Routine Analysis Of Drugs Research Articles

Development and Validation of FTIR Spectroscopic Method for the Quantitative Estimation of Lornoxicam in Tablet Dosage Form.

Lornoxicam is available in the market as a solid dosage form, particularly tablets. Liquid chromatography has been the recommended assay procedure for lornoxicam in various pharmacopeias. While going through the literature, it was observed that for the analysis of lornoxicam, no FTIR based method was found. The present study involves the development of a novel, rapid, superior, labor-free, non-destructive, and economic FTIR-based analytical technique for the analysis of lornoxicam in both bulk and tablet formulations. International Council validated the method for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines Q2A and Q2B in accordance with the United States Food and Drug Administration (USFDA) guidance and by United States Pharmacopoeia (USP). FTIR based method is extremely precise, quite linear, very accurate, and has adequate ruggedness. The developed method can be of potential use in the routine quantitative drug analysis of lornoxicam in the pharmaceutical industry for quality control purposes.

The use of an RP-HPLC-UV method for the analysis of oxcarbazepine in the presence of its preservatives; stability studies and application to human plasma samples

AbstractA simple, sensitive, selective, accurate and precise method was developed and fully validated for determination of oxcarbazepine (OXC) in presence of their preservatives and determination of oxcarbazepine (OXC) in human plasma. A reversed phase liquid chromatography (RP-HPLC) with UV detection techniques were applied for separation and quantification of studied drug OXC. Successful separation of the drug from methyl paraben (M.P.), propyl paraben (P.P.) and potassium sorbate (P.ST.) was achieved on a Kromasil C18 column (5 μm particle size, pore size 300 Å, l × I.D. 250 × 4.6 mm). The mobile phase that contain aqueous 0.05M potassium dihydrogen phosphate buffer (pH 7): acetonitrile, (50: 50, %v/v). The method was linear over concentration ranges 5.0–50 μg mL−1 for OXC. Bioanalytical validation of the developed method was carried out according to US-FDA guidelines and revealed a good linear relations over a range of (5.0–50), (0.5–10), (0.05–0.15), and (1.0–10) μg mL−1 for OXC, M.P, P.P, and P.ST, respectively, with a correlation coefficient (R2) of more than 0.999. Limit of detection (LOD) were 1.15, 0.03, 0.01 and 0.04 μg mL−1 for OXC, M.P, P.P, and P.ST, respectively, Intra and inter-day precisions, calculated as percentage relative standard deviation (% RSD), were lower than 2.0%. The developed method can be applied for routine drug analysis, therapeutic drug monitoring and bioequivalence studies through the analysis of plasma samples taken from blood bank.

The sensitive detection of low molecular mass peptide drugs in dried blood spots by solid-phase extraction and LC-HRMS.

Dried blood spot (DBS) technique has become a new popular topic in anti-doping field in recent years due to its advantages of sample stability and easy operation. It can be employed as a supplementary method to routine urine analysis. However, the small volume of DBS samples (usually 10-20 μL) significantly reduces the application value of this technique. Therefore, the development of sensitive detection methods for the analysis of prohibited substances in DBS is particularly important. In this study, based on the characteristics of low molecular mass peptide (LMMP) drugs, systematic optimization strategies were utilized for the first time to establish a sensitive detection method for LMMPs in DBS. Without using DMSO to enhance mass spectrometry ionization efficiency of peptides, the limits of detection (LOD) ranged between 0.05 and 3.74ng/mL, significantly better than the previously reported method (0.5-20ng/mL). This method was validated according to the guidelines of the World Anti-Doping Agency (WADA), and corresponding post-administration study was conducted, demonstrating that the method could be applied to routine analysis of LMMP drugs in DBS. Moreover, since DMSO is not involved, this method also has the potential to simultaneously detect both LMMP and small molecular drugs.

A One-Step Green Microwell Spectrophotometric Assay for the Determination of Certain New Chemotherapeutic Drug Formulations.

