Stability-indicating HPTLC Method Research Articles

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.

Red, Green and Blue Model-based Assessment and Principles of White Analytical Chemistry to Robust Stability-Indicating Chromatographic Estimation of Thiocolchicoside and Diclofenac Sodium.

White analytical chemistry (WAC) is a recent approach for evaluating analytical procedures based on their effectiveness in validating results, capacity to be environmentally friendly and economic effectiveness. The detection of diclofenac sodium (DCF) and thiocolchicoside (THC) simultaneously has been established using a WAC-driven stability indicating chromatographic method (SICM). For the concurrent stability study of THC and DCF, the suggested chromatographic technique was developed employing a safe and environmentally acceptable organic solvents. To identify critical analytical method parameters (AMPs) and analytical quality attributes (AQAs), the design of experiments (DoE)-based screening design was applied. For the DoE-based response surface modelling (RSM) of critical AMPs and AQAs, the Box-Behnken design (BBD) was employed. A robust SICM was developed by navigating the analytical design space for simultaneous estimation of THC and DCF. The IR, NMR, and mass spectral data were used to characterize the degradation products. The red, green, and blue (RGB) models were used to evaluate the suggested method's validation effectiveness, greenness power, and economic efficiency with published chromatographic techniques. The effectiveness of the chromatographic method's validation concerning the ICH Q2 (R1) guideline was evaluated using the red model. Analytical greenness (AGREE) evaluation tool and eco-scale assessment (ESA) approach were used to evaluate the green model's methodology. The blue model-based assessment was carried out for comparison of handling of instruments, cost, and time during sample analysis. The red, blue, and green scores of the techniques were averaged to arrive at the white score of the suggested and reported methods. For the concurrent stability study of THC and DCF, the suggested technique was shown to be validated, environmentally friendly, and cost-effective. The suggested approach could be a cost-effective and environmentally friendly analytical technique for determining the stability and monitoring the quality of fixed-dose combinations of THC and DCF. Development of stability-indicating HPTLC method for concomitant analysis of THC and DCF using concepts of DoE and white analytical chemistry.

Development and Validation of a Green Stability-Indicating HPTLC Method for Estimation of Curcumin, Gallic Acid, and Ursolic Acid From Polyherbal Formulation Jatyadi Taila.

Jatyadi taila (JT) is a well-known Ayurvedic wound-healing product, comprising 16 different medicinally important plants, including Curcuma longa, Terminalia chebula, and Jasminum officinale. The proposed work discusses the development and validation of the green and economical stability-indicating HPTLC method for quantification of the key marker phytoconstituents, curcumin (CUR), gallic acid (GA), and ursolic acid (UA), from JT. Quality standard parameters for JT were determined following standard procedures. The marker constituents CUR, GA, and UA were resolved from JT using toluene-ethyl acetate-formic acid (6:2:1, v/v/v) as the mobile phase and subsequently derivatized to estimate UA. The developed plates were subjected to HPTLC-MS analysis. All constituents were subjected to forced degradation to determine the proposed technique's stability-indicating property and the accelerated stability studies of marketed formulation and marker constituents. Greenness evaluation of the method was aided by the AGREE methodology. The Rf values of CUR, GA, and UA were found to be 0.60 and 0.60; 0.27 and 0.28; and 0.74 and 0.77 from reference standard and oil samples respectively, when analyzed at 366 nm, 290 nm, and 366 nm, respectively. HPTLC-MS was carried out to verify the active constituents present in JT. The constituents followed first-order degradation kinetics. The quantity of CUR, GA, and UA in JT was reduced at the end of accelerated stability studies. The developed approach was validated in compliance with the International Conference on Harmonization (ICH) Q2 (R2) guideline. Among the chosen key markers, GA was highly unstable during forced degradation. JT should be stored at a controlled temperature using more protective packaging material to ensure its quality and efficacy. The developed method can be used as a quality control tool for JT as it can be used to determine the stability of the key marker compounds the herbal formulation.

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.

