ICH Q2 R1 Guidelines Research Articles

Advanced Derivative Spectroscopic Method for Estimation of Montelukast and Bilastine in Their Tablet Dosage Form

The present work aims to develop and validate the advanced derivative spectroscopic method for estimation of Montelukast (MON) and Bilastine (BIL) in their tablet dosage form. This tablet dosage form is used for anti-asthmatic and allergic rhinitis. The developed method can be applied for simultaneous estimation of Montelukast and Bilastine in their combined dosage form. For this advanced derivative method, the absorbance at 226.8 nm (ZCP of Montelukast) and 326.4nm (ZCP of Bilastine) was used to estimate Montelukast and Bilastine, respectively. The developed method is validated by ICH Q2R1 guidelines with validation parameters like linearity, LOD, LOQ, accuracy, precision, robustness, ruggedness and assay were performed using this guideline. The method was found to be linear in the concentration range of 2-14 µg/mL for Montelukast (R2 = 0.999) and 4–28 µg/mL for Bilastine (R2 =0.9997). LOD and LOQ found 0.1216 and 0.3686 for MON, and 0.3406 and 1.0320 for BIL. The precision study wasa carried out by comparing on 3 different concentrations and the result of their %RSD was is less than 2%. Robustness study carried out by the change in scanning speed and change in methanol manufacturer and Ruggedness study carried out by different analyst. Assay study was performed using tablet formulation. The developed method was utilized for simultaneous estimation of Montelukast and Bilastine for its tablet dosage form.

LC-MS characterization of acid degradation products of metoclopramide: Development and validation of a stability-indicating RP-HPLC method

The present study aims to develop a simple, accurate and specific stability-indicating RP-HPLC technique for the analysis of metoclopramide in the presence of its stress degradation products and characterization of degradation compounds by LC-MS/MS analysis. As per ICH Q1A-R2 guidelines, the drug was exposed to acid hydrolytic stress condition. Three degradation products were formed for MCP in acid hydrolysis. The liquid chromatography was processed on a Luna C18-(2) 100A,250×4.6mm 5micron column using an isocratic mobile phase consisting of 0.1% formic acid in water-acetonitrile (20:80, v/v) by adjusting the mobile phase at 1 ml/min flow rate with wavelength detection at 273 nm. The developed procedure was applied to LC-MS/MS (liquid chromatography-tandem mass spectrometry) for the characterization of all the degradant components. Total new three degradation compounds were recognized and identified by LC-MS/MS. The developed RP-HPLC technique was validated as per the ICH Q2-R1 guidelines. Limit of detection and limit of quantification values of MCP were evaluated from the linearity graph and were found to be 5.23 µg/ml and 17.44 µg/ml. Accuracy study was established at 80.0, 100.0 and 120.0 µg/ml concentration levels and the findings were found in the range of 98.4% - 101.8%. The linearity of the technique was assessed over the drug concentration range of 50.0 µg/ml to 250.0 µg/ml and the regression equation, slope and correlation coefficient values were found to be y = 10618x + 1623.2, 10618 and 0.9996 respectively. The developed technique was uninterruptedly applied for the quantification of metoclopramide inactive pharmaceuticals.

QUANTITATIVE ANALYSIS OF PAMABROM AND IBUPROFEN IN SYNTHETIC MIXTURE USING 1ST ORDER DERIVATIVE SPECTROSCOPY

Objective: The preliminary goal was to develop and validate 1st order derivative spectroscopic method for quantitative analysis of Pamabrom (PAMA) which is a xanthine diuretic and ibuprofen (IBU) which is a non-steroidal anti-inflammatory agent from its synthetic mixture.
 Methods: Analytical method was developed on Shimadzu double beam spectrophotometer equipped with UV probe 2.42 as software using methanol as solvent. Quantification of PAMA was carried out at zero cross over point of IBU that is 291 nm and for IBU, it was achieved at 278 nm which is zero cross over point of PAMA. Method was validated according to ICH Q2 R1 guidelines.
 Results: Method showed a linear response in the range of 2-12 µg/ml of PAMA and 20-120 µg/ml of IBU. Method was found to be accurate with recovery between 99.7–100.9 % for PAMA and 100.3–100.7 % for IBU. The method was found to be accurate and precise for quantitative analysis of PAMA and IBU.
 Conclusion: The developed method was successfully validated as per ICH Q2 R1 guidelines and was successfully applied for quantitative analysis of a synthetic mixture of PAMA and IBU.

