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Journal of Analytical Methods in Chemistry
Volume 2017, Article ID 1208753, 9 pages
Research Article

UHPLC-PDA Assay for Simultaneous Determination of Vitamin D3 and Menaquinone-7 in Pharmaceutical Solid Dosage Formulation

1Department of Chemistry, Forman Christian College (A Chartered University), Lahore, Pakistan
2NovaMed Pharmaceuticals, Lahore, Pakistan
3Institute of Chemistry, University of the Punjab, Lahore 54590, Pakistan
4Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore 54590, Pakistan

Correspondence should be addressed to Muhammad Imtiaz Shafiq; kp.ude.up@bbi.zaitmi

Received 5 June 2017; Revised 29 June 2017; Accepted 18 July 2017; Published 21 August 2017

Academic Editor: Bengi Uslu

Copyright © 2017 Muhammad Jehangir et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


A newly developed method based on ultrahigh performance liquid chromatography (UHPLC) was optimized for the simultaneous determination of vitamin D3 and menaquinone-7 (MK-7) in tablet formulation in the present study. UHPLC separation of vitamin D3 and MK-7 was performed with ACE Excel 2 C18-PFP column (2 m, 2.1 100 mm) at 0.6 mL min−1 flow rate, whereas the mobile phase consisted of methanol/water (, v/v, phase A) and isopropyl alcohol (99.9%, phase B) containing 0.5% triethylamine. Isocratic separation of both the analytes was performed at 40°C by pumping the mobile phases A and B in the ratio of (v/v, pH, 6.0). Both analytes were detected at a wavelength of 265 nm and the injection volume was 1.0 L. The overall runtime per sample was 4.5 min with retention time of 1.26 and 3.64 min for vitamin D3 and MK-7, respectively. The calibration curve was linear from 5.0 to 100 g mL−1 for vitamin D3 and MK-7 with a coefficient of determination () 0.9981, while repeatability and reproducibility (expressed as relative standard deviation) were lower than 1.46 and 2.21%, respectively. The proposed HPLC method was demonstrated to be simple and rapid for the determination of vitamin D3 and MK-7 in tablets.