Development of validated stability-indicating HPTLC method for the estimation of ulipristal acetate in bulk and dosage form

A Study of Method Development, Validation and Forced Degradation for Quantification of Buprenorphine Hydrochloride in a Microemulsion FormulationDhanashree Arun Mundhey, Vishal V. Rajkondawar, Anwar S. Daud, Nidhi P. Sapkal

Development and Validation of a Stability-Indicating HPTLC Method for the Estimation of Eszopiclone in Pharmaceutical Dosage Forms

Objective: A simple, sensitive, selective, precise repeatable and stability-indicating high-performance thin layer chromatographic method was developed and validated for Eszopiclone in bulk drug and in formulation. Method: Silica gel 60 F-254, TLC precoated aluminium plates was used as the stationary phase for analyzing Eszopiclone and its degradation products, using mobile phase consisting toluene: ethyl acetate: methanol (6: 4: 2 v/v/v). Result: This mobile phase gave compact spots for Eszopiclone with Rf value of 0.52 ± 0.02. Eszopiclone was exposed to hydrolysis, oxidation, neutral and photolytic conditions for conducting stress degradation study. The peak of Eszopiclone and the degradation product was well resolved from each other with a significantly different Rf value. Densitometric estimation of Eszopiclone was performed at 304nm. A good linear plot was obtained in the concentration range of 150-300ng/spot. The method was validated for precision, accuracy (recovery) and robustness study. The limit of detection (LOD) and limit of quantitation (LOQ) was found to be 130ng/spot and 150ng/spot, respectively. Conclusion: The developed HPTLC method can separate Eszopiclone from its degradation products, hence stability studies can be performed using this method.

Intensity of heavy metal accumulation in plants of Silybum marianum L. in conditions of field rotation

Intensity of heavy metal accumulation in plants of Silybum marianum L. in conditions of field rotation

Stability-indicating HPTLC method for the simultaneous detection and quantification of alfuzosin hydrochloride, solifenacin succinate along with four of their official impurities

Stability and impurity profiling of new drug formulations are now receiving great attention in modern pharmaceutical analysis. In this work, a novel stability-indicating HPTLC coupled with densitometric quantification was developed for the simultaneous determination of alfuzosin (ALF) and solifenacin (SOL) along with their degradation products/official impurities. The two drugs were subjected to different stress conditions. ALF was liable to acidic and basic hydrolysis, while SOL was found to be susceptible to basic and oxidative degradation. The obtained degradation products, namely; ALF impurity-D and SOL impurities-A, E and I were then characterized by IR and mass spectrometry. Chromatographic separation was then performed on HPTLC silica plates 60 F254, as a stationary phase, using ethyl acetate: toluene: ethanol: ammonia (5: 2: 3: 0.4, by volume), as a mobile phase. The plates were scanned at 220 nm and visualized by iodine vapor in daylight. Effect of different factors, including the mobile phase composition and the detection wavelengths, were carefully studied to achieve the optimum conditions for chromatographic separation. Calibration curves were constructed over the ranges of 0.8 – 30.0 µg/band for ALF and SOL, 0.5 – 15.0 µg/band for ALF imp-D, 0.5 – 4.0 µg/band for SOL imp-I and 0.75 – 7.50 µg/band for SOL basic degradation. The method was also exploited for assessment of the two drugs’ stability in Solitral® capsules under accelerated storage conditions. The proposed method has many potentials of being simple, economic and selective making it an attractive procedure in analyzing not only the two cited drugs but also their impurities.

Stability-indicating HPTLC method development for determination of arterolane maleate and piperaquine phosphate in combined dosage form

Stability-indicating HPTLC method development for determination of arterolane maleate and piperaquine phosphate in combined dosage form

Study of Intrinsic Stability of Mometasone Furoate in Presence of Salicylic Acid by HPTLC and Characterization, Cytotoxicity Testing of Major Degradation Product of Mometasone Furoate

