Stress Degradation Conditions Research Articles

Identification of major degradation products of Clorsulon drug substance including its degradation pathways by high resolution mass spectrometry and NMR

Clorsulon is an effective anthelmintic drug substance extensively used in the treatment of parasitic infestations in both cattle and sheep. An in-depth investigation of Clorsulon’s degradation products (DPs) was carried out through forced degradation study to comprehend its degradation path. A total of eight degradation products were separated under various stress degradation conditions. Structural elucidation of these DPs was conducted using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS), and their fragmentation patterns were compared to that of the parent compound. Adequate amount of DP-4 was isolated and purified by semi-preparative high-performance liquid chromatography (HPLC) methods. Subsequently, it was examined in detail using both 1D and 2D NMR (nuclear magnetic resonance spectroscopy). Most probable mechanistic pathways for the formation of degradation products under various stress degradation conditions were proposed to better understand the degradation profile. Based on the results of the stress study, Clorsulon drug substance was found to be unstable under photolytic and oxidative conditions. Understanding Clorsulon's degradation pathway is essential for determining shelf-life prediction of finished product, safety and efficacy assurance, and guiding the development of stable, high-quality formulations.

A NOVEL VALIDATED STABILITY INDICATIVE UPLC METHOD FOR RELUGOLIX FOR THE DETERMINATION OF PROCESS-RELATED AND DEGRADATION IMPURITIES

A NOVEL VALIDATED STABILITY INDICATIVE UPLC METHOD FOR RELUGOLIX FOR THE DETERMINATION OF PROCESS-RELATED AND DEGRADATION IMPURITIES

Method validation and characterization of stress degradation products of gefitinib through UPLC-UV/PDA and LC–MS/TOF studies

Gefitinib was subjected to stress degradation conditions of hydrolysis (neutral, acidic and alkaline), oxidation, photolysis and thermal stress, as per the ICH guideline Q1A(R2). A UPLC-UV/PDA based stability-indicating method was developed for the drug and validated as per ICH guidelines. The drug showed significant degradation under acidic/alkaline/neutral hydrolytic, acidic/alkaline/neutral photolytic, thermal and oxidative stress conditions. In total, seven degradation products (I-VII) were formed, which could be separated from the drug by UPLC on a Kinetex® C18 (100 mm × 4.6 mm; 2.6 μ) column using isocratic elution method. Detection wavelength was selected as 257 nm. The developed method was extended to UPLC-MS/TOF studies and accurate mass spectral data was used to identify and characterize the degradation products. Out of the seven degradation products, five products (I-II; V-VII) could be characterized through their MS/TOF spectral data. The degradation pathway of the drug leading to generation of various products was postulated and this has not been reported so far.

A Facile RP-HPLC Strategy for Sensitive Detection and Stability Study of Dacarbazine in API and in Pharmaceutical Dosage Form

The current work reflects a simple, robustic, accurate, precise, rapid and stable RP-HPLC method and validates it for quantifying Dacarbazine in API and its parenteral preparation. it is based on identification of drug in reversed phase mode using HPLC. The mobile phase consist of mixture of 960ml of 0.5% v/v glacial acetic acid, added with 0.94g of n-hexane sulphonic acid sodium salt monohydrated and 40 ml of methanol. The drug was detected at 329nm wavelength. The method was later validated as per guidelines proposed by ICH. The drug was exposed to variable stressed conditions and was analyzed by proposed method. The chromatogram reflects no inference from degradative product, excipients or impurities. However, drug under various stress degradation conditions reflects that it is highly susceptible to oxidative condition as 21.89% of drug was degraded. In acidic and alkaline conditions, the drug was degraded by 14.96% and 11.31% respectively, while thermal and photolytic condition causes least degradation, i.e. 1.08% and 1.15% respectively. The proposed method was found to be accurate, reproducible and linear. Thus can be applicable for the quantitative estimation of Dacarbazine and can be successfully applied in the quality control of API and its parenteral preparation.