The formation of charge-transfer complexes (CTCs) of iodine with five chemotherapeutic drugs used for the treatment of different types of cancer has not been investigated. These drugs are olaparib, seliciclib, vandetanib, dasatinib, and tozasertib. Additionally, these drugs need an appropriate general spectrophotometric assay for their analysis in the dosage forms regardless of the differences in their chemical structures. The aim of this study was the development of a novel microwell spectrophotometric assay (MW-SPA) for one-step determination of these drugs via their interactions with iodine, which resulted in instantaneous production of bright lemon-yellow CTCs. A spectrophotometric study of the CTCs was conducted, and all CTCs were characterized. Site(s) of interaction on each drug were assigned, and the MW-SPA was developed and applied to the analysis of dosage forms. The findings confirmed that the reactions proceeded via CTC formation. Beer's law was obeyed over a general concentration range of 1-6 µg/mL. The LODs and LOQs were in the ranges of 0.5-2.1 and 1.5-6.4 µg/mL, respectively. The proposed MW-SPA demonstrated excellent precisions as the relative standard deviations were < 2.24 and 2.23% for the intra- and inter-assay precision, respectively. Recovery studies demonstrated the accuracy of MW-SPA. Successful determination of all drugs in bulk and tablet forms was achieved using the MW-SPA. The environmental sustainability of the proposed methodology was determined, providing evidence of the assay's alignment with the basis of green analytical chemistry. The high throughput of the assay was documented. In contrast to other existing methods, the MW-SPA described herein was valid for analyzing all drugs at the same wavelength. The assay is useful for routine analysis of drugs in their formulations in QC laboratories.

Cloud‐Enabled Handheld NIR Spectroscopy: A Transformative Approach for Real‐Time Forensic Analysis of Cannabis Specimens

AbstractIn the past few years, there has been significant interest within the forensic community regarding the deployment of portable solutions that provide real‐time results. This article introduces an innovative technology or technology architecture that enables the integration of a handheld device, specifically, Viavi MicroNIR, with a cloud‐based system. This cloud system encompasses a server responsible for data processing and a mobile application acting as a user interface.To demonstrate the transformative impact of this technology on field operators, the analysis of cannabis specimens has been utilized. System's capacity to distinguish between CBD‐type and THC‐type cannabis has been particularly highlighted, along with the remarkable congruence observed between the near‐infrared (NIR) spectra and the reference analytical method involving ultra‐high‐performance liquid chromatography (UHPLC)The article will present the advantages of this application primarily focusing on its potential to alleviate the burden on laboratories by expediting routine illicit drug analysis. Viavi MicroNIR technology provides laboratory personnel with additional time to handle more complex cases, thereby enhancing overall efficiency.

Development and validation of a stability-indicating RP-HPLC method for the determination of fifteen impurities in rivaroxaban

A simple and stability-indicating reverse phase high-performance liquid chromatographic (RP-HPLC) method for the determination of rivaroxaban (RIX) and its related substances was developed. Fifteen impurities of RIX, including three unreported isomers, were identified, synthesized, purified, and confirmed using MS, 1H NMR, 13C NMR, and HSQC spectral methods. This new method offered baseline separation for all monitored impurities, and was fast and reliable when compared to the European Pharmacopoeia method. Optimum separation for RIX and its related impurities was achieved on an octyldecyl silica column (YMC Core C18, 4.6 ×100 mm, 2.7 µm) by using a gradient HPLC method in 38 min. The final method was validated with respect to precision, LOD and LOQ, linearity, accuracy, and robustness. This developed method was suitable for routine quality control and drug analysis of RIX active substance.

A Validated RP-HPLC Method Development and Validation for The Simultaneous Estimation of Nebivolol and Valsartan in Bulk Drug and Pharmaceutical Dosage Form

A novel, simple, accurate and rapid Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method has been developed and validated for the determination of Nebivolol and Valsartan in bulk and tablet dosage form. Gradient elution chromatography has been developed on C18 column (5µm, 4.6mm* 250mm) with a mobile phase consist of pH 4 acetonitrile,water and Methanol (25:35:40) with a flow rate of 1ml/min at detection of 270 nm wavelength. The retention time for Nebivolol and Valsartan was found to be 4.59 and 10.2 respectively. Chromatography parameters were validated as per ICH guidelines and the method can be applicable for routine quantitative analysis of drugs in combined dosage form.

