Method For Estimation Of Metformin Research Articles

Validation of a novel UPLC-MS/MS method for estimation of metformin and empagliflozin simultaneously in human plasma using freezing lipid precipitation approach and its application to pharmacokinetic study

A fast, sensitive one step UPLC ESI-MS/MS method was successfully applied for the simultaneous estimation of two concurrently administrated antidiabetic drugs, Metformin (MET) and Empagliflozin (EMPA) in human plasma. Metformin-d6 (MET-d6) and Empagliflozin-d4 (EMPA-d4) were utilized as internal standards. Extraction of the analytes from the human plasma was performed through acetonitrile precipitation technique followed by freezing the precipitated plasma proteins and lipids to minimize the matrix effect. Chromatographic analysis was developed on Acquity UPLC BEH C18 column (1.7 μm, 2.1 × 50 mm) using isocratic elution mode. A mobile phase of formic acid (0.01 %): acetonitrile (70:30 v/v) with a flow rate of 0.3 mL/min achieved optimum separation. Multiple reaction monitoring (MRM) in positive ion mode, with transitions at (m/z) 130.14 →71.08 for (MET), 451.72 →71.29 for (EMPA), 136.03 →77.02 for (MET-d6), and 455.43 → 75.05 for (EMPA-d4) was used for quantification. The obtained linearity covered the concentration ranges of 10−1500 ng/mL and 2.0–250.0 ng/mL for MET and EMPA, respectively. The run time of the proposed Method didn’t exceed 3.0 min allowing faster analysis and determination of larger number of samples per day without affecting accuracy and sensitivity. The presented chromatographic method could be successfully applied in pharmacokinetics studies and therapeutic monitoring of MET and EMPA in patients’ plasma administrating fixed dose combination of both drug with high reproducibility and ruggedness.

Development and Validation of a Stability-indicating RP-HPLC Method for Estimation of Metformin and Rosuvastatin along with Impurities from a Combined Oral Solid Dosage Form

The objective of current study was to develop and validate a stability-indicating reversed-phase high-performance liquid chromatography for simultaneous quantification of impurities in metformin sustained release-rosuvastatin immediate release combined oral solid dosage form as per ICH guidelines. Chromatographic separation was achieved using a Waters HPLC system with a YMC-Pack ODS-A 100×4.6 mm; 3 µm column. Mobile phase A comprised of 20 mM ammonium acetate:acetonitrile (9:1 v/v) pH 3.5, and mobile phase B, consisted of 20 mM ammonium acetate:acetonitrile:methanol (3:5:2 v/v/v) pH 3.5. The gradient program was set as (T/% B) 0/30, 5/30, 10/50, 30/50, 40/80, 42/30 and 45/30, which pumped at a rate of 0.9 ml/min. About 10 μl of sample solutions were injected and monitored at 240 nm. Column temperature and sample compartment was maintained at 25° and 5°, respectively. Forced degradation studies revealed that the peak threshould was more than the peak purity and no purity flag was observed. Repeatability, intra, and inter-day precision results were well within the tolerable limits. Limits of detection were found to be 0.10, 0.05, 0.15, 0.20 and 0.40 ppm and limits of quantitation as 0.30, 0.15, 0.45, 0.60 and 1.0 ppm for metformin, rosuvastatin, rosuvastatin impurity A, B, and C, respectively. The correlation coefficient of linearity for metformin, rosuvastatin and its impurities were found to be ≥0.999. The method recovery was found between 99 and 102 %. The proposed method appeared to be a simple, rapid, economical, accurate, specific, robust and able to distinguish the two key components from their degradants, impurities, and excipients. Therefore, current method could be applied successfully for stability and routine examination.

A NOVEL STABILITY-INDICATING REVERSE PHASE LIQUID CHROMATOGRAPHIC METHOD FOR THE SIMULTANEOUS ESTIMATION OF METFORMIN AND TENELIGLIPTIN IN PURE AND PHARMACEUTICAL FORMULATIONS

Objective: The present method was proposed to develop a simple, sensitive, rapid, accurate and stability-indicating reverse phase-high performance liquid chromatographic (RP-HPLC) method for the simultaneous estimation of metformin and teneligliptin in pure and pharmaceutical formulations. Methods: The chromatographic separation was done on Discovery [250 mm X 4.6 mm: 5 μm is particle size] using a mobile phase composed of 0.1% orthophosphoric acid buffer: acetonitrile [65:35, v/v], the flow rate is 1 ml/min and the detection was carried out with a photodiode array (PDA) at 260 nm.Results: The retention time of metformin and teneligliptin were found to be 2.517 min and 3.687 min, respectively. Stability indicating studies were conducted under the guidelines of an international conference on harmonization [ICH] Q1A R2 and the developed method was validated as per the guidelines of ICH Q2 R1. The linearity was found in the range of concentration of 125-750 μg/ml and 5-30 μg/ml for metformin and teneligliptin. The lower limit of detection (LOD) and lower limit of quantification (LOQ) was found to be 0.02μg/ml and 0.07 μg/ml for metformin and 0.19 μg/ml and 0.56 μg/ml for teneligliptin, respectively.Conclusion: A novel stability-indicating reverse phase liquid chromatographic method developed for the simultaneous estimation of metformin and teneligliptin. The proposed method was adopted for the routine estimation of metformin and teneligliptin in bulk and pharmaceutical dosage forms.