Background: A successful attempt has been done to develop and validate a simple stability indicating HPTLC method for the estimation of Mometasone furoate (MF) and its degradation product in the presence of Salicylic acid (SA). The degradation product was isolated, characterized and tested for cytotoxicity. Introduction: Mometasone furoate (MF) is chemically 9,21-Dichloro-17α-[(2-furanylcarbonyl) oxy]- 11β-hydroxy-16-α-methylpregna-1,4-diene-3,20-dione, a high potency glucocorticoid. Salicylic acid (SA) has antiseptic, antifungal and keratolytic properties. Combination of MF and SA is available in the market as an ointment and is used for the treatment of skin inflammation, skin diseases, acne, skin redness and other conditions. Till now, there is no scientific documentation on HPTLC method for simultaneous estimation of MF and SA in the topical formulation; stress testing of drugs and determination of degradation products. Methods: Combination of Toluene: Ethyl Acetate: Methanol: Ammonia (6.4:1.5:2.0:0.1) was selected as the mobile phase. Detection was done by UV absorbance mode at wavelength 250 nm. Topical formulation containing MF and SA was analyzed by the developed method. The developed method was validated as per ICH guidelines. The standard drugs were subjected to stress testing like hydrolysis, oxidative, thermal and photolytic degradation. Results: Good separation with Rf values 0.61 ± 0.02 (MF) and 0.21 ± 0.02 (SA) was achieved by optimized chromatographic conditions. The % drug content was found to be 97.41±1.15 and 99.43 ± 0.73 for MF and SA, respectively in a topical formulation. From the results of validation parameters, the developed method was found to be specific, accurate, precise, sensitive and robust. After stress testing, SA was found to be stable under different stress conditions. Whereas, MF was found to be base sensitive and single degradation product was observed and isolated by preparative TLC. It was characterized by LC-MS and LC-MS/MS studies. Isolated degradation product was subjected to cytotoxicity testing on A549 and SiHa cell lines. Conclusion: A simple stability indicating HPTLC method was developed and validated for the estimation of MF and its degradation product in presence of SA. Probable structure of degradation product of MF and probable pathway of degradation was interpreted. Results of cytotoxicity testing showed that the degradation product was more cytotoxic as compared to MF against both the cell lines.

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

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

A Validated Stability-Indicating HPTLC Method for the Estimation of Capecitabine in its Tablet Dosage Form

Background: Capecitabine is an orally available prodrug of 5-flurouracil used in the treatment of breast cancer, metastatic colorectal cancer and stage III colorectal cancer. Various studies have reported the HPLC, HPLC-MS, MS/MS methods for estimation of capecitabine. However, till date HPTLC method for estimation of capecitabine and its validation is not reported in tablet dosage form. Introduction: Presented study deals with the development and validation of stability indicating high performance thin layer chromatography method for the determination of Capecitabine in tablet dosage form. Methods: The method was developed using precoated HPTLC plates with silica gel 60 F254 as stationary phase and toluene-methanol the ratio of 7.5:2.5 v/v as the mobile phase. Capecitabine (RF 0.48 ± 0.03) and its degradation products were well resolved. The wavelength selected for study was 240 nm. The method was linear in the concentration range 50–550 ng/band with a correlation coefficient of 0.994. The repeatability for six samples was 1.25% RSD. The intraday and interday precisions were 1.46-1.71%RSD and 1.31-1.67% RSD, respectively. The accuracy (recovery) was found to be in the range of 99.10-101.23% with LOD and LOQ were found to be 0.650 and 1.765 mg/band. The mean content of drug in tablet dosage form was found to be 101.51% with a % RSD of 1.20. The drug was subjected to stress conditions such as hydrolysis, oxidation, photolysis, and heat. Results: Degradation products produced as a result of the stress conditions did not interfere with the detection of Capecitabine; therefore, the proposed technique can be considered stability-indicating. Capecitabine did not degrade under thermal and photolytic conditions but showed degradation under acidic and alkaline conditions with 15 and 11% decompositions respectively. Conclusion: The developed method was found to be facile, simple, specific, precise, and stabilityindicating. It can be employed for the routine analysis of capecitabine in tablet dosage form.

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.

A Novel Validated Stability-indicating HPTLC Method to Quantitate Forskolin as a Bulk Drug and in a Nanosuspension

