This document describes the development and validation of an analytical method for the quantification of flupirtine maleate using reverse phase high-performance liquid chromatography (RP-HPLC). The method was optimized using a C18 column with a mobile phase of methanol and water (90:10) at a flow rate of 1 mL/min. Validation studies established the method's system suitability, precision, linearity, accuracy and robustness per ICH guidelines. The developed and validated RP-HPLC method can be used for the routine analysis of flupirtine maleate in pharmaceutical formulations.
The document describes the development and validation of an UPLC method for the simultaneous estimation of Emtricitabine, Tenofovir Alafenamide, and Bictegravir in bulk and pharmaceutical dosage forms. The method utilizes an Acquity BEH C18 column with a mobile phase of triethylamine buffer (pH 3.0) and methanol at a 45:55 ratio. Emtricitabine, Tenofovir Alafenamide, and Bictegravir were well separated with retention times of 2.6, 4.3, and 5.2 minutes respectively. The method was optimized and further validated as per ICH guidelines to quantify the drugs in bulk and pharmaceutical formulations.
Stability indicating RP-HPLC method for estimation of dapagliflozin in bulk a...SriramNagarajan19
A simple, specific, accurate, precise and stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) method is developed for estimation of Dapagliflozin (DGF) in bulk and Pharmaceutical dosage form. The method employed, Hypersil BDS C18 250 mm x 4.6 mm, 5 mm column in isocratic mode with mobile phase of 0.1% Ortho phosphoric acid buffer and acetonitrile 50:50% v/v. The flow rate was 1.0 mL min-1 and effluent was monitored at 245 nm using PDA detector. The injection volume was 10 µl and the total runtime was set as 5min. The retention time for DGF was found to be 2.226min.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 a good linear relationship between response and concentration in the range of 25 - 150 µg/ml respectively. The LOD and LOQ values for HPLC method were found to be 0.04 and 0.121 µg/ml respectively. No chromatographic interference from the tablet excipients was found. The proposed method was successfully used for estimation of Dapagliflozin (DGF) in Bulk and Pharmaceutical dosage form.
A newly validated HPLC method development for simultaneous estimation of rito...SriramNagarajan19
The aim of the present work was to develop a isocratict RP-HPLC for simultaneous analysis of ritonavir and lopinavir in tablet dosage form. Method: chromatographic system was optimized using a Agilent XDB C18(150 x 4.6mm,5µm) column with potassium dihydrogen phosphate (pH 4.6) and acetonitrile in the ratio of 45;55, as a mobile phase, at a flow rate of 1.0 ml/min. detection was carried out at 215nm by a photodiode array detector. Result: ritonavir and lopinavir were eluted with retention times of 4.821 and 3.814mins respectively. Beer’s lambert’s law was obeyed over the concentration ranges of 12.5 to 50µg/ml and 50 to 200µg/ml for ritonavir and lopinavir, respectively. Conclusion: the high recovery and low coefficients of variation confirm the suitability of the method for simultaneous analysis of both drugs in a tablet dosage form. Statistical analysis proves that the method is sensitive and significant for the analysis of ritonavir and lopinavir in pure and in pharmaceutical dosage form without any interference from the excipients. The method was validated in accordance with ICH guidelines. Validation revealed the method is specific, rapid, accurate, precise, reliable, and reproducible.
Stability indicating analytical method development and validation for estimat...SriramNagarajan18
Stability indicating analytical method development and validation for estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form using RP-HPLC
This document describes the development and validation of a new reverse phase high performance liquid chromatography (RP-HPLC) method for the estimation of paracetamol in pharmaceutical dosage forms. Some key points:
- An isocratic RP-HPLC method was developed using a mobile phase of acetonitrile and potassium dihydrogen orthophosphate buffer at a ratio of 15:85, pH 2.5.
- The method was validated for parameters such as linearity, accuracy, precision, limit of detection, limit of quantification, and robustness as per ICH guidelines.
- The method showed good linearity in the range of 25-60 μg/ml with a correlation coefficient of 0.999
This document describes the development and validation of a reverse phase HPLC method for the simultaneous estimation of metformin and linagliptin in pure form and pharmaceutical formulations. The method utilizes a C18 column, mobile phase of phosphate buffer and acetonitrile (60:40) at a flow rate of 1 mL/min. Metformin and linagliptin were well separated with retention times of 3.048 and 4.457 minutes respectively. The method was validated per ICH guidelines and showed good linearity, accuracy, precision and recovery for both drugs. The method can be used to simultaneously quantify metformin and linagliptin in tablet formulations.
Method Development and Validation for Estimation of Oral Hypoglycaemic Drug D...ijtsrd
HPLC is a chromatographic technique employed in active compound chemistry and biochemistry to separate a mixture and substances with the goal of identifying, measuring, and purifying the different components of the mixture. Its a much better variety of column and traditional chromatography. The objective of the research work is to develop and validate a simple and accurate reverse phase chromatographic method to estimate amount of drug in dosage form. The developed method successfully can be applied to estimate the amount of Dapagliflozin in tablet dosage form. After oral administration of dapagliflozin, the maximum plasma concentration Concentration max under two hours. High performance liquid chromatographic system was alleviated according to the chromatographic settings. After attaining the steady base line, to verify the system suitability, a single 40 µg ml of standard solution proportional to 100 test concentration of dapagliflozin was injected into the HPLC system. The gradient mobile phase flow rate programming assisted in optimising the lengthy run duration and resolution of sample analysis, making the approach more cost effective and quick. Validation of the developed and optimized HPLC method was carried out according to ICH guidelines with respect to parameters such as linearity, specificity, precision and accuracy. Junaid Ahmed | Himanchal Sharma | Shiva Teotia "Method Development and Validation for Estimation of Oral Hypoglycaemic Drug Dapagliflozinina Tablet Dosage form by the Employment of Rp-HPLC" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-6 , October 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46395.pdf Paper URL : https://www.ijtsrd.com/pharmacy/analytical-chemistry/46395/method-development-and-validation-for-estimation-of-oral-hypoglycaemic-drug-dapagliflozinina-tablet-dosage-form-by-the-employment-of-rphplc/junaid-ahmed
The document describes the development and validation of an UPLC method for the simultaneous estimation of Emtricitabine, Tenofovir Alafenamide, and Bictegravir in bulk and pharmaceutical dosage forms. The method utilizes an Acquity BEH C18 column with a mobile phase of triethylamine buffer (pH 3.0) and methanol at a 45:55 ratio. Emtricitabine, Tenofovir Alafenamide, and Bictegravir were well separated with retention times of 2.6, 4.3, and 5.2 minutes respectively. The method was optimized and further validated as per ICH guidelines to quantify the drugs in bulk and pharmaceutical formulations.
