Oxidative Degradation Conditions Research Articles

FORCED DEGRADATION STUDIES OF NILOTINIB HYDROCHLORIDE: ISOLATION, IDENTIFICATION & CHARACTERIZATION OF IMPURITIES

Nilotinib hydrochloride is a tyrosine kinase inhibitor approved for the treatment of chronic myelogenous leukemia was subjected to forced degradation studies and the samples were analyzed by utilizing the LCMS compatible HPLC methods. Nilotinib Hydrochloride was subjected to thermal, hydrolytic, oxidative, acidic, basic and photolytic degradation conditions as per the regulatory guidelines. The drug was degraded in oxidative, basic and acidic environments and stable in photolytic and thermal conditions. The main degradation impurity components produced through the forced degradation study were isolated for the identification and quantification in presence of these impurities in the stability studies of drug substances as well as drug products. The identified degradation components were separated by mass assisted auto-purification technique and subjected for the characterization by NMR (13C-NMR, 1H-NMR, HMBC and HSQC), HRMS and FT-IR experimentations. Degradation products obtained from oxidative, basic and acidic environments were isolated and identified by the advanced techniques as acid degradation product (DP-1) with molecular mass of 306.11 g/mol, empirical formula C17H14N4O2 with name as 4-methyl-3- (4 -(pyridine -3-yl) pyrimidin -2 -ylamino) benzoic acid. Base degradation product (DP-2) has molecular weight of 241.08 g/mol, molecular formula C11H10F3N3 with name as 3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline.Oxidative degradation product (DP-3) has molecular weight of 545.18 g/mol, molecular formula C28H22F3N7O2 with name as 3-(2-(2-methyl-5-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenylcarbamoyl) phenylamino)pyrimidin-4-yl)pyridine1-oxide.

Efficient degradation of indigo wastewater by one-step electrochemical oxidation and electro-flocculation

Purpose Indirect electrochemical oxidation and electro-flocculation were combined to degrade indigo wastewater. Design/methodology/approach The degradation efficiency of indigo wastewater in single-cell and double-cell were investigated. Based on the previous single factor experiments, the oxidative degradation conditions of indigo wastewater in single cell were optimized by response surface methodology (RSM). The decolorization rate, chemical oxygen demand (COD) removal rate, the contents of flocculation precipitation and indigo were measured and analyzed. Findings The degradation efficiency in single cell was higher than in double cell. The electrolysis conditions were optimized by RSM and the decolorization rate was 99.01% with COD removal rate of 60.34% and conductivity of 89.75 mS/cm. The amount of flocculated precipitation decreased by 53.33% and the indigo increased by 86.34%. The content of Na and S decreased by 12.13 and 6.49%, respectively. The ratio of Fe3+ to Fe2+ in the solution was 4.62:1, indicating that most of the iron dropped on the electrode sheet was converted to Fe3+. Research limitations/implications The one-step electrochemical oxidation and flocculation method with the advantages of simple operation and environmental protection, provided a reference for the actual treatment of dyeing wastewater. Practical implications Combining the electrochemical flocculation and oxidation provided an efficient and practical solution for degradation of indigo wastewater. Originality/value Combining the advantages of electrochemical oxidation and electroflocculation, the application of electrochemistry in printing and dyeing wastewater treatment technology has been expanded.

Forced degradation studies of medroxyprogesterone acetate injectable suspensions (150 mg/ml) with implementation of HPLC, mass spectrometry, and QSAR techniques

Medroxyprogesterone acetate (MPA) injectable products are a key commodity for reproductive health and are available in the global market from a variety of manufacturing sources. Depending on the climatic zone conditions of the destination country for product use, MPA injectables are at risk of exposure to adverse transport and storage conditions. Analytical methods are available that quantify impurity levels in MPA and MPA injectable products, but minimal information is publicly available on the source of impurity and degradation product generation or the safety risk of these compounds. Forced degradation studies were conducted on MPA and MPA injectables to gain a better understanding of potential sources of impurities and degradation products. Furthermore, QSAR analysis was conducted to assess the toxicity risk of known impurities. More impurities were generated under acidic, basic, light, and oxidative forced degradation conditions relative to thermal degradation, however thermal exposure is the most likely adverse condition to be experienced by these products. Even if impurities are present in MPA injectables, QSAR analysis found that known impurities for MPA are apparently no more of a safety risk than MPA.

