Stability-indicating Method Research Articles

Eco-Friendly Stability-Indicating HPLC Method for Related Compounds in Pemetrexed Ditromethamine (Antineoplastic Agent) for Injection.

An eco-friendly analytical technique was developed with the intention of preserving the environment by using green chemistry principles. Pemetrexed is a folate analogue indicated for the treatment of advanced lung cancer. Development of a green stability-indicating HPLC method for the quantification of pemetrexed ditromethamine (PDT) impurities in Active Pharmaceutical Ingredient (API) and parenteral dosage form. Chromatographic separation was achieved using a Zorbax SB C18 column (150 mm × 4.6 mm i.d., 3.5 µ particle size) with perchlorate buffer (pH 3.0 ± 0.1, 50 mM) as mobile phase A and acetonitrile-perchlorate (90 + 10, v/v) buffer as mobile phase B at a flow rate of 0.8 mL/min with a column temperature of 40°C ± 0.5°C. All analytes were well resolved by gradient elution with a total run time of 75 min. The UV detection wavelength was 230 nm. The RP-HPLC method is capable of resolving all the degradation and process impurities for PDT API and parenteral dosage form. The related compounds method was validated in accordance with International conference on harmonization (ICH) Q2(R1) and United states of Pharmacopoeia (USP) <1225> guidelines, and found to be accurate, specific, precise, linear, robust and stability-indicating. The precision and intermediate results were <5% CV for all the impurities. The accuracy for all the impurities was found to be between 90 and 110%. The linearity of regression co-efficient values for all the impurities were found to be more than 0.999. The proposed related compounds method is found suitable for the determination of process and degradation impurities of commercial formulations, stability samples in QC analysis for PDT API, and drug product. The developed liquid chromatographic method greenness and eco-friendliness were assessed using the green analytical procedure index (GAPI) and the analytical greenness (AGREE) tool, and found to be green. A PDT detoxification procedure was also developed to reduce environmental pollution.

Characterization of Degradation Products of Selpercatinib by Mass Spectrometry: Optimization of Stability-Indicating HPLC Method for Separation and Quantification of Process Related Impurities of Selpercatinib

The study aimed to investigate a novel approach by utilizing liquid chromatography in coupled with mass spectrometry (LC-MS) to resolve, analyze and characterize significantly less quantity of stress degradation products (DPs) of selpercatinib along with its process related impurities. The analytes resolved on ZORBAX Eclipse (250 mm) stationary phase that was maintained employing 0.5 M sodium perchlorate at pH 5.4 methanol and acetonitrile in 50:20:30 (v/v) pumped at 0.8 mL/min isocratic flow and 241 nm wavelength. The method produces very high correlate linear curve in 30-240 μg/mL for selpercatinib and 0.03-0.24 μg/mL for its impurities with significantly low detection limit of 0.01 μg/mL for impurities. Selpercatinib pure compound was subjected to stress studies and chromatographic results confirm the formation of four DPs, which were characterized with the interpretation of their mass fragmentation pattern. The DPs were identified as 6-hydroxy-4-(6-(6-((6-methoxypyridin3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile (DP 1), 6′-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)-1,2-dihydro-[3,3′-bipyridin]-5-ol (DP 2), (6-hydroxy-4-(6-(6-(pyridin-3-ylmethyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile (DP 3), 1-amino-6′-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)-1,2-dihydro-[3,3′-bipyridin]-5-ol (DP 4), 6-(2-hydroxy-2-methyl-propoxy)-4-(6-(3-methylpiperazin-1-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile (DP 5) and 6-(2-hydroxy-2-methyl-propoxy)-4-(6-(3-methylpiperazin-1-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile (DP 6). Based on findings, it was concluded that this method was effectively applied for the regular quantification of impurities of selpercatinib in formulations. Additionally, it demonstrated applicability for the identification of both known and unknown impurities of selpercatinib.

