Stability-indicating High-performance Thin-layer Chromatography Research Articles

A novel stability indicating HPTLC method for comparative quantitative estimation of quercetin in herbal preparations

This research introduces a novel stability-indicating HPTLC (High-Performance Thin-Layer Chromatography) method for the comparative quantitative estimation of Quercetin in herbal preparations. Through extensive stability studies, the method is validated, providing a robust framework for evaluating Quercetin's stability in diverse herbal formulations. The developed method ensures accurate and reliable quantification of Quercetin, facilitating comparative analysis across various herbal preparations. Additionally, results reveal that Quercetin degraded under most tested conditions, such as acidic, alkaline, oxidative and neutral conditions underscoring the necessity for intervention by experts to eliminate such challenges and ensure product efficacy and quality in the herbal industry.

Principles of White Analytical Chemistry and Design of Experiments to Stability-Indicating Chromatographic Method for Simultaneous Estimation of Thiocolchicoside and Lornoxicam.

A variety of chromatographic methods have been published for the stability evaluation of thiocolchicoside and lornoxicam. Nevertheless, the development of chromatographic methods requires the use of neurotoxic and teratogenic organic solvents that are detrimental to the environment and harmful to human life. Using the principles of design of experiments (DoE), a novel white analytical chemistry-driven stability-indicating high-performance thin layer chromatographic (SI-HPTLC) method has been developed for the concurrent stability study of thiocolchicoside (THC) and lornoxicam (LNX). To protect the environment and human life, the stability-indicating HPTLC method was developed using safe organic solvents. Potential analytical method risk parameters (AMRPs) and analytical method performance attributes (AMPAs) were screened using the fractional factorial design. The response surface analysis and optimization of critical AMRPs and AMPAs was carried out using full factorial design. Navigation of the method operable design region (MODR) was used to develop the SI-HPTLC technique. The developed method was validated as per ICH (International council for harmonization) Q2(R1) guideline. The developed method's greenness was evaluated using AGREE (Analytical Procedure Greenness) tool and ESA (Eco-Scale Assessment). The Blue (B) model was used to assess the proposed method's cost and time efficiency and user-friendlyness. For the stability studies of THC and LNX, the twelve principles of WAC (White Analytical Chemistry) were used to evaluate the published and proposed chromatographic techniques. As compared to previously published chromatographic techniques for studying the stability of THC and LNX, the suggested approach was found to be more affordable, environmentally friendly, and user-friendly. Development of stability-indicating HPTLC method using a novel white analytical chemistry approach and organic solvents with low toxicity potential. Application of the developed method for analysis of the forced degraded sample and fixed-dose combinations of THC and LNX.

Stability-Indicating HPTLC Method for Determination of Remogliflozin Etabonate and Vildagliptin in Tablets.

A sensitive, selective, precise and stability-indicating high-performance thin-layer chromatographic (HPTLC) method is developed for the simultaneous estimation of remogliflozin etabonate and vildagliptin in the presence of their degradation products. The separation was performed in thin layer chromatography plate precoated with silica gel G60F254. The mobile phase consists of methanol:ethyl acetate:toluene:ammonia (1.5:4:4.5:0.1, v/v/v/v). Detection and quantification are performed with densitometer at 212nm. The Rf values of remogliflozin etabonate and vildagliptin are 0.53 and 0.42, respectively. Degradation studies for the two drugs were carried out in acidic, alkaline, neutral, oxidative, photolytic and thermal stress conditions and analyzed. The suitability of this method for the quantitative determination of the compounds is proved by validation in accordance with the requirements laid down by International Conference on Harmonization (ICH). Linearity was found over the concentration range of 200-1000ng/band with correlation coefficient of 0.9936 for remogliflozin etabonate and 100-500ng/band with correlation coefficient of 0.9912 for vildagliptin. The method was successively applied to tablets containing two drugs and found to have no chromatographic interferences from the tablet excipients. The degradant formed was not interfering with estimation of two drugs makes the method more selective for the purpose intended.