Development and optimization of a stability-indicating chromatographic method for verapamil hydrochloride and its impurities in tablets using an analytical quality by design (AQbD) approach

Development and optimization of analytical methods for quantifying verapamil hydrochloride in its twelve impurities is a challenge for analytical chemists and pharmacists. The aim of this paper was to apply the analytical quality by design (AQbD) and design of experiments (DoE) approaches in the development of a robust, low-cost, regulatory-flexible, fit for purpose high-performance liquid chromatography (HPLC) analytical method to determine VPM and its impurities. A Placket-Burman screening design was adopted to identify the most relevant analytical conditions affecting the chromatographic responses. In addition, a central composite optimization design was used to establish regression models explaining the chromatographic responses as functions of the buffer pH, concentration of ammonium hydroxide in mobile phase, and the injection volume of test solutions. Response surface methodology allowed for establishing the method operable design space, which ensures suitable method performance with respect to all chromatographic responses studied. Optimal chromatographic conditions were achieved using a gradient elution consisting of a mixture of 10 mM ammonium formate buffer pH and acetonitrile, with a flow rate of 0.7 mL/min, and a XSelect CSH C18 100 mm x 4.6 mm 3.5 µm chromatographic column. The optimized analytical method was selective, linear (320–480 µg/mL for VPM and 0.4–4.8 µg/mL for impurities), precise (0.5% and 1.0% for VPM and impurities, respectively), accurate (mean recovery of 101.0% and 102.5% for VPM and impurities, respectively), and robust. In addition, the application of the validated analytical method in forced degradation conditions allowed for studying the degradation profile of verapamil hydrochloride, particularly in acid and oxidative degradation conditions.

Development and validation of a stability indicating high performance liquid chromatography method for trimethobenzamide

A simple, accurate, precise and robust stability indicating RP-HPLC assay method has been developed for the estimation of trimethobenzamide in stress sample. An isocratic separation of trimethobenzamide was achieved on Kromasil 100 C-18 column (250 X 4.6mm, 5μ) with a flow rate of 1.0 ml/min and by using a photodiode array detector to detect the analyte at 213nm. The optimized mobile phase consisted of methanol: ammonium formate (44:56, v/v). The drug was subjected to different forced degradation conditions according to ICH guidelines including acid, base, neutral hydrolysis, oxidation, photolysis and thermal degradation. Degradation products were found only in basic and oxidative degradation conditions. All the degradation products got eluted in an overall analytical run time of 12min. The developed analytical method has been validated according to the ICH guidelines. Response of trimethobenzamide was linear over the concentration range of 0.5-50μg/ mL (r2 = 0.999). Accuracy was found to be in between 94.03% to 100.39%. Degradation products resulting from the stress studies did not interfere with the detection of the analyte.

Identification and Characterization of Forced Degradation Products for Dofetilide using Rapid and Sensitive UPLC-MS/MS Method and HRMS Studies

A simple, rapid and efficient method has been developed and validated using ultra UPLC combined with Q-ToF MS system for recognition and characterization of forced degradation products obtained from dofetilide degradation studies. The dofetilide drug is an antiarrhythmic and belongs to Class III and it was treated with various stress conditions like acidic, basic, oxidative, photolytic and thermal conditions as per ICH guidelines. The main drug shows extensive degradation towards oxidative degradation conditions and single degradation product was identified through chromatogram. The chromatographic separation among main and its impurities were attained through 2.1 × 150, 1.8 μm column from gradient elution using UPLC and its detection at wavelength 230 nm. The validation was performed for the developed method using various parameters like specificity, linearity and robustness studies. Waters Synapt G2 Q TOF system was used and performed MSn studies to establish mass spectral fragmentation pathway for drug and its degradation products and determined accurate masses study. The efficiency of this method was helpful to identify and characterize the drug and degradation products using LC/MSn techniques.