Stability indicating RP-HPLC method for estimation of dapagliflozin in bulk a...SriramNagarajan19
A simple, specific, accurate, precise and stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) method is developed for estimation of Dapagliflozin (DGF) in bulk and Pharmaceutical dosage form. The method employed, Hypersil BDS C18 250 mm x 4.6 mm, 5 mm column in isocratic mode with mobile phase of 0.1% Ortho phosphoric acid buffer and acetonitrile 50:50% v/v. The flow rate was 1.0 mL min-1 and effluent was monitored at 245 nm using PDA detector. The injection volume was 10 µl and the total runtime was set as 5min. The retention time for DGF was found to be 2.226min.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 a good linear relationship between response and concentration in the range of 25 - 150 µg/ml respectively. The LOD and LOQ values for HPLC method were found to be 0.04 and 0.121 µg/ml respectively. No chromatographic interference from the tablet excipients was found. The proposed method was successfully used for estimation of Dapagliflozin (DGF) in Bulk and Pharmaceutical dosage form.
A newly validated HPLC method development for simultaneous estimation of rito...SriramNagarajan19
The aim of the present work was to develop a isocratict RP-HPLC for simultaneous analysis of ritonavir and lopinavir in tablet dosage form. Method: chromatographic system was optimized using a Agilent XDB C18(150 x 4.6mm,5µm) column with potassium dihydrogen phosphate (pH 4.6) and acetonitrile in the ratio of 45;55, as a mobile phase, at a flow rate of 1.0 ml/min. detection was carried out at 215nm by a photodiode array detector. Result: ritonavir and lopinavir were eluted with retention times of 4.821 and 3.814mins respectively. Beer’s lambert’s law was obeyed over the concentration ranges of 12.5 to 50µg/ml and 50 to 200µg/ml for ritonavir and lopinavir, respectively. Conclusion: the high recovery and low coefficients of variation confirm the suitability of the method for simultaneous analysis of both drugs in a tablet dosage form. Statistical analysis proves that the method is sensitive and significant for the analysis of ritonavir and lopinavir in pure and in pharmaceutical dosage form without any interference from the excipients. The method was validated in accordance with ICH guidelines. Validation revealed the method is specific, rapid, accurate, precise, reliable, and reproducible.
Stability indicating analytical method development and validation for estimat...SriramNagarajan18
Stability indicating analytical method development and validation for estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form using RP-HPLC
This document describes the development and validation of a new reverse phase high performance liquid chromatography (RP-HPLC) method for the estimation of paracetamol in pharmaceutical dosage forms. Some key points:
- An isocratic RP-HPLC method was developed using a mobile phase of acetonitrile and potassium dihydrogen orthophosphate buffer at a ratio of 15:85, pH 2.5.
- The method was validated for parameters such as linearity, accuracy, precision, limit of detection, limit of quantification, and robustness as per ICH guidelines.
- The method showed good linearity in the range of 25-60 μg/ml with a correlation coefficient of 0.999
This document describes the development and validation of a reverse phase HPLC method for the simultaneous estimation of metformin and linagliptin in pure form and pharmaceutical formulations. The method utilizes a C18 column, mobile phase of phosphate buffer and acetonitrile (60:40) at a flow rate of 1 mL/min. Metformin and linagliptin were well separated with retention times of 3.048 and 4.457 minutes respectively. The method was validated per ICH guidelines and showed good linearity, accuracy, precision and recovery for both drugs. The method can be used to simultaneously quantify metformin and linagliptin in tablet formulations.
Method Development and Validation for Estimation of Oral Hypoglycaemic Drug D...ijtsrd
HPLC is a chromatographic technique employed in active compound chemistry and biochemistry to separate a mixture and substances with the goal of identifying, measuring, and purifying the different components of the mixture. Its a much better variety of column and traditional chromatography. The objective of the research work is to develop and validate a simple and accurate reverse phase chromatographic method to estimate amount of drug in dosage form. The developed method successfully can be applied to estimate the amount of Dapagliflozin in tablet dosage form. After oral administration of dapagliflozin, the maximum plasma concentration Concentration max under two hours. High performance liquid chromatographic system was alleviated according to the chromatographic settings. After attaining the steady base line, to verify the system suitability, a single 40 µg ml of standard solution proportional to 100 test concentration of dapagliflozin was injected into the HPLC system. The gradient mobile phase flow rate programming assisted in optimising the lengthy run duration and resolution of sample analysis, making the approach more cost effective and quick. Validation of the developed and optimized HPLC method was carried out according to ICH guidelines with respect to parameters such as linearity, specificity, precision and accuracy. Junaid Ahmed | Himanchal Sharma | Shiva Teotia "Method Development and Validation for Estimation of Oral Hypoglycaemic Drug Dapagliflozinina Tablet Dosage form by the Employment of Rp-HPLC" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-6 , October 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46395.pdf Paper URL : https://www.ijtsrd.com/pharmacy/analytical-chemistry/46395/method-development-and-validation-for-estimation-of-oral-hypoglycaemic-drug-dapagliflozinina-tablet-dosage-form-by-the-employment-of-rphplc/junaid-ahmed
Analytical Method Development and Validation of Tolvaptan in Bulk and Tablet ...pharmaindexing
This document describes the development and validation of a reverse phase high performance liquid chromatographic (RP-HPLC) method for the estimation of Tolvaptan in pharmaceutical dosage forms. A C18 column was used with a mobile phase of sodium dihydrogen phosphate and acetonitrile. The method was validated per ICH guidelines and showed good linearity, accuracy, precision, specificity, and robustness. Recovery of Tolvaptan from formulations ranged from 99.74-99.87% indicating the method is accurate for quantifying Tolvaptan in tablets.
