This article describes the development and validation of a simple reverse phase HPLC method for the quantitative analysis of diazepam in pharmaceutical dosage forms. A C18 column with a mobile phase of acetonitrile, methanol and phosphate buffer at a flow rate of 1 mL/min was used to achieve good resolution and retention of diazepam at 6.23 minutes. The method was validated and found to be linear, precise, accurate, sensitive and robust for the analysis of diazepam in tablets with recoveries ranging from 99.4-100.3%.
Simultaneous estimation and validation of atenolol hydrochloro thiazide and l...srirampharma
This document describes the development and validation of a reverse phase HPLC method for the simultaneous estimation of atenolol, hydrochlorothiazide, and losartan K from pharmaceutical tablet dosage forms. The method utilizes a C18 column with a mobile phase of acetonitrile and phosphate buffer at a ratio of 70:30 delivered at 1.2 mL/min. The drugs were detected at 229 nm and provided good resolution and peak symmetry. The developed method was validated in terms of accuracy, precision, linearity, limits of detection and quantification, robustness, and solution stability. The method was applied to a commercial tablet dosage form and found to provide accurate results within 100% of the labeled claim for each drug.
Development and Validation of Stability Indicating Method for Simultaneous Es...iosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Differential spectrophotometric method for estimation and validation of verap...roshan telrandhe
The aimed of current research to development of the simple, rapid and sensitive Differential spectrophotometric method for the estimation of Verapamil in tablet dosage form. In this method two medium was use acid and alkaline and the difference spectrum was calculated. 0.1N HCL and 0.1N NaOH was used in this differential method. The λmax 278, Beers law limits 5-25µg/ml, regression equation Y= 0.024x-0.009, slope 0.024, intercept 0.09, correlation coefficient (r2) 0.998, %RSD <1.5, % Recovery (Tablet) 100.46% was shows the good efficacy and results. This method future scope in quality control of the verapamil in simple, precise and economically and it recommended for the routine drug quality analysis investigation
Development and validation of a stability-indicating HPLC method for the simultaneous determination of Losartan potassium, hydrochlorothiazide, and their degradationproducts
This document presents a method for determining levels of psoralens like psoralen and bergapten in oral solutions of herbal medicines using liquid chromatography. The method involves extracting samples with chloroform, dissolving residues in methanol, and analyzing by HPLC. The method was validated in terms of linearity, accuracy, precision and specificity. Psoralen and bergapten were found to be stable in samples and solutions under various storage conditions. The method provides a fast and reliable means of quantifying psoralen levels in herbal medicines.
This document summarizes the development and validation of a stability-indicating high performance liquid chromatography (HPLC) method for the quantitative determination of carbamazepine in controlled-release tablets. The method uses a Hypersil ODS V column with a mobile phase of water, methanol and methylene chloride. Method validation included tests for accuracy, precision, robustness and linearity according to ICH guidelines. The results of the validation tests confirmed the method is accurate, precise and robust for the quantitative analysis of carbamazepine in controlled-release tablets.
Spectrophotometric Determination of Drugs and Pharmaceuticals by Cerium (IV) ...IOSR Journals
Simple, sensitive, accurate, and precise spectrophotometric methods for quantitative determination of drugs, viz., Darifenacin (DAR), Esmolol Hydrochloride (ESM), Montelukast Sodium (MON), Sildenafil citrate (SIL),Terbinafine (TER) and Tramadol Hydrochloride (TRA) were developed. The method of each drug depends upon oxidation of drugs by Ce (IV) (Excess) and estimating the amount of unreacted Ce (IV) by amaranth dye at 523nm. The calibration curves obeyed Beer’s law over the concentration range of 1.4-7.0 μg ml-1 (DAR), 2-14 μg ml-1 (ESM), 2-10 μg ml-1 (MON), 20-70 μg ml-1 (SIL), 3-21 μg ml-1 (TER) & 2-14 μg ml-1 (TRA). The methods have been validated in terms of guidelines of ICH and applied to analysis of pharmaceuticals.
Stability indicating method development and validation for the simultaneous e...pharmaindexing
Stability indicating method development and validation for the simultaneous estimation of rabeprazole sodium and ketorolac tromethamine in bulk and synthetic mixture by RP-HPLC
Simultaneous estimation and validation of atenolol hydrochloro thiazide and l...srirampharma
This document describes the development and validation of a reverse phase HPLC method for the simultaneous estimation of atenolol, hydrochlorothiazide, and losartan K from pharmaceutical tablet dosage forms. The method utilizes a C18 column with a mobile phase of acetonitrile and phosphate buffer at a ratio of 70:30 delivered at 1.2 mL/min. The drugs were detected at 229 nm and provided good resolution and peak symmetry. The developed method was validated in terms of accuracy, precision, linearity, limits of detection and quantification, robustness, and solution stability. The method was applied to a commercial tablet dosage form and found to provide accurate results within 100% of the labeled claim for each drug.