Development and Validation of UV-Spectrophotometric Method for Estimation of Metformin in Bulk and Tablet Dosage Form

Development and Validation of UV-Spectrophotometric Method for Estimation of Metformin in Bulk and Tablet Dosage Form

A NOVEL STABILITY-INDICATING REVERSE PHASE LIQUID CHROMATOGRAPHIC METHOD FOR THE SIMULTANEOUS ESTIMATION OF METFORMIN AND TENELIGLIPTIN IN PURE AND PHARMACEUTICAL FORMULATIONS

Objective: The present method was proposed to develop a simple, sensitive, rapid, accurate and stability-indicating reverse phase liquid chromatographic method for the simultaneous estimation of metformin and teneligliptin in pure and pharmaceutical formulations. Methods: The chromatographic separation was done on Discovery [250 mm X 4.6 mm: 5 μm is particle size] using a mobile phase composed of 0.1% orthophosphoric acid buffer: acetonitrile [65:35, v/v], the flow rate is 1 ml/min and the detection was carried out at 260 nm.Results: The retention time of metformin and teneligliptin were found to be 2.517 min and 3.687 min, respectively. Stability indicating studies were conducting under the guidelines of an international conference on harmonization [ICH] Q1A R2 and the developed method was validated as per guidelines of ICH Q2 RI. The linearity was found in the range of concentration of 125-750 μg/ml and 5-30 μg/ml for metformin and teneligliptin. The detection of limit and quantification of limit was found to be 0.02 μg/ml and 0.07 μg/ml for metformin and 0.19 μg/ml and 0.56 μg/ml for teneligliptin, respectively.Conclusion: A novel stability-indicating reverse-phase liquid chromatographic method for the simultaneous estimation of metformin and teneligliptin. The proposed method was adopted for the routine estimation of metformin and teneligliptin in bulk and pharmaceutical dosage forms.

Development of validated analytical method for estimation of Metformin and Pioglitazone in pharmaceutical dosage form

A RP-HPLC method was developed using Comosil RP-C18 (4.6 x 150mm, 5m) as stationary phase with younglin (S.K.) isocratic system UV detector in a gradient mode with mobile phase comprising of Acetonitrie : Pott. Dihydrogen Phosphate (adjusted pH-2.5 using OPA). 0.7ml/min flow rate and monitoring of effluent were done at 254.0 nm for MET (Metformin) and PIOG(Pioglitazone) estimation in combined dosage form.2.1 min for MET and 7.53 min for PIOG retention times were found. The dynamic range of linearity 50- 250 g/mL for MET and 3- 15 g/mL for PIOG were exhibited for the assay. The linear calibration curves were found over the entire range linearity (r 2 = 0.996 for MET and r 2 = 0.995 for PIOG) and 99.81 % for MET and 99.33 % for PIOG mean % recovery was found with % RSD was NMT 2 for both estimations which fully agrees by system suitability in good agreement with labeled claimed of formulation. The % RSD for Intra & Inter-Day Precision was NMT than 2 for both drugs. The developed method was accurate, precise, rugged and linear as per ICH guidelines

New Validated Stability Indicating RP-HPLC Method for Simultaneous Estimation of Metformin and Alogliptin in Human Plasma

A validated new stability indicating RP-HPLC method for the quantitative determination of metformin and alogliptin in human plasma was developed as per US-FDA guidelines. The drug was spiked in the plasma and extracted with mobile phase by precipitation method. The extracted analyte was injected into X-Terra C18 (4.6 × 150 mm, 3.5 μm, Make: ACE) or equivalent, maintained at 25°C temperature and effluent was monitored at 235 nm. The mobile phase was consisted of sodium dihydrogen ortho phosphate [pH 4.0]:acetonitrile [HPLC Grade] (70:30 v/v). The flow rate was maintained at 1.0 mL/min. The calibration curve for metformin and alogliptin was linear from 300.0 to 700.0 μg/mL (r2=0.997) and 7.5 to 17.5 μg/mL (r2=0.998) respectively. The inter-day and intra-day precision was found to be within limits. The Lower limit of quantification (LLOQ) for metformin and alogliptin were 5.936 and 1.983 μg/mL respectively. The average % recovery for metformin and alogliptin were 100.17 and 99.40-99.55% respectively and reproducibility was found to be satisfactory. This RP-HPLC method is suitable for determining the concentration of metformin and alogliptin in human plasma and it can applied for routine analysis for determination of the metformin and alogliptin from dosage form during pharmacokinetic study.