Stability-indicating Thin-layer Chromatographic Method Research Articles

Simultaneous Determination of Three 5-HT3Receptor Antagonists Accompanied by Stability Study Using Thin-Layer Chromatography–Densitometry

A thin-layer chromatographic (TLC)—densitometric method for the separation and simultaneous determination of 3 5-HT3 receptor antagonists, namely, tropisetron hydrochloride (TRP), granisetron hydrochloride (GRN), and ondansetron hydrochloride dihydrate (OND) was developed. Densitometric measurements were done at 285 nm for TRP and 305 nm for both OND and GRN using the reflectance—absorbance mode. Separation was carried out on silica gel TLC plates using chloroform—methanol—ammonia (10 m) (8:2:0.1, V/V) as the mobile phase. The three drugs have been separated with RF values of 0.23 ± 0.01, 0.63 ± 0.01, and 0.76 ± 0.01 for TRP, GRN, and OND, respectively. The limits of detection and quantification for linear regression analysis of the studied drugs ranged from 2.79 to 12.05 and from 8.45 to 36.50 ng per band, respectively, while for polynomial regression, the limits of detection and quantification for the studied drugs ranged from 3.40 to 15.51 and from 10.31 to 46.99 ng per band, respectively. The developed method was applied for determination of the studied drugs in pharmaceuticals and human plasma with good precision and accuracy. Stability-indicating thin-layer chromatographic method for the determination of OND in the presence of its degradation products under different conditions and subsequent kinetic study had been successfully applied.

Quality assessment and stability study of compounded furosemide syrup

Compounded oral preparations, made with use of manufactured products as sources of active pharmaceutical ingredients, are characterized by short beyond-use-dates due to their instability.Aim: The aim of this study is to investigate the physical, chemical and microbiological stability of compounded furosemide syrups within a 30 days period.Methods: Batches of 5 mg/ml compounded syrups, using furosemide substance and commercial tablets (two brands) as sources of active pharmaceutical ingredients, were stored in the dark at 5±3 °C and 23±2 °C and examined at days 0, 7, 15, 23, and 30 for changes in physical (pH, formation of colour, gas, odour and changes in viscosity), chemical and microbial stability. A stress test was conducted in order to distinguish signs of chemical instability using a stability-indicating thin-layer chromatographic method. Bacterial inoculation of these samples were examined for microbial stability based on the total aerobic microbial count (TAMC <100), the total combined yeasts/moulds count (TYMC <10) and absence of Escherichia coli.Results: Throughout the storage period the investigated syrups showed no extra spots on the chromatogram, no significant changes in pH, colour, odour, gas formation, viscosity. On day 30, the content (≥99.3%) of furosemide, total aerobic microbial count (< 102), total combined yeasts/moulds count (<101) in studied samples were within acceptable limits. Stressed samples showed presence of extra and diminished spots.Conclusions: Extemporaneous syrups of furosemide substance and dispersed furosemide tablets, stored in glass bottles in the dark at 5±3 °C and 23±2 °C, was found to be physically, chemically and microbiologically stable for at least 30 days

Stability-indicating high-performance thin-layer chromatographic method for simultaneous estimation of the active pharmaceutical ingredients metolazone and spironolactone

This work represents the validation of a stability-indicating thin-layer chromatographic technique for the simultaneous estimation of metolazone (METO) and spironolactone (SPIRO) from marketed formulation (tablets). Thin-layer chromatography was performed using precoated silica gel plate 60 F254 using ethyl acetate—chloroform—GAA (5:5:0.1 v/v) as the mobile phase for the separation of METO and SPIRO. The stability study forms an integral part of the formulation development process, and its use is also encouraged by various guidelines. Stress study was performed on active pharmaceutical ingredients (APIs) as well as on formulation for establishing a stability-indicating thin-layer chromatographic method for both drugs. The APIs were subjected to change under various environmental conditions such as pH, temperature, oxidation, etc. to determine their effect on the stability of drugs. The developed method was able to resolve drugs and their degradation products formed under the aforementioned conditions. The...