Development and Validation of Stability Indicating Method for Simultaneous Es...iosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Differential spectrophotometric method for estimation and validation of verap...roshan telrandhe
The aimed of current research to development of the simple, rapid and sensitive Differential spectrophotometric method for the estimation of Verapamil in tablet dosage form. In this method two medium was use acid and alkaline and the difference spectrum was calculated. 0.1N HCL and 0.1N NaOH was used in this differential method. The λmax 278, Beers law limits 5-25µg/ml, regression equation Y= 0.024x-0.009, slope 0.024, intercept 0.09, correlation coefficient (r2) 0.998, %RSD <1.5, % Recovery (Tablet) 100.46% was shows the good efficacy and results. This method future scope in quality control of the verapamil in simple, precise and economically and it recommended for the routine drug quality analysis investigation
Development and validation of a stability-indicating HPLC method for the simultaneous determination of Losartan potassium, hydrochlorothiazide, and their degradationproducts
This document presents a method for determining levels of psoralens like psoralen and bergapten in oral solutions of herbal medicines using liquid chromatography. The method involves extracting samples with chloroform, dissolving residues in methanol, and analyzing by HPLC. The method was validated in terms of linearity, accuracy, precision and specificity. Psoralen and bergapten were found to be stable in samples and solutions under various storage conditions. The method provides a fast and reliable means of quantifying psoralen levels in herbal medicines.
This document summarizes the development and validation of a stability-indicating high performance liquid chromatography (HPLC) method for the quantitative determination of carbamazepine in controlled-release tablets. The method uses a Hypersil ODS V column with a mobile phase of water, methanol and methylene chloride. Method validation included tests for accuracy, precision, robustness and linearity according to ICH guidelines. The results of the validation tests confirmed the method is accurate, precise and robust for the quantitative analysis of carbamazepine in controlled-release tablets.
Spectrophotometric Determination of Drugs and Pharmaceuticals by Cerium (IV) ...IOSR Journals
Simple, sensitive, accurate, and precise spectrophotometric methods for quantitative determination of drugs, viz., Darifenacin (DAR), Esmolol Hydrochloride (ESM), Montelukast Sodium (MON), Sildenafil citrate (SIL),Terbinafine (TER) and Tramadol Hydrochloride (TRA) were developed. The method of each drug depends upon oxidation of drugs by Ce (IV) (Excess) and estimating the amount of unreacted Ce (IV) by amaranth dye at 523nm. The calibration curves obeyed Beer’s law over the concentration range of 1.4-7.0 μg ml-1 (DAR), 2-14 μg ml-1 (ESM), 2-10 μg ml-1 (MON), 20-70 μg ml-1 (SIL), 3-21 μg ml-1 (TER) & 2-14 μg ml-1 (TRA). The methods have been validated in terms of guidelines of ICH and applied to analysis of pharmaceuticals.
Stability indicating method development and validation for the simultaneous e...pharmaindexing
Stability indicating method development and validation for the simultaneous estimation of rabeprazole sodium and ketorolac tromethamine in bulk and synthetic mixture by RP-HPLC
Stability indicating analytical method development and validation for estimat...SriramNagarajan18
Stability indicating analytical method development and validation for estimation of Sacubitril and Valsartan in bulk and pharmaceutical dosage form using RP-HPLC
Here are the key IR frequencies identified in the sample that match the reference standard of propafenone:
- C-C stretch at 1186 cm-1
- C=C stretch at 1651 cm-1
- C-H stretch (symmetric) at 2939 cm-1
- C-H bend at 1328 cm-1
- CH2 stretch (symmetric) at 1369 cm-1
- CH2 bend at 1485 cm-1
- CH3 bend at 1398 cm-1
- C-O stretch at 1100 cm-1
- C=O stretch at 1695 cm-1
- N-H stretch at 3417 cm-1
The IR spectrum
This document describes a spectrofluorimetry method for determining nalidixic acid concentrations in human plasma. The method involves extracting nalidixic acid from plasma samples using chloroform and analyzing the samples using a spectrofluorimeter. The method was used to analyze plasma samples from a study comparing the bioavailability of a new generic nalidixic acid tablet formulation to a standard brand name formulation. Results of in vitro tests like dissolution and assay showed the new generic formulation was comparable to the standard. Pharmacokinetic analysis of plasma concentration data from subjects who received the formulations found no significant differences between the products.
Shraddha roll no- 7 -m. pharm final presentation ---rbvrr college of pharmacysaathiyaa
The document describes the development and validation of an analytical method for ziprasidone using reverse phase high performance liquid chromatography. The method utilizes a Sunsil C18 column with a mobile phase of water and methanol (55:45) at a flow rate of 1 mL/min. Ziprasidone was detected at 261 nm with a retention time of 3.082 minutes. The method was validated for specificity, precision, linearity, accuracy, limit of detection, limit of quantification and robustness as per ICH guidelines. The developed and validated method can be used for the analysis of ziprasidone in bulk and pharmaceutical formulations.