Stability studies of simvastatin and fenofibrate and degradants identificatio...Mehar Raghavendra YEGGINA
1. A stability indicating RP-HPLC method was developed for the simultaneous quantification of simvastatin and fenofibrate.
2. The method used a C18 column with a mobile phase of acetonitrile and ammonium acetate buffer at pH 4.3 to achieve separation of the drugs from their degradation products.
3. The drugs were subjected to stress conditions and the degraded products were identified using LC-MS to prove the stability indicating capability of the developed method.
This document summarizes the presentation of a research article on the development and validation of an RP-HPLC method for the simultaneous determination of metformin and evogliptin in pharmaceutical formulations. The method was developed using a C18 column with an isocratic mobile phase of methanol and water at pH 3. Both drugs showed good separation and were detected at 254 nm. The method was validated and found to be linear, precise, accurate, robust, and specific for quantifying the drugs in tablets within 3 minutes. The method can be applied for routine quality control analysis of metformin and evogliptin combinations.
Method Development and Validation for Simultaneous Estimation of Dasatinib an...YogeshIJTSRD
The document describes the development and validation of an isocratic reverse phase HPLC method for the simultaneous quantification of Dasatinib and Erlotinib in pharmaceutical formulations. The method was optimized using a response surface methodology with a central composite design to evaluate the effects of varying the methanol percentage, pH, and flow rate on the separation. The optimized conditions provided good resolution of Dasatinib, Erlotinib and the internal standard within 9 minutes. The method was validated as per ICH guidelines and successfully applied to the analysis of commercial tablet and capsule formulations containing the two drugs.
Development and validation of a stability indicating RP-HPLC method for estim...BRNSSPublicationHubI
This document describes the development and validation of a reverse-phase high-performance liquid chromatography (RP-HPLC) method for the quantitative analysis of daclatasvir in pharmaceutical formulations. The method utilizes a C18 column, mobile phase of acetonitrile and 0.1% formic acid buffer at a ratio of 40:60, and detection wavelength of 305 nm. Standard and sample solutions of daclatasvir were prepared and analyzed using the optimized method. The results demonstrate that the developed RP-HPLC method is accurate, precise, sensitive and stability-indicating for the analysis of daclatasvir in pharmaceutical dosage forms.
This document describes the development and validation of an RP-HPLC method for the quantification of flunarizine dihydrochloride. Key steps included selecting the instrument and optimizing conditions such as the column, mobile phase, and detection wavelength. The method was then validated based on parameters like accuracy, precision, specificity, detection limit, quantitation limit, and linearity range according to ICH guidelines. The proposed RP-HPLC method was found to be simple, accurate, precise and cost-effective for the analysis of flunarizine dihydrochloride in drug substances and pharmaceutical formulations.
A novel validated stability Indicating RP-HPLC Method Development for the est...Naveen Chennamaneni
Best reserch paper A novel validated stability Indicating RP-HPLC Method Development for the estimation of Certinib in its bulk and finished Dosage form as per ICH Guidelines
DESIGN OF EXPERIMENTS TO DEVELOP AND VALIDATE NEW ANALYTICAL METHODS FOR QUAN...Pranjali837829
Estimation of Lapatinib, an Anticancer drug by HPLC (High-Performance Liquid Chromatography). This research would help treat women suffering from Breast Cancer.
RP-HPLC Assay Method Validation for the estimation of new Anti-retroviral dru...SriramNagarajan15
A reverse phase HPLC method was developed and validated for the quantification of lamivudine in bulk and tablet formulations. The method used an ODS column with a mobile phase of acetate buffer and acetonitrile (50:50) at a flow rate of 1.5 mL/min. Lamivudine had a retention time of 1.85 minutes when detected at 272 nm. The method was linear over a concentration range of 10-50 μg/mL with a correlation coefficient of 0.999. Accuracy and precision studies demonstrated recoveries between 98-102% and %RSD below 2%, respectively. The method was found to be robust, specific, and suitable for the routine analysis of lamivudine in
The document describes the development and validation of a reverse phase HPLC method for the estimation of the anti-retroviral drug lamivudine in bulk and tablet formulations. An ODS column with a mobile phase of acetate buffer and acetonitrile was used to achieve separation of lamivudine. The method was validated as per ICH guidelines and was found to be linear, precise, accurate and robust. The developed method can be used for the routine analysis of lamivudine in pharmaceutical dosage forms.
The document describes the development and validation of a reverse phase HPLC method for the estimation of the anti-retroviral drug lamivudine in bulk and tablet formulations. An ODS column with a mobile phase of acetate buffer and acetonitrile was used to achieve separation of lamivudine. The method was validated as per ICH guidelines and was found to be linear, precise, accurate and robust. The developed method can be used for the routine analysis of lamivudine in pharmaceutical dosage forms.
RP-HPLC Assay Method Validation for the estimation of new Anti-retroviral dru...SriramNagarajan15
A Reverse phase HPLC method was developed for estimation of the Lamivudine in bulk and tablet formulation by using ODS column (250mm×4.6mm, 5µm) and Acetate buffer: acetonitrile (50:50) as mobile phase, at a flow rate of 1.5ml/min. The detection was carried at the 272nm the retention time of the Lamivudine is 1.850. The developed method was validated for the various parameters as per the ICH guidelines like accuracy precision, linearity and range, Robustnes. Linearity was obtained in the concentration range of 10µg/ml to 50µg/ml with correlation coefficient of 0.999. The accuracy of the method was assessed by recovery studies at three different concentration levels. The percentage recovery of Lamivudine was found to be in the range of 98% -102%. The method was found to be precise as indicated by the repeatability, inter-day, intra-day analysis, showing %RSD less than 2. Key words: RP-HPLC, Lamivudine, Pharmaceutical dosage form.
