Analytical Method Development and Validation for the Analysis of Donepezil Hydrochloride and Its Related Substances Using Ultra Perfomance Liquid Chromatography

2017 ◽  
Vol 9 (5) ◽  
Author(s):  
Mahalingam V. ◽  
Kalaivani V. ◽  
Somanathan T. ◽  
Vijayabaskar S.
Keyword(s):  
Analytical Method ◽  
Method Development ◽  
Degradation Products ◽  
Gradient Elution ◽  
Stress Conditions ◽  
Base Hydrolysis ◽  
Forced Degradation ◽  
Equilibration Time ◽  
Elution Time ◽  
Donepezil Hydrochloride

A novel, economic and time-efficient reverse-phase ultra-performance liquid chromatographic (RP-UPLC) method has been developed for the analysis of Donepezil hydrochloride in the presence of both impurities and degradation products generated by forced degradation. When Donepezil hydrochloride was subjected to acid hydrolysis, oxidative, base hydrolysis, photolytic, and thermal stress, degradation was observed only in oxidative and base hydrolysis. The drug was found to be stable to other stress conditions. Successful chromatographic separation of the drug from impurities formed during synthesis and from degradation products formed under stress conditions was achieved on a Waters Acquity C18, 50 mm x 2.1mm, 1.7µ particle size column, UV detection at 286nm and a gradient elution of Trifluoroacetic acid, Acetonitrile and methanol as mobile phase. The method was validated for specificity, precision, linearity, accuracy, robustness and can be used in quality control during manufacture and for assessment of the stability samples of Donepezil hydrochloride. Total elution time was about 6 min and equilibration time of about 2 min which allowed analysis of more than 100 samples per day. The analytical method discussed in British Pharmacopeia was pH sensitive and not compatible to LC-MS analysis but the method reported in this study is more compatible to LC-MS which will be more suitable to perform LC-MS.

Analytical Method Development and Validation for the Analysis of Nepafenac and its related substances using Ultra Performance Liquid Chromatography

2021 ◽  
pp. 4178-4184
Author(s):  
T. S. Balaji ◽  
Gavaskar. D ◽  
Somanathan T.
Keyword(s):  
Analytical Method ◽  
Method Development ◽  
Degradation Products ◽  
Gradient Elution ◽  
Ultra Performance Liquid Chromatography ◽  
Base Hydrolysis ◽  
Forced Degradation ◽  
Equilibration Time ◽  
Elution Time ◽  
Related Substances

A novel, economic and time-efficient reverse-phase ultra-performance liquid chromatographic (RP-UPLC) method has been developed for the analysis of Nepafenac in the presence of both impurities and degradation products generated by forced degradation. When Nepafenac was subjected to acid hydrolysis, oxidative, base hydrolysis, photolytic, and thermal stress, observed degradation only in oxidative and base hydrolysis. The drug was found to be stable to other stress conditions. Various method development trails were performed for the separation of drug from impurities. However, best chromatographic separation was achieved on a Waters Acquity CSHC18, 100mm x 2.1mm, 1.7µ particle size column, UV detection at 245nm, a gradient elution of Ammonium formate (pH 4.0), mixture of organic solvents (Acetonitrile, Methanol) as mobile phase for drug, its impurities and it was captured. The method was validated for specificity, precision, linearity, accuracy, robustness and can be used in quality control during manufacture and for assessment of the stability samples of Nepafenac. Total elution time was about 6.5 min and equilibration time of about 1.5 min which allowed analysis of more than 100 samples per day. The analytical method discussed in United states Pharmacopoeia was pH sensitive and compatible to LC-MS analysis.

scholarly journals Development and Validation of a Stability-Indicating Assay Method for Simultaneous Determination of Perindopril and Indapamide in Combined Dosage Form by Reversed-Phase High-Performance Liquid Chromatography

2010 ◽  
Vol 93 (1) ◽  
pp. 108-115 ◽  
Author(s):  
Hitesh Jogia ◽  
Umesh Khandelwal ◽  
Tripti Gandhi ◽  
Sukhdev Singh ◽  
Darshana Modi
Keyword(s):  
Simultaneous Determination ◽  
Stress Testing ◽  
Degradation Products ◽  
Stress Conditions ◽  
Base Hydrolysis ◽  
Assay Method ◽  
Forced Degradation ◽  
Solution Stability ◽  
Stability Indicating