DEVELOPMENT AND VALIDATION OF UV-VISIBLE SPECTROPHOTOMETRIC METHOD FOR DETERMINATION OF DULOXETINE

Objective: The aim of the present work was to develop and validate a simple UV spectroscopic method for the determination of duloxetine, which is a thiophene derivative and a selective neurotransmitter reuptake inhibitor for serotonin, norepinephrine, and to lesser degree dopamine.
 Methods: The UV Spectrophotometric analysis was performed using Shimadzu UV-1800 and Shimadzu UV-1700 spectrophotometer by using solvent system acetonitrile and water in the ratio of 8:2. Detection was performed at a wavelength of 290 nm. Method validation was carried out according to ICH Q2R1 guidelines by taking the parameters linearity, accuracy, precision, ruggedness, and robustness, LOD and LOQ.
 Results: The UV Spectrophotometric method was found linear in the range of 10-50 μg/ml. The method was rugged and robust with % relative standard deviation less than 2. The extraction recoveries were found to be higher than 99% in all experimental conditions.
 Conclusion: Based upon the performance characteristics, the proposed method was found accurate, precise and rapid and suitable for the determination of Duloxetine for routine analysis.

HILIC TECHNIQUE BASED HPCL ANALYSIS OF CAPTOPRIL, GLUTAMIC ACID AND TAURINE FROM VARIOUS MATRICES

A simple and rapid chromatographic method using HILIC technique has been developed for simultaneous estimation of captopril, glutamic acid and taurine from their admixture as well as from biological matrices. Determination was carried out on Luna HILIC column (4.6 mm x 250 mm, 5 μm) using a mobile phase of acetonitrile: water (70: 30 % V/V) with 0.1 % acetic acid. Flow rate was 1 mL/min with detection at 237 nm. All these analytes have shown linearity over concentration range of 20-140 µg/mL and 0.2 to 12.8 µg/mL with retention time of 2.67, 4.05 and 7.85 and 2.87, 4.25 and 8.09 min for taurine, glutamic acid and captopril in admixture analysis and bio analysis, respectively. The methods were validated using ICH Q2 R1 and USFDA guidelines. The developed method was found to be accurate, precise sensitive and selective for simultaneous estimation of these from their admixture and from biological matrices.

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF CURCUMIN AND CYCLOSPORINE BY RP-HPLC

Objective: The present work was undertaken with an aim to develop and validate a rapid reverse-phase high-performance liquid chromatography (RP-HPLC) method for the estimation of curcumin and cyclosporine in the capsule dosage form.
 Methods: The RP-HPLC method for the simultaneous estimation of curcumin and cyclosporine was developed using Agilent (Infinity 1260) HPLC system and Eclipse XDB-C18 (4.6 x 150 mm i.d., 5µ) stationary phase. The optimized mobile phase comprised of acetonitrile: water: methanol (50: 10: 40 v/v/v) pumped at a flow rate of 0.5 ml/min. Separation of drugs was achieved in an isocratic mode and elution was monitored using PDA detector at 214 nm. The method was validated as per ICH-Q2R1 guidelines.
 Results: Retention time of the curcumin and cyclosporine were found to be 3.073 min and 6.373 min with the correlation coefficient (R2) of 0.9993 and 0.998 respectively. The response of curcumin and cyclosporine was found linear in the concentration range of 8-48 μg/ml and 4-24 μg/ml respectively. The percent recovery values were found in the range of 97-103% indicating satisfactory accuracy of the method. The percent relative standard deviation (% RSD) values for the precision study was less than 2 which suggest that the method is precise.
 Conclusion: The proposed method was found accurate, precise and specific for the determination of curcumin and cyclosporine in bulk as well as in capsule dosage form. Thus, the present method can be used for routine analysis and quality control of curcumin and cyclosporine in bulk and capsule dosage form.