Development and Validation of Reverse Phase Liquid Chromatography Method for ...IOSR Journals
This document describes the development and validation of a reverse phase liquid chromatography method for the estimation of losartan in bulk drug samples. The method utilizes an Acquity BEH C18 column with a mobile phase of buffer and acetonitrile at a ratio of 50:50 delivered isocratically at 0.3 mL/min. Losartan was detected at 230 nm. The method was validated per ICH guidelines and found to be linear, precise, accurate, specific and stability-indicating for the quantification of losartan in the range of 25-75 μg/mL. The method validation shows the method is suitable for the routine analysis of losartan in bulk drug materials.
Simultaneous estimation of meclizine and nicotinic acid by using RP-HPLCpharmaindexing
This document describes the development and validation of a reverse phase high performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of Meclizine and Nicotinic acid. The method utilizes a mobile phase of acetonitrile and potassium dihydrogen phosphate buffer with a flow rate of 1.0 mL/min. The retention times were 3.01 minutes for Meclizine and 6.07 minutes for Nicotinic acid. The method was validated and found to be accurate, precise, specific, linear, robust, sensitive and able to quantify the drugs in tablet dosage forms without interference from other components.
Analytical Method Development and Validation of Metformin Hydrochloride by us...ijtsrd
A simple and reproducible method was developed for Metformin MET by Reverse Phase High Performance Liquid Chromatography RP HPLC . Metformin was separated on C18 column 4.6x250mm, particle size 5µm , using combination of phosphate buffer with pH of 3.0 and Methanol at the UV detection of 238nm. Isocratic elution of phosphate buffer with pH of 3.0 and Methanol was used as a mobile phase with various ratios and flow rates, eventually 30 70 v v phosphate buffer with pH of 3.0 and Methanol was being set with the flow rate of 1mL min. The statistical validation parameters such as linearity, accuracy, precision, inter day and intra day variation were checked, assay studies of Metformin were within 98 to 102 indicating that the proposed method can be adoptable for quality control analysis of Metformin. Mr. Nilesh Nikam | Dr. Avish Maru | Dr. Anil Jadhav | Dr. Prashant Malpure ""Analytical Method Development and Validation of Metformin Hydrochloride by using RP-HPLC with ICH Guidelines"" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd22812.pdf
Paper URL: https://www.ijtsrd.com/pharmacy/analytical-chemistry/22812/analytical-method-development-and-validation-of-metformin-hydrochloride-by-using-rp-hplc-with-ich-guidelines/mr-nilesh-nikam
Analytical Method Development and Validation of Tolvaptan in Bulk and Tablet ...pharmaindexing
This document describes the development and validation of a reverse phase high performance liquid chromatographic (RP-HPLC) method for the estimation of Tolvaptan in pharmaceutical dosage forms. A C18 column was used with a mobile phase of sodium dihydrogen phosphate and acetonitrile. The method was validated per ICH guidelines and showed good linearity, accuracy, precision, specificity, and robustness. Recovery of Tolvaptan from formulations ranged from 99.74-99.87% indicating the method is accurate for quantifying Tolvaptan in tablets.
Stability studies of simvastatin and fenofibrate and degradants identificatio...Mehar Raghavendra YEGGINA
1. A stability indicating RP-HPLC method was developed for the simultaneous quantification of simvastatin and fenofibrate.
2. The method used a C18 column with a mobile phase of acetonitrile and ammonium acetate buffer at pH 4.3 to achieve separation of the drugs from their degradation products.
3. The drugs were subjected to stress conditions and the degraded products were identified using LC-MS to prove the stability indicating capability of the developed method.
This document summarizes the presentation of a research article on the development and validation of an RP-HPLC method for the simultaneous determination of metformin and evogliptin in pharmaceutical formulations. The method was developed using a C18 column with an isocratic mobile phase of methanol and water at pH 3. Both drugs showed good separation and were detected at 254 nm. The method was validated and found to be linear, precise, accurate, robust, and specific for quantifying the drugs in tablets within 3 minutes. The method can be applied for routine quality control analysis of metformin and evogliptin combinations.
Method Development and Validation for Simultaneous Estimation of Dasatinib an...YogeshIJTSRD
The document describes the development and validation of an isocratic reverse phase HPLC method for the simultaneous quantification of Dasatinib and Erlotinib in pharmaceutical formulations. The method was optimized using a response surface methodology with a central composite design to evaluate the effects of varying the methanol percentage, pH, and flow rate on the separation. The optimized conditions provided good resolution of Dasatinib, Erlotinib and the internal standard within 9 minutes. The method was validated as per ICH guidelines and successfully applied to the analysis of commercial tablet and capsule formulations containing the two drugs.
Development and validation of a stability indicating RP-HPLC method for estim...BRNSSPublicationHubI
This document describes the development and validation of a reverse-phase high-performance liquid chromatography (RP-HPLC) method for the quantitative analysis of daclatasvir in pharmaceutical formulations. The method utilizes a C18 column, mobile phase of acetonitrile and 0.1% formic acid buffer at a ratio of 40:60, and detection wavelength of 305 nm. Standard and sample solutions of daclatasvir were prepared and analyzed using the optimized method. The results demonstrate that the developed RP-HPLC method is accurate, precise, sensitive and stability-indicating for the analysis of daclatasvir in pharmaceutical dosage forms.
This document describes the development and validation of an RP-HPLC method for the quantification of flunarizine dihydrochloride. Key steps included selecting the instrument and optimizing conditions such as the column, mobile phase, and detection wavelength. The method was then validated based on parameters like accuracy, precision, specificity, detection limit, quantitation limit, and linearity range according to ICH guidelines. The proposed RP-HPLC method was found to be simple, accurate, precise and cost-effective for the analysis of flunarizine dihydrochloride in drug substances and pharmaceutical formulations.
A novel validated stability Indicating RP-HPLC Method Development for the est...Naveen Chennamaneni
Best reserch paper A novel validated stability Indicating RP-HPLC Method Development for the estimation of Certinib in its bulk and finished Dosage form as per ICH Guidelines
DESIGN OF EXPERIMENTS TO DEVELOP AND VALIDATE NEW ANALYTICAL METHODS FOR QUAN...Pranjali837829
Estimation of Lapatinib, an Anticancer drug by HPLC (High-Performance Liquid Chromatography). This research would help treat women suffering from Breast Cancer.