Development of a new validated stability-indicating high-performance thin-layer chromatography method for determination of dosulepin HCl in bulk and marketed formulation with characterization of its degradants by liquid chromatography–Mass spectrometry

Aim: Dosulepin hydrochloride is a tricyclic antidepressant. It acts by inhibition of the reuptake of biogenic amine and increasing available neurotransmitter levels at the synaptic cleft. The objective of this work is to develop a new validated stability-indicating high-performance thin-layer chromatography (HPTLC) method for the determination of dosulepin hydrochloric acid (HCl) in bulk and marketed formulation and characterization of the degradation products by liquid chromatography–mass spectrometry (LC–MS). Materials and Methods: The HPTLC method development of dosulepin HCl was performed on aluminum plates precoated with silica gel 60 F254 using dichloromethane: toulene: methanol: glacial acetic acid (GAA) (4:4:2:0.2 v/v) with densitometric detection at 220 nm. The method validation was done using linearity, precision, accuracy, and robustness parameter. Results: Linearity range for dosulepin HCl was found in between 150 and 900 ng/band correlation coefficient was 0.996. The % relative standard deviation for accuracy and precision was found <2%. The developed and validated HPTLC method is simple, accurate, precise, and specific. Conclusion: This study concludes that the dosulepin HCl undergoes degradation to different extent under different stress conditions. From the peak purity profile studies, it was confirmed that the peak of the degradation product was not interfering with the peak of drugs. Major acidic and oxidation degradation products were isolated and characterized by LC–MS and probable degradation pathway for dosulepin HCl was determined.

Simultaneous ultra-sensitive analysis of tamsulosin hydrochloride and tolterodine tartrate binary mixture in their dosage form via high-performance thin-layer chromatography with fluorimetric detection

A simple, sensitive, selective, and stability-indicating high-performance thin-layer chromatography method with fluorescence detection was developed for the analysis of a mixture of tamsulosin hydrochloride and tolterodine tartrate, used in the management of benign prostatic hyperplasia in their laboratory-prepared binary mixture and dosage form. The separation was employed on thin-layer chromatography silica gel 60 aluminum sheets. As developing system, ethyl acetate–n-hexane–diethylamine (9:3:1, V/V) was used. Tamsulosin and tolterodine had retention factors (RF) of 0.40 ± 0.03 and 0.85 ± 0.03, respectively. A 225 nm excitation wavelength was used for the fluorescence detection. Linearity was in the range of 10.0–200.0 and 100.0–900.0 ng/band for tamsulosin and tolterodine, respectively. The method was validated successfully according to the International Council for Harmonisation guidelines. Our suggested method for determining tamsulosin and tolterodine in their dosage forms and in the presence of their degradation products could be conveniently applied in quality-control laboratories.

A Greener Stability-Indicating High-Performance Thin-Layer Chromatography Approach for the Estimation of Topiramate

Despite various reported analytical methods for topiramate (TPM) analysis, greener analytical approaches are scarce in literature. As a consequence, the objective of the current research is to design a normal-phase stability-indicating high-performance thin-layer chromatography (SI-HPTLC) methodology for TPM analysis in marketed tablet dosage forms that is rapid, sensitive, and greener. TPM was derivatized densitometrically and analyzed at 423 nm in visible mode with anisaldehyde-sulfuric acid as the derivatizing agent. The greener SI-HPTLC technique was linear in the 30–1200 ng band−1 range. In addition, the suggested SI-HPTLC methodology for TPM analysis was simple, rapid, cheaper, precise, robust, sensitive, and environmentally friendly. The greener SI-HPTLC method was able to detect TPM along with its degradation products under acid, base, and oxidative degradation conditions. However, no TPM degradation was recorded under thermal and photolytic stress conditions. TPM contents in commercial tablet dosage forms were recorded as 99.14%. Using 12 different principles of green analytical chemistry, the overall analytical GREEnness (AGREE) score for the greener SI-HPTLC method was calculated to be 0.76, confirming the proposed normal-phase SI-HPTLC method’s good greener nature. Overall, these results demonstrated that the suggested SI-HPTLC technique for TPM measurement in pharmaceutical products was reliable and selective.