Stability indicating eco-friendly quantitation of terbutaline and its pro-drug bambuterol using quantitative proton nuclear magnetic spectroscopy

Quantitative 1H-NMR became an increasingly important issue in pharmaceutical analytical chemistry. This study used NMR spectroscopy to assay the bronchodilator drug terbutaline sulfate and its pro-drug bambuterol hydrochloride in pure form and pharmaceutical preparations. The technique proceeded using deuterium oxide (D2O) as an 1H-NMR solvent and phloroglucinol anhydrous as an internal standard (IS). Comparatively, to the phloroglucinol signal at 5.9 ppm, the resulting quantitative signals of the studied drugs were corrected. The terbutaline singlet signal at 6.3 ppm was chosen for quantification, while the bambuterol quantitative singlet signal was at 2.9 ppm. The two drugs were rectilinear over the concentration range of 1.0–16.0 mg/mL. LOD values were 0.19 and 0.21 mg/mL while LOQ values were 0.58 and 0.64 mg/mL for terbutaline and bambuterol respectively. The developed method has been validated according to the International Conference of Harmonization (ICH) regarding linearity, accuracy, precision, specificity, and robustness. A greenness profile assessment was applied, and the method proved to be green. The method enables the assay of the two drugs in pure drug and pharmaceutical preparations. The method also enables the assay of the two drugs in the presence of each other; thus, it is considered a stability-indicating method where terbutaline is an acid degradation product of bambuterol.

Combination of a propofol emulsion with alpha-2 adrenergic receptor agonists used for multimodal analgesia or sedation in intensive care units: a physicochemical stability study

ObjectivesTo assess the physicochemical stability of the combination of a propofol emulsion with an alpha-2 (α2) adrenergic receptor agonist (α2A; clonidine or dexmedetomidine) under conditions mimicking routine practice in an...

Origanum majorana L. polyphenols: in vivo antiepileptic effect, in silico evaluation of their bioavailability, and interaction with the NMDA receptor.

Introduction: Epilepsy is a chronic brain disease characterized by repeated seizures and caused by excessive glutamate receptor activation. Many plants are traditionally used in the treatment of this disease. This study aimed to evaluate the bioavailability of a polyphenolic extract obtained from Origanum majorana L. (OMP) leaves, as well as its antiepileptic activity and its potential mechanism of action. Methods: We have developed and validated a simple, rapid, and accurate stability-indicating reversed-phase liquid chromatographic method for the simultaneous determination of caffeine and quercetin in rat plasma. The OMP antiepileptic effect was evaluated with pilocarpine-induced seizures, and a docking method was used to determine the possible interaction between caffeic acid and quercetin with the N-methyl-D-aspartate (NMDA) receptor. Results and Discussion: Both compounds tested showed low bioavailability in unchanged form. However, the tested extract showed an anticonvulsant effect due to the considerably delayed onset of seizures in the pilocarpine model at a dose of 100mg/kg. The molecular docking proved a high-affinity interaction between the caffeic acid and quercetin with the NMDA receptor. Taken together, OLP polyphenols demonstrated good antiepileptic activity, probably due to the interaction of quercetin, caffeic acid, or their metabolites with the NMDA receptor.

LC/Q-TOF-MS-based structural characterization of enasidenib degradation products and establishment of a stability-indicating assay method: Insights into chemical stability.

Enasidenib (EDB) is an orally active selective mutant isocitrate dehydrogenase-2 enzyme inhibitor approved by the U.S. Food and Drug Administration to treat acute myeloid leukemia. It lacks a reported forced degradation study and a stability-indicating assay method (SIAM). This study addresses this gap by establishing a degradation profile in accordance with the International Council for Harmonisation Q1A and Q1B (R2) guidelines and developing a validated SIAM for EDB. EDB was exposed to forced degradation under various conditions (hydrolytic, photolytic, oxidative, and thermal stress). Degradation samples were analyzed using high-performance liquid chromatography on an Agilent ZORBAX Eclipse Plus C18 column with a mobile phase consisting of 0.1% formic acid in Milli-Q water and acetonitrile at a flow rate of 1mL/min and detection at 270 nm. Liquid chromatography-quadrupole time-of-flight-high-resolution mass spectrometry (LC/Q-TOF HRMS) was used for the identification and characterization of degradation products. Nitrosamine risk assessment was conducted using a modified nitrosation assay procedure (NAP) test due to the presence of a secondary amine group in the drug, which is liable to forming nitrosamine drug substance-related impurities (NDSRI). The drug exhibited significant degradation under acidic, basic, photolytic, and oxidative conditions in the solution state. A total of nine degradation products (DP) were formed (DP-I, DP-III, and DP-IV: acidic conditions; DP-I and DP-III: basic conditions; DP-II, DP-V, DP-VI, and DP-VII: oxidative stress; and DP-VII, DP-VIII, and DP-IX: photolytic conditions), which were separated and identified using reversed-phase high-performance liquid chromatography and characterized using liquid chromatography-tandem mass spectrometry. The mechanism behind the formation of EDB degradation products has been discussed, and this study was the first to develop a degradation pathway for EDB. In addition, the possibilities of NDSRI formation for EDB were studied using a modified NAP test, which can contribute to the risk assessment of the drug. Forced degradation studies were conducted by establishing a SIAM for EDB. All the degradation products were characterized by mass spectral data obtained using LC/Q-TOF-HRMS.