RP-HPLC Assay Method Validation for the estimation of new Anti-retroviral dru...SriramNagarajan15
A reverse phase HPLC method was developed and validated for the quantification of lamivudine in bulk and tablet formulations. The method used an ODS column with a mobile phase of acetate buffer and acetonitrile (50:50) at a flow rate of 1.5 mL/min. Lamivudine had a retention time of 1.85 minutes when detected at 272 nm. The method was linear over a concentration range of 10-50 μg/mL with a correlation coefficient of 0.999. Accuracy and precision studies demonstrated recoveries between 98-102% and %RSD below 2%, respectively. The method was found to be robust, specific, and suitable for the routine analysis of lamivudine in
The document describes the development and validation of a reverse phase HPLC method for the estimation of the anti-retroviral drug lamivudine in bulk and tablet formulations. An ODS column with a mobile phase of acetate buffer and acetonitrile was used to achieve separation of lamivudine. The method was validated as per ICH guidelines and was found to be linear, precise, accurate and robust. The developed method can be used for the routine analysis of lamivudine in pharmaceutical dosage forms.
The document describes the development and validation of a reverse phase HPLC method for the estimation of the anti-retroviral drug lamivudine in bulk and tablet formulations. An ODS column with a mobile phase of acetate buffer and acetonitrile was used to achieve separation of lamivudine. The method was validated as per ICH guidelines and was found to be linear, precise, accurate and robust. The developed method can be used for the routine analysis of lamivudine in pharmaceutical dosage forms.
RP-HPLC Assay Method Validation for the estimation of new Anti-retroviral dru...SriramNagarajan15
A Reverse phase HPLC method was developed for estimation of the Lamivudine in bulk and tablet formulation by using ODS column (250mm×4.6mm, 5µm) and Acetate buffer: acetonitrile (50:50) as mobile phase, at a flow rate of 1.5ml/min. The detection was carried at the 272nm the retention time of the Lamivudine is 1.850. The developed method was validated for the various parameters as per the ICH guidelines like accuracy precision, linearity and range, Robustnes. Linearity was obtained in the concentration range of 10µg/ml to 50µg/ml with correlation coefficient of 0.999. The accuracy of the method was assessed by recovery studies at three different concentration levels. The percentage recovery of Lamivudine was found to be in the range of 98% -102%. The method was found to be precise as indicated by the repeatability, inter-day, intra-day analysis, showing %RSD less than 2. Key words: RP-HPLC, Lamivudine, Pharmaceutical dosage form.
Development and Validation of Reverse Phase Liquid Chromatography Method for ...IOSR Journals
This document describes the development and validation of a reverse phase liquid chromatography method for the estimation of losartan in bulk drug samples. The method utilizes an Acquity BEH C18 column with a mobile phase of buffer and acetonitrile at a ratio of 50:50 delivered isocratically at 0.3 mL/min. Losartan was detected at 230 nm. The method was validated per ICH guidelines and found to be linear, precise, accurate, specific and stability-indicating for the quantification of losartan in the range of 25-75 μg/mL. The method validation shows the method is suitable for the routine analysis of losartan in bulk drug materials.
Simultaneous estimation of meclizine and nicotinic acid by using RP-HPLCpharmaindexing
This document describes the development and validation of a reverse phase high performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of Meclizine and Nicotinic acid. The method utilizes a mobile phase of acetonitrile and potassium dihydrogen phosphate buffer with a flow rate of 1.0 mL/min. The retention times were 3.01 minutes for Meclizine and 6.07 minutes for Nicotinic acid. The method was validated and found to be accurate, precise, specific, linear, robust, sensitive and able to quantify the drugs in tablet dosage forms without interference from other components.
Analytical Method Development and Validation of Metformin Hydrochloride by us...ijtsrd
A simple and reproducible method was developed for Metformin MET by Reverse Phase High Performance Liquid Chromatography RP HPLC . Metformin was separated on C18 column 4.6x250mm, particle size 5µm , using combination of phosphate buffer with pH of 3.0 and Methanol at the UV detection of 238nm. Isocratic elution of phosphate buffer with pH of 3.0 and Methanol was used as a mobile phase with various ratios and flow rates, eventually 30 70 v v phosphate buffer with pH of 3.0 and Methanol was being set with the flow rate of 1mL min. The statistical validation parameters such as linearity, accuracy, precision, inter day and intra day variation were checked, assay studies of Metformin were within 98 to 102 indicating that the proposed method can be adoptable for quality control analysis of Metformin. Mr. Nilesh Nikam | Dr. Avish Maru | Dr. Anil Jadhav | Dr. Prashant Malpure ""Analytical Method Development and Validation of Metformin Hydrochloride by using RP-HPLC with ICH Guidelines"" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd22812.pdf
Paper URL: https://www.ijtsrd.com/pharmacy/analytical-chemistry/22812/analytical-method-development-and-validation-of-metformin-hydrochloride-by-using-rp-hplc-with-ich-guidelines/mr-nilesh-nikam
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Date: May 29, 2024
Tags: Information Security, ISO/IEC 27001, ISO/IEC 42001, Artificial Intelligence, GDPR
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Chapter wise All Notes of First year Basic Civil Engineering.pptxDenish Jangid
Chapter wise All Notes of First year Basic Civil Engineering
Syllabus
Chapter-1
Introduction to objective, scope and outcome the subject
Chapter 2
Introduction: Scope and Specialization of Civil Engineering, Role of civil Engineer in Society, Impact of infrastructural development on economy of country.
Chapter 3
Surveying: Object Principles & Types of Surveying; Site Plans, Plans & Maps; Scales & Unit of different Measurements.
Linear Measurements: Instruments used. Linear Measurement by Tape, Ranging out Survey Lines and overcoming Obstructions; Measurements on sloping ground; Tape corrections, conventional symbols. Angular Measurements: Instruments used; Introduction to Compass Surveying, Bearings and Longitude & Latitude of a Line, Introduction to total station.
