RP-HPLC

To develop and validate simple, rapid, linear, accurate, precise and economical reverse phase-high-performance liquid chromatography (RP-HPLC) method for of stress degradation study of Teneligliptin. The separation and quantization were achieved on Inertsil C18(250 mm × 4.6ID,5 um ). The mobile phase selected was Methanol: Water (90:10) at a flow rate of 0.8 ml/min and detection of analytes was carried out at 248nm at pH 3. The method exhibited good linearity over the range of 10–50 µg/mL. The drug is freely soluble in organic solvents Methanol. The drug was identified in terms of solubility studies and on the basis of melting point done by capillary tube method. The drug which when subjected to thermal, photolytic, oxidative, and acidic stress degraded into many degradation products. In most of the cases, the degradation rate was seen to be directly proportional to the amount of stress applied. The thermal stress was increased by increasing the incubation temperature, the faster the degradation took place. The values of LOD were found to be  0.956  ug/ml for TNG  and the calculated LOQ values were found to be 0.171 ug/ml.

Teneligliptin hydrobromide hydrate is a new FDA approved drug for treatment of Diabetes Mellitus. Very few methods have been reported for its identified degradation products and their effects on human. A simple, rapid, precise and accurate stability indicating RP-HPLC method was developed and validated for identification of Teneligliptin hydrobromide hydrate and its degradants on Kromacil C18 column using pH 5.5 phosphate buffer and methanol (75:25v/v) as a mobile phase in isocratic mode of elution at a flow rate 1.2 ml/min. The column effluents were monitored by a variable wavelength UV detector at 270 nm. The method was validated as per ICH guidelines. Forced degradation studies of Teneligliptin hydrobromide hydrate were carried out under acidic, basic, neutral, peroxide, photo and thermal conditions. Degradation was observed in basic and neutral stress samples, but not in acid, peroxide, photo and thermal stress samples.

The aim of the research study was the development and validation of a simple, rapid, accurate and precise reversed-phase high performance liquid chromatography (RP-HPLC) stability-indicating method for the determination of aceclofenac in bulk and pharmaceutical dosage forms. The RP-HPLC studies was performed on the instrument Jasco HPLC system with Jasco UV 2010 photo diode array detector, ODS C18 RP-column (Intersile 250 mm × 4.6 mm; i.d. 10 μm), Rheodyne injection syringe with 20µL loop volume and windows based chrompass software was used for separation. The isocratic elution was performed using the mobile phase ratio of methanol: water (65:35 v/v) and UV detection wavelength at 263 nm. The overall run time of the analysis was 20 minutes and the flow rate was 1.0 mL/min. The RP-HPLC method developed for analysis of aceclofenac was validated as per the ICH guidelines with respect to specificity, selectivity, linearity, accuracy, precision and robustness. The linearity for developed method was observed in the concentration range of 5-50 μg/mL with the correlation coefficient (r2) of 0.9992. The percentage accuracy of aceclofenac ranged from 99.40 to 100.79%. The relative standard deviation for inter-day precision was lower than 2.0%. The assay of aceclofenac was determined in tablet dosage form was found to be within limits. Aceclofenac was subjected to stress conditions such as neutral, acidic, alkaline, oxidation, and photolysis degradations as per ICH guidelines. The results of degradation studies revealed that the drug degraded a maximum (32.68%) in acidic conditions followed by alkaline conditions (15.05%). The drug was found to be resistant towards neutral, acidic and photolytic degradation conditions.

A linear, precise, accurate and simple reversed-phase high performance liquid chromatography (RP-HPLC) bioanalytical method was developed and validated for determination of sofosbuvir in human plasma (K2EDTA) using ledipasvir as an internal standard. Sofosbuvir and ledipasvir (ISTD) were separated from plasma using solid phase extraction technique and The chromatographic separation was accomplished by using a Zorbax eclipse XDB C18 Column (250×4.6 mm i.d, 5 μm particle size) with 1% orthophosphoric acid (pH6.4) and Acetonitrile in the ratio of 30:70% v/v, as mobile phase in an isocratic elution mode pumped at a flow rate 0.7 mL/min and the column oven temperature was maintained at 25?C. The wavelength selected for quantitation was 254 nm. Retention times of sofosbuvir and internal standard were found to be 3.8min and 7.4min, respectively. The standard curves were found to be linear in the range of 30.566-2000.381 ng/ml for sofosbuvir in human plasma. This method performed an intra-day and inter-day precision within the range of 2.44–11.14 and 5.01–8.70%, respectively. Additional intra-day and inter-day accuracy was within the range of 87.99–105.93% and 94.97–99.18% respectively. Total percentage mean recovery of sofosbuvir from spiked plasma was found to be 86.54%. All the validated parameters were found to be within the Acceptance limit.

This paper portrays the advancement and approval of a solidness showing fluid chromatographic strategy for measure of desvenlafaxine in unadulterated and in plans utilizing X-Terra RP C18, 250x4.6 mm, 5 µm particle size column with detection at wavelength 226nm. In the present investigation the mobile phase comprising of sodium dihydrogen orthophosphate buffer (pH 4.0) and acetonitrile in the proportion of 60:40%/v with the Flow rate of 1.0ml/min and column temperature at surrounding temperature uncovered the better resolution and affectability for desvenlafaxine. The maintenance time of desvenlafaxine peak was about 3.632min independently. The created approach be accepted according to ICH regulations which incorporate framework appropriateness, particularity, linearity, exactness, accuracy, vigor, toughness, affectability, cutoff of detection and measurement contemplates and the outcomes were incorporated in this paper. Keywords: Desvenlafaxine, stability indicating method, RP-HPLC, ICH regulations.