Stability-indicating thin-layer chromatographic method for determination of darunavir in complex darunavir–β-cyclodextrin in the presence of its degradation products

A method was developed to identify degradation products of the complex darunavir–β-cyclodextrin.

Development of Stability-Indicating Methods for Cefquinome Sulphate

The degradation behavior of cefquinome sulphate in alkaline medium at different temperatures was investigated using both first derivative spectrophotometric and HPLC methods. The drug degradation was found to be pH and temperature dependant. The pH-rate profile indicated a first order dependence of Kobs on [OH-] at pHs ranging between 9 and 11. Arrhenius plot obtained at pH 10 was linear between 65° and 100°C. The estimated activation energy of the hydrolysis was found to be 21.1 kcal mol-1. Stability-indicating thin-layer chromatographic method for the separation of the drug and its alkaline hydrolysis product has been developed.

Stability-indicating densitometric determination of some angiotensin II receptor antagonists in presence of their degradation products

Summary A simple, selective, precise, and stability-indicating thin-layer chromatographic method has been developed and validated for analysis of some angiotensin II receptor antagonists (AIIRAs), namely, Losartan potassium (Los-K), Irbesartan (Irb), and Candesartan cilexetil (Cand) in the bulk drug and in pharmaceutical formulations (tablets). The method was based on using TLC plates pre-coated with silica gel G 60 on aluminum sheets as stationary phase and the development system was performed using chloroform:methanol (9:1) giving well separated and compact spots for all the studied drugs (RF values 0.41–0.53). The separated spots were characterized by viewing under the UV lamp, then visualized as orange spots by spraying with Dragendorff's reagent and measured by densitometry. Under the optimum chromatographic conditions, linear relationships were obtained between response and concentrations of each studied drug with high correlation coefficients (0.9985–0.9994). Good accuracy and precision were successfully obtained for the analysis of tablets containing each drug alone or combined with diuretic drug hydrochlorothiazide (HCTZ). No interferences could be observed from the co-formulated HCTZ, commonly encountered excipients present in tablets as well as the degradation products. The results were compared successfully with reported methods and can be used as a stability-indicating assay.

Design Expert-Supported Development and Validation of High-Performance Thin-Layer Chromatographic Stability-Indicating (HPTLC) Method: an Application in Quantitative Analysis of Ropinirole in the Bulk Drug and in Marketed Dosage Forms

A simple, precise and stability-indicating thin-layer chromatographic method for estimation of ropinirole HCl was developed and validated as per ICH guidelines. The mobile phase consisted of acetone–cyclohexane–6 M ammonia solution (8:5.5:0.5, v/v/v). Scanning the drug was done at 250 nm. Compact spots for ropinirole HCl were found at an Rf value of 0.51 ± 0.002. The linear regression analysis data for the calibration plots showed good linear relationship with R2 0.9976 ± 0.0011 in the working concentration range of 100–3,000 ng spot−1. The method was validated for precision, accuracy, ruggedness, robustness, specificity, recovery, limit of detection (LOD), and limit of quantitation (LOQ). The LOD and LOQ were 12.89 and 42.53 ng spot−1, respectively. The drug was subjected to degradation; the peaks of degradation products were well resolved from the standard with significantly different Rf values. Hence, this method can be used for quality control assay of ropinirole.

APPLICATION OF A STABILITY-INDICATING TLC METHOD FOR THE QUANTITATIVE DETERMINATION OF DEXKETOPROFEN TROMETAMOL IN PHARMACEUTICAL DOSAGE FORMS

Objective: A sensitive, selective, precise and stability-indicating thin layer chromatographic method for analysis of Dexketoprofen trometamol, both as a bulk drug and in formulation, was developed and validated. Method: The method employed TLC aluminum plates precoated with silica gel 60F-254 as the stationary phase and the solvent system consisted of toluene:ethyl acetate:glacial acetic acid (6:2:0.2 v/v/v). This system was found to give compact spots for Dexketoprofen trometamol (Rf value of 0.45 ± 0.02). Dexketoprofen trometamol was subjected to acid and alkali hydrolysis, oxidation, photo, and neutral degradation. Result: The peak of the degradation product was well-resolved from the pure drug and had significantly different Rf value. Densitometric analysis was carried out in the absorbance mode at 255 nm. The method was validated for precision, robustness, and recovery. Conclusion: As the method can separate the drug from its degradation products, it can be used to monitor stability.

