Stability-indicating Assay Method Research Articles

Testosterone Therapy for Late-Onset Hypogonadism: A Clinical, Biological, and Analytical Approach Using Compounded Testosterone 0.5-20% Topical Gels.

Testosterone is integral to men's sexual and overall health, but there is a gradual decline in the ageing male. The topical administration of testosterone is a valuable option as a supplement (replacement) therapy to alleviate hypogonadal symptoms. The clinical efficacy of a compounded testosterone 5% topical gel was assessed retrospectively in a male patient in his seventies by evaluating the laboratory testing of the serum total testosterone and the results of a validated androgen deficiency questionnaire. After treatment, the patient's hypogonadal symptoms improved and the serum total testosterone level achieved was considered clinically optimal. The skin permeation of the testosterone topical gel (biological testing) was evaluated in vitro using the Franz finite dose model and human cadaver skin, and it is shown that testosterone can penetrate into and through ex vivo human skin. Testosterone therapy is often prescribed for extended periods, and consequently, it is crucial to determine the beyond-use date of the compounded formulations. The analytical testing involved a valid, stability-indicating assay method for compounded testosterone 0.5% and 20% topical gels. This multidisciplinary study shows evidence supporting topically applied testosterone's clinical efficacy and the compounded formulations' extended stability. Personalized, topical testosterone therapy is a promising alternative in current therapeutics for hypogonadal patients.

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 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.

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.

Investigation of aclidinium bromide degradation by stability-indicating HPLC methods, characterization of impurities by NMR, and identification of degradation products by LC-MS

Aclidinium bromide (ACL) is a long-acting muscarinic receptor antagonist used for the long-term treatment of chronic obstructive pulmonary disease (COPD). The aim of this study was to investigate the degradation of aclidinium bromide under stress and stability testing conditions, for which we developed and validated the first stability-indicating, specific, precise, accurate, and robust assay and related substances HPLC methods. Nine of the compounds used as reference standards were synthesized and fully characterized by 1H and 13C NMR, MS, and FTIR techniques. Two of these molecules, namely ACL-dimer and ACL-desphenyl, are novel compounds and reported herein for the first time. Hydrolysis of aclidinium resulted in major degradation via the formation of ACL-desDTG and DTGA metabolites. ACL-desphenyl and phenol were observed only under oxidative conditions at very low levels (< 0.10%), while ACL-hydroxy, known as a metabolite of aclidinium and confirmed by LC/QDa and LC/Q-TOF m/z data, formed under oxidative stress-testing conditions, UV light, and daylight. The identification of two impurities formed only when aclidinium bromide was treated with hydrogen peroxide was done by LC/QDa and LC/Q-TOF studies and the novel structures were proposed as ACL-bromophenoxy and ACL-bromothiophenyl formed via bromination of the phenyl and thiophene ring on the aclidinium, respectively. The stress and photostability testing studies showed that the aclidinium bromide drug substance is not sensitive to elevated temperature (105 °C, 10 days), slightly sensitive to daylight and UV-radiation, and it showed significant degradation under all hydrolysis and oxidizing conditions. The related substances HPLC method reported herein is also capable to monitor the potential genotoxic impurity 3-bromopropoxybenzene (3-BPB), a class 2 impurity according to ICH M7, and ensures that the impurity remains below the threshold of toxicological concern (TTC) limit, making it safe for patients.

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.

Analytical Method Development, Validation and Forced Degradation Study of Dapagliflozin by RP-HPLC.