Abstract An approach of forced degradation study was successfully applied for the development of a stability-indicating assay method for simultaneous determination of perindopril and indapamide in a formulation in the presence of its degradation products. The method showed adequate separation of perindopril and indapamide from their associated main impurities and degradation products. Separation was achieved on an XTerra<sup/> RP18, 5 µm, 150 4.6 mm id column at 55°C by using the mobile phase NaH2PO4 buffer (pH 2.0; 0.005 M)acetonitrile (75 + 25, v/v ) at a flow rate of 1 mL/min and UV detection at 215 nm. Comprehensive stress testing of perindopril and indapamide was carried out according to the International Conference on Harmonization (ICH) guideline Q1A (R2). The specificity of the method was determined by assessing interference from the placebo and by stress testing of the drug (forced degradation). The drug was subjected to acid hydrolysis, base hydrolysis, oxidation, dry heat, and photolysis to apply stress conditions. There were no other coeluting, interfering peaks from excipients, impurities, or degradation products due to variable stress conditions, and the method was specific for determination of perindopril and indapamide in the presence of degradation products. The method was validated in terms of linearity, precision, accuracy, specificity, robustness, and solution stability. The linearity of the proposed method was investigated in the range of 2456 µg/mL (r2 = 0.9993) for perindopril and 7.517.5 µg/mL (r2 = 0.9992) for indapamide. Degradation products produced as a result of stress studies did not interfere with the detection of perindopril and indapamide, and the assay can thus be considered stability indicating.

scholarly journals Characterization and toxicity evaluation of degradation products of febantel

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Anand A. Mahajan ◽  
Amey M. Marathe ◽  
Suvarna S. Jarande ◽  
Raghuvir Pissurlenkar ◽  
Vandana T. Gawande
Keyword(s):  
High Performance ◽  
Degradation Products ◽  
Gradient Elution ◽  
Stress Conditions ◽  
Array Detector ◽  
Forced Degradation ◽  
Photodiode Array Detector ◽  
Positive Mode ◽  
In Silico Toxicity ◽  
The Stability

Abstract Background The aim of the present work was to determine potential toxicity of degradation products of febantel generated under different stress conditions mentioned in guideline Q1A (R2) laid down by International Council for Harmonization (ICH). The stability behavior of febantel was studied by subjecting it to hydrolytic, oxidative, photolytic and thermal forced degradation conditions. Results Five degradation products (DPs) were observed which were resolved using high-performance liquid chromatography (HPLC) and characterized by LC-MS/MS using positive mode of electrospray ionization. The chromatographic separation was carried out on Hypersil® BDS C18 (150 × 4.6 mm, 5 μm) column. Optimum resolution was obtained using ammonium formate buffer (10 mM, pH 3.5) and acetonitrile programmed in gradient elution mode at 281.0 nm using photodiode array detector. Conclusion The drug was found susceptible to degradation under all the stress conditions except thermal and oxidative stress. Five major unknown degradation products DP–I, DP–II, DP–III, DP–IV, and DP–V generated under photolytic, alkali, and acidic stress condition were identified and characterized by LC-MS/MS. The drug and identified degradation products were screened for prediction of in-silico toxicity using software viz. Swiss ADME, OSIRIS Property Explorer and Pro Tox II which indicated overall no toxicological concerns. Graphical abstract

A New, Rapid and Simple RP-HPLC Method for Stability Quantification of a Protease Inhibitor in Tablets

2021 ◽  
Author(s):  
Rochele Cassanta Rossi ◽  
Josué Guilherme Lisbôa Moura ◽  
Vanessa Mossmann ◽  
Patrícia Weimer ◽  
Pedro Eduardo Fröehlich
Keyword(s):  
Quality Control ◽  
Protease Inhibitor ◽  
Degradation Products ◽  
Gradient Elution ◽  
Hplc Method ◽  
Array Detector ◽  
Forced Degradation ◽  
Control Analysis ◽  
Quality Control Laboratory ◽  
The Stability