Development and validation of a simple stability-indicating TLC method for the determination of levamisole in pharmaceutical tablet formulation

Asimple, selective, precise, and stability-indicating thin-layer chromatographic (TLC) method has been established and validated for analysis of levamisole (LEV) in pharmaceutical dosage. Normal phase precoated aluminum TLC silica gel 60F254 plates were used as stationary phase and MeOH-toluene-chloroform (14:36:50 v/v/v) was used as mobile phase to determine LEV in pharmaceutical ingredients. The system was found to give compact spot for LEV (RF = 0.30). Determination was performed in absorbance mode at 223 nm. Regression analysis data for the calibration plots indicated good linear relationships between response and concentration over the range 60–2000 ng per spot and 0.998 correlation coefficient. The slope and intercept of the calibration plot were 4.946 and 267.6, respectively. The recovery of 88.44–102.57% with 2.02% inter-day and 1.91% intra-day relative standard deviations (RSDs) shows the validity of the method. Further, the limit of detection (LOD) and the limit of quantification (LOQ) were 2.2 ...

Application of a stability-indicating TLC method for the quantitative determination of Levocetirizine in pharmaceutical dosage forms

A sensitive, selective, precise and stability-indicating thin layer chromatographic method of analysis for Levocetirizine both as a bulk drug and in formulation was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60 F-254 as the stationary phase and the solvent system consisted of ethyl acetate: methanol: ammonia (9: 2.5: 1.5 v/v/v). This system was found to give compact spots for Levocetirizine (Rf value of 0.50 ± 0.02). Levocetirizine was subjected to acid and alkali hydrolysis, oxidation, photo and neutral degradation. The peak of the degradation product was well resolved from the pure drug and had significantly different Rf value. Densitometric analysis of Levocetirizine was carried out in the absorbance mode at 230 nm. The linear regression analysis data for the calibration plots showed a good linear relationship over the concentration range of 200-1200 ng/spot. The mean values of the correlation coefficient, slope and intercept were 0.9904 ± 1.53, 2.8899 ± 0.998 and 802.6 ± 0.982 respectively. The method was validated for precision, robustness and recovery. The limit of detection (LOD) and limit of quantitation (LOQ) was 100 and 200 ng/spot respectively. Statistical analysis showed that the method is repeatable and selective for the estimation of Levocetirizine. Since the method can separate the drug from its degradation products, it can be used to monitor stability. Keywords: Degradation; Levocetirizine; TLC; stability-indicating.

Stability-Indicating TLC Method for the Determination of Dutasteride in Pharmaceutical Dosage Forms

A simple, sensitive, selective, precise and stability-indicating thin-layer chromatographic method for determination of dutasteride both as a bulk drug and as pharmaceutical tablets was developed and validated as per the International Conference on Harmonization guidelines. The method employed thin-layer chromatography aluminium plates precoated with silica gel 60F254 as the stationary phase and the mobile phase consisted of acetonitrile:methanol:dichloromethane in the ratio of 2.0:1.0:2.0, v/v/v. This solvent system was found to give compact spots for dutasteride (Rf value of 0.64 ± 0.02). Densitometric analysis of dutasteride was carried out in the absorbance mode at 244 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r = 0.9943 with respect to peak area in the concentration range of 100–600 ng per band. The method was validated for precision, accuracy, ruggedness and recovery. The limits of detection and quantitation were 7.54 and 22.85 ng per band, respectively. Dutasteride was subjected to acid and alkali hydrolysis, oxidation, photo degradation, dry heat and wet heat treatment. The drug undergoes degradation under acidic, basic conditions, photolytic, oxidative and upon wet and dry heat treatment. The degraded products were well separated from the pure drug. The statistical analysis proves that the developed method for quantification of dutasteride as bulk drug and from pharmaceutical tablets is reproducible and selective. As the method could effectively separate the drug from its degradation products, it can be employed as stability-indicating.