STABILITY-INDICATING METHOD DEVELOPMENT AND VALIDATION OF ITRACONAZOLE AND TERBINAFINE HCL IN BULK AND PHARMACEUTICAL TABLET DOSAGE FORM

Objective: The objective of the present work was to develop and validate the stability-indicating method for the simultaneous estimation of itraconazole and terbinafine HCl in bulk and pharmaceutical tablet dosage form by reversed-phase high-performance liquid chromatography (HPLC). This combination of drugs is not reported for simultaneous HPLC analysis as of now.
 Methods: The analysis of the developed method was carried on Shimadzu LC Prominence-i 2030 model with Lab Solution software and the separation was done on Shim-pack C18 GIST (250 mm×50 mm, 5 μm) column with a flow rate of 1.2 ml/min and run time of 12 min. The injection volume was 10 μl and mobile phase consisted of acetonitrile and 0.1% triethylamine in the ratio of 90:10 and 225 nm was used as a detection wavelength.
 Results: The retention time was found to be 3.464 min and 8.705 min for itraconazole and terbinafine HCl, respectively. The calibration curve was found to be linear and r2 values were 0.9989 and 0.9995 for itraconazole and terbinafine HCl, respectively.
 Conclusion: The stability-indicating method was developed by subjecting itraconazole and terbinafine HCl marketed formulation to various stress conditions such as acidic, basic, oxidative, thermal, and water hydrolysis degradation conditions and the degraded product peaks were well resolved from sample peaks.

STABILITY-INDICATING HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY METHOD FOR SIMULTANEOUS ESTIMATION OF FORMOTEROL FUMARATE DIHYDRATE AND FLUTICASONE PROPIONATE IN BULK DRUG AND PHARMACEUTICAL DOSAGE FORM

Objective: The objective of the present work was to develop validated stability-indicating high-performance thin-layer chromatographic method for simultaneous estimation of formoterol fumarate dihydrate (FFD) and fluticasone propionate (FP) in bulk drug and pharmaceutical dosage form.
 Methods: Pre-coated silica gel aluminum plates 60 F-254 were used as stationary phase. The mixture of toluene:ethyl acetate:formic acid (98%) (6:4:0.1; v/v/v) was used as a mobile phase. The densitometric quantification was carried out at 233 nm. The method was validated according to the ICH guidelines. The specificity and stability indicating the capability of the method were proven though degradation studies. Both drugs were subjected to acid (0.1N HCl) and base (0.1N NaOH) hydrolysis, oxidation (3% v/v H2O2), photolytic, and neutral degradation conditions.
 Results: The selected mobile phase resolved peaks of FFD and FP with Rf values 0.27±0.10 and 0.64±0.10, respectively. Determination coefficients of calibration curves were found to be 0.998 and 0.999 in the range of 1–3.5 μg/spot and 10–60 μg/spot for FFD and FP with an accuracy of 99.09% for FFD and 99.20% for FP. The degradation products of FFD and FP were resolved from the pure drug with significant differences in their retention factor values.
 Conclusion: The developed method is simple, accurate and can be successfully applied for quantification of FFD and FP in bulk drug and pharmaceutical dosage form, contributing to improve the quality control and assure the therapeutic efficacy.

Development of a validated spectrofluorimetric method for assay of sotalol hydrochloride in tablets and human plasma: application for stability-indicating studies

AbstractThe native fluorescence of sotalol hydrochloride (SOT) was used as a basis for establishing a new method of analysis for SOT in tablets and spiked human plasma. The fluorescence of SOT in water was measured at 310 nm when excited at 235 nm. The detection limit (LOD) was 0.37 ng/mL and the quantification limit (LOQ) was 1.08 ng/mL. The proposed method offers high sensitivity which permits determination of SOT, even if present in a very small amount, in human plasma. The obtained results were successfully compared to that of a reference pharmacopeial method and statistical analysis proved a good agreement between the results of both methods. Further investigation of the SOT stability upon exposure to various stress conditions, such as acidic, alkaline, oxidative and photolytic degradation conditions was also performed. The kinetics of acidic, alkaline and oxidative degradation of the drug showed a pseudo first order degradation reaction. A proposal of the degradation pathway was suggested and confirmed by developing a thin layer chromatographic method used for separation of SOT and its acidic and alkaline degradation products.