Stability-Indicating Analytical Method Development Using Quality by Design (QbD) Approach for Simultaneous Estimation of Budesonide and Levosalbutamol.

Budesonide is a corticosteroid; levosalbutamol is bronchodilator. In combination, they are used in the treatment of asthma and chronic obstructive pulmonary disease. The present article portrays the development of a stability-indicating high-performance thin-layer chromatography (HPTLC) method for the simultaneous estimation of budesonide and levosalbutamol using design of experiment (DOE) principles. DOE was applied in development part in which a Box-Behnken design was used to study the effect of factors on the Rf value of the drugs. Twenty-five experimental runs were performed to optimize the chromatographic conditions. Aluminum sheets precoated with silica gel 60 F254 were used as the stationary phase. The optimized mobile phase composition was found to be toluene-ethyl acetate-methanol-ammonia (4:1.8:1.8:0.2) and saturation time 20 min, quantified by densitometric analysis at 231 nm. Moreover, the drugs were subjected to acid and alkali hydrolysis, oxidation, thermal, and photodegradation. The drugs undergo degradation under mainly acidic and basic conditions. Also, the degraded products were well resolved from the pure drug with significantly different Rf values. Linearity was performed in the ranges of 224-1120 ng/band for budesonide and 280-1400 ng/band for levosalbutamol. The method was validated according to the International Conference on Harmonization guidelines. This approach is useful to expedite method development and optimization activities in analytical laboratories. The experimental data revealed that the volume of toluene and methanol in the mobile phase significantly affected the Rf value of both the drugs, and ammonia significantly affected the Rf value of levosalbutamol.

Stability-indicating high-performance thin-layer chromatography method for the simultaneous estimation of emtricitabine and tenofovir alafenamide fumarate

Stability-indicating high-performance thin-layer chromatography method for the simultaneous estimation of emtricitabine and tenofovir alafenamide fumarate

A rapid and highly sensitive stability-indicating high-performance thin-layer chromatography technique for the determination of tedizolid phosphate with a classical univariate calibration

A rapid and highly sensitive stability-indicating high-performance thin-layer chromatography technique for the determination of tedizolid phosphate with a classical univariate calibration

A rapid and sensitive stability-indicating green RP-HPTLC method for the quantitation of flibanserin compared to green NP-HPTLC method: Validation studies and greenness assessment

A rapid and sensitive stability-indicating green reversed-phase high-performance thin-layer chromatography (SI-G-RP-HPTLC) method was established and validated for the detection of flibanserin (FBN) compared to the stability-indicating green normal-phase high-performance thin-layer chromatography (SI-G-NP-HPTLC) method. The G-RP-HPTLC-densitometry analysis of FBN was conducted using acetone/water (80:20, v/v) solvent system as the mobile phase, while, the G-NP-HPTLC-densitometry analysis of FBN was conducted using ethyl acetate/methanol (95:5, v/v) solvent system. The quantitation of FBN was carried out at λmax = 204 nm for both of the techniques. The SI-G-RP-HPTLC and SI-G-NP-HPTLC methods were found to be linear in the range of 100–1600 and 200–1600 ng/band, respectively. The system suitability parameters for both of the methods were found to be acceptable for the quantitation of FBN. The SI-G-RP-HPTLC technique was found to be more rapid, accurate, precise, and sensitive for the determination of FBN compared to the SI-G-NP-HPTLC technique. The SI-G-RP-HPTLC technique was observed as more selective and stability-indicating for the determination of FBN in the presence of its degradation products. The FBN contents of commercial film-coated tablets were found to be 98.76% using the SI-G-RP-HPTLC technique, while, the FBN contents of commercial film-coated tablets were recorded as 96.28% using the SI-G-NP-HPTLC technique. The greenness of both of the green methods was assessed using AGREE software which utilized 12 principles of green analytical chemistry. The eco-scales of SI-G-RP-HPTLC and SI-G-NP-HPTLC methods were predicted as 0.86 and 0.80, respectively. The obtained eco-scale values for both of the methods suggested excellent greenness of both of the methods. Based on these observations and validation studies, the SI-G-RP-HPTLC methodology was considered as superior over the SI-G-NP-HPTLC technique for pharmaceutical analysis. Accordingly, the SI-G-RP-HPTLC technique could be utilized for the quantitation and stability evaluation of FBN in commercial products containing FBN.

