Multivariate optimization of liquid chromatographic conditions for determination of dapagliflozin and saxagliptin, application to an in vitro dissolution and stability studies (original) (raw)
Related papers
Current Issues in Pharmacy and Medical Sciences
A simple, fast, and highly selective RP-HPLC method was developed for the determination of Dapagliflozin (DAP) and Saxagliptin (SAX) in API and tablet dosage form. The separation was done using a Xterra RP18 (4.6×150 mm, 5 μm particle size) column with Acetonitrile: water (60:40). The isocratic elution mode at a flow rate of 1 mL/min, and the analytes were measured at 248 nm. The retention time for DAP and SAX were about 2.091 and 3.249 min, respectively. Calibration curves were found to be linear in the ranges of 100-500 μg/ml for DAP and 50-250 μg/ml for SAX, with correlation coefficients of 0.9998. The detection and quantification values for DAP was 3.0 and 9.98 μg/ml and SAX was 3.02 and 10 μg/ml respectively.
International Journal of Pharmacy and Pharmaceutical Sciences, 2019
Objective: To develop and validate a sensitive liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS/MS) technique for the quantification of dapagliflozin and saxagliptin in plasma by linagliptin as internal standard. Methods: Chromatography was achieved on hypersil C18 (50 mmx4 mm) 5 µ analytical column with 0.1% formic acid and acetonitrile (25:75 V/V) as mobile phase at 0.7 ml/min flow rate. Dapagliflozin, saxagliptin, and linagliptin were detected at m/z 409.14/135.0, m/z 316.2/180.13 and m/z 472.54/456.21 respectively. Drugs and internal standard were extracted by LLE (liquid-liquid extraction). Results: Developed technique was validated over 0.5-1500.0 ng/ml linear concentration range for dapagliflozin and 2.00-2000.0 ng/ml for saxagliptin. This method established with intra-batch and inter-batch precision within 2.44-8.12% and 1.25-7.14 % for dapagliflozin and 1.84-7.5 % and 1.02-6.00 % for saxagliptin. This method established with intra-batch and inter-batch accuracy for dapagliflozin within 98.86-103% and 96.98-102 % respectively and for saxagliptin within 98.05-109.06 % and 97.00-104.00 % respectively. Conclusion: Both dapagliflozin and saxagliptin were stable during three freeze-thaw cycles, long term and bench-top stability studies. The developed method was useful for the routine analysis of dapagliflozin and saxagliptin simultaneously in plasma samples.
International Journal of Research in Pharmaceutical Sciences
The present study is aimed to develop a linear, precise, and accurate RP-HPLC (Reverse Phase High-Performance Liquid Chromatography) method for the determination of dapagliflozin in the formulation. The method was accomplished on a C18 column (250×4.6mm; 5µm), & Samples were eluted using acetonitrile: water (40:60%v/v) delivered at a flow rate of 1.0ml/min with a chromatographic run time of 10 min. The eluents were observed utilizing a UV detector with a wavelength set at 277nm. The method that was developed resulted in the retention of dapagliflozin at 7.029minutes. Dapagliflozin through current method has shown linearity (r2 > 0.999) over the concentration range of 1-16 µg/ml. The percentage recovery was observed to be within the limits of 98-102%, demonstrating the accuracy of the method. Limit of detection (LOD) and limit of quantification (LOQ) were qualified at 0.049µg/ml and 0.1485µg/ml, respectively. A Linear, precise, accurate, simple, and rapid RP-HPLC method has been d...
A new, simple, fast, selective, precise and accurate RP-HPLC method was developed and validated for the estimation of Dapagliflozin from bulk and marketed formulations. The proposed method was developed by HPLC Waters ODS C18 column, 5m, 25cmx4.6mm i.d, Separation Module with UV detector connected to D Elite 2000 software with an injection volume of 20 μl was injected and eluted with a mobile phase composition of Buffer (Potassium hydrogen orthophosphate & pH adjusted to 4.2 with orthophosphoric acid) and Methanol in a ratio of 65:35. Mobile phase is pumped at a flow rate of 1.0 ml/min and detected by UV detector at 225nm. Ambient column temperature has maintained. The retention time of Dapagliflozin was found to be 2.93 min. Linearity was observed in the concentration range of 20-100 μg/ml for Dapagliflozin with correlation coefficient 0.999. The proposed method was found to be precise and reproducible with %RSD of 0.42 for dapagliflozin. Percent recovery obtained in the range of 99.25 to 101.16% w/w for dapagliflozin. The LOD & LOQ were found to be 0.003 & 0.009 µg/ml respectively.The method was validated according to the ICH guidelines with respect to specificity, linearity, accuracy, precision and robustness. The results of the stress studies indicated the specificity of the method that has been developed. Dapagliflozin was stable in both oxidation & acidic stress conditions. The result shows the developed method is yet another suitable method for assay & stability which can help in the analysis of Dapagliflozin in different formulations.
A Review on Analytical Methods of Dapagliflozin: An Update
INTERNATIONAL JOURNAL OF PHARMACEUTICAL QUALITY ASSURANCE
co-transporter offer. It is used in patients with type 2 diabetes. It is administered as tablets. It has several analytical papers for estimation of active pharmaceutical ingredient (API) or drug formulation by reverse phase-high performance liquid chromatography (RP-HPLC) and ultraviolet spectroscopy (UV). It is very challenging to use of chemicals, drugs, and solvent of separation methods used in the pharmaceutical product to green chemistry. This review mostly used dihydrogen phosphate buffer and other toxic reagents for estimation and these agents harm instruments, as well as, environment and a lot of waste so that novel analytical techniques for quantifying and defining dapagliflozin should be built as easy as possible and secure for the individual and the community. This review pays attention to the critical condition of physicochemical, properties, action, and aims to focus on different analytical methods for the estimation of dapagliflozin in pharmaceutical formulations.