Multiple Method Development and Validation for Simultaneous Estimation of Chl...ijtsrd
A simple, precise and accurate multiple analytical method has been developed for the simultaneous estimation of Chlorzoxazone and Nimesulide in bulk and tablet formulations by reversed-phase liquid chromatographic and UV-Visible spectrophotometric techniques. The chromatographic separation was achieved on C18 analytical column. A mixture of Methanol 0.1 Ortho-phosphoric acid 75 25 was used as mobile phase, at a flow rate of 1mL min and detection wavelength at 295 nm. The retention time of Chlorzoxazone and Nimesulide was found to be 4.69 and 5.45 min respectively. The linear dynamic ranges for HPLC were from 2-10 µg mL and for simultaneous equation method, derivative spectroscopy, Q-ratio Absorbance method, Dual wavelength it was 10-30 µg mL for both Chlorzoxazone and Nimesulide. The percentage recovery obtained for Chlorzoxazone and Nimesulide were 100.93 and 102.19 respectively for RP-HPLC, 9.7 and 100.1 for simultaneous equation method of CZ and NIM respectively, 99.97 and 99.78 for derivative spectroscopy of CZ and NIM respectively, 101.37 and 99.48 for Q-ratio Absorbance method of CZ and NIM respectively, 100.13 and 99.96 for dual wavelength method of CZ and NIM respectively. The validation of the proposed methods were carried out for linearity, accuracy, precision, limit of detection, limit of quantitation and robustness. The developed method can be used for routine quality control analysis of titled drugs in combination in tablet formulation. Swetha Yarramsetti | A. Elphine Prabahar | Rama Rao Nadendla "Multiple Method Development and Validation for Simultaneous Estimation of Chlorzoxazone and Nimesulide in Bulk and Pharmaceutical Dosage Form" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-2 , February 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21503.pdf
Paper URL: https://www.ijtsrd.com/pharmacy/analytical-chemistry/21503/multiple-method-development-and-validation-for--simultaneous-estimation-of-chlorzoxazone-and--nimesulide-in-bulk-and-pharmaceutical-dosage-form/swetha-yarramsetti
Method Development and Validation of Clopidogrel Bisulphate by Reverse Phase-...SriramNagarajan15
A new, simple sensitive, rapid, accurate and precise RP-HPLC method was developed for the estimation of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Clopidogrel bisulphate was chromatographed on a reverse phase C18column (150 mm x 4.5 mm, i.d 5μm) in a mobile phase consisting of acetonitrile and phosphate buffer (pH: 3.0) in the ratio of 60:40 % v/v. The mobile phase was pumped at a flow rate of 1 ml/min with detection at 224 nm. The detector response was linear in the concentration of 50-150 μg /ml. The limit of detection and limit of quantitation was found to be 1.3 and 4.2 µg/ml, respectively. The intra and inter day variation was found to be less than 2%. The mean recovery of the drug from the solution was 99.79%. The proposed method is simple, fast, accurate, precise and reproducible hence, it can be applied for routine quality control analysis of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Key words: Clopidogrel bisulphate, RP-HPLC, Validation, Accuracy, Precision.
1. A new RP-HPLC method was developed and validated for the estimation of Clopidogrel bisulphate in bulk drug and pharmaceutical formulations.
2. The method involved using a C18 column, mobile phase of acetonitrile and phosphate buffer at a ratio of 60:40, and detection at 224 nm.
3. The method was found to be linear, precise, accurate, specific, robust and suitable for the routine analysis of Clopidogrel bisulphate in quality control labs.
Validated HPLC Method for Assay and Content Uniformity Testing of Roflumilast...Ratnakaram Venkata Nadh
Roflumilast is a selective enzyme inhibitor of phosphodiesterase-4. This drug is recommended for treatment of patients suffering from
chronic-obstructive-pulmonary-disease with chronic-bronchitis. Roflumilast is not official in pharmacopoeia and the reported methods
are having high chromatographic run times. A short run time HPLC method was developed for assay and content uniformity testing to
determine the roflumilast in blend and tablets. The mobile phase consists of 10 mM sodium dihydrogen phosphate monohydrate buffer
and acetonitrile in the ratio of 45:55 v/v. The HPLC method was developed using accucore-C18 150 × 4.6 mm, 4 μm column with a flow
rate of 1.0 mL min-1, 215 nm wavelength and 10 μL injection volume with run time of 5 min. The method linearity was proved between
5.02-40.17 μg mL-1 and obtained correlation-coefficient value is 1.0000. The mean recovery of roflumilast was 100.6%. The stability
indicating nature was established and performed the validation by considering ICH Q2 (R1) recommendations.
Diazo coupling for the determination of selexipag by visible spectrophotometryRatnakaram Venkata Nadh
Aim and Objective: The aim and objective of this study were to develop a spectrophotometric method for the assay of selexipag (selective IP prostacyclin receptor agonist indicated for the treatment of pulmonary arterial hypertension) in pure and pharmaceutical formulations so that it will be an alternative quantitative method to chromatographic methods which require large quantities of organic solvents, where some are with hazardous and toxic properties. Materials and Methods: The method is based on the diazo coupling of selexipag with diazotized p-nitroaniline in alkaline medium to form a stable green-colored and water-soluble azo dye with a maximum absorption at 510 nm. Optimization of reaction conditions was carried out to get highly sensitive and stable colored complex. Results and Discussion: Beer’s law is obeyed over the concentration range of 2–12 μg/mL with a molar absorptivity of 3.33 × 104 L/mol/cm. The limit of detection was 0.35 μg/mL and limit of quantification was 1.0 μg/mL. The results demonstrated that the procedure is accurate, precise, and reproducible (relative standard deviation <2%). Conclusions: This method was tested and validated for various parameters according to the current ICH guidelines.
Validated UV Method Development for the Simultaneous Estimation of Rabeprazol...pharmaindexing
This document describes the development and validation of a UV spectrophotometric method for the simultaneous estimation of Rabeprazole sodium and Cinitapride in tablet formulations. Wavelengths of 284.5 nm and 267 nm were selected for analysis of Rabeprazole sodium and Cinitapride, respectively. The method was validated according to ICH guidelines and showed good linearity, precision, and accuracy for simultaneous analysis of the two drugs without interference from excipients. The developed UV method provides a simple, accurate, and cost-effective approach for quality control testing of Rabeprazole sodium and Cinitapride in combined tablet dosage forms.