A Rapid and reliable thin-layer chromatographic method for the simultaneous estimation of celecoxib and diacerein in their binary mixture using nanosilica gel plate

A novel, rapid, and sensitive stability-indicating high-performance thin-layer chromatography method was developed and validated for the simultaneous determination of celecoxib (CEL) and diacerein (DIA) in bulk and a capsule binary mixture. The complete separation was achieved by the proposed method on nanosilica gel F254 plates using n-hexane‒ethyl acetate‒tetrahydrofuran‒glacial acetic acid (7:3:1:0.8, V/V) as the mobile phase. The chromatographic bands were visualized using short-wave ultraviolet light, and the amount of CEL and DIA was determined by scanning densitometry at 254 nm using the peak area. The RF values were 0.59 and 0.37 for CEL and DIA, respectively. From regression plots, it was found that good linearity was achieved in the concentration ranges of 20.00–320.00 ng/band and 5.00–80.00 ng/band with mean percentage recoveries 99.86 ± 0.70 and 99.64 ± 0.96 for CEL and DIA, respectively. The proposed method was validated according to the International Conference on Harmonisation guidelines. The statistical analysis of the results revealed high accuracy and precision, the suggested procedures could be used as a stability-indicating assay for the simultaneous determination of CEL and DIA in bulk drug and their capsule binary mixture in the presence of their degradation products.

Validated Stability-Indicating HPLC and HPTLC Methods for the Determination of Ritonavir in Bulk Powder and in Capsules

Two sensitive and reproducible methods were described for the quantitative determination of the antireteroviral drug ritonavir in the presence of its degradation products. The first method was based on high performance liquid chromatographic (HPLC) separation of the drug from its stress degradation products with the use of a reversed phase Agilent Eclipse XDB-C18 column (5 µm, 4.6 × 150 mm) and a mobile phase consisting of acetonitrile : 0.05 M phosphoric acid (55 : 45, v/v) at a flow rate of 1.0 mL/min. The retention time of the drug was found to be 4.82 ± 0.002 min. Quantification was achieved with diode array detection (DAD) at 210 nm based on peak area and a linear calibration curve in the concentration range of 1-500 µg/mL. The proposed method made use of diode array detection as a tool for peak purity and identification. The second method involved a high performance thin layer chromatographic (HPTLC) separation followed by densitometric measurement of the spots at 240 nm. The separation was carried out on Fluka TLC aluminium sheets of silica gel with fluorescent indicator (254) nm and the mobile phase was acetonitrile - water (1 : 2, v/v), adjusted to pH 5.0 using 1 M orthophosphoric acid solution . The proposed procedure gave compact spots for ritonavir (retention factor, Rf = 0.41 ± 0.014). The linear regression equation was generated by least-squares treatment of the calibration data in the range of 0.8-12.5 µg/spot. The reliability and analytical performance of the proposed methods, including linearity, range, precision, accuracy, detection and quantitation limits, were statistically validated. The proposed methods were applied to Norvir capsules and no chromatographic interference was observed. When ritonavir was subjected to stress conditions; according to ICH guidelines, the proposed methods could effectively separate the drug from its degradation products, and were thus considered as good stability-indicating procedures.

A Validated Stability-Indicating HPTLC Assay for Determination of 10-Hydroxy-2-Decenoic Acid Content in Royal Jelly Products Using Robust Regression Methods.

