ICH-recommended Stress Conditions Research Articles

Identification, Separation and Characterization of Acidic and Oxide Degradation of Atazanavir Sulphate Under ICH-Recommended Stress Conditions by HPTLC with MS/ TOF

Identification, Separation and Characterization of Acidic and Oxide Degradation of Atazanavir Sulphate Under ICH-Recommended Stress Conditions by HPTLC with MS/ TOF

Development and validation of a stability-indicating assay method for determination of metronidazole benzoate in bulk: Spectroscopic approach

A new, simple, rapid, accurate, sensitive and cost effective spectrophotometric stability indicating assay method was developed for the quantification of metronidazole benzoate in bulk and validated according to ICH guidelines using different parameters. In the present study an attempted has been made to develop an effective method which will surpass the disadvantages associate with other reported methods like tedious in use, less sensitive and costly etc. The assay method was based on the forced degradation study of metronidazole benzoate under different ICH recommended stress conditions like hydrolytic, oxidation, thermal, and photolysis. Drug shown maximum absorption at 322nm and a regression coefficient value (R2) of 0.999 indicates linearity followed within the concentration range of 5µg/ml to 50µg/ml. During forced degradation study, metronidazole was significantly degraded in alkaline conditions whereas mild degradation occurs in acidic, neutral and oxidative stress. The drug was stable to dry heat and photolytic degradation conditions. Statistical validation result of the developed method indicates that it was a simple, precise, reproducible, selective, specific and accurate method for analysis of Metronidazole and could be successfully adopted to estimate the drug in bulk and formulations.

Stability Indicating Isocratic HPLC Method for Bilastine and Characterization of Forced Degradation Products by LC-MS/MS

Abstract: The present study aims to develop and validate a simple, precise, accurate, stability indicating and isocratic reversephase high-performance liquid chromatography (RP-HPLC) method for estimating Bilastine in bulk and synthetic mixture. Bilastine from its degradation products were well separated and estimated on Discovery C8 column (250 mm x 4.6 mm, 5μm) using methanol: 0.1% ortho-phosphoric acid (55:45 %v/v) as a mobile phase and detection was performed at 276 nm by PDA detector. The degradation of Bilastine was studied under different ICH recommended stress conditions. The developed stability-indicating method was validated for system suitability, linearity, accuracy, precision, robustness, detection limit and quantitation as per ICH guidelines. The method was linear over 25-150 μg/ml. Mathematically computed Limit of Detection and Limit of Quantitation were found to be 0.19μg/ml and 0.57μg/ml, respectively. It was discovered that Bilastine degrades under acid and oxidation conditions. By using LC-MS/MS analysis, the structure of Bilastine breakdown products created amid acidic and oxidative settings was elucidated. A complete fragmentation pathway of Bilastine was established to elucidate the degradation products' structures using LC–MS/MS. The elemental concentrations of oxidation products and their fragmentation products were studied using the acquired mass values. The overall data was utilized to describe the degradation of products and their fragmentation process. The developed method can be used for routine analysis of Bilastine in dosage form as it can estimate of Bilastine in presence of excipients and degradation products.

Stability Profiling of Amodiaquine under stress Degradation Conditions

Degradation Product is an impurity which results from a chemical change in the drug substance during storage and/or manufacture of the new drug product due to some environmental changes or by reaction with an excipient and/or the immediate container closure system. Impurity development may be either during formulation or upon aging of both API’S and formulated API’S in medicines. The objective of the current investigation was to study the stress degradation behaviour of Amodiaquine under different ICH recommended stress conditions by HPLC method. The anti-malarial drug Amodiaquine, when subjected to stress degradation conditions of hydrolysis, oxidation, photolysis and thermal decomposition prescribed by ICH guideline Q1A (R2), it is found that, the drug Amodiaquine (API) was stable. The method was established using column C18 Hypersyl BDS (250* 4.6) mm, 5µ, mobile phase as monobasic potassium phosphate PH 2.5 with orthophosphoric acid and methanol. Method was isocratic and composition 22: 78 (A: B), at a flow rate of 1.2ml/ min. To facilitate the development of analytical methodology, in order to obtain a better understanding of active pharmaceutical ingredient (API) and drug product (DP) stability, and also to provide information about degradation pathways and degradation products, this process is very useful. These types of studies will provide information about the degradation products that could form during storage and transportation.