Development of a validated stability-indicating HPLC- DAD method for dasabuvir and the characterization of its degradation products using LC-QToF-MS/MS

A stability-indicating HPLC-DAD method was developed and validated for the simultaneous determination of dasabuvir and its degradation products in the pharmaceutical formulation. The proposed method utilized a Symmetry® C18 (4.6 x 75 mm, 3.5 µm) column, and the mobile phase consisted of an isocratic elution of formic acid (0.1%) and acetonitrile (55:45, v/v), at a flow of 1 mL min-1; analytes were detected at 244 nm. Dasabuvir was submitted to different stress degradation conditions, such as acidic, alkaline, neutral, thermal, oxidative and photolytic, and the structural elucidation of degradation products was performed using LC-QToF-MS/MS. The HPLC-DAD stability-indicating method was validated for selectivity, linearity, limit of detection and quantification, accuracy, precision and robustness, according to ICH guidelines. Dasabuvir produced two degradation products (DP1 and DP2) from the alkaline stress conditions, which were characterized in negative ion mode. Dasabuvir was linear in the range 9.78 to 136.92 µg mL-1, and DP and DP were linear in the range 2.9 to 20.2 µg mL-1 and 1.3 to 14.9 µg mL-1, respectively. The 1 2 recovery ranged between 99.16 and 100.86%, while precision ranged from 1.02 to 2.89%. As the method can effectively separate the dasabuvir from its degradation products and quantitate them, it may be employed as a stability-indicating method for the pharmaceutical formulation.

Comprehensive study on degradation profile of firocoxib and structural elucidation of its key degradation products

Firocoxib is widely used in veterinary medicine as a non-steroidal analgesic and anti-inflammatory drug substance. Herein, a comprehensive study on the degradation profile of firocoxib was performed through force degradation studies to understand its degradation profile and characterize its major degradation products (DPs). Firocoxib drug substance was subjected to acidic, alkaline, oxidation (H2O2, KMnO4, and K2Cr2O7), thermal (solid and solution state), and photolytic (solid and solution state) stress degradation, as recommended in the ICH guidelines. Firocoxib and its major DPs were adequately separated by investigational HPLC method, which utilized a HALO C18 (100 × 2.1 mm, 2.0 µm) column. Mobile phase-A for the HPLC method is composed of 0.1% formic acid in water and mobile phase-B acetonitrile. A total of six major DPs were observed for firocoxib drug substance under these stress degradation conditions. Structural elucidation of the DPs performed using liquid chromatography-high resolution mass spectrometry and comparison of their fragmentation profile with that of the parent compound. For further structural confirmation of two major DPs, DP-2 and DP-6 were isolated and purified from the stressed samples using a preparative HPLC and analyzed by comprehensive nuclear magnetic resonance (NMR) spectroscopy studies. Most probable mechanistic pathways for the formation of DPs of firocoxib under various stress degradation conditions were postulated to understand its degradation profile. Based on the results from forced degradation, firocoxib was found to be quite stable under basic and thermal conditions, and somewhat unstable under acidic, oxidative, and photolytic conditions. The results of this study should facilitate quality monitoring and establish a stability profile of firocoxib drug substance and drug products. These results may also assist in the design and development of new formulations made with firocoxib drug substance with desired shelf life.

LC and LC-MS/MS studies for identification and characterization of degradation products of d-tubocurarine chloride

Forced degradation studies of d-tubocurarine (DTC) was carried out in hydrolytic (acidic, alkaline and neutral), thermal, photolytic and oxidative degradation conditions as per International Conference on Harmonization (ICH) guideline Q1A (R2). The present study revealed that DTC is highly sensitive to oxidative degradation conditions even at room temperature whereas the drug was found to be stable in hydrolytic, photolytic and thermal stress conditions.Separation of DTC and its four degradation products (DPs) (DP-I to DP-IV) formed during stress degradation conditions were achieved on Waters Acquity CSH C18 (1.7 µm, 2.1 mm × 100 mm) column using gradient elution with a mobile phase consisting of Eluent-A: 0.1 % Formic acid Eluent-B: acetonitrile achieved successfully. The detection was carried out at 210 nm wavelength and the flow rate was kept at 0.3 mL/min with a 5 μL injection volume.Also, a highly sensitive and robust HRMS/MS/TOF method was established for the identification and characterization of four DPs formed during the stress study. ESI +ve mode was used throughout the study for ionization of all the DPs. The degradation pathway was also established in the study that is never reported earlier.