STABILITY INDICATING METHOD DEVELOPMENT AND VALIDATION OF METFORMIN AND ERTUGLIFLOZIN BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH PDA DETECTION AND ITS APPLICATION TO TABLET DOSAGE FORM

Objective: A sensitive, rapid, accurate, and precise stability indicating high-performance liquid chromatography method was developed and validated for the simultaneous estimation of metformin (MET) and ertugliflozin (ERT).
 Methods: The planned chromatographic method was developed using Kromasil C18 column using 0.1 M sodium dihydrogen phosphate methanol (50:50, by volume, pH 4.0) as mobile phase system followed by peak area response measurement at 238 nm. The developed method was validated by means of ICH guidelines about system suitability, linearity, sensitivity, selectivity, accuracy, precision, robustness, and specificity.
 Results: Calibration curves of MET and ERT are linear from 250 to 750 μg/ml and 3.75 to 11.25 μg/ml, respectively. Relative standard deviation is <2.0% and recovery is ̴ 100%. Successfully, the developed method was applied for the simultaneous determination of MET and ERT in tablets and to study degradation of MET and ERT in acidic, alkaline, oxidation, thermal, and photolytic degradation conditions. No obstructions from additional common excipients of tablets and degradants were observed.
 Conclusion: The results recommended method suitability for the analysis of MET and ERT in quality control laboratories.

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.

Optimization of a forced degradation study of atorvastatin employing an experimental design approach

This study involved the optimization of experimental conditions for the forced degradation of atorvastatin employing the experimental design (DoE) approach, as a scientific multifactorial strategy. Using 2n full factorial design, stress conditions of oxidative, hydrolytic and thermal degradation were optimized to obtain a targeted level of atorvastatin degradation. Atorvastatin and all related and degradation products were separated on Poroshell 120 EC C18 50 ´ 3.0 mm 2.7 μm, using 10 mM ammonium formate and acetonitrile as mobile phases in the gradient mode. The impurity structures were confirmed by the direct hyphenation of a liquid chromatograph to an ion trap mass spectrometer with a heated electrospray ionization interface.This study highlights the multifold benefits of implementing the DoE concept, which provides a better understanding of the significant factors responsible for degradation and ensures a successful way to achieve degradation, thereby replacing the trial and error approach used in conventional forced degradation studies.

Stability Indicating UV Spectrophotometric Method for Estimation of Omeprazole and Its Application to Content Uniformity Testing

Omeprazole is a most commonly used antiulcer agent in clinical practices. A least time consuming efficient and simple ultraviolet spectrophotometric method for the assay of omeprazole has been developed. The assay was based on the ultraviolet absorbance maxima at about 217.80 nm wavelength of omeprazole using sodium hydroxide as the solvent. In the present study comprehensive stress degradation was carried out according to ICH Q1 (R2) guidelines. The drug was subjected to acidic (0.1N HCl), basic (0.1N NaOH), oxidative (1% H2O2), photolytic and thermal degradation conditions. The developed UV spectrophotometric method showed high degradation under acidic condition, moderate degradation under basic, photolytic and thermal conditions. But it was relatively stable under oxidative conditions. The pathway for degradation has been proposed. The method was validated for Linearity, Accuracy, Precision, Specificity, Ruggedness and Robustness. The method shows good linearity in the range of 2-20μg/ml. The LOD and LOQ were found to be 0.1 μg/ml and 1.1 μg/ml. The %RSD was found to be with in limit i.e. less than 2%.The mean recovery of placebo was 100.68%. It can be concluded that the developed procedure is valid and can be applicable for determination of content uniformity for available brands of omeprazole. This method is applicable for the daily routine quality control quantitative analysis of omeprazole.