Stability indicating HPLC Method Coupled with Fluorescence Detection for the Determination of Cyproheptadine Hydro-chloride in Its Tablets. Studies on Degradation Kinetics

A sensitive, simple and rapid stability-indicating HPLC method using fluorescence detection was developed for the quantitative determination of the antihistaminic drug cyproheptadine hydrochloride (CYP). Good chromatographic separation was achieved using Shimadzu VP-ODS column (150 mm x 4.6 mm i.d., 5 μm particle size) and fluorescence detection at 410 nm after excitation 280 nm. Mobile phase consisting of methanol and 0.02 M phosphate buffer in the ratio of 65:35, v/v at pH 4.5 was pumped at a flow rate of 1 mL/min. Xipamide (XIP) was used as an internal standard (IS). The method was successfully applied to the analysis of CYP in its commercial tablets and the results were in good agreement with those obtained with the official USP method. The application of the proposed method was extended to stability studies of CYP after exposure to different ICH recommended stress conditions, such as acidic, alkaline, oxidative and photolytic degradation. Moreover, the method was utilized to investigate the kinetics of the oxidative degradation of CYP. The apparent first-order rate constant, half-life time, and activation energies of the degradation process were calculated. A proposal for the degradation pathway was presented.

Investigation of ethosuximide stability under certain ICH-recommended stress conditions using a validated stability-indicating HPLC method

An HPLC investigation of the chemical stability of ethosuximide was conducted under certain ICH-recommended stress conditions.

Study of forced degradation behavior of pramlintide acetate by HPLC and LC–MS

Pramlintide acetate (Symlin®), a synthetic analogue of the human hormone amylin. It was approved in March 2005 as a subcutaneous injection for the adjunctive treatment of patients who have type 1 or 2 diabetes mellitus. The objective of current investigation was to study the degradation behavior of pramlintide acetate under different ICH recommended stress conditions by HPLC and LC–MS. Pramlintide acetate was subjected to stress conditions of hydrolysis (acidic or alkaline), oxidation, photolysis and thermal decomposition. Extensive degradation products were observed under the hydrolysis, oxidation or thermal stress conditions, while minimal degradation was found in the photolytic conditions. Successful separation of drug from the degradation products was achieved by the validated chromatography (RP-HPLC and SCX-HPLC) methods. Subsequent to isolation, the molecular weight of each component was determined by LC–MS. The LC–MS m/z values and fragmentation patterns of 4 impurities matched with the predicted degradation products of pramlintide acetate.

Degradation Kinetics of Olopatadine HCL Using a Validated UV-Area under Curve Method

The objectives of this investigation were to establish a validated UV-area under curve (AUC) method for assay of Olopatadine HCL and to study the degradation behaviour of the drug under different ICH-recommended stress conditions. The spectral measurements were performed using AUC; 275 ↔ 320 nm with wavelength maxima at 298 nm in 0.9% w/v sodium chloride as solvent. The method was validated for linearity, precision, accuracy, robustness, specificity, and sensitivity, using sample of the bulk drug. Linear regression analysis data for the calibration plots showed a good linear relationship between response and concentration in the range of 5-100μg/mL. The drug was subjected to mild and moderate forced degradation and the degradation constant was calculated using linear as well as non-linear fit model for all the conditions. The drug was found to be less stable in acidic and photolytic degradation conditions. Photolytic degradation constants of Olopatadine in dry and wet state were 0.002488 and 0.003583 % min-1, respectively. The effect of deliberate changes in solvent strength and upper and lower wavelength were observed on AUC using 33 fractional factorial designs. The overall % RSD for robustness studies was 1.15. The utility of the method was verified by analyzing the drug in bulk form and a marketed formulation, the assay was found to be 99.04-101.5% and 99.15-99.27%, respectively. These results indicate the method can be successfully used for routine analysis of Olopatadine HCL in the bulk drug and in pharmaceutical dosage forms.