Chromatographia, 2020
A simple, rapid, and accurate stability-indicating reverse phase HPLC-DAD method was developed and validated for the simultaneous determination of empagliflozin, dapagliflozin, and canagliflozin (Gliflozins). Optimum chromatographic separations among gliflozins in the presence of matrices and degradation products have been achieved within 7 min by using Hypercil™ C 18 column (25 × 4.6 mm, 5 μm) with acetonitrile and 0.1% formic acid buffer, pH 3.7 (60:40 v/v) as the mobile phase at a flow rate of 1 mL min −1. The proposed method was performed at 230 nm for empagliflozin and dapagliflozin while detection of canagliflozin was carried out at 290 nm. Analytical performance of the method was thoroughly validated in accordance with the ICH guidelines with respect to system suitability, linearity, accuracy, precision, specificity, robustness, detection and quantification limits. Regression analysis showed good correlations with regard to R 2 ≥ 0.997 over the concentration range of 4-160 µg mL −1 for gliflozins. The LOD was found to be 0.07 µg mL −1 , 0.12 µg mL −1 and 0.29 µg mL −1 for empagliflozin, dapagliflozin and canagliflozin, respectively. Method development was established keeping in view the structure of drugs, their pK a values, ionizability of drugs, pH values of buffer system and buffer capacity. Peak purities of gliflozins in stress tests are less than 1.5, which further confirms no co-elution of degradation products. The proposed method is suitable for routine quality control analysis of gliflozins and to carry out stability studies.
International Journal of Applied Pharmaceutics, 2017
Objective: To develop precise, accurate and reproducible stability assay method by RP-HPLC for estimation of dapagliflozin in API and pharmaceutical dosage form. Methods: The adequate separation was carried using agilent C18 (4.6 ml (millimeter)*150,5 µm (micromiter), mixture of acetonitrile: di-potassium hydrogen phosphate with pH-6.5 adjusted with OPA (40:60 %v/v) as a mobile phase with the flow rate of 1 ml/min (milliliter/minute) and the effluent was monitored at 222 nm (nanometer) using photo diode array detector. The retention time of dapagliflozin API and dapagliflozin tablet were 3.160 min (minute) and 3.067 min (minute) respectively. Results: Linearity for dapagliflozin was found in the range of 50-150µg/ml (microgram/milliliter) (R 2 = 0.99) respectively. The accuracy of the present method was evaluated at 50 %, 100% and 150%. The % recoveries of dapagliflozin API and tablet were found to be in the range of 99.00-99.99 % and 98.50-99.99 % respectively. Precision studies were carried out and the relative standard deviation values were less than two. The method was found to be robust. Conclusion: The proposed method was found to be specific, accurate, precise and robust can be used for estimation of dapagliflozin in API and Pharmaceutical dosage form.
International Journal of Pharmacy and Pharmaceutical Sciences, 2019
Objective: To develop a new simple, selective and precise high-performance thin-layer chromatographic method for determination of Dapagliflozin (DAPA) in bulk and tablet dosage form. Methods: The present study describes the development and validation of a High-performance thin-layer chromatographic method for DAPA. The chromatographic separation was carried out on Merck precoated silica gel aluminum plate 60 F254 Results: The chromatographic condition showed a compact band with the retention factor for dapagliflozin as 0.21±0.004. The method was validated as per ICH guidelines for linearity, accuracy, precision and robustness. Response was found to be linear in the concentration range of 400 ng/band to 1200 ng/band with linear regression value of 0.9953 with respect to peak area and concentration value. The LOD and LOQ were found to be 1.2083 ng/band and 3.6616ng/band. The percentage assay was found to be 100±0.05. using Chloroform: Methanol (9:1v/v) as mobile phase. Quantitative determination of drug was carried out by densitometric scanning of plates at 223 nm using Camag TLC Scanner. Conclusion: This method under statistical analysis proved a selective, repeatable and accurate analysis of the drug. This method can be used for quantitative analysis of dapagliflozin in the bulk drug and in tablets as the run time is very minimal.
— A New HPLC method for the simultaneous estimation of Saxagliptine and Dapagliflozin in their combine dosage form was developed and validated as per the ICH guidelines. The method involves separation on XTerra C 18 column (150mm x 4.6mm x5µm particle size). The optimized mobile phase consists of phoaphate buffer (pH 4) and Acetonitrile (50:50v/v) with a flow rate of 1ml/min and UV detection at 225nm. Retention time was 2.1min (Saxagliptine), 2.8min (Dapagliflozin). Linearity was observed in the range of 20-60µg/ml for Saxagliptine and 10-120µg/ml for Dapagliflozin with correlation coefficients (r 2 =0.999). The percentage recoveries of Saxagliptine and Dapagliflozin were in the range of 99.99-100.50% which was with in the acceptance criteria. The percent RSD was NMT 2% which proved the precision of the developed method. The developed method is simple, specific, sensitive, precise and accurate and was found suitable for estimation of Saxagliptine and Dapagliflozin in bulk and dosage forms. The developed HPLC method was also found suitable for application in pharmacokinetic studies for the estimation of Saxagliptine and Dapagliflozin in plasma samples.In the real in vivo pharmacokinetic study the biological half-lives (t 1/2) of Saxagliptine and Dapagliflozin estimated by the proposed method are in good agreement with the literature values. The good agreement of the absorption and elimination parameters estimated using HPLC method developed with those of literature values indicated that newly developed HPLC method is suitable and could be used in pharmacokinetic studies.