Advanced forced degradation technique for simultaneous estimation of azelnidipine and telmisartan implementing AQbD approach in its tablet dosage form

For simultaneous estimation of Azelnidipine and Telmisartan in pharmaceutical dosage form, a simple, rapid, precise and reproducible RP-HPLC method was developed. Separation was achieved on a Phenomanax Luna C18; 5 μm, 150 mm × 4.6 mm i.d. column using water and methanol 50: 50 (v/v) as mobile phase and detected at 256 nm using PDA detector. Azelnidipine and Telmisartan were exposed to thermal, photolytic, hydrolytic and oxidative stress conditions and stressed samples were analyzed by the proposed method. Linearity, precision, accuracy, specificity and robustness were studied as reported in the International Council for Harmonization guidelines. Linearity of the proposed method was in the range of 4-12 μg mL-1 (r2 = 0.998) for Azelnidipine and 20-60 μg mL-1 (r2 = 0.997) for Telmisartan. Limits of Detection were 1.041 μg mL-1 and 0.208 μg mL-1, while Limits of Quantitation were 3.155 μg mL-1 and 0.631 μg mL-1 for Azelnidipine and Telmisartan respectively. Degradation products produced as a result of stress studies did not interfere with the detection. The proposed stability studies method for simultaneous estimation was found to be specific, accurate and economical that could be applied for routine analysis of drugs in bulk and multi component formulations in quality control laboratories.

Simultaneous and enantiospecific quantification of primaquine and carboxyprimaquine in human plasma using liquid chromatography-tandem mass spectrometry

BackgroundThe enantiomers of the 8-aminoquinoline anti-malarial primaquine have different pharmacological properties. Development of an analytical method for simultaneous quantification of the enantiomers of primaquine and its metabolite, carboxyprimaquine, will support clinical pharmacometric assessments.MethodsA simple and sensitive method consisting of liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) was developed for simultaneous and enantiospecific determination of primaquine and its metabolite, carboxyprimaquine, in human plasma. Stable isotopes were used as internal standards to compensate for potential interference and matrix effects. Plasma samples (100 µL) were precipitated with 1% formic acid in acetonitrile followed by phospholipid removal solid phase extraction. Primaquine and carboxyprimaquine enantiomers were separated on a Chiralcel OD-3R (150 mm × 4.6 mm; I.D. 3 μm) column using a LC gradient mode. For separation of racemic primaquine and carboxyprimaquine, the LC method was modified and validated using a reverse phase column (Hypersil Gold 100 mm × 4.6 mm; I.D. 3 µm) and a mobile phase composed of 10 mM ammonium acetate buffer, pH 3.5 and acetonitrile in the isocratic mode. Method validation was performed according to regulatory guidelines.ResultsThe calibration range was set to 0.571–260 ng/mL and 2.44–2,500 ng/mL for primaquine and carboxyprimaquine enantiomers, respectively, resulting in a correlation coefficient (r2) ≥ 0.0998 for all calibration curves. The intra- and inter-day assay precisions were < 10% and the accuracy was between 94.7 to 103% for all enantiomers of primaquine and carboxyprimaquine. The enantiospecific method was also modified and validated to quantify racemic primaquine and carboxyprimaquine, reducing the total run time from 30 to 8 min. The inter-, intra-day assay precision of the racemic quantification method was < 15%. The absolute recoveries of primaquine and carboxyprimaquine were between 70 and 80%. Stability was demonstrated for up to 2 years in − 80 °C. Both the enantiomeric and racemic LC–MS/MS methods were successfully implemented in pharmacokinetic studies in healthy volunteers.ConclusionsSimple, sensitive and accurate LC–MS/MS methods for the quantification of enantiomeric and racemic primaquine and carboxyprimaquine in human plasma were validated successfully and implemented in clinical routine drug analysis.

Development of Validated Stability Indicating Method for Estimation of Vandetanib and Characterization of its Degradants by LC-ESI-MS

Abstract: Aim: In the current study, stability indicating high performance liquid chromatography method (RP-HPLC) was developed and validated for the determination of Vandetanib, also its major degradants were identified and characterized by Liquid Chromatography- Tandem Mass spectrophotometric method (LC-ESI-MS). Methods and Materials: This method was developed on Nucleosil 100-5, C18 (250×4.6 mm, 5μm) column by using Methanol: Ammonium acetate buffer as Mobile phase in the ratio, 90:10 v/v, having flow rate of 1 ml/min. The estimation was carried out at 249 nm. Further Vandetanib was subjected to various stress condition like acidic, alkali, oxidative, thermal and photolytic degradation. The degradation pathways for major degradants were idenitifed. Results: The method was developed and validated for linearity, robustness, accuracy, precision, linear regression analysis data which indicates the good linear relationship, correlation coefficient was found 0.992 in the concentration range of 1-10 μg/ml. In the stress results, the degradation of drug in alkaline, as well as acidic medium showed significantly. The product degradation was characterized by the LC-MS technique. Conclusion: The developed method was found to be rapid, sensitive, accurate, precise, and robust for the analysis of Vandetanib by which routine analysis of drugs can be done. Key words: Mass Spectroscopy (MS), Vandetanib, High-Performance Liquid Chromatography (HPLC), Validation, Stress degradation.