Abstract Fosamprenavir calcium is a protease inhibitor widely used in the treatment and prevention of human immunodeficiency virus and acquired immunodeficiency syndrome. This protease inhibitor serves as a prodrug of amprenavir, offering better oral bioavailability. Although this drug was approved by the FDA in 2003, there are few methods established for quantifying the stability for quality control analysis of fosamprenavir-coated tablets. The purpose of the study was to develop and validate a method for determining the stability of fosamprenavir-coated tablets (Telzir®) that may be applied by any quality control laboratory. Chromatographic separation was performed using a Vertical RP-18 column programmed to run a gradient elution with sodium acetate buffer and acetonitrile. Flow rate was 1.2 mL min−1 for a total run time of 15 min. Ultraviolet detection was set at 264 nm and the use of a photodiode array detector in scan mode allowed selectivity confirmation by peak purity evaluation. The analyte peak was found to be adequately separated from degradation products generated during forced degradation studies. Thus, the proposed method was found to accurately indicate stability and was sufficient for routine quantitative analysis of fosamprenavir in coated tablets without interference from major degradation products and excipients.

scholarly journals Comparison of Three Complementary Analytical Techniques for the Evaluation of the Biosimilar Comparability of a Monoclonal Antibody and an Fc-Fusion Protein

2021 ◽  
Vol 9 ◽  
Author(s):  
Alice Demelenne ◽  
Arij Ben Yahia ◽  
Delphine Lempereur ◽  
Jacques Crommen ◽  
Anne-Catherine Servais ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fusion Protein ◽  
Method Development ◽  
Reference Product ◽  
Degradation Products ◽  
Stress Conditions ◽  
Size Exclusion ◽  
Mild Stress ◽  
Molecular Weight Determination ◽  
Fc Fusion Protein

In this work, a monoclonal antibody, adalimumab, and an Fc-fusion protein, etanercept, were studied and compared to one of their biosimilars. Samples submitted to stress conditions (agitation and high temperature) were used for method development. The developed methods were also applied to samples reduced by beta-mercaptoethanol to evaluate their capability to distinguish the expected species. Capillary gel electrophoresis (CGE), reversed-phase liquid chromatography (RPLC), and size-exclusion chromatography (SEC) methods coupled with UV detection were used to analyze the biopharmaceuticals. Their complementarity was investigated. For further molecular weight determination, SEC-multi angle light scattering and RPLC-quadrupole time-of-flight were occasionally used. For adalimumab, a larger amount of fragments and aggregates was observed in the biosimilar compared with the reference product. For etanercept, more related species were found in the reference product. Those three separation techniques showed good complementarity. Indeed, RPLC enabled the separation of hydrophilic and hydrophobic degradation products. CGE provided good selectivity for several adalimumab fragments, and SEC was useful for the analysis of aggregates and certain fragments that cannot be separated by the other approaches. Moreover, those formulations were submitted to mild stress conditions (30°C, 300 rpm for 4 h) that mimic shipping conditions. No additional peak was found under these conditions for the two studied biopharmaceuticals.

scholarly journals RP-HPLC assay method development for Paracetamol and Lornoxicam in combination and characterization of oxidative degradation products of Lornoxicam

2013 ◽  
Vol 19 (4) ◽  
pp. 471-484
Author(s):  
Pritam Jain ◽  
Miketa Patel ◽  
Amar Chaudhari ◽  
Sanjay Surana
Keyword(s):  
Method Development ◽  
Degradation Products ◽  
Oxidative Degradation ◽  
Assay Method ◽  
Forced Degradation ◽  
Control Analysis ◽  
Reverse Phase ◽  
Solution Form ◽  
Forced Degradation Studies ◽  
Degradation Studies

A simple, specific, accurate and precise reverse phase high pressure liquid chromatographic method has been developed for the simultaneous determination of Paracetamol and Lornoxicam from tablets and to characterize degradation products of Lornoxicam by reverse phase C18 column (Inertsil ODS 3V C-18, 250 x 4.6 mm, 5 ?). The sample was analyzed using Buffer (0.02504 Molar): Methanol in the ratio of 45:55, as a mobile phase at a flow rate of 1.5 mL/min and detection at 290 nm. The retention time for Paracetamol and Lornoxicam was found to be 2.45 and 9.40 min respectively. The method can be used for estimation of combination of these drugs in tablets. The method was validated as per ICH guidelines. The linearity of developed method was achieved in the range of 249.09 - 747.29 ?g/mL (r2=0.9999) for Paracetamol and 4.0125 - 12.0375 ?g/mL (r2=0.9999) for Lornoxicam. Recoveries from tablets were between 98 and 102%. The method was validated with respect to linearity, accuracy, precision, robustness and forced degradation studies which further proved the stability-indicating power. During the forced degradation studies lornoxicam was observed to be labile to alkaline hydrolytic stress and oxidative stress (in the solution form). However, it was stable to the acid hydrolytic, photolytic and thermal stress (in both solid and solution form). The degraded products formed were investigated by electrospray ionization (ESI) time-of-flight mass spectrometry, NMR and IR spectroscopy. A possible degradation pathway was outlined based on the results. The method was found to be sensitive with a detection limit of 0.193 ?g/ml, 2.768 ?g/ml and a quantitation limit of 0.638 ?g/ml, 9.137 ?g/ml for lornoxicam and paracetamol, respectively. Due to these attributes, the proposed method could be used for routine quality control analysis of these drugs in combined dosage forms.

