RP-HPLC

A specific and accurate reverse phase high performance liquid chromatographic method was developed for the simultaneous determination Ebastine (EBA) and Montelukast Sodium (MONTE) in Combined dosage forms. The analysis was carried out using Phenomenex  C18, column (250 mm × 4.6 mm id, 5 µm particle size) with Mobile phase consisting of  Methanol : Phosphate buffer (65:35 v/v) pH 5.0+0.05 was  pumped at a flow rate was 1.0 ml/ min and  Quantification was achieved with photodiode array (PDA) detection at 261 nm over the concentration range of 10 - 50 µg/mL for ebastine & Montelukast Sodium  with mean recovery of 100.32 ± 0.85 % and 100.15 ± 0.94 % for Ebastine and Montelukast Sodium, respectively. These methods were found to be simple, sensitive, accurate, precise and economical and applicable for the simultaneous determination of Ebastine & Montelukast Sodium in combined dosage form.

The article, for the first time, reports High performance liquid chromatography [HPLC] method for estimation of Dapoxetine hydrochloride [DAP] and Tadalafil hydrochloride [TAD]. The analytical method was developed for both the drugs as API and for tablet dosage form. The separation of two drugs was achieved on High quality octa decyl silane [C18 250x 4mm i.d] 5μ column. The mobile phase consists of Acetonitrile and phosphate buffer in the ratio of 45:55. Mobile phase flow rate adjusted at 1 ml/min and pH at 4.2 using ortho phosphoric acid. The detection was carried out at a wavelength 254 nm. Retention time was found 4.46 and 10.11 min respectively for TAD and DAP. The method was validated for system suitability, linearity, accuracy, and precision and solution stability as per ICH guidelines. Linearity was obeyed in the range of 8-48 mg/ml and 24- 144 mg/ml with correlation coefficient of 0.997 and 0.998 for TAD and DAP respectively. Recovery studies were found within prescribed limits that was 98.83 for TAD and 98.93 for DAP respectively. Detection and quantification limit were found to be 0.225mg/ml and 0.682mg/ml for TAD and 0.163 mg/ml and 0.494mg/ml for DAP respectively which expresses higher sensitivity of the method. Assay results were found to be 98.52 % and 98.26 % in generic brand whereas 99.03 % and 98.11 in other brand for TAD and DAP respectively.

The manuscript describes validated Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for the simultaneous estimation of Perindopril Erbumine and Indapamide in combined dosage form. The RP-HPLC separation was achieved on Intersil ODS C18 column (250 mm x 4.6 mm i.d., 5 µm particle size) at 40oC using mobile phase Acetonitrile : Acid Phthalate Buffer (pH 3.0 ± 0.05) (50 :50, v/v) at flow rate 1.0 mL/min. Quantification was achieved with photodiode array (PDA) detection at 240 nm over the concentration range of 10 - 50 µg/mL for Perindopril Erbumine and 3 - 15 µg/mL for Indapamide with mean recovery of 100.09 ± 0.52 % and 100.13 ± 0.09 % for Perindopril Erbumine and Indapamide, respectively. These methods were found to be simple, sensitive, accurate, precise and economical and applicable for the simultaneous determination of Perindopril Erbumine and Indapamide in combined dosage form.

A simple, precise, accurate and stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) method is developed for estimation of Levosalbutamol sulphate and Ipratropium Bromide in bulk and nebulizer dosage form. The method employed, with reverse phase Inertsil® 5μ C18 (250 × 4.0 mm) column in an isocratic mode, with mobile phase of acetonitrile: buffer in the ratio 77:23 (%v/v). The flow rate was 1.3 ml/min and effluent was monitored at 210 nm. Retention time was found to be 3.05 min., and 10.59 min. The method was validated in terms of linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ) etc. in accordance with ICH guidelines. Linear regression analysis data for the calibration plot showed that there was good linear relationship between response and concentration in the range of 25 – 150% of the working concentration (r2 > 0.999) respectively. The LOD and LOQ values for were found to be0.72,0.43, 1.24and 0.97 and μg/ml respectively. No chromatographic interference from placebo and degradants were found. The proposed method was successfully used for estimation of Levosalbutamol sulphate and Ipratropium bromide in bulk and nebulizer dosage forms.