In the present study a simple isocratic reverse phase HPLC method was developed for the estimation of rilpivirine in pharmaceutical formulation. The separation was carried out using a column of Zorbax Eclipse XDB-C18, 250x4.6mmi.d with 5micron particle size. The mobile phase comprises of 0.03M di potassium hydrogen orthophosphate with pH adjusted to 2.5 using dilute ortho-phosphoric acid (mobile phase solvent-A) and acetonitrile (mobile phase solvent-B) in the ratio of 15: 85 (v/v).The flow rate was 1.0 ml/min and the effluents were monitored at 284 nm. The retention time was 7.19 min. The detector response was linear in the concentration range of 100-300µg/ml. The respective linear regression equation being Y= 28817.742X-14741.2. The limit of detection (LOD) and limit of quantification (LOQ) for rilpivirine were found to be 0.05µg/ml and 0.15 µg/ml respectively. The assay was found to be 99.85%.The method was validated by determining its accuracy, precision and system suitability. The results of the study showed that the proposed RP-HPLC method is simple, rapid, precise and accurate, which is useful for the routine determination of rilpivirine in its pharmaceutical dosage form.

A simple, economic, selective, precise and accurate High-Performance liquid Chromatographic method used for the estimation of Adenosine in bulk drug. The mobile phase used was of Mixture of Acetonitrile and water in the proportion 5:95 respectively. This Mobile phase was allowed to flow at rate of 0.8ml/min. And this was found to give a sharp peak of Adenosine at a retention time of 3.78 min. Analysis of HPLC for Adenosine was carried out at a wavelength of 256 nm. Linear regression analysis data for the Calibration curve showed a good linear relationship, in concentration range of 50-100ppm and regression coefficient 0.991. The linear regression equation was Y=71258× the developed method was employed with a high degree of precision and accuracy for the analysis of adenosine. The inter and intraday variation was less than 2%. The mean recovery of the drug was 99.39%. The proposed method is simple, fast, accurate, and reproducible hence, it can be applied for routine quality control analysis of Adenosine.

A new, simple, accurate, precise and robust isocratic RP-HPLC method has been developed and subsequently validated for the determination of Regorafenib in pure form and pharmaceutical dosage forms as per ICH guidelines. The separation achieved on a Symmetry C18 Column, 250 mm x 4.6 mm i.d. and 5µm particle size column as a stationary phase and Methanol: Phosphate buffer (pH adjusted to 4.80 with phosphoric acid) in the ratio of 70:30v/v used as mobile phase at a flow rate of 1.0 ml/min. The UV detection was performed at 268nm. The retention time for Regorafenib was found to be 3.544minutes. The detector response was linear in the concentration range of 0-16µg/ml. The respective linear regression equation being Y= 58945.x + 9634 with R2 = 0.999. The percentage of Regorafenib in pharmaceutical dosage form was found to be within the limits. The limit of detection and the limit of quantification were found to be 0.90µg/ml and 2.90µg/ml respectively. The results of the study showed that, the proposed RP-HPLC method was simple, rapid, precise, accurate and stability indicating, which can be used for the routine determination of Regorafenib in pure form and pharmaceutical dosage forms.

A new Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method has been developed for estimation of Paracetamol and Alprazolam in bulk and tablet dosage forms using UV-detector. A RP Cell Pack C18 column (250 mm × 4.6 mm, 5 µ particle size) using acetonitrile and water (80:20 % V/V) as mobile phase by maintaining flow rate of 1 mL/min at 236 nm as detection wavelength. The peaks were eluted at 4.8 and 6.2 mins for Paracetamol and Alprazolam, respectively. The method was validated in accordance with ICH guidelines, the linearity curve for Paracetamol was obtained over the range of 50-175 µg/mL, and it was found to be linear with y = 1961x + 9226 (r2 = 0.999). The linearity curve for Alprazolam was obtained over the range of 0.25-1.5 µg/mL and was found to be linear with y =23328x + 939.3 (r2 = 0.998). The percentage recoveries were found to be 99-101% and 99-102%, respectively. The system suitability parameters such as number of theoretical plates and tailing factor were found to be 7242, 1.56 for PAR and 6755, 1.15 for ALP. Hence the developed RP-HPLC method was found to be simple, accurate, economical, rapid and can be applied for routine analysis of these drugs in their combined formulations.

A stable, simple, accurate, precise, robust and highly selective Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method was developed and validated using ritonavir. Chromatographic separation was achieved using cyber labs, LC 100 separation module, Agilent C18 column at temperature 30°C. Flow rate selected was 1ml/min. Both drugs are identified with UV detector at 256nm. Mobile phase employed was Methanol: Water (50:50), which resulted best   sensitivity. Developed method was validated in terms of linearity, range (25-150µg/ml), precession (correlation coefficient is less than 0.999), robustness, accuracy(recovery was 101.96%) and  under stress conditions drug degradation was less than 10%.The validation of proposed stability indicating method was verified by recovery studies and can be applicable in routine pharmaceutical analysis.