Separation, Identification, and Characterization of Degradation Products of Erlotinib Hydrochloride Under ICH-Recommended Stress Conditions by LC, LC-MS/TOF

The present research work was carried out to determine the stability of erlotinib hydrochloride (ERLO) drug under different stress conditions recommended by International Conference on Harmonization (ICH) guideline Q1A (R2). The stability of the drug was studied under hydrolytic, oxidative, photolytic, and thermal stress conditions. The drug was found susceptible to degradation under acidic, basic, and photolytic stress conditions, while it was stable under neutral, oxidative, and thermal stress conditions. A total of three degradation products (DPs) were formed. Separation was carried out by using high-performance liquid chromatography system (HPLC). Better separation was achieved on Kromasil®(150 mm × 4.6 mm, 5 µm) C18 column using gradient elution program. A mixture of stressed samples was subjected to LC-MS/TOF studies to obtain mass spectral data. The information obtained from accurate mass study was first utilized to build a complete mass fragmentation pathway of the drug, and later it was used for its degradants. Structures were proposed for each fragment based on the best possible molecular formula. Three novel DPs 6-(2-hydroxyethoxy)-7-(2-methoxyethoxy)quinazolin-4-amine, 6,7-bis(2-methoxyethoxy)quinazolin-4-amine, and N-(3-ethynylphenyl)-6, 7-bis (2-methoxyethoxy)-2-oxy-quinazolin-4-amine were identified.

Stability study of the antihistamine drug azelastine HCl along with a kinetic investigation and the identification of new degradation products.

The first stability-indicating HPLC method was developed and validated for azelastine HCl (AZL). The separation of AZL from its degradation products was achieved on a C18 column using acetonitrile-0.04 M phosphate buffer of pH 3.5 (32:68, v/v) as a mobile phase with UV-detection at 210 nm and naftazone as an internal standard. The method was rectilinear over the range of 0.2-20.0 μg mL(-1) with a detection limit of 7.05 ng mL(-1). The degradation behavior of AZL was studied under different ICH-recommended stress conditions along with a kinetic investigation; also, degradation products were identified by mass spectrometry. The method was applied for the quality control and stability assessment of AZL in eye drops and nasal spray. The obtained results were favorably compared with those obtained by a comparison method.

A validated RP-LC method for salmeterol and fluticasone in their binary mixtures and their stress degradation behavior under ICH-recommended stress conditions

Simple, accurate, precise and fully validated analytical methods for the simultaneous determination of salmeterol xinafoate and fluticasone propionate in combined dosage forms have been developed. These drugs were exposed to thermal, photolytic, hydrolytic and oxidative stress conditions, and the stressed samples were detected by the proposed method. Additionally, pK a values of three ionizable drugs (salmeterol xinafoate, fluticasone propionate and thioridazine) were determined using by the dependence of the retention factor on pH of the mobile phase. The effect of the mobile phase composition on the ionization constant was studied by measuring the pK a in different acetonitrile-water mixtures, ranging between 50 and 65% (v/v) using LC-UV method.

Evaluation of agomelatine stability under different stress conditions using an HPLC method with fluorescence detection: application to the analysis of tablets and human plasma

A simple and highly sensitive stability-indicating HPLC method was developed and validated for the determination of the new antidepressant agent, agomelatine (AGM). Separation of AGM from its stress-induced degradation products was achieved on a BDS Hypersil phenyl column (250 mm × 4.6 mm i.d., 5 µm particle size) using methanol-0.05 M phosphate buffer of pH 2.5 (35: 65, v/v) as a mobile phase with fluorescence detection at 230/370 nm. Naproxen was used as an internal standard. The method satisfied all the validation requirements, as evidenced by good linearity (correlation coefficient of 0.9999, over the concentration range 0.4-40.0 ng/mL), accuracy (recovery average 99.55 ± 0.90%), precision (intra-day RSD 0.54-1.35% and inter-day RSD 0.93-1.26%), robustness and specificity. The stability of AGM was investigated under different ICH recommended stress conditions including acidic, alkaline, neutral, oxidative and photolytic. AGM was found to be labile to acidic and alkaline degradation and a kinetic study was conducted to explore its degradation behavior. First-order degradation rate constants and half-life times were calculated in each case. The proposed method was applied for the determination of AGM in tablets and spiked human plasma with mean percentage recoveries of 99.87 ± 0.31 (n = 3) and 102.09 ± 5.01 (n = 5), respectively. Hence, the proposed method was successfully applied for the determination of AGM in human volunteer plasma. The results were compared statistically with those obtained by a comparison HPLC method revealing no significant differences between the two methods regarding accuracy and precision.