Stability Indicating HPTLC Method for Bedaquiline Fumarate

Bedaquiline fumarate is an anti-TB treatment that was approved in 2012 and has a novel mechanism of action for fighting multidrug resistance (MDR). Bedaquiline fumarate, a diarylquinoline having significant bactericidal and sterilising activity, has been licenced by the US Food and Drug Administration,for the treatment of multidrug-resistant tuberculosis (MDR-TB).A simple, sensitive and stability indicating High Performance Thin Layer Chromatography method has been developed and validated for estimation of Bedaquiline fumarate as per ICH guidelines. Method was developed with silica gel 60 F254TLC plates using Acetonitrile: Ethyl acetate (7:3v/v) as mobile phase with densitometric detection at 229 nm. This system was found to give compact bands for Bedaquiline fumarate at (Rf 0.53 ±0.01). The linearity was observed in the range of200-1000 ng/band. Bedaquiline was exposed to different stress degradation conditions like hydrolysis at different pH, oxidation, photolytic and thermal degradation. The proposed method can used for routine quantitative determination of Bedaquiline.

Development and validation of capillary zone electrophoresis and high-performance liquid chromatography methods for the determination of oral anticoagulant edoxaban in pharmaceutical tablets.

The incidence of thrombotic complications in SARS-CoV-2 infections has become a global concern; thus, anticoagulants are an integral part of the treatment. Edoxaban (EDX) is an oral anticoagulant suitable for pharmacologic thromboprophylaxis. Herein, two novel analytical methods for EDX determination in tablets are developed and validated using capillary zone electrophoresis (CZE) and high-performance liquid chromatography (HPLC). Operating conditions such as the electrolyte's concentration and pH value, injection time, volume, and the capillary temperature, were optimized. The methods were successfully validated by establishing the linearity, intra- and inter-day precisions (relative standard deviation [%]), accuracy, and robustness. Adequate separation of excipients and degradation products of EDX generated by stress degradation conditions demonstrated the stability-indicating capability of the methods. The analytical procedures were linear in the range of 25-125µg/ml (r>0.999), with the limits of detection and quantification of 3.26 and 10.87µg/ml for CZE and 0.740 and 2.78µg/ml for HPLC. Although both methodologies are suitable for determining EDX in tablets, CZE provides a greener alternative due to low-cost analysis using less organic solvents and minimizing waste generation.

DEGRADATION KINETICS OF CARVEDILOL PHARMACEUTICAL DOSAGE FORMS (TABLETS) THROUGH STRESS DEGRADATION STUDY

Objective: The aim of the research work to monitor impurities profiling and degradation kinetics of Carvedilol Pharmaceutical Dosage Form (Tablets) through stress degradation study.
 Methods: To study impurity profiling and degradation kinetics Chromatographic condition used as, Inertsil ODS 3V column (150 mm x 4.6 mm, 5μm) with mobile phase consisting Mobile phase-A (Water, Acetonitrile and Trifluroacetic acid in the ratio of 80:20:0.1 v/v/v respectively and pH adjusted to 2.0 with dilute potassium hydroxide solution) and Mobile phase-B (Water and acetonitrile in the ratio of 100:900 v/v respectively) delivered at flow rate of 1.0 ml min-1 and the detection wavelength 240 nm. The column compartment temperature maintained at 40 °C.
 Results: Stress degradation study conducted using Acid, Alkali, Oxidation, Humidity, Thermal and Photolytic stress degradation conditions. Known, unknown and degradant impurities nature and degradation kinetics in different stressed degradation conditions were monitored through stability indicated method reverse phase HPLC method. Carvedilol molecule found sensitive to Oxidation and Alkali conditions. Impurity-A significantly increased from its not detected level.
 Carvedilol molecule found stable in Acid, Humidity, Thermal and Photolytic stress degradation condition. In all stressed conditions, mass balance was found between 95% to 105% and peak purity of carvedilol peak was found pure.
 Conclusion: Stress degradation study is required to know the degradation pathway of product and to prove the stability indicating nature of the analytical method. Study provide information pertaining to the intrinsic stability of drug product.