A new MS compatible HPLC-UV method for Teicoplanin drug substance and related impurities, part 1: Development and validation studies

Teicoplanin is a glycopeptide antibiotic prepared by fermentation from cultures of Actinoplanes teichomyceticus, used as drug of last resort for the treatment of bacterial infections in humans. This study, which is the first in a series of two parts, describes the development of a LC method for the separation of Teicoplanin drug substance and its related impurities compatible with MS detection.The separation conditions for Teicoplanin were set on a LiChrospher 100 RP-18 column under gradient elution with a mobile phase composed of ammonium formate 25 mM at pH 6.00 and ACN. The new method was shown equivalent in terms of selectivity to the one reported in the European Pharmacopoeia Teicoplanin monograph, and was validated according to ICH Q2 R1 guidelines for the drug substance assay. The new method offers similar performance to the compendial one but has the advantage of being fully compatible with MS and it can be proposed as a useful tool also for controlling the quality of Teicoplanin fermentation batches and the occurrence of potential impurities.

Validated UV Spectrophotometric Methods for the Estimation of Letrozole in Solid Dosage Forms

The main intent of this research is to develop two new simple, novel, cost efficient and precise UV spectrophotometric methods for estimation of letrozole in pure and pharmaceutical dosage forms. For estimation of letrozole based on the measurement of UV light absorption, the spectras of letrozole were scanned efficiently and it exhibits maximum absorption wavelength (λmax) at 240nm for Method A and 245nm for Method B. The analysis was carried out against phosphate buffer pH 6.8 (Method A) and phosphate buffer pH 3.8 (Method B) respectively. The drug shows linear response commencing from 0.5-20 µg/mL for both methods A & B. The regression equation was found as Y= 0.099X + 0.079 (Method A) and Y= 0.1359x + 0.0048 (Method B), and correlation coefficient were 0.999 (Method A) and 0.9999 (Method B). The method validation was accomplished as per the regulation of ICH Q2R1 guidelines for linearity, accuracy, and precision studies. These reported methods have good reproducibility with percentage RSD less than one. These methods were extended towards the formulation and there was no interruption from excipients in the formulation. Hence, these proposed methods can be effectively employed for evaluation of letrozole in quality control as well as routine analysis work in pharmaceutical industries.

A validated stability indicating RP-HPLC method for the quantification of Canagliflozin

A simple, authentic and stability indicating high performance liquid chromatographic method for determination of Canagliflozin in bulk and pharmaceutical formulations was developed and validated as per ICH Q2 R1 Guidelines, A C18 Column (250mm length×4.6 mm diameter x 5 μm particle size) with a mobile phase consisting of Acetonitrile: 1-octanesulphonic acid in a ratio of 70:30 v/v was employed for the chromatographic study. A flow rate of 1.0 mL/min with an injection volume of 20 μL was selected for this study and the proposed method was validated with different parameters such as Linearity, Precision, Accuracy, Robustness, Ruggedness, Limit of Detection (LOD) and Limit of Quantification (LOQ). Canagliflozin was eluted at 3.4 ± 0.5 min and detected at 245 nm. The method is linear over the concentration range of 10-100 μg/mL with correlation co-efficient r = 0.9997. The plate count and tailing factor was found 5398 and 1.05 respectively. The LOD and LOQ were found to be 0.0170 μg/mL and 0.1705 μg/mL respectively. The percentage recovery was achieved in the range of 98-102%, which was within the acceptance criteria. Developed method was employed to determine the amount of Canagliflozinpresent in dosage form (Sulisent). The stabilityof the method was demonstrated by forced degradation studies of drug in which it was degraded under conditions of hydrolysis (acidic and alkaline), oxidation, photolytic and thermal stress as per ICH guideline Q1A (R2).