Stability indicating rp hplc method for the determination of candisartansrirampharma
This document describes the development of an RP-HPLC method for the determination of candesartan in pure and pharmaceutical formulations. The method utilizes a C18 column with a mobile phase of phosphate buffer and acetonitrile in a 30:70 ratio at a flow rate of 1.2 mL/min. Candesartan was detected at 255 nm with a retention time of 3.089 minutes. The method was validated per ICH guidelines and demonstrated specificity, linearity from 20-60 μg/mL, precision, accuracy, ruggedness and robustness. The method was found to separate candesartan from degradation products formed under various stress conditions, indicating it is stability-indicating and can be used to conduct
Analytical Method Development and Validation of Dutasteride and Tamsulosin Hc...SriramNagarajan15
A simple,specific, sensitive,precise and reproducible Reverse Phase High Performance liquid Chromatography method has been developed for simultaneous estimation of Dutasteride and Tamsulosin Hcl. Dutasteride and Tamsulosin is Anti-hyperplasia and Anti-hypertensive drug.The determinationwas carried out byusingsymmetryC-18columnwith Methanol:0.1M Monobasic potassiumdihydrogenphosphate buffer(75:25) Adjusted the pH to 2.5 with Ortho phosporic acid as the mobile phase and with the detection wavelength of274 nmrespectively.The flow rate is 0.7 ml/min.TheRetentiontime of Dutasteride,Tamsulosin Hcl was 2.218 minand 6.599 min respectively.Linearityforthe Dutasteride and Tamsulosin Hcl were found inthe rangeof 25-75µgmand 20-60µgm respectively.The limitof quantificationforbothdrugs wasfound to be30,24µg respectively.The recoveries of Tamsulosin and Dutasteride were found to be inthe range of 99.81-99.90 %and98.00-102.00%, respectively. The proposed method was validated suitably and canbeused for routine analysis. The degradation studies indicated Dutasteride and Tamsulosin Hcl to besusceptible to neutralhydrolysis, while Dutasteride and Tamsulosin Hcl showed degradation inacid, H2O2,photolytic and inpresenceof UV radiation.The degradation productsof Dutasteride andTamsulosin Hcl inacidic and photolytic conditions were well resolved from the pure drug with significant differences in the irretention time values. This method can be successfully employed forsimultaneous quantitative analysis of Dutasteride and Tamsulosin Hcl in formulations.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Analytical method development and validation for the estimation of quinapril ...SriramNagarajan19
A simple and selective LC method is described for the determination of Quinapril and Tolcapone tablet dosage forms. Chromatographic separation was achieved on a c18 column using mobile phase consisting of a Mixed Phosphate buffer (KH2PO4 +K2HPO4): Acetonitrile 40:60, with detection of 239 nm. Linearity was observed in the range 50 - 150 µg /ml for Quinapril (r2 =0.995) and 62.5- 187.5µg /ml for Tolcapone (r2 =0.999) for the amount of drugs estimated by the proposed methods was in good agreement with the label claim.
The proposed methods were validated. The accuracy of the methods was assessed by recovery studies at three different levels. Recovery experiments indicated the absence of interference from commonly encountered pharmaceutical additives. The method was found to be precise as indicated by the repeatability analysis, showing %RSD less than 2. All statistical data proves validity of the methods and can be used for routine analysis of pharmaceutical dosage form.
Presentation given by Steve Capes, Cambridgeshire Community Archives Network, at "Building a Heritage Network" conference in Stornoway, 29 October 2013
Mainstreaming Open Educational Practice in a Research University: Prospects a...Liz Masterman
This document summarizes a study on mainstreaming open educational practice at a research university. The study used interviews and frameworks of openness to examine academics' recognition of open values, the influence of disciplines and organizational culture on teaching practices, and what constitutes optimal engagement with open education. Key findings included that research-informed teaching was well-suited to open practices by openly sharing research outputs and insights. However, barriers included the privileging of research over teaching at research universities. The conclusion discussed balancing open outreach with use of open educational resources, reciprocity, and fitting open approaches to institutional objectives of equipping students for the future.
Experiences in building a PaaS Platform - Java One SFO 2012Jagadish Prasath
The document discusses Oracle's Platform as a Service (PaaS) offering and service orchestration capabilities. It describes how PaaS simplifies Java application deployment through automatic service provisioning and association. Key features include simplified single-click deployment, automatic scaling of services, and standards-based development for multiple cloud deployment models.
Stability indicating analytical method development and validation for estimat...SriramNagarajan18
Stability indicating analytical method development and validation for estimation of Sacubitril and Valsartan in bulk and pharmaceutical dosage form using RP-HPLC
Here are the key IR frequencies identified in the sample that match the reference standard of propafenone:
- C-C stretch at 1186 cm-1
- C=C stretch at 1651 cm-1
- C-H stretch (symmetric) at 2939 cm-1
- C-H bend at 1328 cm-1
- CH2 stretch (symmetric) at 1369 cm-1
- CH2 bend at 1485 cm-1
- CH3 bend at 1398 cm-1
- C-O stretch at 1100 cm-1
- C=O stretch at 1695 cm-1
- N-H stretch at 3417 cm-1
The IR spectrum
This document describes a spectrofluorimetry method for determining nalidixic acid concentrations in human plasma. The method involves extracting nalidixic acid from plasma samples using chloroform and analyzing the samples using a spectrofluorimeter. The method was used to analyze plasma samples from a study comparing the bioavailability of a new generic nalidixic acid tablet formulation to a standard brand name formulation. Results of in vitro tests like dissolution and assay showed the new generic formulation was comparable to the standard. Pharmacokinetic analysis of plasma concentration data from subjects who received the formulations found no significant differences between the products.