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  

A simple, precise, accurate and stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) method is developed for estimation of Levosalbutamol sulphate and Budesonide in bulk and suspension for inhalation dosage form. The method employed, with reverse phase Inertsil® 5μ C18 (250 × 4.0 mm) column in an isocratic mode, with mobile phase of acetonitrile: buffer in the ratio 40:60 (%v/v). The flow rate was 0.8 ml/min and effluent was monitored at 266 nm. Retention time was found to be 3.16 min., 17.94 min. and 20.90 min. The method was validated in terms of linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ) etc. in accordance with ICH guidelines. Linear regression analysis data for the calibration plot showed that there was good linear relationship between response and concentration in the range of 25 – 150% of the working concentration (r2 > 0.999) respectively. The LOD and LOQ values for were found to be 0.43, 0.72, 0.97 and 1.24 µg/ml respectively. No chromatographic interference from placebo and degradants were found. The proposed method was successfully used for estimation of Levosalbutamol sulphate and Budesonide in bulk and suspension for inhalation dosage forms.

A specific, accurate and precise reverse phase high performance liquid chromatographic method was developed and validated for the estimation of vardenafil in bulk and pharmaceutical formulation. A Hypersil BDS C 8, 5μ column having 250 x 4.6 mm internal diameter in isocratic mode with mobile phase containing buffer and acetonitrile (60:40, v/v) was used. The flow rate was 1.0 ml/min and effluents were monitored at 240.0 nm over the concentration range of 50-250 µg/ml. The retention time obtained for vardenafil was 5.4 min. The method was validated for linearity, accuracy, repeatability, precision, specificity (in terms of acid, alkali, peroxide and thermal degradation), limit of detection, limit of quantification and ruggedness. Chromatographic peak purity data demonstrated specificity of the method to estimate vardenafil in presence of degradation products.  Limit of detection and limit of quantification were found 0.0125 µg/ml and 0.025 µg/ml respectively. Accuracy in terms of recovery of vardenafil from tablet formulation was found to be above 98 % indicating that vardenafil can be estimated from tablet dosage form without interference from the excipients. The validation data proves the potential utility of the proposed method for the quantitative determination of vardenafil in tablet formulation.

A simple, rapid, accurate, precise and reproducible reverse phase high performance liquid chromatographic method has been developed for the estimation of Risperidone and Trihexyphenidyl hydrochloride was determined using reversed-phase liquid chromatography method using ODS Hypersil C18 column (250 mm × 4.6 mm id, 5μm as a stationary Phase and Methanol : Acetonitrile : Acetate Buffer (pH 4.0) (70 : 20 : 10, v/v/v) as a mobile phase pumped at a flow rate of 1.0 ml/min. Quantification was achieved with ultraviolet detection at 214 nm over concentration ranges of 2-20 μg/ml for Risperidone and 1-10 μg/ml for Trihexyphenidyl hydrochloride with mean accuracy 101.02 ± 0.19 and 101.3 ± 0.38 %, for Risperidone & Trihexyphenidyl hydrochloride respectively. The method was successively applied to tablet dosage forms as no chromatographic interferences from the tablet excipients were observed. The method retained its accuracy and precision when the standard addition technique was applied.

A simple, rapid and precise Reverse Phase High Performance Liquid Chromatographic method was developed for simultaneous estimation of Eperisone hydrochloride and Diclofenac sodium in pharmaceutical dosage form by reverse phase Pinnacle DB C-18 column (250 mm, 4.6 mm, and 5 μm). The sample was analyzed using 50mM ammonium acetate buffer containing 0.2% triethylamine (pH-4.0 adjusted with glacial acetic acid): Acetonitrile (40:60, v/v), as a mobile phase at a flow rate of 1.0 ml/min. and detection at 273 nm. The retention time for Eperisone hydrochloride and Diclofenac sodium was found to be 3.07 min and 5.56 min, respectively. The linearity of developed method was achieved in the range of 10-100 μg/ml for Eperisone hydrochloride and 10-100 μg/ml for Diclofenac sodium. The method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantitation, robustness and ruggedness as per ICH guidelines.

A simple, precise and accurate RP-HPLC method was developed and validated for rapid assay of Dosulepin tablet dosage form. Isocratic elution at a flow rate of 1mL/min was employed on a symmetry Chromosil C18 (250x4.6mm, 5µm in particle size) at ambient temperature. The mobile phase consisted Methanol: Acetonitrile: 0.01M Phosphate buffer in the ratio of 55:20:25 (v/v/v). The UV detection wavelength was 230nm and 20 μL sample was injected. The retention time for Dosulepin was 3.46min. The percentage RSD for precision and accuracy of the method was found to be less than 2%. The method was validated as per the ICH guidelines. The method was successfully applied for routine analysis of Dosulepin tablet dosage form and bulk drug.