A Validated RP–LC Method for Salmeterol and Fluticasone in Their Binary Mixtures and Their Stress Degradation Behavior under ICH-Recommended Stress Conditions

A Validated RP–LC Method for Salmeterol and Fluticasone in Their Binary Mixtures and Their Stress Degradation Behavior under ICH-Recommended Stress Conditions

Validated specific densitometric method for simultaneous estimation of telmisartan and atorvastatin in presence of degradation products formed under ICH-recommended stress conditions

Validated specific densitometric method for simultaneous estimation of telmisartan and atorvastatin in presence of degradation products formed under ICH-recommended stress conditions

A Validated Stability-Indicating RP-LC Method for the Simultaneous Determination of Amlodipine and Perindopril in Tablet Dosage Form and Their Stress Degradation Behavior Under ICH-Recommended Stress Conditions

A stability-indicating RP-LC assay method was developed for the simultaneous determination of the cardiovascular drugs amlodipine and perindopril in the presence of degradation products generated from forced decomposition studies. The developed method is applicable for the determination of related substances in bulk drugs and simultaneous assay in a tablet pharmaceutical dosage form. Separation of the drugs and their degradation products was obtained using an RP Waters Spherisorb ODS1 column (250 x 4.6 mm id, 5 pm particle size) with the mobile phase acetonitrile-water (30 + 70, v/v) containing 15 mM phosphoric acid. The pH of the mobile phase was adjusted to 5.0. A flow rate of 1.2 mL/min was used for the separations, with detection at 215 nm. The chromatographic separation was performed at a column temperature of 45 degrees C. Atenolol was chosen as the internal standard. Amlodipine and perindopril were exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions, and the stressed samples were analyzed by the proposed method. Degradation studies showed that both compounds were degraded under these stress conditions. The method was found to be stability-indicating and can be used for the routine analysis of amlodipine and perindopril in the studied combined tablet dosage form.

Validated high-performance thin-layer chromatographic method for quantitation of fluvoxamine in the presence of degradation products formed under ICH recommended stress conditions

This paper describes a sensitive, selective, precise, and stability-indicating high-performance thin-layer chromatographic method for the determination of fluvoxamine as a bulk drug and in formulation. The method uses aluminium plates precoated with silica gel 60F-254 as the stationary phase and solvent system ethyl acetatetoluene-methanol-ammonia 7:2:1:0.5 (v/v/v/v). Fluvoxamine was subjected to acid and alkali hydrolysis, oxidation, and photodegradation. The peaks of the degradation products were well resolved from that of the pure drug and had significantly different RF values. The linear regression analysis data for the calibration plots showed a good linear relationship over a concentration range of 100–1000 ng spot−1. The mean values of the correlation coefficient, slope, and intercept were 0.9992 ± 0.02, 9.8493 ± 0.347, and 386.8 ± 0.71, respectively. Statistical analysis showed that the method is repeatable, selective and can separate the drug from its degradation products and can be used to monit...

Study of the forced degradation of isoconazole nitrate in bulk drug and cream formulations

Bulk drug and cream formulations of isoconazole were subjected to the following ICH-recommended stress conditions: hydrochloric acid (0.1 M), sodium hydroxide (0.1 and 1 M), and hydrogen peroxide (3%). Photostability was monitored for 60 h in a UV chamber. Analysis of drug degradation was performed using an HPLC method that had been validated for linearity, range, precision, accuracy, limits of detection and quantification, and selectivity and robustness in accordance with the criteria established by Resolution 899 of the Brazilian National Health Surveillance Agency (ANVISA). When isoconazole cream was exposed to a light source, the isoconazole content decreased slightly, but no degradation products were formed as indicated by the absence of additional peaks in the HPLC chromatogram. The drug was found to be unstable in both the bulk form and in the cream formulation when subjected to alkaline hydrolysis conditions, exhibiting reductions of ca. 43 and 70% of the initial content, respectively. Different sample solutions that had been subjected to alkaline degradation indicated, through subsequent pH adjustments and UV analysis, the presence of an isosbestic point at 275 nm, suggestive of a reversible phenomenon in alkaline media. Reversibility by bringing the pH back to the initial value may be a valuable tool for identifying real degradation processes. In contrast to the drug's instability under alkaline conditions, isoconazole was found to be stable under both acid hydrolysis and oxidative conditions.