A novel, reproducible, precise, accurate and dependable high-performance thin-layer chromatographic technique was developed and authenticated as per International Conference on Harmonization guiding principle for quantitative evaluation of forskolin as a bulk drug and in a nanosuspension. Chromatographic plates 60 F-254 pre-coated with silica gel and a solvent system of ethyl acetate:hexane:formic acid (7:2.9:0.1 v/v) was used for the development of the plates. Densitometric scanning of the developed plates in the reflectance mode at 555 nm using a Camag TLC scanner after exposing the plates to anisaldehyde-sulphuric acid reagent, gave a compact spot for forskolin (Rf=0.46±0.012). In accordance to the ICH guidelines, the method was validated for linearity, precision (intra- and inter-day), limit of quantification and limit of detection, robustness and accuracy. Good linear relationship was observed for the calibration plots with linear range from 100-1000 ng/spot and a coefficient of correlation r2=0.994. Limit of detection and limit of quantification were found to be 10.83 and 36.12 ng/spot, respectively. The method appeared to be well suited for determining forskolin content in the developed nanoformulation. Forskolin was found to be susceptible to acid and alkali hydrolysis, oxidation, photochemical degradation with UV and sunlight. Statistical analysis confirmed that the process developed is precise, repeatable, accurate, and suitable for the analysis of forskolin in a formulation and could be used also for analysis of forskolin in plasma and other biological fluids. The ability of the developed method to separate the drug from its degradation products effectively confirmed that it could be a stability indicating method.

A Validated Stability-indicating HPTLC Method for the Determination of Roflumilast in Tablets and Application to Accelerated Stability Studies

A high performance thin layer chromatography method was developed for roflumilast on silica gel 60F254 thin layer chromatography plates using mobile phase comprising of toluene:ethyl acetate (7:3% v/v) with detection set at 254 nm on a densitometer. The Rf value of roflumilast was 0.54±0.02. Percent recovery of the drug from tablet formulation was carried out by standard addition method and was found to be close to 100 and percent relative standard deviation was less than 2%, which indicated good accuracy and precision of the method. Roflumilast was subjected to acid, alkali and neutral hydrolysis, oxidation and thermal and photolytic degradation. Degradation studies indicated roflumilast to be susceptible to acid and alkaline hydrolysis and oxidation. The degradation product was well resolved from the pure drug with significant differences in Rf values. The developed validated method was applied to formulation, content uniformity test and accelerated stability studies of roflumilast. The rate of hydrolysis was found to increase with increase in the strength of hydrochloric acid. It was observed that temperature had a major effect over acid hydrolysis of roflumilast compared to varying the strength of hydrochloric acid. Further this study can be extended for the determination of shelf life of roflumilast formulations.

Degradation Kinetics Study of Alogliptin Benzoate in Alkaline Medium by Validated Stability-Indicating HPTLC Method

A rapid, sensitive, and stability-indicating high-performance thin-layer chromatographic method was developed and validated to study degradation kinetics of Alogliptin benzoate (ALG) in an alkaline medium. ALG was degraded under acidic, alkaline, oxidative, and thermal stress conditions. The degraded samples were chromatographed on silica gel 60F254-TLC plates, developed using a quaternary-solvent system (chloroform-methanol-ethyl acetate-triethyl amine, 9+1+1+0.5, v/v/v/v), and scanned at 278 nm. The developed method was validated per International Conference on Harmonization guidelines using validation parameters such as specificity, linearity and range, precision, accuracy, LOD, and LOQ. The linearity range for ALG was 100-500 ng/band (correlation coefficient = 0.9997) with an average recovery of 99.47%. The LOD and LOQ for ALG were 9.8 and 32.7 ng/band, respectively. The developed method was successfully applied for the quantitative estimation of ALG in its synthetic mixture with common excipients. Degradation kinetics of ALG in an alkaline medium was studied by degrading it under three different temperatures and three different concentrations of alkali. Degradation of ALG in the alkaline medium was found to follow first-order kinetics. Contour plots have been generated to predict degradation rate constant, half-life, and shelf life of ALG in various combinations of temperature and concentration of alkali using Design Expert software.

Development and Validation of Stability-Indicating HPTLC Method for Estimation of Naratriptan Hydrochloride in Its Pharmaceutical Dosage Form and Its Content Uniformity Testing.