Development and Validation of A UV Spectroscopic Method to Estimate Etoricoxib in Bulk and Tablet Formulation

This present UV Spectrophotometric method developed using methanol as a solvent is simple, rapid, specific, precise and sensitive to estimation Etoricoxib in bulk in day to day analysis. The method was validated as per ICH Q2R1 guideline. Etoricoxib is the newest addition to the group of Non-Steroidal Anti-Inflammatory Drugs - highly selective COX-2 inhibitor. The solvent used in the entire method development and validation was methanol. The maximum wavelength of absorption was found to be 234nm. Beer’s law was obeyed in the concentration range of 1 to 11µg/mL with correlation coefficient of 0.9986. The method was precise with an RSD of0.42%the LOD and LOQ were found to be 0.09µg/ml and 0.2µg/ml respectively. The method was validated for precision, linearity, accuracy and robustness and all parameters were found to be satisfactory which proves that this method can be used for routine analysis of Etoricoxib.

Development and Validation of an Analytical Method for Related Substances in N-acetyl–L-cysteine Effervescent Tablets by RP-HPLC

Background: The reported chromatographic methods for N-acetyl cysteine [NAC] are reverse phase HPLC and ion pair chromatography [IPC] for related substances test in bulk and in formulations. No reported stability indicating methods for the estimation of related substances in NAC effervescent formulation was found in literature. Objective: The present work was aimed at developing a selective, sensitive and reproducible stability indicating high-performance liquid chromatographic method for the quantitative determination of known, unknown impurities, degradation impurities and processrelated impurities of NAC effervescent formulation. Method: A reversed phase ion pair chromatographic method was developed employing Cadenza C18 column as the stationary phase and 0.01M octane sulphonate [pH 2.20], methanol and acetonitrile in the ratio 90:8:2 as the mobile phase. A gradient programme was followed with a run time of 55 minutes. 0.3 M hydrochloric acid was selected as the optimum diluent. The performance of the method was validated according to the ICHQ2R1 guidelines. Results: The method was found to be linear from 1.5 to 25μg/ml for impurities A, C and D and from 2.0 to 25 μg/ml for impurity B. The official impurities C and D were mapped in all stress conditions. Additionally, impurity B was also seen in acidic conditions. Conclusion: The results from the study demonstrate that the method is suitable for evaluating the stability of NAC effervescent tablet.

Quantitation and biospecific identification of virus-like particles of human papillomavirus by capillary electrophoresis

Capillary electrophoresis (CE) for HPV-VLP quantitation is a very interesting alternative technique compared to those currently used in viral analysis, such as SDS-PAGE, Western blot or protein assay that are destructive and semi-quantitative or non specific. In this study, the quantitative performance of the CE method was evaluated. A main issue in virus quantitation is the absence of reference material. Therefore, the concentration of a HPV16-VLP sample produced in the laboratory was determined using ELISA with Gardasil®, after adjuvant dissolution, as reference material and conformational H16.V5 antibody. HPV16-VLP concentration was found to influence particles electrophoretic mobility until a plateau was reached for concentrations ≤ 50µgml−1. As zeta potential is directly proportional to the electrophoretic mobility, it was measured at different HPV-VLP concentrations and the results were in complete accordance with the measured electrophoretic mobilities. The concentration dependence of the electrophoretic mobility could be explained by an overlap of the electrical double layers of adjacent particles. The HPV16-VLP peak identity was demonstrated unequivocally by the study of HPV16-VLP/H16.V5 antibody complex formation using affinity CE. Finally, the CE method was successfully validated following the ICH Q2R1 guidelines. To overcome the sample heterogeneity issue, a well-designed sample preparation was used. Considering sample complexity, validation results were satisfactory with maximum repeatability and intermediate precision RSD of 12.2% and a maximum relative bias of 1.4%.