Methodical Analysis of an Absolute Efficacious Ayurveda Medicine-- Amruthotharam

Amruthotharam/Amrutottaram Kashayam is one of such preparation which takes care of metabolic disorder through inflammation. The formulation composed of three ingredients-- Tinospora cordifolia, Terminalia chebula and Zingiber officinale. Amrutottaram acts by treating indigestion and neutralization of AMA, thereby helps reduce inflammation and fever by virtue of its immune-modulatory function providing ingredients. The antiinflammatory properties prove to be significant in an array of pathologies like fevers, swelling, systemic inflammation or local inflammations, painful pathologies associated with muscular stiffness, neurogenic pain and thereby useful in any pathology pertaining to any system. The metabolic syndrome is a group of risk factors that together enhance chance of developing cardiovascular disease. The dried drugs were extracted separately by soxhlet extraction process. The present work involved the detailed study of drugs in dried form which can be used in routine analysis of drug. As, the formulation contains Terminalia chebula in major amount, gallic acid was considered as marker compound. The formulation was quantified for the content of gallic acid by HPLC method.

SIMULTANEOUS SPECTROPHOTOMETRIC DETERMINATION OF PHENYLEPHRINE HYDROCHLORIDE AND NAPHAZOLINE HYDROCHLORIDE IN EYE DROPS BY CHEMOMETRIC TECHNIQUES AND ARTIFICIAL NEURAL NETWORK

Three multivariate calibration-prediction techniques, partial least squares (PLS), principal component regression (PCR) and artifi cial neural networks (ANN), have been applied without separation in the spectrophotometric multi-component analysis of phenylephrine hydrochloride and naphazoline hydrochloride. A set of 25 synthetic mixtures of phenylephrine hydrochloride and naphazoline hydrochloride has been evaluated to determine the predictability of PLS, PCR and ANN. The absorbance data matrix was obtained by measuring zero-order absorbances between 230-300 nm at intervals of 3 nm. The suitability of the models was determined on the basis of root mean square error (RMSE), root mean squared cross validation error (RMSECV) and root mean squared prediction error (RMSEP) values of calibration and validation data. The results showed a very good correlation between true values and the predicted concentration values. Therefore, the methods developed can be used for routine drug analysis without chemical pre-treatment.

Method Development and Validation for the Estimation of Etizolam and Propranolol Hydrochloride in bulk and Combined Dosage Forms by Simultaneous and Derivative Spectroscopic Methods

Accurate, simple, sensitive and rapid economic UV spectroscopic methods were developed for the estimation of Etizolam and Propranolol Hydrochloride in bulk and combined dosage form. The present study deals with the UV spectroscopic method development and validation for the Simultaneous Equation method and First Derivative method of Etizolam and Propranolol Hydrochloride in bulk and combined dosage form at determined wavelength of Etizolam and Propranolol Hydrochloride at 244nm and 288nm for Simultaneous Equation method and 234nm and 289nm for First Derivative Method. The linearity range for Etizolam and Propranolol Hydrochloride was 1-5µg/ml and 10-50µg/ml, and exhibit good correlation coefficient of Etizolam and Propranolol Hydrochloride was 0.9877 and 0.9977 for Simultaneous Equation method and 0.9872 and 0.9977 for First Derivative method, respectively and excellent mean recovery (98-102%). The precision was found to be within limit (%RSD &lt;2). Comparatively First Derivative method is more sensitive than Simultaneous Equation method. The methods were validated statistically and parameters like linearity, precision, accuracy, specificity and assay was studied according to ICH guidelines and can be applicable in determination of both drugs in routine quality control analysis of drugs in bulk and combined dosage form.

METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS DETERMINATION OF AZELNIDIPINE AND TELMISARTAN IN TABLET DOSAGE FORM BY RP- HPLC

A simple, accurate and precise method for the simultaneous determination of azelnidipine and telmisartan in bulk drug and pharmaceutical dosage has been developed by RPHPLC method. Separation was performed on a Hyperchrom ODS C18 HPLC Column (250*4.6mm) column and Buffer 0.05M Potassium dihydrogen orthophosphate (KH₂PO₄) Buffer (pH-4.0): Methanol (60:40) as a mobile phase, at a flow rate 1ml/min and UV detection wavelength 215 nm. The calibration of the method was performed by concentration range of 20-60μg/ml for telmisartan and 40-120 μg/ml for azelndipine. The validation of proposed method was carried out for accuracy, precision, ruggedness, specificity for both alzenidipine and telmisartan the method can be used for routine quality analysis of titled drug in tablet formulation.

Thin Layer Chromatographic Detection of Dicyclomine Hydrochloride Extracted from Whole Blood

Dicyclomine Hydrochloride belongs to a class of synthetic anticholinergics. It is regularly used for the treatment of spasmodic pain, spasmodic dysmenorrhea, and renal, ureteric and biliary colic. This drug is easily available in the market and can be abused for suicidal purposes. Its unsupervised use can lead to abuse, accidental poisoning along with the occasional report of intentional suicidal poisoning. Hence its analysis is very important for medicolegal purposes.Biological samples of choice for routine qualitative analysis of drug is liver, stomach content, whole blood, and urine. Routine toxicological procedures employ highly sophisticated state of the art instruments like High Performance Liquid Chromatography (HPLC), Gas Liquid Chromatography (GLC) and Liquid ChromatographyMass Spectrophotometer (LC-MS) for the estimation of drug from biological and non-biological matrices but the present paper deals with an attempt to analyse the drug extracted from the biological sample of interest using a simple, feasible and optimal method with the help of Thin Layer Chromatography (TLC). The study involves the extraction of the drug from matrix (blood), using Liquid-Liquid Extraction method of alkaline plasma followed by its separation and identification by Thin Layer Chromatography. The present paper describes a simple, economical, reproducible sample extraction, clean-up and detection method that can be easily attempted in a laboratory. Keywords: Whole blood; Dicyclomine; Extraction; Analysis; Drug.

Potentiometric determination of mepivacaine hydrochloride local anesthetic drug in pharmaceutical and biological fluids using ion selective electrode

The objective of this work is to determine mepivacaine hydrochloride (MPVHC) as local anesthetic drug. Modified carbon paste ion-selective electrode was used for the determination of MPVHC in different dosage forms and biological fluids. The study depends on potentiometric titration of MPVHC drug using modified carbon paste (MCPE) as end point indicator electrode. The effects of the paste composition, different conditioning parameters and foreign ions on the electrode performance were investigated and response time of the electrode has been studied. The results obtained showed that carbon paste electrode modified with sodium tetraphenyl borate-MPV ion pair gives the highest potential break at the end point, Nernstian slope, wide concentration range and lower detection limit than the reinckate-, phosphotungestic- and phosphomolybdic-MPV ion pairs. The fabricated electrode obtained Nernstian response of 57.71±0.67 mV decade-1 in the concentration range of 3.1×10-7 – 1.0×10-1 mol L-1 for MCPE electrode. The electrode was found to be operating within the pH range of 3.0–8.0 and exhibited a fast response time (about 14 s), low detection limit of 3.1×10-7 mol L-1 and long lifetime (98 days). The electrode showed high selectivity to the MPVHC drug in the presence of different organic, inorganic and amino acids. The electrode was successfully applied for the determination of MPVHC in pharmaceutical preparations and biological fluids (urine and plasma) with high percent recovery and low standard and relative standard deviation values. The results obtained applying this potentiometric electrode are comparable with British pharmacopeia. The method validation parameters were optimized and the method can be applied for routine analysis of MPVHC drug.

Consistency of a dialyzable leucocyte extract manufactured at GMP facilities by nuclear magnetic resonance spectroscopy

The present work describes the development and validation of a first report including several non-invasive NMR schemes to identify parameters as local chemical environments, homo- and heteronuclear site-specific spin correlations, diffusion coefficient-dependent polydispersity indexes and quantification of identified peptide entities that composes a commercial human Dialyzable Leucocyte Extract (DLE), Transferon, an oral liquid formulation of low-molecular-weight peptides. The above parameters were useful indicators to verify reproducibility, consistency and homogeneity among the DLE batches manufactured at Good Manufacturing Practice (GMP) facilities and for batch-releasing purposes in a quality control laboratory. The results showed that peptide identity of the DLE is represented with both high reproducible one-dimensional proton spectra and diffusion coefficient distributions that predicts in turn a weight-average molecular weight of around 6.7-7.4 kDa and a mean polydispersity index of 1.13. The obtained NMR peptide fingerprint of the analyzed DLE allowed to i) confirm its structural homogeneity by line-shape analysis, ii) identify and quantify its peptide content within the total solution with qNMR methods iii) to confirm the robustness of the technique as a feasible alternative for routine analysis of Natural or non-Natural Complex Drugs, such as DLEs.