Worldwide, it is projected that 285 million individuals have diabetes, and by 2030, this number is expected to climb to 438 million. About 90% of cases of diabetes mellitus are type 2 (T2DM). Insulin sensitizers, such as metformin and thiazolidinediones; insulin secretagogues, such as sulfonylureas and glinides; dipeptidyl peptidase 4 (DPP-4) inhibitors; glucosidase inhibitors, or oral combination therapy are currently available treatments for type 2 diabetes. Some of these drugs exhibit serious limitations; thus, it is crucial to design an innovative therapy that is efficient and depends on a new channel. In the current work, a stability-indicating reverse phase HPLC (RP-HPLC) technique was developed and subsequently validated for the detection of dapagliflozin in its API. The stability-indicating HPLC method for assay included the use of Kromasil 100-5-C8 (100 mm × 4.6 mm) column, UV detector 224 nm, mobile phase composition involving a mixture of acetonitrile:water (52:48), and a flow rate of 1.0 mL/min. ICH guidelines were followed for the method's validation. To assess the method's specificity and stability in showing characteristics, stress degradation studies were carried out. The working standard solution of dapagliflozin was exposed to 1 and 2 N HCl by refluxing 1 and 2 N NaOH with 30% hydrogen peroxide by volume and UV radiation in order to conduct a degradation study. All system suitability parameters were determined to be within the intended ranges, and the drug's retention duration was discovered to be 1.67 minutes. It was also investigated as to how the drug degraded under various circumstances. The drug was discovered to be stable under situations of photolytic, thermal, neutral, alkaline, and oxidative deterioration. The developed stabilityindicating HPLC technique was validated in accordance with ICH Q2 recommendations, and the validation parameters, such as linearity, precision, and robustness, were achieved within the approved standards. It may be concluded that this method is stability-indicating and specific, and it can be successfully applied to analyze tablet dosage forms containing dapagliflozin.

A Different Perspective on the Characterization of a New Degradation Product of Flibanserin with HPLC-DAD-ESI-IT-TOF-MSn and its Pharmaceutical Formulation Analysis with Inter-Laboratory Comparison.

Flibanserin (FLB) was firstly synthesized as an antidepressant drug; however, due to its enhancing effects on sexual activity, it was approved for treatment of hypoactive sexual desire disorder in women in 2015. The aim of this study was to develop a new and fully validated HPLC method for analysis of FLB in pharmaceutical formulations besides its degradation products, and identification of possible formation mechanisms by using HPLC-DAD-ESI-IT-TOF-MSn. The HPLC separation was achieved in an Supelco Ascentis® Express series phenyl hexyl column (100 × 4.6 mm, ID 2.7µm). The mobile phase was acetonitrile-ammonium acetate solution (50:50, v/v, 10 mM, pH 5.4) mixture, which was pumped at the rate of 0.5 mL/min. Chromatography, detection and structural identification was performed by using LCMS-IT-TOF instrument (Shimadzu, Japan). 1-(2-(4-(3-hydroxy-5-(trifluoromethyl)phenyl)piperazine-1-yl)ethyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one is proposed as a novel degradation product, with a mass of 407.1695, and a formula of C20H21F3N4O2 with a margin of error about 0.001 ppm. The developed method is applicable with 98% accuracy within 2.5-50.0 µg/mL range. The LOD and LOQ was about 500 ng/mL and 1.50 µg/mL, respectively. The transferability and variation between laboratories were tested by inter-laboratory comparison and evaluated with one-way analysis of variance. A novel FLB degradation product, which was produced under oxidative forced degradation condition was observed and identified for the first time; in addition, the formation kinetics of the degradation product, besides decomposition of FLB was studied. Furthermore, an inter-laboratory comparison was carried out and application of the proposed method on pseudo Addyi® sample was tested using both instrument configurations. A novel stability indicating assay method was developed and fully validated according to ICH (Q2)R1 for the analysis of FLB in the pharmaceutical preparations. A new degradation product was identified in the oxidative forced degradation condition and characterized using HPLC-DAD-ESI-IT-TOF-MS3. Moreover, the possible mechanism and the formation kinetic of the degradation product were revealed. In addition, the developed method was transferred to another LC-PDA instrument for inter-laboratory comparison. Finally, the current method was applied to pseudo formulation of Addy® in both instruments and ANOVA was applied for evaluation.

Green and Sustainable Analytical Chemistry-Driven Chromatographic Method Development for Stability Study of Apixaban Using Box-Behnken Design and Principal Component Analysis.

Apixaban (APX) is a novel anti-coagulant drug approved by USFDA. According to referred literature, numerous chromatographic methods such as RP-HPLC and high-performance thin-layer chromatography have been published for the stability study of APX. But these chromatographic methods have been developed using toxic organic solvents that are hazardous to the environment and unsafe for analysts. Hence, green and sustainable analytical chemistry-driven chromatographic method has been developed for the stability study of APX using safe organic solvents for the safety of analysts and the protection of the environment. APX was subjected to forced degradation for the development of a stability-indicating assay method. The method development was carried out by the implementation of chemometric and DoE approaches for minimizing solvent wastage. Principal component analysis was applied for the identification of critical method risk variables (MRVs) and method performance attributes. DoE-based response surface modelling was applied for the optimisation of critical MRVs. The greenness profile scales of published and developed chromatographic methods have been assessed by NEMI and AGREE methods for the estimation of APX. The developed method was found to be more eco-friendly and robust than the published chromatographic methods for the estimation of APX.