Development and Validation of a Stability-Indicating Liquid Chromatographic Method with Mass-Spectrometric Detection for the Analysis of Apixaban and Determination of Production Related Impurities

Apixaban is a novel anticoagulant drug acting as a direct, selective and reversible inhibitor of coagulation factor Xa. The purpose of this work was to develop and validate a sensitive, stability-indicating LC-MS method for the analysis of apixaban and estimation of production-related substances in the active pharmaceutical ingredient (API) and pharmaceutical dosage forms. The analyses were carried out on a Macherey-Nagel Nucleodur C18 column (250 × 4.6 mm, 5 μm particle size) with a mobile phase containing acetonitrile and water in gradient program at a flow rate of 0.8 mL/min, the column oven temperature 25°C, and the UV detection tuned to 270 nm. Apixaban was subjected to stress conditions of oxidative, acid, base, hydrolytic, thermal and photolytic degradation, and found susceptible to acid and base hydrolysis conditions, while it was stable under thermal, photolytic, and oxidative degradation conditions. The degradation products were well resolved from the main peak and its impurities, proving the stability-indicating power of the proposed method. The proposed method was validated as per International Conference on Harmonization (ICH) guidelines. The method is suitable for practical routine analysis and it has been successfully applied to analyze apixaban in eliquis brand preparations. All analytical results were acceptable according to the pharmaceutical requirements.

Comprehensive Assessment of Degradation Behavior of Simvastatin by UHPLC/MS Method, Employing Experimental Design Methodology.

This manuscript describes comprehensive approach for assessment of degradation behavior of simvastatin employing experimental design methodology as scientific multifactorial strategy. Experimental design methodology was used for sample preparation and UHPLC method development and optimization. Simvastatin was subjected to stress conditions of oxidative, acid, base, hydrolytic, thermal, and photolytic degradation. Using 2n full factorial design degradation conditions were optimized to obtain targeted level of degradation. Screening for optimal chromatographic condition was made by Plackett–Burman design and optimization chromatographic experiments were conducted according to Box-Behnken design. Successful separation of simvastatin from the impurities and degradation products was achieved on Poroshell 120 EC C18 50 × 3.0 mm 2.7 μm, using solutions of 20 mM ammonium formate pH 4.0 and acetonitrile as the mobile phase in gradient mode. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines. Validation results have shown that the proposed method is selective, linear, sensitive, accurate, and robust and it is suitable for quantitative determination of simvastatin and its impurities. Afterwards, the degradation products were confirmed by a direct hyphenation of liquid chromatograph to ion-trap mass spectrometer with heated electrospray ionization interface. This study highlights the multiple benefits of implementing experimental design, which provides a better understanding of significant factors responsible for degradation and ensures successful way to achieve degradation and can replace the trial and error approach used in conventional forced degradation studies.

Development and Validation of a Novel Stability-Indicating Reversed-Phase Ion-Pair Chromatographic Method for the Quantitation of Impurities in Marbofloxacin Tablets

A stability-indicating isocratic reversed-phase ion-pair chromatographic method was designed for the separation of impurities in the presence of degradation products. Marbofloxacin tablets and a placebo were exposed to the stress conditions of oxidative, acid, base, humidity, thermal, and photolytic degradation. Significant and moderate degradation was observed in acidic and oxidative stress conditions, respectively. The degradation products were well resolved from the main peak and its impurities, thus proving the stability-indicating analytical method. The method was developed by using an XTerra RP18 3.5 μm (150 × 4.6 mm) column, with the mobile phase containing a mixture of buffer (pH 2.5)-methanol-glacial acetic acid (77 + 23 + 0.5, v/v). The flow rate of the mobile phase was 1.2 mL/min, with a column oven temperature of 40°C and a detection wavelength of 315 nm. The proposed method met Veterinary International Conference on Harmonization requirements and was successfully used for impurity quantitation in marbofloxacin tablets.

STABILITY STUDY AND VALIDATED REVERSED PHASE LIQUID CHROMATOGRAPHIC METHOD FOR THE DETERMINATION OF TIROFIBAN HYDROCHLORIDE IN PRESENCE OF TYROSINE AS A PROCESS IMPURITY

Tirofiban hydrochloride was subjected to the degradation under conditions of hydrolysis (acidic and alkaline degradation), oxidative, thermal and photolytic degradation as prescribed by ICH. A simple and precise liquid chromatographic method has been developed and validated for the simultaneous determination of tirofiban hydrochloride monohydrate (TIR) and its synthetic starting material; tyrosine (TRS). All the chromatographic separations were achieved on Zorbax SB C18, 250 mm×4.6 mm i.d., 5μm column at a flow rate of 1 mL min−1. Isocratic elution based on 0.1 M phosphate buffer (pH 3) - acetonitrile (70:30, v/v ) with UV detection at 227 nm was applied. For the stability study separation of TIR from its degradation products was achieved using 0.1 M phosphate buffer (pH 3) - acetonitrile (72:28, v/v ) with UV detection at 210 nm. Method validation parameters namely, linearity, accuracy and precision were found to be acceptable over the concentration ranges of 10-250 μg mL-1 for TIR and 1-70 μg mL-1 for TRS. The minimum detection limits were 1.76 μg mL-1 for TIR and 0.13 μg mL-1 for TRS. The optimized method was validated and proved to be specific, robust and accurate for the quality control of the cited drug in drug substance and drug product.

