 0.9996. LOD and LOQ was found to be 1.0 and 3.0. The developed RP HPLC method was applied to EFX in tablet dosage form and results were found to be in agreement with the label claim.

A simple, specific, accurate, precise and sensitive RP- HPLC method has been developed for the rapid estimation of Alogliptin in bulk and its formulations. The chromatographic separation was carried on Phenomenex Gemini-NX-5 µm C18(2) 110A, LC Column 250 x 4.6 mm, using Acetonitrile:1-octasulphonoic acid (0.005mM) at pH-5 [60:40] (v/v) as mobile phase, at a flow rate of 1.0 ml/min.  The detection was carried out at 220 nm and drug eluted with a retention time of 3.48 min. Beer’s law was obeyed in the concentration range of 2-10?g/ml with correlation coefficient 0.9995. 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 Alogliptin in bulk and tablets dosage form. Thus it can be employed for the routine analysis.

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.

A rapid and sensitive stability indicating RP-HPLC method was developed for simultaneous estimation of salbutamol, carbocisteine and theophylline in combined tablet formulations. Chromatography was carried out on a Discovery HS C18 HPLC Column at 35 °C (250 x 4.6 mm; 5m) by eluting with a mobile phase consisting of a 50:50 v/v mixture of acetonitrile and 0.1 % orthophosphoric acid in water at a flow rate of 1.0 mL/ min. The detection wavelength was set at 215 nm. Accuracy was assessed by using standard addition method. The developed HPLC method was validated with respect to precision, specificity, accuracy, linearity and robustness. Forced degradation studies on the formulation were conducted by adopting the proposed method to assess the stability of the analytes under acid, base, peroxide, thermal and photolytic conditions and suitability of the method to resolve the degradation products.

The objective of this work was to develop a simple, sensitive, accurate, precise and reproducible high performance liquid chromatography (HPLC) method for the determination of salicylic acid in pharmaceutical dosage forms. Shimadzo Prominance model L20 AD HPLC system equipped with SPD 20A UV-Vis detector was used for the analysis. The separation was done on RESTEX allure C18 column (3 μm, 15 cm × 4.6 mm), for an isocratic elution a mixture of water, methanol and glacial acetic acid (65:35:1, v/v) mobile phase at a wavelength of 254 nm. The flow rate was 1.0mL/min. The RP-HPLC method developed for analysis of salicylic acid was validated with respect to specificity, selectivity, linearity, accuracy, precision and robustness as per the ICH guidelines. The retention time of salicylic acid was 7.575 min. The linearity was established over the concentration ranges of 50-350 μg/mL with correlation coefficients ( r2) 0.999.  The percentage accuracy of salicylic acid ranged from 99.76 -101.66%. The relative standard deviation values for intra-day and inter-day precision was lower than 2.0% and the assay result was found to be in the range 99.57-101.32%. Salicylic acid was subjected to stress conditions such as neutral, acidic, alkaline, oxidation and photolysis degradations as per ICH guidelines. The degradation studies revealed that the drug was found to degrade maximum (1.67%) in alkaline degradation conditions and was highly resistant towards neutral, acidic, oxidative and photolytic degradation conditions.

A rapid, simple, selective, and specific reverse phase high performance liquid chromatography (RP-HPLC) method was developed and validated for estimation of Naproxen and Sumatriptan from tablet using spiked human plasma. The chromatographic separation was performed on Phenomenex Luna C18 column (5μm, 25cmx4.6mm id)  with a mobile phase comprised of Acetonitrile: Methanol: phosphate buffer pH 6 (50:10:40 v/v), at a flow rate of 1.0ml/min. The calibration curve was linear in the range of 1-3 µg/ml. The developed method was found to accurate and sensitive. Results of recovery studies prove the extraction efficiency. Stability data indicated that Sumatriptan and Naproxen was stable in plasma after three freeze thaw cycles and upon storage at −20°C for 30 days.

A simple, specific, quick, isocratic Reversed Phase High Performance Liquid Chromatographic method was developed and validated for the analysis of Noscapine. RP-HPLC method was developed on a Symmetry C-8 (4.6 × 150 mm), 3.5 µm particle, reversed-phase column. The mobile phase was 0.1% octane sulphonic acid (pH- 3): acetonitrile, 40:60 (v/v) at a flow rate of 0.8 ml/min. and the eluate was monitored at 260 nm. The retention time of the drug was found to be 2.314 min. The method was linear over the range of 4-8 μg/ml with a regression coefficient of 0.999 and validated with respect to accuracy, precision, linearity, and specificity, limit of detection and limit of quantization as per the guidelines of International Conference for Harmonization (ICH). This method can be used in the industries for determination of Noscapine to analyze the quality of formulation without interference of the excipients.

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.

This present study was undertaken with an objective to develop & validate a simple, precise, cost effective, sensitive & fast RP- HPLC method for the analysis of Ranolazine. A Shimazu HPLC system with Luna 5µm C18 is employed for the analysis using Methanol: Acetonitrile(50:50, v/v) as mobile phase. Signal from Ranolazine is detected at 227nm by UV Spectrophotometer. The total retention time was 5 min with a flow rate of 1.0 ml/min. % 0f RSD values for precision is found to be 0.798%.The limits of detection (LOD) and quantification (LOQ) were 0.616 and 1.86, respectively. As per ICH guidelines the proposed method is fully validated and found to be linear over a workable drug concentration, accurate, precise and robust. This fast, simple and inexpensive method is suitable for research laboratories & also for quality control analysis in pharmaceutical industries.