Validated stability-indicating RP-HPLC assay method for fesoterodine fumarate in pharmaceutical products

The goal of the current study was to develop a precise, sensitive and validated reverse phase HPLC method for determination of fesoterodine fumarate (FST). The current investigation was performed for a stability indicating method in order to obtain both assay and impurity profiles of FST. All the experiment was done on HPLC Shimadzu Prominence 20A having stainless steel ACE5 C18 column with a particle size of 5µm and a dimension of 4.6 mm X 150 mm. The mobile phase was in gradient mode with mobile phase A (0.03M ammonium acetate buffer : acetonitrile : methanol (55:30:15 v/v) pH 3.8) and mobile phase B (water) . The flow rate was 1.2 mL/min and the wavelength 208 nm was selected for detection. This method was validated for linearity and range, accuracy, precision and robustness in accordance with ICH requirements. The retention time of fesoterodine was found as 11.0 min. The main degradation product 5-hydroxymethyltolterodine (5-HMT; active metabolite of FST) and the other process impurities; aldehyde, benzylated hydroxy, tolterodine ester, diester impurities were detected easily at the retention times 3.8, 13.5, 16.7, 21.5, 37.1 minutes, respectively. It was observed that the retention times of the main peak and its impurities were not affected and the selectivity of the method was proved under the applied stress conditions. With this developed method, FST and its impurities can easily be detected. It is also stable, simple, accurate, and completely validated RP-HPLC technique for the analysis of FST and related impurities that meets with ICH standards.

Quality by Design based pareto charts responses Evaluation approach for a Validated Stability-indicating RP-HPLC assay Method for sunitinib and its process related impurities in Oral Dosage Forms

For sunitinib (SUN) and its associated impurities, a simple and rapid stability-indicating liquid chromatographic assay technique was developed. The SUN related impurities in the completed oral dose forms were detected and assessed utilizing the high-pressure liquid chromatography with help of analytical quality by design (AQbD) approach. Impurity quantification necessitates a more comprehensive approach to analytical technique generation. The pareto charts evaluation technique, that is dependent on quality by design, enables for the evaluation of many analytical aspects and their consequences with a small series of investigations. A spiked sample mixture was separated to six specified known SUN impurities as well as undefined degradation products with significant chromatographic resolution. The separation was undertaken on a column of C18 (150 mm x 4.6 mm, 3.5 µm) with a mobile phase flow volume of 1.0 ml in a minute in a gradient elution manner. The mobile phase component A was composed of 20 mM KH2PO4 (pH 7.0), while the mobile phase component B was acetonitrile. Compound detection was conducted out at 268 nm, with the column temperature kept fixed at 40 oC. Stress degradation samples were subjected to oxidation, acid, base, thermal, and photolysis consistent with the endorsements of “International Conference on Harmonization” (Q2) methodology. The established method for SUN and its associated impurities assessment was validated as stability indicating, precise, robust, specific, rugged, and accurate.

Development and Validation of Stability Indicating RP-HPLC Method for Quantitative Estimation of Enzalutamide in Enzalutamide Capsules Dosage Form