AN INNOVATIVE STABILITY INDICATING HPLC METHOD WITH IMPURITY PROFILING OF NIRAPARIB-AN ANTICANCER DRUG IN PHARMACEUTICAL FORMULATIONS

A novel and sensitive HPLC method has been developed, and subsequently validated for the analysis of Niraparib and its impurity 1, Impurity 1 Acyl Glucuronide, and impurity 2 in formulations. The separation of Niraparib and its impurities was achieved on Inertsil 3V ODS (250mm × 4.6mm, 5μm) as stationary phase, methanol, 20mM ammonium formate and 0.05% of triethylamine in 80:15:5 (V/V) at pH 5.5 (adjusted with formic acid) at 1.0 mL/min isocratic flow and 246 nm wavelength. In the optimized conditions, the retention time was observed at 8.10, 11.01, 13.50, and 5.18 min respectively for Niraparib, impurity 1, Impurity 1 Acyl Glucuronide, and impurity 2. The method was confirmed to be sensitive with a detection limit of 0.015, 0.045, and 0.024 µg/mL for impurity 1, Impurity 1 Acyl Glucuronide, and impurity 2 respectively. Good linear correlation was observed in the concentration level of 15–120 µg/mL for Niraparib and 0.15 – 1.20 µg/mL for the impurities studied. Stress degradation studies were carried for evaluating the effectiveness of the method for the separation of stress degradation compounds along with known impurities. based on the results, it can be confirmed that the method was stable that can resolve the degradation compounds and the analytes in the study. The % degradation of Niraparib was confirmed to be less than 10% in all the stress degradation conditions studied. Hence the method can effectively be utilized for the routine analysis of Niraparib and its impurity 1, Impurity 1 Acyl Glucuronide, and impurity 2 in the formulation as well as stability studies.

Stability-Indicating Determination of Tedizolid Phosphate in the Presence of its Active Form and Possible Degradants.

Tedizolid phosphate is an antibiotic prodrug that is metabolized into tedizolid which is used against various resistant bacterial strains. In this study, tedizolid phosphate was subjected to stress degradation conditions, namely, hydrolysis (neutral, acidic and alkaline), thermal, oxidative and photolytic ones. The prodrug was stable toward thermal and photolytic stress conditions, while it showed significant degradation upon applying oxidative and hydrolytic conditions. Two suggested chromatographic methods are described for separation and determination of tedizolid phosphate from the resulted degradation products. The first method is HPLC using Waters Xselect HSS C18 (250×4.6mm, 5μm) analytical column and mobile phase composed of phosphate buffer (50mM, pH6.5):acetonitrile (70:30, %v/v) pumped at flow rate of 1.0mL/min with UV-detection at 300nm. The second method is a TLC coupled with densitometric quantitation, precoated silica TLC-plates as a stationary phase and a mobile phase of methanol:butanol:ethyl acetate:ammonia (33%, w/v) (60:20:20:10,%v/v) were used. The chromatographed plates were scanned at 300nm. The linearity was confirmed over concentration range of 1-100μg/mL and 1-12μg/band for HPLC and TLC-densitometric methods, respectively. Both methods were found to be suitable for determination of tedizolid phosphate in pure form and in its pharmaceutical formulations.

Development and Validation of a Stability-Indicating HPLC Method for Assay of Milbemycin Oxime and Estimation of Its Related Compounds

A stability-indicating reversed-phase HPLC method for assay of milbemycin oxime (MO) and estimation of its related compounds has been developed and validated as per VICH and ICH method validation guidelines. The stability-indicating capability of this method has been demonstrated by adequate separation of all process-related impurities including all potential degradation products of milbemycin oxime (MO) that were generated using stress degradation conditions that are prescribed in ICH guideline. The stability-indicating capabilities of this method were further challenged by analyzing aged stability samples of MO active pharmaceutical ingredient (API). Chromatographic separation of MO and its related compounds was achieved using an isocratic elution at a flow rate of 0.5 mL/min on a Supelco Ascentis Express C18 column (100 mm × 3.0 mm, 2.7 µm particle size, 90 A pore size) maintained at 50 °C. The isocratic mobile phase was composed of 30% v/v of 0.05% phosphoric acid in aqueous solution and 70% v/v of a mixture of methanol and acetonitrile (6:4, v/v). UV detection at 244 nm was employed to monitor the analytes.