Application of a Stability-Indicating Thin-Layer Chromatographic Method to the Determination of Tenatoprazole in Pharmaceutical Dosage Forms

A sensitive, selective, precise, and stability-indicating thin-layer chromatographic (TLC) method was developed and validated for the determination of tenatoprazole both as a bulk drug and in formulation. The method uses TLC aluminum plates precoated with Silica Gel 60F-254 as the stationary phase and the solvent system toluene-ethyl acetate-methanol (6 + 4 + 1, v/v/v). This system gave compact spots for tenatoprazole (Rf value of 0.34 +/- 0.02). Tenatoprazole was subjected to acid and alkali hydrolysis, oxidation, and photodegradation. The peaks of the degradation products were well-resolved from that of the pure drug and had significantly different Rf values. Densitometric analysis of tenatoprazole was performed in the absorbance mode at 306 nm. The linear regression analysis data for the calibration plots showed a good linear relationship over the concentration range of 100-1500 ng/spot. The mean values of the correlation coefficient, slope, and intercept were 0.9989 +/- 1.42, 10.27 +/- 0.965, and 4894.2 +/- 1.24, respectively. The method was validated for precision, robustness, and recovery. The limit of detection and limit of quantitation were 50 and 100 ng/spot, respectively. Statistical analysis showed that the method is repeatable and selective for estimation of tenatoprazole. Because the method can separate the drug from its degradation products, it can be used to monitor stability.

Selective Densitometric Analysis of Cephalosporins Using Dragendorff’s Reagent

A simple, selective, precise, and stability-indicating thin-layer chromatographic method has been developed and validated for analysis of the cephalosporins cefpodoxime proxetil, ceftriaxone sodium, ceftazidime pentahydrate, cefotaxime sodium, cefoperazone sodium, cefazolin sodium, and cefixime in the bulk drug and in pharmaceutical formulations. TLC was performed on aluminium sheets precoated with silica gel G 60F254 as stationary phase. The mobile phases chosen for development gave compact spots for all the drugs (RF values 0.43–0.60). The separated compounds were visualized as orange spots by spraying with Dragendorff’s reagent. Linear regression analysis data for the calibration plots revealed good linear relationships between response and amounts of the drugs with correlation coefficients ranging from 0.9977 to 0.9998 and determination coefficients ranging from 0.9954 to 0.9996 over the concentration ranges 5–25 μg per spot for cefpodoxime proxetil, ceftriaxone sodium, and ceftazidime pentahydrate and 10–50 μg per spot for cefotaxime sodium, cefoperazone sodium, cefazolin sodium, and cefixime. The method was validated for precision, recovery, and robustness. Limits of detection and quantitation for the drugs ranged from 0.35 to 2.48 and from 1.07 to 7.50 μg per spot, respectively. The method was successfully applied to analysis of the drugs in their pharmaceutical dosage forms with good precision and accuracy. The method can also be used as a stability-indicating assay.

Quantitative analysis of repaglinide in tablets by reversed-phase thin-layer chromatography with densitometric UV detection

A simple, rapid, and stability-indicating thin-layer chromatographic method has been developed for quantitative determination of repaglinide in tablets. Analysis was performed on RP-8 TLC plates with acetonitrile-pH 6.0 phosphate buffer, 60 + 40 (% v/v ), as mobile phase. Detection and quantification were performed by classical densitometry at the wavelength of maximum absorption of repaglinide, 225 nm. A calibration plot constructed in the range 0.6–3.6 μg/10 μL was linear with a good correlation coefficient ( r = 0.998 ± 0.001, mean ± SD , n = 5). Limits of quantitation and detection of repaglinide were 0.27 μg/10 μL and 0.08 μg/10 μL, respectively. Instrumental precision established at three concentrations of the drug ranged from 3.92 to 0.97% for the lowest and highest concentrations of repaglinide, respectively. The mean intra-day and inter-day variability, including three concentrations of repaglinide, were 1.93 and 2.25% ( n = 9), respectively. Recovery from model mixtures, at three levels of addit...