Objectives: A precise, accurate and selective stability-indicating reverse phase high performance liquid chromatographic assay method has been developed for the quantitative estimation of Enzalutamide in Enzalutamide capsules dosage form. Materials and Methods: The separation was achieved by using a stationary phase Waters X-Bridge Shield RP18 (150 x 4.6 mm, 3.5μ) and the mobile phase consisted of perchloric acid buffer and acetonitrile in the proportion of (20:80 volume/volume). The run velocity was 1.2 mL/min. Enzalutamide was identified using UV detector at the wavelength of 210 nm. Column oven temperature 25°C and sample cooler temperature 25°C and infused quantity 20 μL, run time was 15 min. Results: As there is no meddling flanked by blank and placebo at the retention time of Enzalutamide. Degradation study results were shown significant degradation was observed in acid and oxidation (peroxide) stress condition. Hence it can be concluded that Enzalutamide is sensitive to acid and oxidation. To obtain system precision, a study was conducted with six replicate injections. %RSD was estimated from the peak areas of Enzalutamide establish to be 0.55% correspondingly. The relative standard deviation for method exactitude was establish to be 0.55%. The suggested HPLC technique was linear over the range of 100.6-301.8 μg/mL, the correlation coefficient was establish to be 0.9999. The accuracy studies were shown as % recovery for Enzalutamide 50% to150% level. The limit of % recovered revealed is in the assortment of 98 and 102% and the consequences obtained were establish to be within the limits. Hence the technique was establish to be accurate. The solution steadiness of the standard and samples are stable upto 48 hr on a bench top and refrigerator (2-8°C). The method is robust for changes like flow rate and column oven temperature. Performed the filter validation for sample solution 0.45 μm PVDF and 0.45 μm Nylon filterers are suitable for filtration. The method has validated as per ICH guidelines and all the validation parameters are satisfy the ICH Q2 specification acceptance limits. Conclusion: The developed method was validated for an assortment of parameters as per ICH guidelines like accuracy, precision, linearity, specificity, system suitability, solution stability and robustness. The consequences obtained were within the acceptance criteria. So, it can be concluded that the urbanized technique is simple, precise, cost-effective, eco-friendly, and safe and can be successfully employed for the routine analysis of Enzalutamide in bulk and pharmaceutical dosage forms.

Determination of Olmesartan in Bulk and Pharmaceutical Dosage Forms through the Development and Validation of Stability-indicating RP-HPLC Method.

Angiotensin II type 1 (AT 1) receptor antagonist (angiotensin receptor blocker [ARB]) called Olmesartan medoxomil (OLM) prevents angiotensin II from acting on the renin-angiotensin-aldosterone pathway, which is a crucial factor in the development of hypertension. OLM is reported to rapidly hydrolyze into its active metabolite, Olmesartan, in plasma after oral treatment. The objective of the ongoing study was to develop an easy-to-use, precise, and reliable RP-HPLC method for the determination of Olmesartan in bulk as well as pharmaceutical dosage forms. The stability indicating HPLC method for assay includes the use of Kromasil 100-5-C8 (100 mm × 4.6 mm) column, UV detector 265 nm, and mobile phase composition was a mixture of Acetonitrile: water (70:30) and flow rate of 1.0 mL/min. ICH guidelines were followed in the method's validation. To assess the method's specificity and stability in showing characteristics, stress degradation studies were carried out. The working standard solution of Olmesartan was exposed to 0.1 N HCl at room temperature, 0.1 N NaOH at room temperature, 30 percent hydrogen peroxide by volume, and UV radiation in order to conduct a degradation study. The retention periods of the drug were found to be 1.36 and 1.47 min for standard and sample solutions, respectively. The degradation behaviour of drug under different conditions was studied. The drug was found susceptible to acidic, alkaline and oxidative conditions while it was found stable in photolytic condition. The developed stability-indicating RP-HPLC method for assay was validated as per ICH Q2 guidelines and the validation parameters such as accuracy, precision and specificity were obtained within the accepted criteria. It may be concluded that this method is stability-indicating and specific and can successfully be applied to analyze tablet dosage form containing Olmesartan.

A Stability-Indicating Assay for Tetrahydrocurcumin-Diglutaric Acid and Its Applications to Evaluate Bioaccessibility in an In Vitro Digestive Model

A simple and reliable ultra-high-performance liquid chromatographic (UHPLC) method was developed and validated for determination of tetrahydrocurcumin diglutaric acid (TDG) and applied for evaluation of its bioaccessibility. The analytical method was validated to demonstrate as a stability-indicating assay (SIA) according to the ICH Q2(R1) guidelines under various force degradation conditions including thermal degradation, moisture, acid and base hydrolysis, oxidation, and photolysis. The developed chromatographic condition could completely separate all degradants from the analyte of interest. The method linearity was verified in the range of 0.4-12 μg/mL with the coefficient of determination (r2) > 0.995. The accuracy and precision of the method provided %recovery in the range of 98.9-104.2% and %RSD lower than 4.97%, respectively. The limit of detection and quantitation were found to be 0.25 μg/mL and 0.40 μg/mL, respectively. This method has been successfully applied for the bioaccessibility assessment of TDG with the bioaccessibility of TDG approximately four fold greater than THC in simulated gastrointestinal fluid. The validated SIA method can also benefit the quality control of TDG raw materials in pharmaceutical and nutraceutical development.