Development of a novel quality by design-enabled stability-indicating HPLC method and its validation for the quantification of nirmatrelvir in bulk and pharmaceutical dosage forms.

A systematic and novel quality by design-enabled, rapid, simple, and economic stability-indicating HPLC method for quantifying nirmatrelvir (NMT) was successfully developed and validated. An analytical target profile (ATP) was established, and critical analytical attributes (CAAs) were allocated to meet the ATP requirements. The method used chromatographic separation using a Purosphere column with a 4.6 mm inner diameter × 250 mm (2.5 μm). The analysis occurred at 50°C with a flow rate of 1.2 mL/min and detection at 220 nm. A 10 μL sample was injected, and the mobile phase consisted of two components: mobile phase A, containing 0.1% formic acid in water (20%), and mobile phase B, containing 0.1% formic acid in acetonitrile (80%). The diluent was prepared by mixing acetonitrile and water at a 90:10 v/v ratio. The retention time for the analyte was determined to be 2.78 min. Accuracy exceeded 99%, and the correlation coefficient was greater than 0.999. The validated HPLC method was characterized as precise, accurate, and robust. Significantly, NMT was found to be susceptible to alkaline, acidic, and peroxide conditions during forced degradation testing. The stability-indicating method developed effectively separated the degradation products formed during stress testing, underlining its effectiveness in stability testing and offering accuracy, reliability, and sensitivity in determining NMT.

Application of analytical quality by design (AQbD) for stability-indicating method development for quantification of nevirapine and its degradation products

Development of stability-indicating method by liquid chromatography are always a challenge for analytical researchers. A traditional way is by varying one chromatographic parameter at a time assessing the responses from each test. Recent international guidelines encourage the implementation of systematic approaches in analytical development, such as Analytical Quality by Design (AQbD). The aim of this paper was to develop, using the AQbD approach, an efficient and selective stability-indicating method by high performance liquid chromatography for nevirapine and its related substances. The Analytical Target Profile (ATP) and the Critical Method Attributes (CMA) were determined. Through the risk assessment studies, the high-risk analytical conditions were considered as Critical Method Parameters (CMP) and they were screened and optimized by Design of Experiment (DoE). The Method Operable Design Region (MODR) was defined using Monte Carlo Simulation. A control strategy was established by the definition of the system suitability of the analytical procedure. Optimal chromatographic conditions were achieved using an isocratic elution consisting of a mixture of 5 mM ammonium formate buffer pH 5.4/acetonitrile/methanol (60:20:20), with a flow rate of 1.0 mL/min, oven temperature of 30 °C, injection volume of 5 µL and an ACE C18 250 mm x 4,6 mm x 5 µm chromatographic column. Finally, analytical validation and forced degradation studies confirmed the performance of the analytical method. From the estimation of measurement uncertainty, strategies were outlined so that the method is fit for purpose throughout its life cycle. Additionally, guard bands were defined, changing the specification for nevirapine quantification from 90 % to 100 % to the acceptance zone of 92.1–107.9 %. The AQbD method development provided strong justifications for the selection of important parameters for nevirapine method during regulatory filling. Furthermore, with the pass/fail decision rule using guard band, restricted acceptance zones were defined, reducing the risk to the consumer of taking a drug product whose batch was inadequately approved.

A simple Stability-Indicating UV Spectrophotometric Method developed and Validation for Detection of Paracetamol in (liquid, solid) Dosage form and Biological fluid: Application and Comparison

A simple Stability-Indicating UV Spectrophotometric Method developed and Validation for Detection of Paracetamol in (liquid, solid) Dosage form and Biological fluid: Application and Comparison

A fast, sensitive, greener, and stability-indicating HPLC method for the standardization and quantitative determination of chlorhexidine acetate in commercial products

A fast, sensitive, greener, and stability-indicating HPLC method for the standardization and quantitative determination of chlorhexidine acetate in commercial products

Stability-indicating HPLC method optimization using quality by design with design of experiments approach for quantitative estimation of organic related impurities of Doripenem in pharmaceutical formulations