Levelling: Instrument used Object of levelling, Methods of levelling in brief, and Contour maps.
Chapter 4
Buildings: Selection of site for Buildings, Layout of Building Plan, Types of buildings, Plinth area, carpet area, floor space index, Introduction to building byelaws, concept of sun light & ventilation. Components of Buildings & their functions, Basic concept of R.C.C., Introduction to types of foundation
Chapter 5
Transportation: Introduction to Transportation Engineering; Traffic and Road Safety: Types and Characteristics of Various Modes of Transportation; Various Road Traffic Signs, Causes of Accidents and Road Safety Measures.
Chapter 6
Environmental Engineering: Environmental Pollution, Environmental Acts and Regulations, Functional Concepts of Ecology, Basics of Species, Biodiversity, Ecosystem, Hydrological Cycle; Chemical Cycles: Carbon, Nitrogen & Phosphorus; Energy Flow in Ecosystems.
Water Pollution: Water Quality standards, Introduction to Treatment & Disposal of Waste Water. Reuse and Saving of Water, Rain Water Harvesting. Solid Waste Management: Classification of Solid Waste, Collection, Transportation and Disposal of Solid. Recycling of Solid Waste: Energy Recovery, Sanitary Landfill, On-Site Sanitation. Air & Noise Pollution: Primary and Secondary air pollutants, Harmful effects of Air Pollution, Control of Air Pollution. . Noise Pollution Harmful Effects of noise pollution, control of noise pollution, Global warming & Climate Change, Ozone depletion, Greenhouse effect
Text Books:
1. Palancharmy, Basic Civil Engineering, McGraw Hill publishers.
2. Satheesh Gopi, Basic Civil Engineering, Pearson Publishers.
3. Ketki Rangwala Dalal, Essentials of Civil Engineering, Charotar Publishing House.
4. BCP, Surveying volume 1
Level 3 NCEA - NZ: A Nation In the Making 1872 - 1900 SML.pptHenry Hollis
The History of NZ 1870-1900.
Making of a Nation.
From the NZ Wars to Liberals,
Richard Seddon, George Grey,
Social Laboratory, New Zealand,
Confiscations, Kotahitanga, Kingitanga, Parliament, Suffrage, Repudiation, Economic Change, Agriculture, Gold Mining, Timber, Flax, Sheep, Dairying,
A Visual Guide to 1 Samuel | A Tale of Two HeartsSteve Thomason
These slides walk through the story of 1 Samuel. Samuel is the last judge of Israel. The people reject God and want a king. Saul is anointed as the first king, but he is not a good king. David, the shepherd boy is anointed and Saul is envious of him. David shows honor while Saul continues to self destruct.
2. Das, et al.: Development and validation of flupirtine maleate by reverse phase HPLC
IJPBA/Apr-Jun-2019/Vol 10/Issue 2 96
in tablet dose structure. This present investigation
reports for the first time quantitation of the drug
by RP-HPLC in tablet dose form. The proposed
strategyisapprovedaccordingtotheICHrules.[17-20]
MATERIALS AND METHODS
Method development involves the evaluation and
optimizationofvariousstagesofsamplepreparation,
chromatographic separation, and detection.
Optimization of various parameters was performed
to develop a selective and sensitive method for
analysis of flupirtine maleate on RP-HPLC using
photodiode array detector (PDAD).[21-36]
Materials
Orthophosphoric acid, HPLC grade methanol,
triethylamine, methanol, and chloroform were
purchased from Rankem New
Delhi, India. All
the chemicals and solvents of HPLC grade were
used without any additional purification. All the
solutions were arranged in HPLC grade water.
Market formulation of Ketoflam SR (LUPIN Ltd.
Kartholi, Bari Brahmana, in Jammu, India) was
procured from the local drug store. Specified all
solutions were filtered through 0.22 µ.
Instrumentation
It is the binary LC system 515 equipped with
W2998 PDA detector. 20 µl Rheodyne loop
injector and Empower 2 data station software were
used. Separation was performed on C18 column.
Chromatographic data were processed using
EMPOWER 2 Software. Methanol:water (90:10)
was selected as the mobile phase for dissolution.
Flupirtine maleate standard preparation
About 25 mg of working standard of flupirtine
maleate was weighed accurately and transferred
it into 100 ml dry volumetric flask. The drug was
dissolved completely into 25 ml of methanol and
make up the volume.
Sample preparation
Tablets were accurately weighed and calculated
for average weight. The tablets were crushed to
powder form and weighed immediately with the
help of weighing balance and transferred into a 100
ml dry volumetric flask. Then 25 ml of methanol
were added into a 100 ml volumetric flask and
shaken for 10 min and made up the volume up to
the mark with methanol.
Chromatographic parameters
Column: C18 (150 mm × 25.4 mm)
Flow rate: 1 ml/min
Injection volume: 20 µl
Run time: 20 min
Mobile phase: Methanol:Water (90:10)
VALIDATION OF PROPOSED METHOD
System suitability
System suitability analysis is an important part of
many analytical procedures. The tests are created
on the idea that the equipment, electronics,
analytical operations, and samples to be analyzed
constitute an integral system that can be evaluated
as such. System suitability test parameters to be
established for a particular procedure depend on
the type of procedure being validated.[37-44]
System
suitability was demonstrated by preparation of
standard solution, which was included in ICH
guideline. Here chromatographs were same for
HPLC system for all 5 replicates; the peak area of
analytewasrecordedforthesereplicatedinjections.
The tailing factor and percentage relative standard
deviation (RSD) were evaluated for analyte peaks.
The results for the above development method are
tabulated in Table 1.