Shraddha roll no- 7 -m. pharm final presentation ---rbvrr college of pharmacysaathiyaa
The document describes the development and validation of an analytical method for ziprasidone using reverse phase high performance liquid chromatography. The method utilizes a Sunsil C18 column with a mobile phase of water and methanol (55:45) at a flow rate of 1 mL/min. Ziprasidone was detected at 261 nm with a retention time of 3.082 minutes. The method was validated for specificity, precision, linearity, accuracy, limit of detection, limit of quantification and robustness as per ICH guidelines. The developed and validated method can be used for the analysis of ziprasidone in bulk and pharmaceutical formulations.
Multiple Method Development and Validation for Simultaneous Estimation of Chl...ijtsrd
A simple, precise and accurate multiple analytical method has been developed for the simultaneous estimation of Chlorzoxazone and Nimesulide in bulk and tablet formulations by reversed-phase liquid chromatographic and UV-Visible spectrophotometric techniques. The chromatographic separation was achieved on C18 analytical column. A mixture of Methanol 0.1 Ortho-phosphoric acid 75 25 was used as mobile phase, at a flow rate of 1mL min and detection wavelength at 295 nm. The retention time of Chlorzoxazone and Nimesulide was found to be 4.69 and 5.45 min respectively. The linear dynamic ranges for HPLC were from 2-10 µg mL and for simultaneous equation method, derivative spectroscopy, Q-ratio Absorbance method, Dual wavelength it was 10-30 µg mL for both Chlorzoxazone and Nimesulide. The percentage recovery obtained for Chlorzoxazone and Nimesulide were 100.93 and 102.19 respectively for RP-HPLC, 9.7 and 100.1 for simultaneous equation method of CZ and NIM respectively, 99.97 and 99.78 for derivative spectroscopy of CZ and NIM respectively, 101.37 and 99.48 for Q-ratio Absorbance method of CZ and NIM respectively, 100.13 and 99.96 for dual wavelength method of CZ and NIM respectively. The validation of the proposed methods were carried out for linearity, accuracy, precision, limit of detection, limit of quantitation and robustness. The developed method can be used for routine quality control analysis of titled drugs in combination in tablet formulation. Swetha Yarramsetti | A. Elphine Prabahar | Rama Rao Nadendla "Multiple Method Development and Validation for Simultaneous Estimation of Chlorzoxazone and Nimesulide in Bulk and Pharmaceutical Dosage Form" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-2 , February 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21503.pdf
Paper URL: https://www.ijtsrd.com/pharmacy/analytical-chemistry/21503/multiple-method-development-and-validation-for--simultaneous-estimation-of-chlorzoxazone-and--nimesulide-in-bulk-and-pharmaceutical-dosage-form/swetha-yarramsetti
Method Development and Validation of Clopidogrel Bisulphate by Reverse Phase-...SriramNagarajan15
A new, simple sensitive, rapid, accurate and precise RP-HPLC method was developed for the estimation of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Clopidogrel bisulphate was chromatographed on a reverse phase C18column (150 mm x 4.5 mm, i.d 5μm) in a mobile phase consisting of acetonitrile and phosphate buffer (pH: 3.0) in the ratio of 60:40 % v/v. The mobile phase was pumped at a flow rate of 1 ml/min with detection at 224 nm. The detector response was linear in the concentration of 50-150 μg /ml. The limit of detection and limit of quantitation was found to be 1.3 and 4.2 µg/ml, respectively. The intra and inter day variation was found to be less than 2%. The mean recovery of the drug from the solution was 99.79%. The proposed method is simple, fast, accurate, precise and reproducible hence, it can be applied for routine quality control analysis of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Key words: Clopidogrel bisulphate, RP-HPLC, Validation, Accuracy, Precision.
1. A new RP-HPLC method was developed and validated for the estimation of Clopidogrel bisulphate in bulk drug and pharmaceutical formulations.
2. The method involved using a C18 column, mobile phase of acetonitrile and phosphate buffer at a ratio of 60:40, and detection at 224 nm.
3. The method was found to be linear, precise, accurate, specific, robust and suitable for the routine analysis of Clopidogrel bisulphate in quality control labs.
Validated HPLC Method for Assay and Content Uniformity Testing of Roflumilast...Ratnakaram Venkata Nadh
Roflumilast is a selective enzyme inhibitor of phosphodiesterase-4. This drug is recommended for treatment of patients suffering from
chronic-obstructive-pulmonary-disease with chronic-bronchitis. Roflumilast is not official in pharmacopoeia and the reported methods
are having high chromatographic run times. A short run time HPLC method was developed for assay and content uniformity testing to
determine the roflumilast in blend and tablets. The mobile phase consists of 10 mM sodium dihydrogen phosphate monohydrate buffer
and acetonitrile in the ratio of 45:55 v/v. The HPLC method was developed using accucore-C18 150 × 4.6 mm, 4 μm column with a flow
rate of 1.0 mL min-1, 215 nm wavelength and 10 μL injection volume with run time of 5 min. The method linearity was proved between
5.02-40.17 μg mL-1 and obtained correlation-coefficient value is 1.0000. The mean recovery of roflumilast was 100.6%. The stability
indicating nature was established and performed the validation by considering ICH Q2 (R1) recommendations.