Validated Stability−Indicating LC Method for Estimation of Amoxicillin Trihydrate in Pharmaceutical Dosage Forms and Time-Dependent Release Formulations

The objective of this study was to establish a validated stability-indicating LC method for routine analysis of amoxicillin trihydrate in bulk drug samples, different pharmaceutical formulations, and degradation kinetics of the drug under different ICH recommended stress conditions. Chromatographic separation was achieved by a Capacel Pak C18 column with 50:50% v/v methanol-0.02 M phosphate buffer as mobile phase having pH 3.5 and flow rate of 1.0 ml/min; with UV absorbance at 229 nm. The method was validated for system suitability, linearity, precision, accuracy, robustness, specificity and sensitivity. The drug was subjected to stress degradation by exposure to acid and alkaline hydrolysis, oxidation, and photodegradation. It was observed that peaks of all degradation products were well resolved from the pure drug with significantly different retention times, which indicated the specificity and stability-indicating properties of the method. When the utility of the method was verified by analysis of the drug in marketed formulations and in-house time-dependent release tablet formulations, the assay was found to be 99.6–100.4%. Statistical analysis proves that the method is repeatable, selective, and accurate for the estimation of amoxicillin trihydrate in bulk drug samples and also in pharmaceutical formulations.

HPLC AND LC-MS STUDIES ON STRESS DEGRADATION BEHAVIOR OF LEVOCETIRIZINE AND DEVELOPMENT OF A VALIDATED SPECIFIC STABILITY-INDICATING METHOD

The objective of the current investigation was to study the degradation behavior of levocetirizine under different ICH recommended stress conditions using HPLC and LC-MS and to establish a validated stability-indicating method. The drug was subjected to stress conditions of hydrolysis, photolysis, and thermal decomposition. Extensive degradation was found to occur in photolytic stress conditions. Mild degradation was observed in acidic, alkaline, and neutral conditions. The drug was stable to thermal stress condition. Successful separation of drugs from degradation products formed under stress conditions was achieved on a C-18 Supelco® Column using water-acetonitrile (50:50%v/v) as the mobile phase. The flow rate was 1.0 mL/min and the detection wavelength was 230 nm. The degradant products were characterized by LC-MS. The method was validated with respect to linearity, precision, accuracy, specificity, and robustness. The developed method efficiently separated the drug and degradation product in actual samples.

Study of the Degradation Kinetics of Carvedilol by Use of a Validated Stability-Indicating LC Method

The objectives of this investigation were to establish a validated stability-indicating LC method for assay of carvedilol and to study the degradation behaviour of the drug under different ICH-recommended stress conditions. Chromatographic separation was achieved on a C18 column with 55:45 (%, v/v) acetonitrile–0.02 m phosphate buffer, pH 3.5, as mobile phase at a flow rate of 1.0 mL min−1; detection was by UV absorbance at 242 nm. The method was validated for linearity, precision, accuracy, robustness, specificity, and sensitivity, with the bulk drug. The drug was subjected to forced degradation and peaks of all the degradation products were well resolved from that of the pure drug, with significantly different retention times, which indicates the specificity and stability-indicating properties of the method. First-order degradation kinetics of carvedilol were observed under acidic and alkaline conditions. When the utility of the method was verified by analysis of the drug in marketed tablets and a nano-emulsion formulation, the assay was found to be 98.60–99.61 and 99.52–99.87, respectively. These results indicate the method can be successfully used for routine analysis of carvedilol in the bulk drug and in pharmaceutical dosage forms.