CRITICAL REVIEW OF CHROMATOGRAPHIC AND SPECTROPHOTOMETRIC METHODS AND APPLICATION OF UV- AUC TECHNIQUE FOR SIMPLE, RAPID, QUANTITATIVE ESTIMATION OF ATAZANAVIR SULFATE

Spectrophotometric analysis has proven to be one of the most referred analytical techniques available; as the enlarged globalization of the pharmaceutical market has reduced the availability of quality pharmaceuticals at minimal cost. Authors have made an attempt for development and validation of simple UV- spectrophotometric area under curve (UV -AUC) method for quantitative estimation of atazanavir sulfate in bulk and in a pharmaceutical formulation. To enhance accuracy, additional attempts were made for the measurement of uniform area around the wavelength maximum (248 nm) and in between the designated wavelengths (238 nm and 260 nm). The mixture of methanol and water in a proportion of 30:70 v/v was selected as a solvent system. Efficient correlation between the concentration and corresponding response was maintained by measuring the uniform area under the peak obtained at the wavelength maximum. The calibration plot for the method was constructed by plotting the values for AUC obtained for the concentration range of 6 - 36 μg/mL with a correlation coefficient of 0.999. Developed method was validated on the basis of ICH Q2 R1 guideline. The method depicted good reproducibility with percent RSD less than 2.0. Estimation of atazanavir sulfate by aforementioned method in bulk and capsules was carried out efficiently.

STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS DETERMINATION OF VILDAGLIPTIN AND METFORMIN IN PHARMACEUTICAL DOSAGE FORM

Objective: The present study was aimed to develop a rapid, accurate, linear, sensitive and validate stability-indicating high performance liquid chromatographic [RP-HPLC] method for determination of vildagliptin and metformin in pharmaceutical dosage form.Methods: The chromatographic separation was performed on kromasil-C18 column [4.5 x 250 mm; 5 µm] using a mobile phase consisting of 0.05 mmol potassium dihydrogen phosphate buffer: acetonitrile [80:20 v/v], [pH adjusted to 3.5 using orthophosphoric acid]. The flow rate is 0.9 ml/min and the detection was carried out at 263 nm.Results: The chromatographic condition, the peak retention time of metformin and vildagliptin were found to be 2.215 min and 2.600 min respectively. Stress testing was performed in accordance with an international conference on harmonization [ICH] Q1A R2 guidelines. The method was validated as per ICH Q2 R1 guidelines. The calibration curve was found to be linear in the concentration range of 5-17.5 µg/ml and 50-175 µg/ml for vildagliptin and metformin. The limit of detection and quantification was found to be 0.0182 µg/ml and 0.0553 µg/ml for vildagliptin and 0.4451 µg/ml and 1.3490 µg/ml for metformin respectively.Conclusion: A new sensitive, simple and stability indicating reverse-phase high-performance liquid chromatography [RP-HPLC] method has been developed and validated for the determination of vildagliptin and metformin. The proposed method can be used for routine determination of vildagliptin and metformin.

Simultaneous Spectrophotometric Determination of Linagliptin and Metformin Hydrochloride in Combined Tablet Dosage Form.

Simple, precise and economical UV Spectrophotometric methods have been developed for the simultaneous estimation of Linagliptin and Metformin in bulk and pharmaceutical dosage forms. The simultaneous equation (Vierodt's Method), which is based on measurement of absorption at 227nm and 232nm i.e. λmax of Linagliptin and Metformin respectively. Linearity was observed in the concentration range of 1–11μg/ml for Linagliptin and 3–13 μg/ml. The accuracy of methods was assessed by recovery studies and was found to be withinrange of 9899% for both Linagliptin and Metformin. The developed methods were validated with respect tolinearity, accuracy (recovery), and precision. The method can be employed for estimation of pharmaceuticalformulations with no interference from any other excipients and diluents. The results were validated statistically as per ICH Q2 R1 guidelines and were found to be satisfactory.