Diazo coupling for the determination of selexipag by visible spectrophotometryRatnakaram Venkata Nadh
Aim and Objective: The aim and objective of this study were to develop a spectrophotometric method for the assay of selexipag (selective IP prostacyclin receptor agonist indicated for the treatment of pulmonary arterial hypertension) in pure and pharmaceutical formulations so that it will be an alternative quantitative method to chromatographic methods which require large quantities of organic solvents, where some are with hazardous and toxic properties. Materials and Methods: The method is based on the diazo coupling of selexipag with diazotized p-nitroaniline in alkaline medium to form a stable green-colored and water-soluble azo dye with a maximum absorption at 510 nm. Optimization of reaction conditions was carried out to get highly sensitive and stable colored complex. Results and Discussion: Beer’s law is obeyed over the concentration range of 2–12 μg/mL with a molar absorptivity of 3.33 × 104 L/mol/cm. The limit of detection was 0.35 μg/mL and limit of quantification was 1.0 μg/mL. The results demonstrated that the procedure is accurate, precise, and reproducible (relative standard deviation <2%). Conclusions: This method was tested and validated for various parameters according to the current ICH guidelines.
Validated UV Method Development for the Simultaneous Estimation of Rabeprazol...pharmaindexing
This document describes the development and validation of a UV spectrophotometric method for the simultaneous estimation of Rabeprazole sodium and Cinitapride in tablet formulations. Wavelengths of 284.5 nm and 267 nm were selected for analysis of Rabeprazole sodium and Cinitapride, respectively. The method was validated according to ICH guidelines and showed good linearity, precision, and accuracy for simultaneous analysis of the two drugs without interference from excipients. The developed UV method provides a simple, accurate, and cost-effective approach for quality control testing of Rabeprazole sodium and Cinitapride in combined tablet dosage forms.
Stability indicating rp hplc method for the determination of candisartansrirampharma
This document describes the development of an RP-HPLC method for the determination of candesartan in pure and pharmaceutical formulations. The method utilizes a C18 column with a mobile phase of phosphate buffer and acetonitrile in a 30:70 ratio at a flow rate of 1.2 mL/min. Candesartan was detected at 255 nm with a retention time of 3.089 minutes. The method was validated per ICH guidelines and demonstrated specificity, linearity from 20-60 μg/mL, precision, accuracy, ruggedness and robustness. The method was found to separate candesartan from degradation products formed under various stress conditions, indicating it is stability-indicating and can be used to conduct
Analytical Method Development and Validation of Dutasteride and Tamsulosin Hc...SriramNagarajan15
A simple,specific, sensitive,precise and reproducible Reverse Phase High Performance liquid Chromatography method has been developed for simultaneous estimation of Dutasteride and Tamsulosin Hcl. Dutasteride and Tamsulosin is Anti-hyperplasia and Anti-hypertensive drug.The determinationwas carried out byusingsymmetryC-18columnwith Methanol:0.1M Monobasic potassiumdihydrogenphosphate buffer(75:25) Adjusted the pH to 2.5 with Ortho phosporic acid as the mobile phase and with the detection wavelength of274 nmrespectively.The flow rate is 0.7 ml/min.TheRetentiontime of Dutasteride,Tamsulosin Hcl was 2.218 minand 6.599 min respectively.Linearityforthe Dutasteride and Tamsulosin Hcl were found inthe rangeof 25-75µgmand 20-60µgm respectively.The limitof quantificationforbothdrugs wasfound to be30,24µg respectively.The recoveries of Tamsulosin and Dutasteride were found to be inthe range of 99.81-99.90 %and98.00-102.00%, respectively. The proposed method was validated suitably and canbeused for routine analysis. The degradation studies indicated Dutasteride and Tamsulosin Hcl to besusceptible to neutralhydrolysis, while Dutasteride and Tamsulosin Hcl showed degradation inacid, H2O2,photolytic and inpresenceof UV radiation.The degradation productsof Dutasteride andTamsulosin Hcl inacidic and photolytic conditions were well resolved from the pure drug with significant differences in the irretention time values. This method can be successfully employed forsimultaneous quantitative analysis of Dutasteride and Tamsulosin Hcl in formulations.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
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Chlorothiazide is an organic compound used as a diuretic and antihypertensive. It acts in the distal convoluted tubule by blocking sodium-chloride co-transport. High performance liquid chromatography is described as an effective method for quantitatively analyzing chlorothiazide. Standard solutions of chlorothiazide and an internal standard are separated on an octadecyl column and measured by digital integration of peak areas to generate a calibration curve for quantification. The method is shown to accurately analyze chlorothiazide in simulated drug formulations.
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2. Author's personal copy
j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 1 4 0 e1 4 4
Available online at www.sciencedirect.com
journal homepage: www.elsevier.com/locate/JOPR
Original Article
Simple RP-HPLC method for estimation of diazepam in tablet
dosage form
A. Sruthi, P. Tejaswi, N. Thanuja, D. Sudheer Kumar, P. Vivek Sagar*
Department of Pharmaceutical Analysis, Care College of Pharmacy, Warangal, Andhra Pradesh, India
article info abstract
Article history: A simple reverse phase HPLC method was developed and validated for the determination
Received 11 September 2012 of diazepam present in pharmaceutical dosage form. A Hypersil ODS C-18 column
Accepted 4 November 2012 (250 Â 4.6 mm, packed with 5 microns) is used as stationary phase. An isocratic mode with
mobile phase consisting of acetonitrile, methanol and 1% phosphate buffer (pH-3) in ratio
Keywords: of 18:58:24 (v/v/v) at a flow rate of 1 ml/min and effluent was monitored at 232 nm.
