Title:A Bioanalytical Method Using High-performance Liquid Chromatography-mass Spectrometry for Determining Empagliflozin and Linagliptin in Human Plasma: Application in Bioequivalence Pharmacokinetic Study
Volume: 20
Issue: 9
Author(s): Rana Said*, Basel Arafat and Tawfiq Arafat
Affiliation:
- Pharmacological and Diagnostic Research Centre (PDRC), Faculty of Pharmacy, Al-Ahliyya Amman University,
Amman, Jordan
Keywords:
Empagliflozin, linagliptin, LC-MS/MS, protein precipitation, validation, pharmacokinetic, bioequivalence.
Abstract:
Background and Objectives: A combination of empagliflozin and linagliptin in a fixed
dosage was employed for treating individuals with a diagnosis of type 2 diabetes mellitus. A rapid,
accurate, and sensitive liquid chromatography-tandem mass spectrometry method was devised and
validated for simultaneous measuring empagliflozin and linagliptin levels in human plasma. This
method provides a good analytical tool for bioequivalence and pharmacokinetic studies.
Methods: The separation was conducted employing a C8 column using a mobile phase consisting
of acetonitrile (ACN, 2.5mM) and ammonium chloride (55:45). Optimal detection of the analytes
and their deuterated internal standards was accomplished through electrospray ionization in the
positive mode.
Results: Validation of standard curve concentrations linearity was carried out within the ranges of
1.500 – 500.000 ng/mL for empagliflozin and 0.050 – 7.000 ng/mL for linagliptin. Both drugs
showed intra-batch and inter-batch precision (CV%) of less than 3.7%. The stability of the drugs
was confirmed under various storage conditions, proving suitability for routine laboratory analysis.
Conclusion: This validated method is appropriate for pharmacokinetic studies and large-scale
analysis with high precision and accuracy.