A new, simple, stability-indicating high-performance thin-layer chromatography method was developed for the quantification of 10-hydroxy-2-decenoic acid (10-HDA) in some royal jelly products marketed in Egypt. The used solvent system was chloroform:acetic acid (10:1, v/v) and the bands were measured densitometrically at 210nm. First- and second-derivative treatments of the data were performed. The present study shows a comparison between three statistical regression methods for handling data: parametric, nonparametric and weighted regression (WR) methods. The developed methods were validated as per International Conference on Harmonization guidelines. To validate the stability-indicating power of the developed analytical method, the royal jelly standard was subjected to forced degradation studies including the effect of hydrolysis, oxidation, photolysis and dry heat. It was found that derivative treatment of the chromatographic response data gives improved quantitation and sensitivity of the chromatographic signals. Weighted regression of the response data is found to be advantageous over the use of both parametric and nonparametric regression models. This was shown by a great enhancement in the accuracy and precision in the analysis of 10-HDA in royal jelly products. The % recovery in case of WR was 99.92±0.16, while % recovery in case of nonparametric and parametric regressions were 99.56±0.25 and 98.63±0.65, respectively.

Novel Stability-Indicating High-Performance Thin Layer Chromatography (HPTLC) Method Development and Validation for Estimation of Daclatasvir Dihydrochloride in Pharmaceutical Dosage Form

Daclatasvir dihydrochloride (DTDH) is a direct-acting antiviral agent used against the hepatitis C Virus. Different analytical methods were reported for the determination of DTDH in pharmaceutical dosage form, including spectrophotometry, high-performance liquid chromatography (HPLC), and stability- indicating HPLC method but to date, no HPTLC method has been reported. A novel, simple, sensitive, selective, precise, and accurate stability-indicating high-performance thin-layer chromatographic (HPTLC) method has been developed for the quantification of Daclatasvir dihydrochloride (DTDH) in pharmaceutical formulation. Chromatography separation was carried out on pre-coated silica gel G60 F254 HPTLC plates using chloroform: methanol (9:1, v/v) as the mobile phase. Scanning and densitometric analysis were done at 316 nm. The band of DTDH were obtain at RF= 0.48±0.02. The calibration curve was plotted in the concentration range of 400– 900 ng/band and the method was found linear with r = 0.995. The LOD and LOQ were found to be 95 ng/band and 313.5 ng/band respectively. The percentage recovery was found to be 97.31-101.81 %. The developed was able to separate peaks of all the degradation products formed in ICH-prescribed stress conditions with a significant difference in their RF values. The developed method was validated according to ICH guidelines and results were found to be within acceptance criteria. The developed stability-indicating method successfully evaluated the DTDH in pharmaceutical tablet dosage form and assay results showed good recovery when statistically compared with the reversed-phase high-performance liquid chromatography (RP-HPLC) method.

Design of Experiment Approach for the Development and Validation of a Stability-Indicating High-Performance Thin-Layer Chromatography Method for the Estimation of Metformin Hydrochloride and Ursodeoxycholic Acid in Pharmaceutical Dosage Form

In the present study, a design of experiment (DoE) approach was used to optimize chromatographic conditions for the development of a high-performance thin-layer chromatography (HPTLC) method for the simultaneous estimation of metformin hydrochloride (MET) and ursodeoxycholic acid (URSO) in pharmaceutical dosage form. The critical factors were identified using a Taguchi design, and after identification of critical factors, optimization was done using Box–Behnken design (BBD). BBD was used to optimize the compositional parameters and to evaluate the main effect, interaction effects, and quadratic effects of the mobile phase compositions, development distance, and saturation time on the retardation factor (RF) of both drugs. HPTLC separation was performed on aluminum plates pre-coated with silica gel 60 F254 as the stationary phase, using toluene-ethanol-acetone-formic acid (4.5:2:2.5:0.85, V/V) as the mobile phase at a wavelength of 234 nm and 700 nm for MET and URSO, respectively. A sharp and well-resolved peak was obtained for MET and URSO at RF values of 0.19 and 0.80 min, respectively. The calibration curve was in the range of 5000–40000 ng per spot and 1500–12000 ng per spot for MET and URSO, with r2 = 0.984 and r2 = 0.980, respectively. The method was validated for linearity, accuracy, precision, limit of detection, limit of quantification, and specificity. To provide a better visualization of the statistically significant factors derived from the statistical analysis, the perturbation plot and response surface plot for the effect of independent variables on the RF of MET and URSO were evaluated. Stability study was performed under different stress conditions such as acid and alkali hydrolysis, oxidation, and temperature. The developed method was able to resolve drugs and their degradation products formed under the afore-mentioned conditions.