Optimization of Robust HPLC Method for Quantitation of Ambroxol Hydrochloride and Roxithromycin Using a DoE Approach.

The aim of this work was to develop and optimize a robust HPLC method for the separation and quantitation of ambroxol hydrochloride and roxithromycin utilizing Design of Experiment (DoE) approach. The Plackett-Burman design was used to assess the impact of independent variables (concentration of organic phase, mobile phase pH, flow rate and column temperature) on peak resolution, USP tailing and number of plates. A central composite design was utilized to evaluate the main, interaction, and quadratic effects of independent variables on the selected dependent variables. The optimized HPLC method was validated based on ICH Q2R1 guideline and was used to separate and quantify ambroxol hydrochloride and roxithromycin in tablet formulations. The findings showed that DoE approach could be effectively applied to optimize a robust HPLC method for quantification of ambroxol hydrochloride and roxithromycin in tablet formulations. Statistical comparison between results of proposed and reported HPLC method revealed no significant difference; indicating the ability of proposed HPLC method for analysis of ambroxol hydrochloride and roxithromycin in pharmaceutical formulations.

473. Qualification of a p24 ELISA Assay for Quantitation of Total Lentiviral Vector Concentration

Lentiviral vectors (LVV) are efficient gene transfer vehicles used in a growing list of gene therapy applications that are currently being evaluated clinically. We are conducting clinical trials in subjects with β-thalassemia major, severe sickle cell disease, and childhood cerebral adrenoleukodystrophy, which employ autologous CD34+ cells transduced with LVV encoding a corrective transgene, βA-T87Q-Globin or ABCD1. Essential to LVV process development is the availability of reliable analytical tools to characterize the purified LVV product and in-process samples. Total lentiviral vector particle titers can be estimated by quantitation of HIV-1 p24 antigen concentration using commercial ELISA kits. In support of late-stage process characterization, it is essential to qualify the assay and demonstrate its suitability to test in-process samples in different matrices. Following ICH Q2 R1 guidelines, assay accuracy, repeatability, intermediate precision, quantitation limit, linearity, and linear range were evaluated. Additionally, qualification tests for specificity and dilutional linearity were performed on in-process samples and buffers to assess potential matrix interference. The results of these tests were used to define appropriate dilutions and acceptance criteria for p24 ELISA in-process samples. The assay has shown to be linear between 400 pg/mL and 25 pg/mL, while maintaining accuracy for p24 detection with the limit of quantitation being 25 pg/mL. To demonstrate assay repeatability, various concentrations within the linear range were tested resulting in coefficients of variation below 5%. The coefficient of variation across different days and different operators was below 8% for all concentrations tested, indicating consistent assay precision. Spike recovery studies have shown consistent recovery of spiked p24 antigen (73-126%), demonstrating assay specificity and lack of matrix interference in all in-process matrices tested while maintaining dilutional linearity. To conclude, the p24 ELISA assay used for LVV quantitation has shown precision, repeatability, specificity and accuracy, within an established linear quantitation range, adequate to support late-stage process characterization activities.

Development and Validation of a Stability-Indicating Reverse Phase HPLC-PDA Method for Determination of Canagliflozin in Bulk and Pharmaceutical Dosage Form