RP-HPLC-PDA Method Development, Validation and Stability Studies of the Novel Antineoplastic Drug Combination - Decitabine and Cedazuridine

Aim: The aim of our present work was to develop and validate a reverse phase high-performance liquid chromatography (RP-HPLC) method for the simultanious determination of Decitabine (DEC) and Cedazuridine (CED).
 Methodology: The developed method was further applied to observe the degradation of analytes under the influence of different forced degradation conditions. Analytes were resolved on C18, 250 x 4.6 mm, particle size 5 µm Xterra column, using a mobile phase combination of 0.1% Ortho Phospharic Acid buffer pH 6.5: Methanol (40:60v/v) with flow rate of 1mL/min and injection volume of 10 µL. Quantification was carried out with PDA detector at an isosbestic point of 220 nm with a linear calibration curve in the concentration range of 35-175 μg/mL for DEC and 100-500 μg/mL for CED.
 Results: Validation of the developed method was performed as per ICH guidelines viz., linearity, accuracy, precision, and robustness. The limits of detection (LOD) and the limits of quantification (LOQ) for CED were found to be 2.69 µg/mL and 8.15 µg/mL respectively. LOD and LOQ for DEC 1.55 µg/mL and 4.68 µg/mL respectively. Moreover, validated method was applied to study the degradation profile of analytes under various stress degradation conditions.
 Conclusion: The proposed method was found to be sensitive, specific and was successfully applied for the simultaneous estimation of Decitabine (DEC) and Cedazuridine (CED) in bulk drug, and tablets.

Development and Validation of a Stability Indicating LC-MS/MS Method for the Determination of Clenbuterol HCl.

Clenbuterol hydrochloride (CLT), β2 adrenergic agonist is used as a bronchodilator in the therapeutic treatment of asthma. It is important to know the stability behaviour of the drug in different degradation conditions as per ICH Q1A (R2) guidelines for safety and efficacy purpose. The main objective of the study is to develop and validate stability indicating LC-MS/MS method for the determination of Clenbuterol HCl. The separation was achieved using Phenomenex Gemini NX C18 (250*4.6 mm, 5 μ) column and the mobile phase consisting of ammonium acetate buffer (5 mM), 0.15% triethylamine (TEA), pH 7.5 with acetic acid: methanol (70:30, v/v) at flow rate 1 ml/min. The detection was done using PDA detector at 245 nm. The validation was performed as per ICH Q2 (R1) guideline. The drug was subjected to stress degradation conditions as per ICH Q1A (R2) guidelines. The significant degradation was observed in acidic (8.78%) and sunlight (liquid) (9%) condition while no degradation was observed in neutral, basic, oxidation and thermal condition. The drug and its degradation products were characterized using LC-MS/MS and the proposed degradation mechanism was communicated. The developed method was found to be stability-indicating, simple, specific, selective, sensitive, linear, accurate, robust and precise and used as a routine analysis in quality control laboratory.

Preservatives Stability: Stability Study of Parabens under Different Conditions of Stress Degradation

Preservatives are used in a wide field of application to maintain the quality of products. They are used to deal with the chemical, physical and microbiological problems and the constraints of the development of several formulations. In this work, we tested the stability of antimicrobial preservatives which are the parabens under different stress degradation conditions to evaluate their degree of effectiveness. The tested parabens were incubated in different solutions at different pH, Temperatures, light conditions, and presence or absence of sucrose in solution (create a medium similar of syrups). The HPLC was used for the assay, by a validated method for the parabens assay and the statistical analysis of the data is carried out by JMP software. The results show a direct influence of temperature and pH on the level of parabens, while the influence of light remains negligible. The increase in temperature gives a degree of immunity of parabens levels, especially with the extreme values of pH. This study is one of the first studies of forced decomposition of parabens carried out under the various conditions suggested. The results give an idea of the stability profile of the tested parabens and suggest a model of the conditions of conservation and use of these products in different domains and under different conditions.