Validated stability-indicating HPTLC method for cefixime and azithromycin with preparative isolation, identification, and characterization of degradation products

Stability-indicating High-Performance Thin-Layer Chromatography (HPTLC) method for simultaneous estimation of cefixime trihydrate and azithromycin dihydrate was developed. Both the drugs were subjected to different stress conditions recommended by International Conference on Harmonization (ICH) guideline Q1A (R2). Forced degradation was carried out for hydrolytic, oxidative, photolytic, and thermal degradation conditions. Cefixime was susceptible for degradation under all stress conditions showing four degradation products (CI–IV). However, azithromycin formed only one degradation product (AI) under acid hydrolysis. Aluminum plates precoated with silica gel 60F254 were used as the stationary phase while mixture of ethyl acetate–methanol–acetone–toluene–ammonia (1:5:7:0.5:0.5, v/v) was used as mobile phase. Detection wavelength used was 235 nm for CEFI and CI–IV. AZI and AI were detected by post development derivatization, spraying with sulfuric acid–ethanol (1:4, v/v) followed by heating at 100 °C for 5 min. Degradation products were isolated by preparative HPTLC and characterized by MS/MS. The developed method was validated for linearity, precision, accuracy, specificity, and robustness and has been successfully applied in the analysis of these drugs in tablet dosage form.

Analytical quality-by-design approach for the stability study of thiocolchicoside by eco-friendly chromatographic method

A robust and eco-friendly stability-indicating high-performance thin-layer chromatography (HPTLC) method was developed for the stability study of thiocolchicoside using analytical quality-by-design approach. Full factorial design was used for screening potential variables affecting method development. Box–Behnken design was used subsequently for investigation of the main, interactive, and quadratic effects of these variables on response. Four potential variables were selected on the basis of scientific knowledge for the development of a method for the stability study of thiocolchicoside. The selected potential factors, namely, volume of water (mL), saturation time (min), migration distance (mm), and volume of mobile phase (mL) were screened by 24 full factorial designs by selecting resolution as a critical method attribute. Pareto chart analysis showed that 3 variables, namely, volume of water (mL), saturation time (mm), and migration distance (mm), out of 4 potential variables were significantly affecting the response variable (resolution). Optimization with response surface methodology further clarified the relationship between critical variables and resolution using Box—Behnken design. The experimental design model was found to be quadratic, and the design space was developed on the basis of suggested model for optimization of critical method variables for maximum desirable resolution and for the development of a control strategy of the HPTLC method for the stability study of thiocolchicoside. The developed method was validated for linearity, range, specificity, precision, accuracy, limit of detection and limit of quantification as per the International Conference on Harmonization guidelines (ICH) Q2 (R1). The developed method was applied for the estimation of thiocolchicoside in its pharmaceutical dosage forms. The degradation products formed in acidic and alkaline media were isolated and characterized by their infrared (IR), nuclear magnetic resonance (NMR), and mass spectral data.