Method precision
Method precision was confirmed by preparing six
samples as per the test method representing a single
batch. The assay and precision of the methods
were determined. The precision of the method was
evaluated by computing the percentage RSD of
Figure 1: Structure of flupirtine
3. Das, et al.: Development and validation of flupirtine maleate by reverse phase HPLC
IJPBA/Apr-Jun-2019/Vol 10/Issue 2 97
the assay result.[45-50]
The result precision studies
are tabulated in Tables 2 and 3. Intermediate
precision (Ruggedness): The ruggedness of an
tampering method is the stratum of reproducibility
of test results obtained by the wringer of the same
samples under a variety of conditions. Intraday
and interday precision for the ripened method
was measured in terms of percentage RSD. The
experiments were repeated 3
times a day for
intraday precision and on three variegated days
for interday precision. The concentration value
for both intraday and interday precision was
calculated. Finally, the midpoint of percentage
RSD is calculated.
Linearity
Linearity should be calculated by graphical
inspection of a plot of signals as a function
of analyte concentration or content. If there
is a linear relationship, test results should be
evaluated by appropriate statistical methods,
for example, by calculation of a regression line
by the method of least squares in addition, an
analysis of the deviation of the actual data points
from the regression line may also be helpful for
evaluating linearity. Linearity is the ability of the
method to obtain the test results which are directly
proportional to the concentration of analyte in
the sample. The linearity of detector response for
flupirtine maleate was demonstrated by preparing
solutionofworkingstandardasperthemethodover
the range of 50%–150% of target concentration.
These solutions were injected into the system and
the peak area of analyte was recorded. A graph
of concentration versus peak area of analyte was
plotted.
Accuracy
Accuracy has to be described as percent regaining
by the assay of known added amount of analyte
in the sample or as the difference between the
mean and the accepted true value together with
the confidence intervals.[11,12]
The parameters
provide information about the regaining of
the drug from sample and effect of matrix, as
regaining are likely to be excessive as well as
incomplete. The accuracy of test method was
demonstrated by preparing recovery samples
(i.e., spiking placebo with known quantities of
levels of 50%, 100%, and 150%), the recovery
samples were prepared in triplicate at each level.
The above samples were chromatographed and
the percentage recovery for the amount added
was estimated.
Robustness
The evaluation of robustness should be considered
during the development phase and depends on the
type of procedure under study. If measurements are
susceptible to variations in analytical conditions,
the analytical conditions should be suitably
controlled, or a precautionary statement should
be included in the procedure.[13-16]
The robustness
of the method was determined by performing
the assay in triplicate by deliberately alternating
parameters. Here, the peak area of analyte in
sample solution and standard solution should not
differ by more than 2.0% from initial peak area
Table 1: Linearity study of flupirtine maleate
Linearity
level (%)
Concentration of flupirtine
maleate in mcg/ml (ppm)
Peak area of
flupirtine maleate
60 150 332,190
80 200 453,216
100 250 620,176
120 300 753,150
140 350 920,570
Table 2: System suitability data
Injection number Peak areas FlupirtineMeleate
1 620,357
2 622,535
3 621,375
4 623,012
5 621,032
Average 621,662
%RSD 0.17
Tailing factor 0.5
%RSD: Percentage relative standard deviation
Table 3: Intraday precision data
Serial number Precision Percentage assay of
FlupirtineMeleate
1 Precision set – 1 98.6
2 Precision set – 2 100.2
3 Precision set – 3 99.4
4 Precision set – 4 100.4
5 Precision set – 5 98.9
6 Precision set – 6 99.7
Average 99.5
%RSD 0.8
%RSD: Percentage relative standard deviation
4. Das, et al.: Development and validation of flupirtine maleate by reverse phase HPLC
IJPBA/Apr-Jun-2019/Vol 10/Issue 2 98
for the accepted storage time. Hence sample and
standard were stable for 36 hours
RESULTS AND DISCUSSION
Identification tests
Physical appearance
The physical appearance of flupirtine maleate was
noted by visual observation. It appeared as white
colored powder and off-white.
Melting point
The melting point of flupirtine maleate was
found to be 176–179°C which complies with
literature.
Solubility
Flupirtine maleate was found to slightly soluble
in water, freely soluble in methanol, and sparingly
soluble in ethanol.
Analytical method development by RP-HPLC
method
Method development
The chromatographic separation was achieved
on C18 (150 mm × 25.4 mm) analytical column
with the mobile phase consisting ratio mixture
of methanol and OPA0.2% (90:10) at a flow
rate of 1 ml/min when injection volume 20 µl
and run time 20 min at detector wavelength of
254 nm.
Trial 1 and Trial 2
In trial first, chromatographic separation
was achieved on C18
(150
mm × 25.4
mm).
Analytical column with the mobile phase of
methanol and OPA 0.2% (80:20) at a flow rate of
1ml/min at a detection wavelength of 256 mm.
Peak was broad and not symmetrical. In trial
second, chromatographic separation was achieved
on C18 (150 mm × 25.4 mm). Analytical column
with the mobile phase of methanol and OPA 0.2%
(70:30) at a flow rate of 1 ml/min at the detection
wavelength of 256 mm. Peak was not symmetrical.
In trial second, chromatographic separation was
achieved on C18 (150 mm × 25.4 mm).Analytical
column with the mobile phase of methanol and
OPA 0.2% (90:10) at a flow rate of 1 ml/min at
the detection wavelength of 256
nm. Peak was
symmetrical.
The observations are presented in Table 4,
Figures 2-4.
Optimization of chromatographic conditions
The effect of chromatographic conditions on the
instrument response creates a situation where one
has to compromise between different experimental
variables to achieve the best chromatographic
separations. The resolution and sensitivity of the
method were obtained at 254 nm, and the mobile
phase flow rate was 1 ml/min. The retention time
of flupirtine maleate was 9.061 when PH at 3.
Figure 2: Spectra 1 of flupirtine maleate
Table 4: Observation
Observation
number
RT Area Percentage
area
Height
1 9.061 604,001 100.00 18,836
2 9.060 603,876 100.00 18,832
3 9.061 603,783 100.00 18,830
RT: Retention time
5. Das, et al.: Development and validation of flupirtine maleate by reverse phase HPLC
IJPBA/Apr-Jun-2019/Vol 10/Issue 2 99
System suitability
System suitability was demonstrated by
preparation of standard solution, which was
included in ICH guideline. Here chromatographs
were same for HPLC system for all 5 replicates;
the peak area of analyte was recorded for these
replicated injections. The tailing factor and RSD
were evaluated for the analytes peak.