Objective: To develop and validate simple, authentic and stability indicating high performance liquid chromatographic method for determination of Canagliflozin in bulk and pharmaceutical formulations as per ICH Q2 R1 Guidelines. Methods: A C 18 Column (250×4.6 mm, 5 μm particle size) with a mobile phase consisting of Acetonitrile: orthophosphoric acid in a ratio of 55:45 v/v was employed for the chromatographic study. A flow rate of 1 ml/min with an injection volume of 20 μL was selected for this study and the proposed method was validated with different parameters such as Linearity, Precision, Accuracy, Robustness, Ruggedness, Limit of Detection (LOD) and Limit of Quantification (LOQ). Results: The separation was achieved at a temperature of 30oC and the eluents were observed by photo diode array detector set at 290 nm. A linear range of 1-6 μg/ ml with a correlation coefficient of 0.998 unfolds good linear relationship between area and concentration in calibration curve. The retention time obtained was at 6.29 min. The LOD and LOQ were found to be 0.41 μg/ml and 1.24 μg/ml respectively. A recovery of Canagliflozin in tablet formulation was observed in the range of 99.6-99.8%. Percentage assay of Canagliflozin tablets (INVOKANA®) was found to be 99.92%. The stability of the method was demonstrated by forced degradation studies of drug in which it was degraded under conditions of hydrolysis (acidic and alkaline), oxidation, photolytic and thermal stress as per ICH guideline Q1A (R2). Conclusion: The proposed method is definite, meticulous and reproducible and can be used for routine analysis of Canagliflozin in bulk and pharmaceutical dosage form. Key words: Canagliflozin, High Performance Liquid Chromatography, Method development, Stability, Validation.

Development and Validation of Stability indicating UV Spectroscopic Method for Determination of Canagliflozin in Bulk and Pharmaceutical Dosage Form

Objective: To develop and validate simple, definite, stability indicating UV spectroscopic method for determination of Canagliflozin in bulk and pharmaceutical formulations as per ICH Q2 R1 Guidelines. Methods: Canagliflozin was subjected to different stress conditions as per ICH guidelineQ1A (R2). A stability-indicating UV Spectrophotometric method has been developed for analysis of the drug in the presence of the degradation products and is validated with different parameters such as Linearity, Precision, Repeatability, Limit of Detection (LOD), Limit of Quantification (LOQ), Accuracy, Robustness and Ruggedness. It involved a 2-h study in which methanol and distilled water were used as solvents. Results: Canagliflozin in methanol shows maximum absorbance at 290 nm. Beer’s law was obeyed in the concentration range of 5-10 mcg/mL. The LOD and LOQ were found to be 0.084 mcg/ml and 0.255 mcg/ml respectively. A recovery of Canagliflozin in tablet formulation was observed in the range of 80.00-120.00%. Percentage assay of Canagliflozin tablets (INVOKANA®) was found to be more than 99%. Degradation of Canagliflozin was found to occur in acid, alkaline, hydrogen peroxide and photolytic conditions where as it was found to be thermally stable. The amount of degraded drug was calculated by taking absorbance at 290 nm. Conclusion: The proposed method is definite, meticulous, reproducible and can be used for routine analysis of Canagliflozin in bulk and pharmaceutical dosage form. Key words : Canagliflozin, Method development, Validation, Ultraviolet Spectroscopy, Forced degradation.

Simultaneous UV Spectrophotometric Methods for Estimation of Cefixime Trihydrate and Ofloxacin in Bulk and Tablet Dosage Form

Simple, precise, economical, fast and reliable two UV methods have been developed for the simultaneous estimation of Cefixime Trihydrate and Ofloxacin in bulk and pharmaceutical dosage form. Method A is Absorbance maxima method, which is based on measurement of absorption at maximum wavelength of 287 nm and 296 nm for Cefixime Trihydrate and Ofloxacin respectively. Method B is area under curve (AUC), in the wavelength range of 265–301 nm for Cefixime Trihydrate and277-320nm for Ofloxacin. Linearity for detector response was observed in the concentration range of 5–25μg/ml for Cefixime Trihydrate and 5–25 μg/ml for Ofloxacin. The accuracy of the methods was assessed by recovery studies and was found to be 98.83% and 100.12% for Cefixime Trihydrate and 102.71% and 99.01% Ofloxacin by using method A and B respectively. The developed method was validated with respect to linearity, accuracy (recovery), precision and specificity. The results were validated statistically as per ICH Q2 R1guideline and were found to be satisfactory. The proposed methods were successfully applied for the determination of for Cefixime Trihydrate and Ofloxacin in commercial pharmaceutical dosage form.