Validated stability-indicating high-performance liquid chromatography and thin-layer chromatography methods for the determination of zopiclone in pharmaceutical formulation

Two novel, sensitive, and selective stability-indicating chromatographic methods were described for the analysis of zopiclone (ZOP) in the presence of its degradation products, namely, 7-oxo-6,7-dihydro-5H-pyrrolo[3,4-b]pyrazin-5-yl-4-methylpiperazine-1-carboxylate (hydrolytic DEG) and 5H-pyrrolo[3,4-b]pyrazine-5,7(6H)-dione (oxidative DEG), in drug substance and product. The first method was an isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) using Inertsil ODS3 (250 × 4 mm, 5 μm) column. Upon using HPLC, the run time could be reduced, and actually, the solvents consumption decreased. Quantification was achieved by detection wavelength at 237 nm, based on peak area. Chromatographic separation was performed over the range of 1–10 μg mL−1 with limits of detection (LOD) and quantification (LOQ) of 0.18 and 0.55 μg mL−1 and a mean recovery of 99.98 ± 0.55. The analysis was achieved at 30°C using a mixture of acetonitrile and water (50:50 v/v) as the mobile phase. The second method was thin-layer chromatography (TLC) applied for the separation and analysis of zopiclone in the presence of its alkaline, acidic, and oxidative degradation products. Chromatography was performed on silica gel 60 F254 plates with ethyl acetate–methanol–ammonia 33% (17:2:1 v/v) as the mobile phase. Successful resolution was observed with significant difference in the RF values, followed by densitometric measurement at 303 nm. Evaluation was carried out over the range of 0.1–2 μg per spot with a mean recovery of 100.52% ± 0.24. The developed methods were successfully applied to the analysis of ZOP in bulk powder, laboratory-prepared mixtures containing different percentages of its degradation products, and pharmaceutical dosage form. The degradation products were separated by HPLC as well as identified by TLC, infrared (IR), and mass spectrometry (MS) to confirm its structures and elucidate degradation pathway. The developed methods were validated as per the International Conference on Harmonization (ICH) guidelines. The results obtained by the proposed methods were statistically compared with the reported methods revealing high accuracy and good precision.

Stability-Indicating HPTLC Method for Estimation of Benazepril in Bulk and Tablet Dosage Form

A stability-indicating high-performance thin-layer chromatography method for determination of benazepril both in bulk and pharmaceutical formulation has been developed and validated as per ICH guidelines. The benazepril was analyzed on pre-coated silica gel GF254 plates using Carbon tetrachloride: Methanol: Triethylamine (7.6: 2.4: 0.1 v/v) as the mobile phase. The concentration of benazepril was determined by densitometry at 240 nm. The method was found to be linear, with r2 = 0.998±0.3347 in the concentration range 200 - 1200 ng per spot. The LOD and LOQ were 19.48 ng and 59.03 ng per spot respectively. Benazepril was subjected to forced degradation like acid and alkali hydrolysis, oxidation, photo and thermal degradation. The drug was found to degrade under acidic, basic, oxidative and thermal conditions. The degraded product was well resolved from the pure drug with significantly different Rf value. In validation, it was found that the method is repeatable, selective and accurate for the estimation of benazepril. The proposed developed stability-indicating HPTLC method can be applied for identification and quantitative determination of benazepril in bulk drug and pharmaceutical formulation.

Stability-indicating high-performance thin-layer chromatographic determination of ondansetron in pure form and pharmaceutical formulations

A new, simple, precise, accurate, stability-indicating, and rapid high-performance thin-layer chromatography (HPTLC) method was developed and validated for the determination of ondansetron hydrochloride in pure form and pharmaceutical formulations. The method used Merck HPTLC aluminum plates precoated with silica gel 60 F254 as the stationary phase. The mobile phase consisted of chloroform–methanol–ethyl acetate (7:2:1, v/v). This system was found to give a compact spot of ondansetron (RF value of 0.67 ± 0.011); ondansetron was subjected to acid and alkali hydrolysis, oxidation, and photodegradation. The wavelength of TLC scanner was set at 302 nm for both detection and quantitation. The calibration curves were linear over the range of 25–500 ng spot−1. The limit of detection was 4.9 ng spot−1, and the limit of quantitation was 14.7 ng spot−1. The proposed analytical method was validated according to the International Conference on Harmonization (ICH) guidelines, and the results were acceptable. The propo...