Doripenem (DM), an antibiotic used for kidney, lung, and urinary infections, underwent the development of a robust high-performance liquid chromatography (HPLC) technique using the quality-by-design approach. This method, integrating design-of-experiments, detected organic impurities in both drug substances and formulated products. Operating with a C18 analytical column in a binary gradient mode with potassium phosphate buffer (pH 6.1) and acetonitrile as mobile phases, the method ran at a flow rate of 1.20 ml/minutes, with an injection volume of 20-μl and a column temperature of 45°C. Stability testing under various conditions, including hydrolysis, oxidation, heat, humidity, and light exposure, confirmed the method’s reliability without interference. Validation studies, compliant with ICH guidelines, revealed quantitation limits of 0.006%, linearity between 0.060 and 1.800 μg/ml (R2 &gt; 0.999), and recoveries ranging from 96.8% to 99.1%. This HPLC method was effectively used for stability assessment in quality control testing, assessing doripenemic acid, doripenem assay, and organic impurities.

Liquid Chromatographic Peak Purity Assessments in Forced Degradation Studies: An Industry Perspective

This review article discusses scientific rationales and current best practices in the pharmaceutical industry for performing chromatographic peak purity assessments (PPA). These activities are associated with the development and validation of liquid chromatographic (LC) stability-indicating analytical methods applicable to regulatory submissions of small-molecule drug candidates. The discussion includes a comprehensive overview of the PPA-related regulatory and scientific landscape and common industry approaches to obtain PPA results, as well as the strengths and weaknesses of PDA-facilitated ultraviolet (UV) PPA and other PPA techniques.

Isolation, identification and structural characterization of forced degradation products of mobocertinib using LC-MS/MS and NMR

The main objective of this study is to develop and validate an LC-MS/MS stability-indicating method for the isolation, identification, and characterization of forced degradation products of mobocertinib under various stress conditions, utilizing LC-MS/MS and NMR techniques. Four degradation products (DP1, DP2, DP3 and DP4) were isolated. Degradation under acidic and oxidative conditions provides DP1 and DP4, respectively. Both DP2 and DP3 were obtained under basic conditions. Chromatographic development and separation were achieved using an ODS C18 column (Chemsil) with a constant flow rate of 1.0 ml/min. The mobile phase consisted of 40 % water containing 0.1 % formic acid in methanol, and detection was performed at a wavelength of 225 nm. The application of LC-MS/MS technology was utilized to separate the above degradation products. The LC-MS/MS method has validated which includes specificity, linearity, LOD and LOQ, precision, accuracy and robustness. The chemical structure of the degradation products were characterized by 1H NMR and mass spectroscopy. The above listed degradation impurities are novel and not listed in the literature. In conclusion, this study provides vital information about mobocertinib’s chemical stability and helpful in further drug development process.

Quality Control Analysis of Neomycin, Gramicidin and Triamcinolone in Ointment Formulation using HPLC Methodology: Methodology Development, Validation and Application

The combination of gramicidin (GMN), a bactericidal agent; neomycin (NMN), an aminoglycoside; and triamcinolone (TAA), a glucocorticoid, is used for treating eye infections that are accompanied by inflammation. The contents of GMN, NMN, and TAA in eye ointments must be monitored for quality. In this research, a quick, selective, and robust stability-indicating HPLC method has been developed for the concurrent assay of GMN, NMN, and TAA in bulk and ointment formulations. Analysis was performed using ODS, a250mm × 4.6mm column and mobile phase having 0.1% aqueous formic acid-acetonitrile with a ratio of 20:80 v/v; PDA analysis was performed at 225nm. The elution times for GMN, NMN, and TAA were 2.597min, 4.440min, and 3.251 min, respectively. The GMN, NMN, and TAA linear ranges were 0.625 µg/ml to 3.75 µg/ml, 6.25 µg/ml to 37.50µg/ml and 2.5µg/ml to 15.0µg/ml, respectively. The method is precise with 0.68-0.96% (GMN), 0.52-1.02% (NMN), and 0.26-0.62% (TAA) RSD values. The method is accurate with 98.75-101.25% (GMN), 98.61-100.33% (NMN), and 98.67-100.23% (TAA) recovery values. In degradation investigation, the degradant’s peak elution times are different from the elution times of GMN, NMN, and TAA. Thus, the proved specificity and stability indicating the power of the method. Finally, this developed analytical approach was efficaciously applied to a commercial ointment formulation containing afixed dose of GMN, NMN, and TAA, demonstrating its usefulness for quality control and degradation investigations on GMN, NMN, and TAA.

Development of stability-indicating assay method and liquid chromatography-quadrupole-time-of-flight mass spectrometry-based structural characterization of the forced degradation products of alpelisib.

