Original Article
STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE
SIMULTANEOUS DETERMINATION OF VILDAGLIPTIN AND METFORMIN IN
PHARMACEUTICAL DOSAGE FORM
RAMESH JAYAPRAKASH
*
, SENTHIL KUMAR NATESAN
Department of Pharmaceutical Analysis, JKK Munirajah Medical Research Foundation’s-Annai JKK Sampoorani Ammal College of
Pharmacy, Komarapalayam, Namakkal-DT, Tamilnadu, India
Email: rameshj1974@gmail.com
Received: 19 Nov 2016 Revised and Accepted: 17 Jan 2017
ABSTRACT
Objective: The present study was aimed to develop a rapid, accurate, linear, sensitive and validate stability-indicating high performance liquid
chromatographic [RP-HPLC] method for determination of vildagliptin and metformin in pharmaceutical dosage form.
Methods: The chromatographic separation was performed on kromasil-C18 column [4.5 x 250 mm; 5 µm] using a mobile phase consisting of 0.05
mmol potassium dihydrogen phosphate buffer: acetonitrile [80:20 v/v], [pH adjusted to 3.5 using orthophosphoric acid]. The flow rate is 0.9
ml/min and the detection was carried out at 263 nm.
Results: The chromatographic condition, the peak retention time of metformin and vildagliptin were found to be 2.215 min and 2.600 min
respectively. Stress testing was performed in accordance with an international conference on harmonization [ICH] Q1A R2 guidelines. The method
was validated as per ICH Q2 R1 guidelines. The calibration curve was found to be linear in the concentration range of 5-17.5 µg/ml and 50-175
µg/ml for vildagliptin and metformin. The limit of detection and quantification was found to be 0.0182 µg/ml and 0.0553 µg/ml for vildagliptin and
0.4451 µg/ml and 1.3490 µg/ml for metformin respectively.
Conclusion: A new sensitive, simple and stability indicating reverse-phase high-performance liquid chromatography [RP-HPLC] method has been
developed and validated for the determination of vildagliptin and metformin. The proposed method can be used for routine determination of
vildagliptin and metformin.
Keywords: Metformin, Method validation, RP-HPLC, Stability indicating method, Vildagliptin
© 2016 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/ijpps.2017v9i3.16233
INTRODUCTION
Vildagliptin [fig. 1] is chemically [S]-1-[N-[3-hydroxy-1-adamantyl]
glycyl] pyrrolidine-2-carbonitrile, is a potent di-peptidyl peptidase
IV [dip-IV] inhibitor, a drug for the treatment of diabetes. DPP-IV
inhibitor represents a new class of oral antihyperglycemic agents to
treat patients with type 2 diabetes [1-4].
Metformin HCl [fig. 2] chemically 3-[diaminomethylidene]-1,1-
dimethylguanidine HCl is an orally administered biguanide widely
used in the treatment of type 2 [non-insulin dependent] diabetes
mellitus [5-6].
Literature survey revealed that RP-HPLC [7-16] have been reported
for the estimation of vildagliptin and metformin.
To the best of our knowledge, no stability indicating reverse-
phase high-performance liquid chromatography [RP-HPLC]
method has been reported for vildagliptin and metformin. The
present work involves stress degradation as per ICH Q1A [R2]
and Q1B [17-18] for the developing new, simple, sensitive
stability indicating RP-HPLC method; the method was validated
as per the ICH guidelines Q2 R1 [19].
The reported methods were not cost effective due to use costly
solvents and in some methods the retention time vildagliptin was
found to be more [7-16]. So in the present work a simple, precise,
sensitive and stability indicating method was developed by using
low-cost solvent acetonitrile with buffer in the ratio of 80:20,
detected by using a photodiode array detector which was highly
sensitive to detect at a lower concentration. The developed method
was used for estimation of vildagliptin and metformin in
pharmaceutical dosage form.
Fig. 1: Chemical structure of vildagliptin
Fig. 2: Chemical structure of metformin Hcl
MATERIALS AND METHODS
Chemicals and reagents
Vildagliptin was provided as a gift sample by Aurobindo
pharmaceuticals limited [Hyderabad, India]. Metformin HCl was
provided as a gift sample by Ranbaxy laboratories limited [Gurgaon,
India]. Acetonitrile [HPLC grade] purchased from E-Merck
specialities private. Limited [Mumbai, India], and Potassium
International Journal of Pharmacy and Pharmaceutical Sciences
ISSN- 0975-1491 Vol 9, Issue 3, 2017