Original Article
A VALIDATED STABILITY INDICATING ULTRA-PERFORMANCE LIQUID CHROMATOGRAPHIC
METHOD FOR SIMULTANEOUS DETERMINATION OF METFORMIN HYDROCHLORIDE AND
EMPAGLIFLOZIN IN BULK DRUG AND TABLET DOSAGE FORM
N. MADANA GOPAL
1*
, C. SRIDHAR
2
1
Research Scholar, Rayalaseema University, Kurnool. Andhra Pradesh, India,
2
Department of Pharmaceutical Analysis, Sri Padmavathi
School of Pharmacy, Tirupathi, Andhra Pradesh, India
Email: madanapharma@gmail.com
Received: 02 Feb 2017, Revised and Accepted: 17 Apr 2017
ABSTRACT
Objective: To develop a simple, precise, accurate, method was developed and validated for analysis of metformin hydrochloride (MET) and
empagliflozin in (EMPA) in bulk and tablet dosage form.
Methods: The method used a reverse phase column, dikma C18 (50×2.1 mm, 1.8 μ), a mobile phase comprising of phosphate buffer (pH-3):
methanol (30:70 v/v) flow rate of 1.0 ml/min and a detection wavelength of 240 nm using a photodiode array detector. The proposed method was
validated for various parameters like linearity, precision, accuracy, robustness, ruggedness, detection, quantification limits, stability studies,
formulation analysis as per International Conference on Harmonization (ICH) guidelines.
Results: The retention time was found to be 1.189 min and 1.712 min for MET and EMPA respectively. The proposed method was found to be
having linearity in the concentration range of 500-2500 μg/ml for MET (r
2
=0.989) and 5-25 μg/ml for EMPA (r
2
=0.994), respectively. The mean %
recoveries obtained were found to be 100.35-100.48% for MET and 99.80-101.30% for EMPA respectively. Stress testing which covered acid, base,
peroxide, photolytic and thermal degradation was performed on under test to prove the specificity of the method and the degradation was achieved.
The developed method has been statistically validated according to International Conference on Harmonization (ICH) guidelines.
Conclusion: Thus, the proposed method can be successfully applied for the stability indicating the simultaneous determination of MET and EMPA in
bulk and combined tablet dosage form and in the routine quality control analysis.
Keywords: Empagliflozin, Metformin hydrochloride, Reversed-phase ultra-performance liquid chromatography, Validation
© 2017 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijap.2017v9i3.17441
INTRODUCTION
Metformin (MET) hydrochloride is chemically named as (3-(diamino
methylidene)-1, 1-dimethylguanidine hydrochloride. It has a
molecular formula of C4H12ClN5 and the molecular weight is 165.62
g/mol. MET is an oral anti-hyperglycemic agent (Type 2 diabetes)
belongs to a class of biguanides and useful for treating non-insulin
dependent diabetes mellitus. MET decreases blood glucose levels by
decreasing hepatic glucose production, decreasing intestinal
absorption of glucose, and improving insulin sensitivity by
increasing peripheral glucose uptake and utilization. These effects
are mediated by the initial activation by MET of AMP-activated
protein kinase, a liver enzyme that plays an important role in insulin
signaling, whole body energy balance, and the metabolism of glucose
and fats [1-5].
Empagliflozin (EMPA) is chemically named as 1-chloro-4-[b-D-
glucopyranos-1-yl]-2-[4-([S]-tetrahydrofuran–3–yl-oxy) benzyl]-
benzene (fig. 2). It has a molecular formula of C23H27ClO7 and
molecular weight is 450.912 g/mol. Empagliflozin is an orally
administered selective sodium glucose cotransporter-2 (SGLT-2)
inhibitor, which lowers blood glucose in people with type 2 diabetes
by blocking the reabsorption of glucose in the kidneys and
promoting the excretion of excess glucose in the urine [6, 8]. The
sodium glucose cotransporter 2 (SGLT2), located in the proximal
tubule of the nephron, is estimated to facilitate-90% of this
reabsorption [9, 10]. It is a potent and selective competitive
inhibitor of the SGLT2 protein. Sodium-glucose co-transporter 2
(SGLT2) inhibitors offer an insulin-independent mechanism for
improving blood glucose levels since they promote urinary glucose
excretion (UGE) by inhibiting glucose reabsorption in the kidney. In
addition to glucose control, SGLT2 inhibitors are associated with
weight loss and blood pressure reductions and do not increase the
risk of hypoglycemia [9].
Fig. 1: Metformin (MET) hydrochloride structure from pub chem
Fig. 2: Empagliflozin (EMPA) structure from pubchem
International Journal of Applied Pharmaceutics
ISSN- 0975-7058 Vol 9, Issue 3, 2017