The US Food and Drug Administration and the European Medicines Agency approved alpelisib in 2019 for the treatment of metastatic breast cancer. A thorough literature review revealed that a stability-indicating analytical method (SIAM) is not available for the quantification of alpelisib and its degradation products (DPs). In this study, per the comprehensive stress study recommended by the International Council for Harmonisation (ICH), alpelisib was exposed to hydrolysis, oxidation, photolysis, and thermal stress. Degradation of the drug was observed under hydrolysis, oxidative, and photolysis conditions, whereas the drug was stable under thermal stress condition. We developed a SIAM for the separation of alpelisib and its major DPs that were formed under different stress conditions. The validation of the developed method was performed per ICH Q2(R1) guidelines. Five DPs were identified and characterized. Structure elucidation of all DPs was performed with the modern characterization tool of liquid chromatography-quadrupole time-of-flight mass spectrometer (LC-Q-TOF-MS/MS). The degradation pathway of the drug and its mechanisms were outlined, and in silico toxicity prediction was performed using the ProTox-II tool.

Characterization of Forced Degradation Products of Netarsudil: Optimization and Validation of a Stability-Indicating RP-HPLC Method for Simultaneous Quantification of Process-Related Impurities.

The aim of this study is to examine resolution, identification, and characterization of forced degradation products of netarsudil by liquid chromatography-tandem mass spectrometry by validating a simple and sensitive high-performance liquid chromatography method for the resolution, identification, and quantification of two process-related impurities in netarsudil. Chromatographic separation was accomplished on a ZORBAX Eclipse XDB C18 (250 x 4.6 mm; 5 µ id) column at room temperature as the stationary phase and 257 nm as the detector wavelength with the mobile phase consisting of acetonitrile, methanol, and pH 4.6 phosphate buffer in 45:35:20 (v/v) at 1.0 mL/min flow rate in isocratic elution. The method reported very sensitive detection limits of 0.008 µg/mL for impurity 1 and 0.003 µg/mL for impurity 1. The method produces a calibration curve linear in the concentration level of 25-200 for netarsudil and 0.025-0.2 µg/mL for impurities. The proposed method gives acceptable results for other validation parameters such as accuracy, precision, ruggedness, and robustness. The drug was subjected to various stress conditions such as acid, base, peroxide, and thermal and ultraviolet light to investigate the stability-indicating ability of the method. Considerable degradation was observed in stress studies, and the degradation products were well resolved from process-related impurities. The characterization of degradation products was performed on the basis of collision-induced dissociation mass spectral data, and the possible structures of the six degradation compounds of netarsudil were proposed. The outcomes of other validation studies were likewise satisfactory and proven adequate for the regular analysis of netarsudil and its process-related impurities in bulk drug and pharmaceutical dosage forms and can also be applied for the evaluation of the stress degradation mechanism of netarsudil.

Stability-Indicating Liquid Chromatography Method for Rifaximin and LC-MS Characterization of its Potential Degradants

Rifaximin (RFX) is a structural analog of rifampin that has been shown to be a gastrointestinal-selective antibiotic with broad-spectrum antibacterial properties, a pronounced safety profile, and limited drug interactions. Crohn’s disease, diarrhea, hepatic encephalopathy, irritable bowel syndrome, and traveler’s diarrhea caused by non-invasive diarrheagenic Escherichia coli, can all be treated with rifaximin. FDA has designated RFX as an orphan drug for the treatment of hepatic encephalopathy. There is a substantial need for analytical quality monitoring of Rifaximin to ensure the safety and efficacy of treatment. To generate degradants, forced degradation study of the drug was carried out in acid, alkali, oxidative, thermal, and photolytic stress conditions. The optimum separation of the drug, and its degradants was achieved on an Inertsil ODS-3V C18 column (250 X 4.6 mm X 5 μm) under gradient elution conditions using, 10 mM potassium dihydrogen orthophosphate (pH 5.0 ± 0.05) as mobile phase A, and acetonitrile as mobile phase B, at a flow rate of 1.0 mL/min. The run time was 41 minutes and the detection wavelength was 258 nm. The method was validated in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human use Guidelines. The drug was stable in neutral hydrolysis, photo and dry heat degradation conditions. The percentage degradation observed during acid, alkali hydrolysis and oxidative stress conditions were 70.46, 15.11 and 24.18, respectively. The degradants were investigated by LC-MS which indicated the presence of four (m/z 784.2, 744.5, 784.3, 753.8), three (m/z 744.5, 784.3, 753.8) and five (m/z 772.4, 838.4, 744.5, 753.8, and 801.9) degradants in acid, alkali and oxidative stress conditions, respectively. The structures of degradants were elucidated using Liquid chromatography–mass spectrometry (LC-MS) and Liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses.