Acceptance criteria
Tailing factor for analyte peak should not be 2.0.
Percentage RSD of five replicate standard
injections should not be 2.0. The results are
shown in Table 2.
Method precision
The precision of an analytical procedure expresses
the closeness of agreement (degree of scatter)
between a series of measurements obtained from
multiple sampling of the same homogeneous
sample under the prescribed conditions. Precision
may be considered at three levels: Repeatability,
intermediate precision, and reproducibility.
Method precision was demonstrated by preparing
six samples as per the test method representing
a single batch. The assay of these samples was
determined, and the precision of the method was
evaluated by computing the percentage RSD of
the assay results. The results of the precision study
are tabulated in Table 3.
Acceptance criteria
Percentage RSD for assay of six preparations
should not be 2.0.
Intermediate precision (ruggedness)
The ruggedness of test method was demonstrated
by carrying out a precision study in six replicates of
the assay on a single batch sample on two different
days, using two different columns [Tables 1,5,6],
and on two different instruments, as per the matrix
tabulated in Table 7-9. Linearity is the ability of the
method to obtain the test results which are directly
proportional to the concentration of analyte in
the sample. The linearity of detector response for
flupirtine meleate was demonstrated by preparing the
solution of flupirtine maleate working standard over
the range 50%–150% of target concentration [Tables
1, 8-10, and Figure 5]. These solutions were injected
into the system and the peak area of analyte was
recorded. A graph of concentration versus peak area
Figure 3: Spectra 2 of flupirtine maleate
Figure 4: Spectra 3 of flupirtine maleate
6. Das, et al.: Development and validation of flupirtine maleate by reverse phase HPLC
IJPBA/Apr-Jun-2019/Vol 10/Issue 2 100
of analyte was plotted [Table 11]. The correlation
coefficient between concentration and peak area and
y-intercept of the correlation plot were evaluated.
The observations are tabulated in Table 12.
CONCLUSION
It is concluded that RP-HPLC method is
successfully utilized for the estimation of
flupirtine maleate. The validation data obtained
in the developed method indicated that proposed
developed RP-HPLC method with PDA detection
is simple, sensitive, accurate, more precise,
Table 5: Interday precision data
Serial number Precision Percentage assay of
Flupirtine Maleate
1 Precision set – 1 99.3
2 Precision set – 2 100.2
3 Precision set – 3 98.7
4 Precision set – 4 99.5
5 Precision set – 5 100.5
6 Precision set – 6 100.6
Average 99.8
%RSD 0.7
%RSD: Percentage relative standard deviation
Table 6: Comparison between interday and intraday
precision
Parameters Percentage assay of flupirtine maleate
Intraday precision 99.5
Interday precision 99.8
Difference 0.3
%RSD 0.2
%RSD: Percentage relative standard deviation
Table 7: Validation parameters of linearity by
reversed‑phase high‑performance liquid chromatography
method for flupirtine maleate
Parameters Flupirtine Maleate
Linearity range 150–350 ppm
Regression equation 115486 + 2933 X
Slope 2933
Intercept 115,486
Correlation coefficient 0.999
Table 8: Recovery at 50% level
Sample Amount of
flupirtine
maleate
spiked (mg)
Amount of
flupirtine
maleate
recovered (mg)
Percentage
recovery
1 12.35 12.50 98.8
2 12.35 12.45 99.1
3 12.35 12.55 98.4
Average 98.7
%RSD 0.3
%RSD: Percentage relative standard deviation
Table 9: Recovery at 100% level
Sample Amount of
flupirtine
maleate
spiked (mg)
Amount of
flupirtine
maleate
recovered (mg)
Percentage
recovery
1 Equivalent to 25 25.45 98.2
2 Equivalent to 25 25.25 99.0
3 Equivalent to 25 25.50 98.0
Average 98.4
%RSD 0.4
%RSD: Percentage relative standard deviation
Table 10: Recovery at 150% level
Sample Amount of
flupirtine
maleate
spiked (mg)
Amount of
flupirtine
maleate
recovered (mg)
Percentage
recovery
1 38.40 38.75 99.0
2 38.40 38.55 99.6
3 38.40 38.70 99.2
Average 99.2
%RSD 0.8
%RSD: Percentage relative standard deviation
Table 11: Robustness data for standard solution
Time (in h) Peak area of
flupirtine maleate
Percentage deviation
from the initial area
00 621,032 ‑
6 609,945 −1.7
12 630,335 1.4
18 619,025 −0.3
24 622,110 0.2
30 619,530 −0.2
36 610,120 −1.8
%RSD 1.1
%RSD: Percentage relative standard deviation
Table 12: Robustness data for sample solution
Time (in h) Peak area of
flupirtine maleate
Percentage deviation
from initial area
00 604,530 ‑
6 613,025 1.4
12 600,253 −0.7
18 603,975 0.1
24 600,290 −0.7
30 613,876 1.5
36 602,520 −0.3
%RSD 1.9
%RSD: Percentage relative standard deviation
7. Das, et al.: Development and validation of flupirtine maleate by reverse phase HPLC
IJPBA/Apr-Jun-2019/Vol 10/Issue 2 101
and less time consuming and can be usefulfor
routine determination of flupirtine maleate.
Method development involves the evaluation
and optimization of various stages of sample
preparation, chromatographic separation, and
detection. Optimization of various parameters was
performed to develop a selective and sensitive
method for analysis of flupirtine maleate on RP-
HPLC using PDAD. All the system suitability
parameters were within the limit, and a sharp peak
with better resolution and purity was obtained
with the developed method. Recovery studies
are between the ranges of 98.0% and 120% with
a RSD at each level of 2.0%, which proves
that the method is accurate for the estimation of
flupirtine maleate over the range 50%–150% of
target concentration. In precision studies, the low
percentage RSD has been observed on the assay
value which indicates that method is precise
while the intraday precision studies, assay result
obtained on two different analysts, on two different
days were found to be within an acceptable limit,
which shows that the test method is rugged.
The above HPLC analysis method is, therefore,
recommended to use for routine analysis.
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