DETERMINATION OF METFORMIN AND SITAGLIPTIN IN HEALTHY HUMAN VOLUNTEERS' BLOOD PLASMA AND ITS BIOEQUIVALENCE STUDY UNDER FASTING CONDITION

DIBYA DAS; DHIMAN HALDER; ANIRBANDEEP BOSE; TAPAN KUMAR SHAW; CHIRANJIT SAHA; PINTU KUMAR DE; HIMANGSHU SEKHAR MAJI; TAPAN KUMAR PAL

doi:10.22159/ijap.2022v14i6.45140

Authors

DIBYA DAS Department of Pharmaceutical Technology, JIS University, Kolkata, India https://orcid.org/0000-0002-8788-3795
DHIMAN HALDER Department of Pharmaceutical Technology, Bioequivalence Study Centre, Jadavpur University, Kolkata, India https://orcid.org/0000-0002-8694-8243
ANIRBANDEEP BOSE TAAB Biostudy Services, Jadavpur, Kolkata, India https://orcid.org/0000-0002-9488-9503
TAPAN KUMAR SHAW Department of Pharmaceutical Technology, JIS University, Kolkata, India https://orcid.org/0000-0001-8585-7641
CHIRANJIT SAHA TAAB Biostudy Services, Jadavpur, Kolkata, India https://orcid.org/0000-0001-5942-0390
PINTU KUMAR DE Department of Pharmaceutical Technology, JIS University, Kolkata, India https://orcid.org/0000-0002-9898-7506
HIMANGSHU SEKHAR MAJI Department of Pharmaceutical Technology, JIS University, Kolkata, India https://orcid.org/0000-0003-0640-0374
TAPAN KUMAR PAL Department of Pharmaceutical Technology, Bioequivalence Study Centre, Jadavpur University, Kolkata, India

DOI:

https://doi.org/10.22159/ijap.2022v14i6.45140

Keywords:

Metformin, Sitagliptin, Type-2 diabetes, Method development and validation, USFDA, EMA, Liquid-liquid extraction, and Bioequivalence studies

Abstract

Objective: Metformin hydrochloride and sitagliptin are the oral anti-hyperglycemic medications used to treat type 2 diabetes and are used in combination to treat patients. In this work, we have developed a bioanalytical method for simultaneous estimation of both the drugs form some formulation and subsequently the validation of the developed method metformin and sitagliptin in human plasma.

Methods: The stability studies were done as per USFDA and EMA guidelines. The sample extraction approach presented here was a straightforward liquid extraction. The linearity range of metformin was 11.72 ng/ml to 3000 ng/ml, and sitagliptin was 4.68 ng/ml. to 1200 ng/ml. For metformin, the LOD was 1.0 ng/ml, and LLOQ was 11.72 ng/ml. and for sitagliptin, the LOD was 0.75 ng/ml, and LLOQ was 4.68 ng/ml. LC-ESI-MS/MS was used to develop and validate this method using the Phenomenex Kinetex C18 column. Milli-Q water containing 10 mmol Ammonium Acetate (pH =3.6) and Acetonitrile containing 0.1% Formic Acid (pH =2.4) as solvent systems for the estimation of Sitagliptin in a single dose. Metoprolol is used as an Internal Standard.

Results: The total chromatographic run time was only 7.0 min, and the elute time of metformin and sitagliptin was 3.94 min and 3.97 min, respectively. Relative Bioavailability was found at 101.14% for Metformin and 96.96% for Sitagliptin. The overall results show that the Cmax, AUC0-t, and AUC0-∞ for metformin and sitagliptin were within the acceptable limit of 80%-125%.

Conclusion: This bioanalytical method was successfully applied in the bioequivalence study. The study design was a randomized, open-label, two treatment, two-period, two sequences, single-dose, crossover bioequivalence study under fasting conditions.

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