Development and Full Validation of a Stability-indicating HPLC Method for the Determination of the Anticancer Drug Temozolomide in Pharmaceutical Form.

In the present study, an accurate, precise and simple method has been developed for the determination of TMZ in its pharmaceutical form by using HPLC. An HPLC method with a DAD was validated according to ICH guidelines. A C18 column (150x4.6 mm. i.d., 5 µm particle size) and an aqueous acetate buffer (0.02 M)-acetonitrile (90:10, v/v) (pH 4.5) as a mobile phase were used. The linear range and LOD value were 5-100 µg/mL and 0.02 µg/mL, respectively. The accuracy of the method was determined using a recovery test and found as 98.8-100.3%. In addition, forced degradation studies of the drug were also performed in bulk drug samples to demonstrate the specificity and stability-indicating. Degradation studies under acidic, basic, oxidative, and thermal degradation conditions were applied. The proposed method could be applied successfully for the determination and identification of the degradation of the drug.

RP-HPLC AND HPTLC STABILITY INDICATING ASSAY METHODS FOR IVERMECTIN IN BULK AND TABLET DOSAGE FORM

Simple, rapid, precise, accurate RP-HPLC and HPTLC methods have been developed and validated for ivermectin in bulk and its marketed formulation. RP-HPLC method for drug was achieved on Grace C18 (250 mm X 4.6 ID, Particle size; 5 μ) column using mobile phase acetonitrile: 10 mM phosphate buffer (95:05 v/v) pH adjusted to 3 with o-phosphoric acid. Detection of drug was done at 245 nm. The retention time was found to be 5.83 min. HPTLC method for ivermectin was accomplished on a precoated silica gel aluminium plate 60F-254 (CAMAG Linomat 5), using toluene: methanol: glacial acetic acid (8:2:0.1 v/v/v) as a mobile phase. The densitometric scanning was performed at 245 nm which showed Rf 0.46 for ivermectin. The stress testing of the drug individually was carried out under acidic, alkaline, oxidation, photo-stability and thermal degradation conditions. The proposed methods were successfully applied for the determination of drug in bulk and its marketed formulation.

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-HPLC METHOD FOR FLUTICASONE FUROATE AND VILANTEROL IN PHARMACEUTICAL FORMULATIONS

A simple, rapid, accurate and stability indicating High Performance Liquid Chromatographic method was developed and validated for the simultaneous determination of vilanterol (VTL) and fluticasone furoate (FFE) in pure and in pharmaceutical formulations using C18 Column and methanol: acetonitrile: 1% sodium perchlorate in the ratio 50:40:10 v/v/v as mobile phase at pH 4.8 and flow rate of 1 mL/min with isocratic elution. The eluted compounds were detected using UV7000 detector at a wavelength of 254nm. The retention times of VTL and FFE were found to be 4.54 and 7.65 min, respectively, with a correlation coefficient of 0.9990 each. The linearity ranges for VTL and FFE were found to be 2.5–15μg/ml and 10–60μg/ml respectively. The LOD and LOQ values were observed to be 0.001μg/mL and 0.02μg/mL for VTL and 0.005μg/mL and 0.07μg/mL for FFE. The percentage recovery was found to be in range of 98.37 – 99.84 % and 98.46 –101.42 % for VTL and FFE, respectively. Both the drugs were subjected to acid, base, hydrolysis, oxidation, photolytic and thermal degradation conditions. The degradation products of VTL and FFE were well resolved from the pure drug with significant differences in their retention time values. This validated method was applied for the simultaneous estimation of VTL and FFE in a commercially available formulation sample.