A quality-by-design evaluated liquid chromatography method development and validation for the separation and quantification of nitroxoline and its impurities.

A novel, isocratic, sensitive, stability-indicating high-performance liquid chromatography method was developed for the separation and quantification of related substances in nitroxoline (NTL). The chromatographic separation has been achieved on Inertsil ODS-3V, (250 × 4.6mm, 5μm) at 240nm using ethylenediamine tetraacetic acid buffer and methanol in the ratio of 60:40 v/v as mobile phase. The performance of the method has been checked as per the International Conference on Harmonization guidelines for specificity, linearity, accuracy, precision, and robustness. Regression analysis showed a correlation coefficient value greater than 0.99 for NTL and its three impurities. The detection limit of impurities was in the range of 0.01% (0.05μg/mL)-0.22% (1.1μg/mL) indicating the sensitivity of the newly developed method. The accuracy of the method was established based on the recovery obtained between 94.7% and 104.1% for all the impurities. The percentage relative standard deviation obtained for the repeatability was less than 4.0% at the specification level for all impurities. Forced degradation was performed to establish the stability-indicating nature of the method and to know about the degradation products, the quality of a drug substance changes with time under the influence of stress conditions. Thus, the proposed method was validated and found to be specific, sensitive, linear, accurate, precise, reproducible, and beneficial for routine usage.

Quality by design tool evaluated green stability-indicating UPLC content determination method for the Olanzapine and Samidorphan dosage form

A green, environmentally friendly, quick, sensitive, cost-effective, stability-indicating, and analyst-friendly UPLC method for determining the content of Olanzapine and Samidorphan tablets formulation was developed since no method was reported in published articles. Waters CHS (50 × 2.1) mm, 1.7 um column, flow rate of 0.3 mL/min, injection volume of 1.0 µL, column temperature of 30 °C, detection wavelength of 232 nm, and a total runtime of 2 min were used to quantify the analytes. The mobile phase was composed of a solution containing 0.01 % di sodium phosphate and acetonitrile in the ratio of (30:70, v/v). The optimised technique has been validated, and the results are met the regulatory criteria. Olanzapine's accuracy results found from 99.5 % to 99.8 %, whereas Samidorphan accuracy results found from 99.4 % to 100.4 %. Olanzapine's precision and intermediate precision %RSD results found from 0.3 % to 0.7 %, whereas Samidorphan found from 0.5 % to 0.7 %. The correlation coefficient value for both analytes was found to be greater than 0.999 for the linearity range from 2.5 µg/mL to 15.0 µg/mL. Solution stability established for 24 hrs and filter compatibility studied. By studying forced degradation, the method stability indicating nature is proven. The method's robustness was assessed by utilising a design expert software tool. The method greenness was evaluated by using National method environmental index (NEMI), Green analytical procedure index (GAPI), Analytical greenness (AGREE) tools and Analytical Eco-Scale. AGREE value found 0.66 and eco-scale value found 81.

Develop and Validate a Stability-Indicating HPLC Assay Method for Paracetamol, and parabens in Pharmaceutical Dosages

The drug's stability, which is a prerequisite for good drug quality, cannot be simply established unless a reliable and trustworthy method for demonstrating stability is applied. Creating such a method is a challenging task. In this work, we developed and validated a reversed-phase stability indicating liquid chromatographic technique with photo diode-array detection to analyze paracetamol (PCM), methylparaben (MP), propylparaben (PP), and their degradants in commercial oral suspension. The compounds and their degradants were separated using a Phenomenex C8 column (250 mm x 4.6 mm, 5 µm) and a mobile phase of pH 4 consisting acetonitrile, potassium di-hydrogen ortho-phosphate, tri-ethylamine, and glacial acetic acid in a 40:58:1.5:0.5 ratio, at 1.2 mL/min. All chemicals separated in within 12 min. The developed method followed the guidelines of the International Conference on Harmonization (ICH) and fulfilled all its requirements in terms of linearity, recovery, reproducibility, detection, and quantitation limits. The dependability of the developed method was shown by recoveries of 98.88% to 101.80% in 102 samples from 34 batches, indicating that it can be used in quality control step for paracetamol, methylparaben, and propylparaben in oral suspension products.