Highly Sensitive Levodopa Determination by Means of Adsorptive Stripping Voltammetry on Ruthenium Dioxide-Carbon Black-Nafion Modified Glassy Carbon Electrode.

A new, highly sensitive Adsorptive Stripping Voltammetric method for levodopa determination was developed. As a working electrode, the glassy carbon electrode (GCE) modified with carbon black (CB), RuO2·xH2O (RuO2) and Nafion was used (CB-RuO2-Nafion GCE). Levodopa signal obtained on the modified electrode was 12 times higher compared to GCE. During research, instrumental parameters were optimized: sampling time ts = 10 ms, waiting time tw = 10 ms, step potential Es = 5 mV and pulse amplitude ΔE = 50 mV. Preconcentration potential Eprec was equal to 0 mV. The best results were obtained in 0.025 M perchloric acid (approx. pH 1.4). Signal repeatability measured on the CB-RuO2-Nafion modified electrode for 0.2 µM of levodopa was equal to 2.1% (levodopa concentration 1 µM, n = 5). Linearity of the method was achieved in the concentration range from 1 to 8 µM. Limit of detection was equal to 17 nM. Recoveries calculated for pharmaceutical products and tap water measurements were in the range 102–105%, which confirms the accuracy of the developed. The applicability of the method was confirmed by analysis of pharmaceutical products and tap water samples. Based on obtained results, it might be concluded that the developed voltammetric method could be a useful tool in routine drug analysis.

Analysis of Total Flavonoids in Herbal Drugs Expressed as Quercetin by Reversed Phase-UHPLC Method

A rapid and accurate Reversed Phase-UHPLC method was developed for determination of gallic acid, quercetin and kaempferol in herbal drugs found in herbal product purchased from local market. A reversed phase chromatographic analysis was carried out under isocratic conditions using column Phenomenex, Prodigy, ODS3, 5.0 μm, 100 A, 250 × 4.6 mm (USA). The mobile phase was methanol and 0.5% phosphoric acid (50:50 v/v). The flow rate was 1.3 mL/min. The injection volume was 20 µL. The detection wavelength (λ max) was 370 nm using a PDA (photodiode array detector). Linearity of the method was established over the concentration ranges of 240 – 960 µg mL-1 for quercetin with a retention time about 7.8 minutes. Correlation coefficients was greater than 0.99. The recovery level of quercetin was 99.94%. The described method was quite suitable for routine analysis of herbal drugs by Reversed Phase-UHPLC Method.The proposed method was used for routine analysis to estimate the selected polyphenols under study in its pure form as well as in some herbal drugs forms.

Development and Validation of Stability-Indicating Reverse Phase-High Performance Liquid Chromatography Method for Simultaneous Determination of Atenolol and Nifedipine in Bulk and Tablet Dosage Form

The new stability-indicating high performance liquid chromatography (HPLC) method has been developed and validated with different parameters for atenolol (ATE) and nifedipine (NIFE) in the combined dosage form. The chromatographic conditions were optimized using a mobile phase of MeOH:OPA (70:30) with a flow rate of 0.7 mL/min. Column (C18) of 4.6 × 250 mm dimension was used as a stationary phase; the particle size capacity of the column was 5 μm. The detection was carried out at 233 nm. The method was validated according to ICH guidelines for linearity, precision, repeatability, the limit of detection (LoD), and limit of quantitation (LoQ). The response was found to be linear in the concentration range of 20 to 100 mcg/mL for ATE and 1 to 5 mcg/mL for NIFE. The developed method shows the minimum quantity of drugs to be identified (LoD) and minimum drug to be quantified (LoQ). The LoD and LoQ were found to be 0.1415 and 0.4289, respectively, for ATE, and 0.1834 and 0.5558, respectively, for NIFE. The method was linear, simple, precise, and accurate and, therefore, suitable for routine analysis of drugs in tablet form. The forced degradation studies were also done through the exposure of analyte solution to four different stress conditions.