RP-HPLC Chromatogram showed a peak of DZP at retention time of 6.23 Æ 0.002 min. The linearity
Diazepam range was found to be 2e20 mg/ml. The method was validated for linearity, accuracy,
Validation precision, limit of detection, limit of quantitation, robustness and ruggedness. Recovery of
Pharmaceutical dosage form DZP was found to be in the range of 99.4e100.3%. The limit of detection and limit of
quantitation for estimation of DZP was found to be 0.898 mg/ml and 2.72 mg/ml, respectively.
Proposed method was successfully applied for the quantitative determination of DZP in
pharmaceutical dosage forms.
Copyright ª 2012, JPR Solutions; Published by Reed Elsevier India Pvt. Ltd. All rights
reserved.
1. Introduction expensive. The present study focused on minimizing these
limitations and to develop a simple precise accurate and
Diazepam (7-chloro-1, 3-dihydro-1-methyl-5-phenyl-2H-1, economic method for estimation of diazepam in tablet dosage
4-benzodiazepin-2-one) is a benzodiazepine (BZD) generally form.
used as hypnotic, anxiolytic and muscle relaxant. Diazepam
(DZP) is also routinely prescribed as the standard first-line
treatment for acute convulsions and prolonged status epi-
lepticus.1 Several methods for the analysis of BZDs have been
reported.2 A number of chromatographic methods, such as
thin-layer chromatography (TLC)3 gas chromatography4e6 and
gas chromatographicemass spectrometry (GCeMS)7,8 have
been used in the analysis of diazepam and other 1,4-benzo-
diazopines. Several high-performance liquid chromato-
graphic (HPLC) methods have also been reported for the
determination of diazepam and other BZDs.9,10 However, all of
these methods have limitations such as long run times and/or
* Corresponding author.
E-mail address: viveksagar.p111@gmail.com (P. Vivek Sagar).
0974-6943/$ e see front matter Copyright ª 2012, JPR Solutions; Published by Reed Elsevier India Pvt. Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jopr.2012.11.029
3. Author's personal copy
j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 1 4 0 e1 4 4 141
2.4. Preparation of working standard solutions
2. Materials and methods
Working standard solutions ranging from 0.5 to 50 mg/ml were
2.1. Chemicals and reagents prepared by appropriate dilutions of the stock with distilled
water.
An analytically pure sample of diazepam was procured as
gift sample from Natco Pharma Ltd. (Hyderabad, India). 2.5. Preparation of sample solution
HPLC grade methanol was procured from E. Merck (Hyder-
abad). Liquid chromatographic grade water was obtained by Twenty tablets of diazepam hydrochloride were weighed and
double distillation and purification through Milli-Q water ground into a fine powder. A quantity of powder equivalent to
purification system. Potassium dihydrogen phosphate (AR 25 mg of diazepam was weighed and transferred into a 25 ml
grade, purity 99.5%) was procured from Qualigens. Tablet volumetric flask and was dissolved in 0.1 N HCl. The volume
formulations VALIUM (Nicholas Piramal India Ltd.) was was made up to the mark with the same. Above solution was
procured from a local pharmacy with labeled amount 5 mg suitably diluted with distilled water. From this stock, appro-
per tablet. priate dilution (10 mg/ml) was prepared. The solution thus
prepared was filtered through 0.45 m membrane filter and the
2.2. Instrumentation & chromatographic conditions resulting filtrate was sonicated for 10 min. After setting the
chromatographic conditions and stabilizing the instrument to
The HPLC analysis was performed on CYBERLAB HPLC obtain a steady baseline, the sample solution was loaded in
equipped with an LCP-100 reciprocating HPLC pump. A the 20 ml fixed e sample loop of the injection port.
manually operating Rheodyne injector with 20 mL sample
loop, a LC-UV 100 ultraviolet detector was used. Chromato- 2.6. Method development
graphic analysis was performed on a Hypersil reversed phase
C-18 column with 250 Â 4.6 mm i.d. and 5 mm particle size. Initial trial experiments were conducted, with a view to select
The mobile phase consist of acetonitrile, methanol, 1% a suitable solvent system for the accurate estimation of the
phosphate buffer (pH-3) in ratio of 18:58:24 (v/v/v) that was set drug and to achieve good retention time. The suitability of the
at a flow rate of 1 ml/min. The mobile phase was degassed mobile phase decided on the basis of the sensitivity of the
and filtered through 0.25 mm membrane filter before pumping assay, time required for the analysis, ease of preparation, and
into HPLC system. The eluent was monitored by UV detection use of readily available cost effective solvents. These include
at 232 nm. methanol-potassium dihydrogen phosphate, methanol-
ammonium acetate, acetonitrile-potassium dihydrogen phos-
2.3. Stock solutions and standards phate, acetonitrile-ammonium acetate, methanol-water. The
mobile phase consisting of acetonitrile, methanol, 1% phos-
Stock solution of diazepam (1 mg/ml) was prepared by phate buffer (pH-3) in ratio of 18:58:24 (v/v/v) that was set at
transferring 25 mg of drug in a 25 ml volumetric flask. The a flow rate of 1 ml/min was found to be optimum and further
drug is dissolved in sufficient amount of 0.1 N HCl and optimized by adjusting pH 3e4 by adding orthophosphoric acid.
finally the volume was made up to the mark with distilled The composition of acetonitrile, methanol, 1% phosphate
water. buffer in ratio of 18:58:24 (v/v/v) with pH-3 gave the best results.
Fig. 1 e Chromatogram of diazepam.
4. Author's personal copy
142 j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 1 4 0 e1 4 4
Table 1 e Linearity of diazepam. Table 2 e Precision.