Green approach for tracking the photofate of ciprofloxacin and levofloxacin in different matrices adopting synchronous fluorescence spectroscopy: a kinetic study.

First derivative synchronous fluorescence spectroscopy (FDSFS) was applied to detect and quantify either ciprofloxacin (CIP) or levofloxacin (LEV) simultaneously with their photodegradation products, where the photolytic pathway for each analyte was found to be pH dependent. Under the guidance of early published articles, the structure of the produced photolytic products could be concluded, and further related to their resultant fluorescence spectra. The proposed method was subjected to full validation procedure which enables its application in investigating the photodegradation kinetics for both drugs. The obtained kinetic parameters were in accordance with previous reports and could be linked to predict the antibacterial activity of the resultant photodegradation products. These facts prove the suitability of the suggested FDSFS to serve as a stability-indicating assay method and to trace the photofate of CIP and LEV in the ecosystem as potential contaminants. Furthermore, the greenness of the suggested analytical methodology was evaluated via 'Green Analytical Procedure Index' (GAPI), which classifies it as an eco-friendly assay. Eventually, no extraction, treatment or preparation steps were needed during all analysis steps, which renders the proposed assay an appealing tool in environmental analysis.

Multiple spectrophotometric determinations of Chlorthalidone and Cilnidipine using propylene carbonate – As a step towards greenness

Analyzing a drug based on its overlapping spectra requires the use of sophisticated equipment and more hazardous solvents, which is detrimental to ecological sustainability. There is a critical need to create a simple, unique, and cost-effective approach for detecting a mixture of compounds in a safer environment. The aim was to develop an eco-friendly, stability-indicating assay method to determine Chlorthalidone (CLD) and Cilnidipine (CIL) in bulk and tablet dosage form using four different Ultra-Violet (UV) spectrophotometric methods. The ratio difference method showed absorbance peaks at 256.01 nm, 220.87 nm, first ratio difference spectra at 267.21 nm, 288.03 nm, and second ratio difference spectra at 309.2 nm, 280.03 nm. The area under curve techniques showed an absorbance range of around 224–232 nm for CIL and 218–227 nm for CLD. Further, the spectral changes have been assessed at various conditions like acid, base, oxidation, and UV radiation, and it has been found that CLD tends to lose its spectral property by more than 45%. The method developed for two drugs has obeyed Beers law with the selected concentration range of 7–13 μg/mL for CLD and 8.75–16.25 μg/mL for CIL. The developed method was finally evaluated using four tools, and the results showed green pictographically representation in the GAPI and score near to 100 for AES and closer to 1 for AGREE indicated that the method was found to be most eco-friendly. The findings were easy to replicate, adoptive with major regulatory requirements, environmentally friendly, fast, and inexpensive to perform. This approach does not require any specific expensive equipment, and it might be inexpensive to use in the future to assess laboratory and commercial mixtures.

Method Greenness Profile Assessment to AQbD-Driven Stability Indicating HPTLC Method for Estimation of Aripiprazole Using Screening Design and Response Surface Modeling.

According to the literature review, organic solvents such as methanol, acetonitrile, toluene, and carbon tetrachloride have been used for the chromatographic analysis of aripiprazole (APZ). The green chemistry approach recommends these organic solvents are unsafe for analysts and the environment and should be avoided or minimized in chromatographic analysis. Hence, the stability-indicating assay method (SIAM) has been developed for the estimation of aripiprazole using safe organic solvents. The quality risk management was started with risk identification, which was followed by risk assessment. By the risk assessment process, seven analytical risk factors (ARFs) were found to be potentially risky for method development. Further risk analysis was done by Taguchi OA design for the study of the main effect of ARF on resolution between the peaks. Design of experiments (DoE)-based response surface modeling (RSM) was performed by central composite design. Method operable design region (MODR) was navigated for resolution between peaks more than 1.0 for risk control. After navigation of the MODR, a risk review was done by validation of the design model for SIAM. Control strategy was set for ARFs and separation was carried out on the precoated aluminum plate with silica gel 60 F254 using ethyl acetate-ethanol (8.0 + 2.0, v/v) as the mobile phase keeping 15 min saturation time. The developed method was validated as per the International Council for Harmonisation (ICH) Q2 (R1) guideline. The developed SIAM was applied for the assay of aripiprazole in its tablet, and results were found in agreement with the labeled claim. The organic solvents ethyl acetate and ethanol used in chromatographic analysis of APZ are recommended as safe organic solvents by the ICH Q3C guidelines. The method greenness profiles of developed and published methods were evaluated by national environmental method index (NEMI) and analytical greenness (AGREE) methods. The developed method was found to be safe and green for chromatographic analysis of APZ. Development of a green, robust, accurate, and precise stability-indicating HPTLC method for estimation of APZ. The quality risk management (QRM) and DoE-based analytical quality by design (AQbD) approach was implemented in support of the green analytical chemistry concept. Estimation of greenness profile of method by NEMI and AGREE methods.