scholarly journals Comprehensive Assessment of Degradation Behavior of Simvastatin by UHPLC/MS Method, Employing Experimental Design Methodology

2018 ◽  
Vol 2018 ◽  
pp. 1-17 ◽  
Author(s):  
Maja Hadzieva Gigovska ◽  
Ana Petkovska ◽  
Jelena Acevska ◽  
Natalija Nakov ◽  
Packa Antovska ◽  
...  
Keyword(s):  
Experimental Design ◽  
Design Methodology ◽  
Method Development ◽  
Ion Trap ◽  
Degradation Products ◽  
Forced Degradation ◽  
Degradation Behavior ◽  
Liquid Chromatograph ◽  
Photolytic Degradation ◽  
Experimental Design Methodology

This manuscript describes comprehensive approach for assessment of degradation behavior of simvastatin employing experimental design methodology as scientific multifactorial strategy. Experimental design methodology was used for sample preparation and UHPLC method development and optimization. Simvastatin was subjected to stress conditions of oxidative, acid, base, hydrolytic, thermal, and photolytic degradation. Using2nfull factorial design degradation conditions were optimized to obtain targeted level of degradation. Screening for optimal chromatographic condition was made by Plackett–Burman design and optimization chromatographic experiments were conducted according to Box-Behnken design. Successful separation of simvastatin from the impurities and degradation products was achieved on Poroshell 120 EC C18 50 × 3.0 mm 2.7μm, using solutions of 20 mM ammonium formate pH 4.0 and acetonitrile as the mobile phase in gradient mode. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines. Validation results have shown that the proposed method is selective, linear, sensitive, accurate, and robust and it is suitable for quantitative determination of simvastatin and its impurities. Afterwards, the degradation products were confirmed by a direct hyphenation of liquid chromatograph to ion-trap mass spectrometer with heated electrospray ionization interface. This study highlights the multiple benefits of implementing experimental design, which provides a better understanding of significant factors responsible for degradation and ensures successful way to achieve degradation and can replace the trial and error approach used in conventional forced degradation studies.

scholarly journals On the Stability and Degradation Pathways of Venetoclax under Stress Conditions

Pharmaceutics ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 639
Author(s):  
Nina Žigart ◽  
Martin Črnugelj ◽  
Janez Ilaš ◽  
Zdenko Časar
Keyword(s):  
High Resolution Mass Spectrometry ◽  
Degradation Products ◽  
Single Agent ◽  
B Cell Lymphoma ◽  
Lymphocytic Leukemia ◽  
Stress Conditions ◽  
Forced Degradation ◽  
Degradation Pathways ◽  
Orally Bioavailable ◽  
The Stability

Venetoclax is an orally bioavailable, B-cell lymphoma-2 (BCL-2) selective inhibitor, used for the treatment of various types of blood cancers, such as chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL). In this study we investigated the degradation of venetoclax under various stress conditions including acidic, basic, oxidative, photolytic and thermolytic conditions. We isolated and identified six of its main degradation products produced in forced degradation studies. The structures of the isolated degradation products were determined by using nuclear magnetic resonance (NMR) spectroscopy, high resolution mass spectrometry (HRMS) and infrared (IR) spectroscopy. Additionally, one oxidation degradation product was identified with comparison to a commercially obtained venetoclax impurity. We proposed the key degradation pathways of venetoclax in solution. To the best of our knowledge, no structures of degradation products of venetoclax have been previously published. The study provides novel and primary knowledge of the stability characteristics of venetoclax under stress conditions. Venetoclax is currently the only BCL-2 protein inhibitor on the market. In addition to single agent treatment, it is effective in combinational therapy, so future drug development involving venetoclax can be expected. A better insight into the stability properties of the therapeutic can facilitate future studies involving venetoclax and aid in the search of new similar therapeutics.

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