RP-HPLC

Vasopressin, nonapeptide, used as an antidiuretic hormone. Very few method has been reported for analysis of Vasopressin from pharmaceutical dosage form. A simple and rapid high performance liquid chromatography (HPLC) method was developed for the quantitative analysis of arginine vasopressin released from polymeric nanoparticles. Chromatographic analysis was performed on an RP C18 column with a mobile phase consisting of acetonitrile and phosphate buffer (13:87 v/v) at a flow rate of 1.6 ml/min at a wavelength of 220 nm,  with a retention time 4.1 min. The method was shown to be specific and linear in the range of 1-50 IU/ml (r2 = 0.9997). Developed method was validated for various evaluation parameters as per ICH guidelines. The method showed no peak interference in presence of formulation excipients. The limit of detection and quantitation were 0.32 and 1.06 IU/ml, respectively. The method was applied to the quantitative analysis of drug to study in vitro drug release from polymeric nanoparticles.

A novel, simple, precise, rapid, reproducible and cost effective RP-HPLC method was developed and validated for the simultaneous estimation of Nadifloxacin and Clobetasol propionate in its pharmaceutical dosage form. The chromatographic separation was carried out using C18 Shim pack XR ODS II (250 mm × 4.6 mm, 5µm) column with mobile phase comprising of Acetonitrile : Water (50:50)(%v/v). Flow rate was maintained 1.0 mL/min and quantitation was carried out using UV detection at 242nm. Retention time of Nadifloxacin and Clobetasol propionate were found to be 2.64 min and 6.19 min respectively. The method was validated by assessing different parameters such as specificity, linearity, precision, accuracy, robustness, LOD and LOQ for the developed method. The linearity range 20-240 µg/mL and 1-12 µg/mL were selected for Nadifloxacin and Clobetasol propionate respectively. The correlation coefficient (r2) for Nadifloxacin and Clobetasol propionate were found to be 0.9995 and 0.9999 respectively. The limit of detection for Nadifloxacin and Clobetasol propionate were found to be 0.029 µg/mL and 0.21 µg/mL. The limit of quantitation for Nadifloxacin and Clobetasol propionate were found to be 0.089 µg/mL and 0.64 µg/ml respectively. Percentage recovery was found to be 99% to 100.56% for Nadifloxacin and 99.37% to 99.79% for Clobetasol propionate. All the validation parameters were with-in the acceptance limit. The relative standard deviation (%RSD) was found to beless than 2% in all the assessed parameters. The developed HPLC method can successfully used for the quantitative estimation of both the drugs in its formulation.

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.

A simple, cheap, fast and accurate Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method was developed and validated for determination of Trifluoperazine Hydrochloride and its degraded products in pharmaceutical dosage form. This method was developed by using an analytical Zodiac C18 Column (250 mmx4.6mm, 5µm) and mobile phase comprises of 70% methanol and 30% acetonitrile. The method was validated and found to be linear, selective, accurate, robust, rugged and precise. The lower Limit of Detection (LOD) and lower Limit of Quantification (LOQ) respectively were 1.50 µg/ml and 5.0 µg/ml.  The methods utilized in this study are the degradation of drug i.e. aqueous, 0.1N HCl, 0.1N NaOH, 3% H2O2, thermal, photolytic and UV light. Developed HPLC method is able to separate all degrading products from any stress condition from drug peak by resolution of more than 2. This developed method can be used for routine analysis for the estimation of Trifluoperazine Hydrochloride in bulk and in tablet dosage form in pharmaceutical industry because it is simple, cheap and accurate.

A new stability indicating RP-HPLC method was described for the assay of olmesartan in market formulations. This developed RP-HPLC method was based on high performance liquid chromatographic (HPLC) separation of olmesartan with the use of a reversed phase HPLC column [Kromasil BDS, C18, 100 x 4.6 mm, 5μ] with mobile phase consisting of .01M KH2PO4 buffer (pH-4.5) and acetonitrile in the ratio of 55:45 %v/v at ambient temperature. The flow rate of the mobile phase was adjusted to 1.0mL/min and the injection volume was 10μL. Detection was performed by photodiode array detector at a wavelength of 257nm and the chromatographic runtime was 8 minutes for the analysis. The reliability and analytical performance of the proposed method, including linearity, range, precision, accuracy, detection and quantitation limits, were statistically validated. The proposed method can be adopted apparently for routine quality control analysis of raw materials, formulations and testing

The article aims at developing simple, fast and effective dissolution method for Dabigatran etexilate mesylate capsules by RP-HPLC and validate as per ICH guidelines. The optimized RP-HPLC method for dissolution studies uses a reverse phase column, Phenomenex Luna C18 (250 X 4.6 mm;5μ), a mobile phase of triethylammonium phosphate buffer (pH 3.0):acetonitrile in the proportion of 40:60 v/v, diluent as 0.01N HCl, flow rate of 1.0ml/min, injection volume as 20µl. and a detection wavelength of 341nm using a UV detector. The optimized dissolution conditions include, 0.01N HCl as dissolution media, apparatus as USP Type 1 Basket, rpm as 100, dissolution media temperature as 37±0.5ºC, dissolution volume as 500ml, dissolution time point as 30mts, working concentration of standard and sample as 5µg/ml and a detection wavelength of 341 nm. The developed method resulted in Dabigatran etexilate exhibiting linearity in the range 1.25-10μg/ml. System precision and intra-day precision is exemplified by relative standard deviation of 1.59% and 2.21% respectively. Method was found to be rugged/inter day precise as %RSD was found to be 3.25. Percentage Mean recovery was found to be greater than 80% at all the three levels by absolute method during accuracy studies. LOD and LOQ for Dabigatran etexilate were found to be 0.05ng/ml and 5ng/ml respectively. Hence it can be concluded that effective dissolution method by RP-HPLC is developed and validated as per ICH guidelines which can be applicable in various pharmaceutical industries.