Novel Stability-Indicating UHPLC Method Development and Validation for the Quantification of Perindopril, Amlodipine and Their Impurities in Pharmaceutical Formulations: Application of QbD Approach

We developed a novel, rapid, sensitive, and stability-indicating UHPLC method for quantification of Perindopril (PER), Amlodipine (AML), and their impurities in pharmaceutical formulations. The optimum chromatographic separation was achieved in gradient elution mode using Agilent SB C18 (50 × 3.0 mm, 1.8 μm) column. A buffer containing 0.1% perchloric acid and acetonitrile was used as a mobile phase component. The flow rate was 0.8 mL per minute and the detection wavelength at 215 nm was optimized. We also set an injection volume of 10 μL and the column thermostat at 60 °C. The findings were interesting: Resolution for PER, AML, and nine impurities is more than 1.5 for any pair of compounds. The proposed method is validated according to the ICH guidelines, and it is accurate, precise, linear, rugged, and robust. Specificity study proves the method efficiency in estimating PER, AML, and their related substances within the same run. The drug product was subjected to various stress degradation conditions and a significant amount of degradation in acid hydrolysis was observed. We proved the method robustness of chromatographic conditions by applying response surface methodology as a tool under a QbD approach and established the design space. This method is found to be stability-indicating and can be applied for analysis in quality control laboratories.

Stability-indicating capillary electrophoresis method for the simultaneous determination of metformin hydrochloride, saxagliptin hydrochloride, and dapagliflozin in pharmaceutical tablets

A capillary electrophoretic method coupled to a diode array detector (CE-DAD) was developed and validated for the simultaneous determination of metformin hydrochloride (MET), the dipeptidyl peptidase-4 (DPP-4) inhibitor saxagliptin hydrochloride (SAX), and the sodium glucose co-transporter (SGLT 2) inhibitor dapagliflozin (DAP). The proposed method was used for the determination of these drugs in binary antidiabetic combinations namely, SAX/MET, combination I, DAP/MET, combination II, and SAX/DAP, combination III. CE separation was performed on a fused silica capillary with background electrolyte consisting of 30 mM phosphate buffer (pH 6.0) with a high voltage of 30 kV, a pressure of 20 mbar, and an injection time of 40 s. The compounds were detected at 203 nm for SAX/DAP and 250 nm for MET. The method was linear in the concentration range of 10–200 µ g/mL (SAX), 1.25–50 µ g/mL (DAP), and 7.5–1000 µ g/mL (MET). Full validation of the proposed method was performed as per the ICH guidelines. The obtained errors and deviation values did not exceed 2% assessing good accuracy and precision, respectively. The stability-indicating potential of the proposed method was proved under different stress-degradation conditions. The proposed method was successfully applied to the analysis of the three binary combinations in their tablets.

Chromatographic methods development, validation and degradation characterization of the antithyroid drug Carbimazole.

The current paper reports the development and validation of stability-indicating HPLC and HPTLC methods for the separation and quantification of main impurity and degradation product of Carbimazole. The structures of the degradation products formed under stress degradation conditions, including hydrolytic and oxidative, photolytic and thermal conditions, were characterized and confirmed by MS and IR analyses. Based on the characterization data, the obtained degradation product from hydrolytic conditions was found to be methimazole-impurity A of Carbimazole as reported by the British Pharmacopeia and the European Pharmacopeia. A stability-indicating HPLC method was carried out using a Zorbax Eclipse Plus CN column (150 × 4.6 mm i.d, 5 μm particle size) and a mobile phase composed of acetonitrile-0.05m KH2 PO4 (20: 80, v/v) in isocratic elution, at a flow rate of 1 mL/min. The method was proved to be sensitive for the determination down to 0.5% of Carbimazole impurity A. Additionally, a stability-indicating chromatographic HPTLC method was achieved using cyclohexane-ethanol (9:1, v/v) as a developing system on HPTLC plates F254 with UV detection at 225 nm. The proposed HPLC and HPTLC methods were successfully applied to Carbimazole® tablets with mean percentage recoveries of 100.12 and 99.73%, respectively.

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.