The present research project involves development and validation of a stability-indicating HPTLC method for the estimation of naratriptan-HCl in their pharmaceutical dosage forms and its content uniformity testing. Naratriptan-HCl was subjected to alkaline, acidic, oxidative, neutral, thermal (dry heat) and photo-degradation conditions. The chromatographic separation was carried out using a precoated silica gel G 60 F254 TLC plate as the stationary phase and dichloromethane-toluene-methanol-triethylamine (4 : 4 : 2 : 1, v/v/v/v) as the mobile phase. The spots of NRT-HCl and its degradation products were detected at 290 nm. The Rf value of NRT-HCl was found to be 0.60 ± 0.02. The linearity was obtained in the range of 100-500 ng/spot. The limit of detection and limit of quantitation were found to be 6.07 ng/spot and 18.41 ng/spot, respectively. The percentage recovery was found in the range of 98.87-99.55%. NRT-HCl was degraded under acidic, alkaline and oxidative conditions while stable under photolytic, neutral and dry heat conditions. The developed method was applied for estimation of naratriptan-HCl in marketed formulations and its content uniformity testing.

Stability-Indicating HPTLC Method for Quantitative estimation of Linagliptin and Metformin Hydrochloride.

Stability-Indicating HPTLC Method for Quantitative estimation of Linagliptin and Metformin Hydrochloride.

Stability-Indicating HPTLC Method for Studying Stress Degradation Behavior of Sulbutiamine HCl.

Sulbutiamine (SUL) is an ester of thiazides with neurotropic action. A new stability indicating HPTLC method has been developed and validated for the determination of SUL in the presence of different degradation products. The drug was subjected to different stress conditions following ICH strategy such as hydrolytic degradation (neutral, alkaline and acidic hydrolysis), oxidation, photodegradation and dry heat degradation. The drug demonstrated degradation under all decomposition conditions except neutral hydrolysis and dry heat, where the drug was completely degraded with 0.1 N NaOH, 1 N HCl and 30% H2O2 while it was partially degradaed by 0.1 N HCl, 3% H2O2 and UV light. Structure elucidation of the resulting degradation products was performed using ESI-Q-MS-MS. A well-defined peak for SUL was obtained at Rf = 0.46 and was completely separated from all obtained degradation products. Chromatographic separation was carried out on HPTLC aluminum plates precoated with silica gel 60 F254 using acetone-methylene chloride-ammonia buffer (pH 8.5 ± 0.2) (7:3:0.5, v/v) as a developing system. Densitometric scanning of the separated peaks was performed at 254 nm. System suitability testing parameters were calculated to ascertain the quality performance of the developed method. The method was validated with respect to USP guidelines regarding accuracy, precision, specificity, robustness and ruggedness. Good correlation coefficients were achieved in the range of 0.4-5.0 µg/band, and the limit of detection and limit of quantitation were found to be 0.11 and 0.33 µg/band, respectively. The utility of the suggested method was verified by application to Arcalion forte® tablets where no interference from additives was found.

Stability-Indicating HPTLC Method for Estimation of Guaifenesin in Bulk and in Pharmaceutical Formulation

A HPTLC/ Densitometric method has been developed for the determination of Guaifenesin (GFN) in bulk and pharmaceutical formulation. The estimation of drug was performed on HPTLC aluminium plates precoated with silica gel 60 RP-18 TLC F254 S using toluene: methanol: ethyl acetate: acetic acid (7:0.8:1.2:0.5 v/v/v/v) as mobile phase. The densitometric quantification for the drug was carried out at 274 nm. GFN obeyed linearity in concentration range 800 – 2800 ng/band with coefficient of correlation 0.999. The Rf for GFN was found to be 0.6 ± 0.02. The proposed method was applied for pharmaceutical formulation and % label claim for GFN was found to be 100.34 ± 1.06. The method was validated for accuracy, precision and ruggedness. Accuracy of the method was checked by recovery studies at three different levels i.e. 80 %, 100 % and 120 %. The % recovery of GFN was found to be in the range of 99.48% - 100.68 %; the % RSD value was less than 2 indicates the accuracy of the method. The method was found to be precise as indicated by the inter-day, intra-day and repeatability analysis; showing % RSD less than 2. The results did not show any statistical difference between operators showing that developed method was rugged. GFN was subjected to acid and alkali hydrolysis, oxidation and thermal degradation. The drug undergoes degradation under acid-base conditions, hydrolysis, oxidation, photo degradation except dry heat degradation. This indicates that the drug is susceptible to acid and base. The degraded product was well resolved from the pure drug with significantly different Rf value. Statistical analysis proves that the method is repeatable, selective and accurate for the estimation of investigated drug. The proposed developed HPTLC method can be applied for the identification and quantitative determination of GFN in bulk drug and pharmaceutical formulation.