A simple, precise and accurate stability indicating RP-HPLC method has been developed and subsequently validated for the simultaneous estimation of Sildenafil citrate and Estradiol valerate in bulk and pharmaceutical formulation. The separation was carried out using C18 column (150mm x 4.6mm, 5μm), mixture of acetonitrile and water 80:20 % v/v as a mobile phase with a flow rate of 1 ml/min and the effluent was monitored at 290 nm using PDA detector. The retention time of Sildenafil citrate and Estradiol valerate were 2.55 min and 5.56 min respectively. The method is linear over the range of 125 - 750 μg/ml and 5 - 30 μg/ml for Sildenafil citrate and Estradiol valerate respectively. The method was found to be precise, accurate and specific during the study. The percentage assay was found to be 100.68 % and 99.58 % for Sildenafil citrate and Estradiol valerate respectively from the tablet formulation. Sildenafil citrate and Estradiol valerate were subjected to stress condition to check the degradation behaviour of them. The drugs undergo degradation under acidic, basic, oxidative and thermal condition. The proposed method enables rapid quantification and simultaneous analysis of both drugs from commercial formulations without any interference of excipients. So, the method can be used for routine analysis of Sildenafil citrate and Estradiol valerate in combined tablet formulation.

A simple, accurate, precise and specific RP-HPLC method has been developed for the simultaneous estimation of Aceclofenac and Esomeprazole Sodium in bulk. Chromatographic analysis was carried out on C18 column ( 250mm × 4.6mm, 5µm). Mobile phase used is a homogenous mixture of ACN: Methanol in the ratio of 50:50 v/v. The detection was carried out at 285nm. The retention times were found to be 3.00 and 4.41 min for Aceclofenac and Esomeprazole Na respectively. Both the drugs showed linearity in the range of 30 – 70mcg/ml. The correlation coefficient was found to be 0.99 and 0.992 for Aceclofenac and Esomeprazole Na respectively. The developed method was validated as per ICH guidelines.

A stability indicating reverse phase High performance liquid chromatography (RP-HPLC) method has been developed and subsequently validated for the simultaneous determination of Atazanavir and Cobicistat in bulk and pharmaceutical formulation. Separation was achieved in isocratic mode with a Kinetex C18 100 A (250 mm x 4.6 mm,  5µ) column and mixture consisting of 0.1% OPA(pH 3) and methanol in 80:20 v/v was used as mobile phase with a flow rate of 1 ml/min, column temperature at 25°C and the run time as 10 mins. UV detection was performed at 239 nm and the sample temperature was maintained ambient. The described method for simultaneous determination of Atazanavir and Cobicistat is linear over a range of 8 μg/ml to 120 μg/ml and 5 μg/ml to 60 μg/ml respectively. The method shows good precision results which were below 2.0%RSD. Limit of Detection (LOD) and Limit of Quantification (LOQ) of Atazanavir and Cobicistat was established and found to be 1.49 and 4.97 μg/ml and 1.13 and 3.77 μg/ ml respectively. The developed method was validated according to ICH guidelines for various parameters.The method is simple, rapid, selective and stability indicating method which would be used for regular stability indicating quality control determinations.

A simple, new, precise, accurate and reproducible RP-HPLC method for simultaneous estimation of mefenamic acid, ethamsylate and tranexamic acid in bulk and pharmaceutical formulations. Separation of mefenamic acid, ethamsylate and tranexamic acid was successfully achieved on a Kromasil C8 (250 mm x 4.6mm x 5µ ) in an isocratic mode utilizing Ammonium acetate buffer and methanol (60:40 v/v) at a flow rate of 1.0 mL/min. The method was validated according to ICH guidelines for linearity, sensitivity, accuracy, precision, specificity and robustness. The response was found to be linear in the drug concentration range of 25-75 mg/mL for mefenamic acid ,25-75 mg/mL for ethamsylate 50-150 mg/mL for tranexamic acid. The correlation coefficient was found to be 0.9997 for both the drugs. The limit of detection (LOD) was 0.158,0.2183 and 0.321 for  mefenamic acid, ethamsylate and tranexamic acid respectively. The limit of quantification (LOQ) was 0.527,0.7278 and 1.071 for mefenamic acid, ethamsylate and tranexamic acid respectively. The relative standard deviation (RSD) of six replicates is less than 2%. This HPLC method is applied successfully to the simultaneous quantitative analysis of mefenamic acid, ethamsylate and tranexamic acid in commercial tablets.