Review on process validation of Amoxicillin Potassium Clavulanate Dry Syrup

Modern medicines for human use are required to comply with specific standards and regulation set forth by the concerned authorities. A simple and sensitive UV spectrophotometric method was developed and validated for the simultaneous determination of Potassium Clavulanate (PC) and Amoxicillin. As complementary of this work, an assay method for amoxicillin and potassium clavulanate mixtures was developed and validated; stress-testing and stability studies of amox/clav mixtures was carried out under specified conditions according to ICH and analyzed by using validated stability-indicating assay and related substances methods. Aim of this study was to test the validation of derivative spectrophotometric method in simultaneous determination the content amoxicillin and clavulanate potassium in dry syrup by derivative spectrophotometric method with zero crossing technique, in buffer phosphate pH 4,4-methanol (91:9) mixture.&#x0D; &#x0D;

A comprehensive forced degradation studies of Cariprazine hydrochloride using LC-HRMS/MS and in silico toxicity predictions of its degradation products

Cariprazine hydrochloride is a second-generation antipsychotic drug and has been approved by the US Food and Drug Administration and the European Medicines Agency. Herein, we have reported a stability-indicating assay method for the Cariprazine hydrochloride and characterization of its major degradation products using LC-HRMS/MS. The drug substance Cariprazine hydrochloride was subjected to acid, base, oxidation, thermal and photolytic stress degradation. A total of five novel degradation products (DP-1 to DP-5) of Cariprazine hydrochloride drug substance were formed under various acid, base and oxidative conditions. In silico toxicity of the degradation products was evaluated using Leadscope prediction platform. We successfully separated Cariprazine Hydrochloride peak from degradation products (DP-1 to DP-5) by using gradient elution on an Inertsil C18 column (150 × 4.6 mm, 5 µm). The drug substance was found to be labile to acidic, alkaline hydrolytic, and oxidative conditions and stable to photolytic and thermal stress conditions. The degradation pathways were delineated by explaining the putative mechanism of degradation in various conditions. The stability-indicating assay method was validated according to the ICH guideline and hence can be used for routine quality control and stability study analysis of Cariprazine hydrochloride in pharmaceutical industries and research laboratories.

Application of Supervised and Unsupervised Learning Approaches for Mapping Storage Conditions of Biopharmaceutical Product-A Case Study of Human Serum Albumin.

The stability of biopharmaceutical therapeutics over the storage period/shelf life has been a challenging concern for manufacturers. A noble strategy for mapping best and suitable storage conditions for recombinant human serum albumin (rHSA) in laboratory mixture was optimized using chromatographic data as per principal component analysis (PCA), and similarity was defined using hierarchical cluster analysis. In contrast, separability was defined using linear discriminant analysis (LDA) models. The quantitation was performed for rHSA peak (analyte of interest) and its degraded products, i.e., dimer, trimer, agglomerates and other degradation products. The chromatographic variables were calculated using validated stability-indicating assay method. The chromatographic data mapping was done for the above-mentioned peaks over three months at different temperatures, i.e., 20°C, 5-8°C and at room temperature (25°C). The PCA had figured out the ungrouped variable, whereas supervised mapping was done using LDA. As an outcome result of LDA, about 60% of data were correctly classified with the highest sensitivity for 25°C (Aq), 25°C and 5-8°C (Aq with 5% glucose as a stabilizer), whereas the highest specificity was observed for samples stored at 5-8°C (Aq with 5% glucose as a stabilizer).