S. No Concentration Peak area S. No Concentration Peak area
(ppm) (ug/ml)
1 2 7569.8 1 6 21709.4
2 4 17892.9 2 6 21627.7
3 6 23612.2 3 6 21939.6
4 8 34517.2 4 6 21710.5
5 10 40051.3 5 6 21337.9
6 12 47631.9 6 6 21929.5
7 14 54925.1 Mean 21709.1
8 16 65942.9 % RSD 1.02
9 18 72965.0
10 20 80440.3
triplicate.11 20 ml of each of standard solutions were injected
into the HPLC system to get the chromatograms. The retention
2.7. Solution stability
time, average peak areas were recorded. Calibration curve was
constructed by plotting average peak area against concentra-
In order to demonstrate the stability of both standard and
tion and regression equation was computed. The linearity
sample solutions during analysis, both solutions were analyzed
range was found to be 2e20 mg/ml. The results were shown in
over a period of 96 h at an interval of 24 h at room temperature.
Table 1. The results show that an excellent correlation exists
The results show that for solutions, the retention time and peak
between peak area and concentration of drug within the
area of diazepam hydrochloride remained unchanged and no
concentration range, regression graph is presented in Fig. 2.
significant degradation within the indicated period, this indi-
cates that both solutions were stable for 72 h.
3.2.2. Precision
The precision of method was ascertained from the peak area
response obtained by actual determination of six replicates of
3. Results and discussion a fixed amount of drug. The percent relative standard devia-
tions were calculated for diazepam and presented in the Table
3.1. Analysis of formulation 2. The precision of the method was found to be 1.02.
The sample solution was injected and a chromatogram was 3.2.3. Accuracy
recorded. The injections were repeated six times and the peak Accuracy of developed method was confirmed by doing
areas were recorded. The amount of drug present in the recovery study as per ICH norms. A known quantity of the
pharmaceutical formulation was calculated using standard pure drug was added to the pre-analyzed sample formulation
calibration curve (concentration in mg/ml was taken on X-axis (10 mg/ml) at three different concentration levels 80%, 100%
and average peak area on Y-axis). Percentage of drug present and 120% by replicate analysis (n ¼ 3). From the recovery study
in each tablet was found to be 100.2. A representative chro- it was clear that the method is very accurate for quantitative
matogram has been given in Fig. 1. estimation of diazepam hydrochloride in tablet dosage form
as all the statistical results were within the range of accep-
3.2. Validation of the assay method tance, 99.4e100.3%, which shows that there is no interference
with excipients. Percentage recovery values were calculated
3.2.1. Linearity and the results were shown in Table 3.
Different concentrations in the range of 0.5e50 mg/ml were
prepared. Each of the levels of concentration was prepared in 3.2.4. Ruggedness
Ruggedness is the degree of reproducibility of the results ob-
tained under a variety of conditions. From stock solution,
solutions containing 14 mg/ml of diazepam hydrochloride
was prepared and analyzed by two different analysts using
same operational and environmental conditions in different
Table 3 e Accuracy.
Amount of Amount of Average %
drug added drug recovered recovery
(ppm) (ppm)
8 7.592 99.4
10 9.98 99.8
12 12.096 100.3
Fig. 2 e Linearity of diazepam.
5. Author's personal copy
j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 1 4 0 e1 4 4 143
Table 4 e Ruggedness. Table 7 e System suitability parameters.
Date % Recovery Drug RT Peak Peak USP plate USP
area height count tailing
Analyst-1 Day-1 100.2%
Analyst-2 Day-2 99.8% Diazepam 6.2266 31432.2 2588 5238.42 1.1
% Deviation 0.4%
accuracy and precision. The requirements for system suit-
ability are usually developed after method development and
Table 5 e Robustness. validation has been completed. The system suitability
parameters like Theoretical plates (N), Resolution (R), Tailing
S. No Wavelength 232 Wavelength 235
factor (T) were calculated and compared with the standard
1 52156.6 54973 values to ascertain whether the proposed RP-HPLC method for
2 52700.8 52955.4 the estimation of diazepam in pharmaceutical formulations
3 50662.6 52965.9 was validated or not. The results were shown in Table 7.
4 50905.7 54751.1
5 51418.4 54821.7
6 50911.8 52886.1
Mean 51459.32 53892.22 4. Conclusion
% RSD 1.57 1.94
A convenient, rapid, accurate, precise and economical RP-
HPLC method has been developed for estimation of diazepam
experimental periods. Percentage recoveries of the replicates
in bulk and tablet dosage form. The assay provides a linear
were calculated. It is checked that the results are reproducible
response across a wide range of concentrations and it utilizes
under differences in, analysts. The results are shown in Table 4.
a mobile phase which can be easily prepared and diluent is
economic, readily available. The proposed method can be
3.2.5. Robustness
used for the routine analysis of diazepam hydrochloride in
The method was found to be robust, although small deliberate
bulk preparations of the drug and, in pharmaceutical dosage
changes in method conditions did have a negligible effect on
forms without interference of excipients.
the chromatographic behavior of the solute. The results
indicate that changing the detector wavelength had no large
effect on the chromatographic behavior of diazepam hydro-
chloride. Even a small change of mobile phase composition
Conflicts of interest
(pH 3 Æ 0.2), did not cause a notable change in the peak area of
All authors have none to declare.
the used drug for this method. The results were presented in
Tables 5 and 6.
references
3.2.6. LOD & LOQ
LOD and LOQ for diazepam were estimated by injecting
a series of dilute solutions with known concentration. The
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1 54247.2 52156.6 5. Fisher LE, Perch S, Bonfiglio MF, Geers SM. Simultaneous
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