Table of Contents
Advances in Pharmaceutics
Volume 2014, Article ID 749823, 6 pages
Research Article

A Rapid, Isocratic HPLC Method for Determination of Insulin and Its Degradation Product

1College of Pharmacy, Department of Pharmaceutical Chemistry, Al-Jouf University, Sakaka, Saudi Arabia
2Faculty of Science, Department of Chemistry, The University of Jordan, P.O. Box 13003, Amman 11942, Jordan
3Al-Taqaddom Pharmaceutical Industries Company, Amman, Jordan

Received 7 April 2014; Revised 2 June 2014; Accepted 10 June 2014; Published 23 July 2014

Academic Editor: Maria J. Morilla

Copyright © 2014 Ahmad Najjar 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.


This paper aimed to develop a simple, sensitive, and rapid chromatographic procedure for the simultaneous analysis of human insulin and its main decomposition product using isocratic RP-HPLC/UV. A column type RP-C18 (100 × 4.6 mm, 3 μm particle size, and pore size 130 Å) was used. o-Nitrophenol was used as internal standard. The eluent consists of 62% KH2PO4 buffer (0.1 M), 26% ACN, and 12% MeOH. The final pH was adjusted to 3.1. The eluent was pumped at a flow rate of 1.0 mL/min and the effluent was monitored using DAD detector at 214 nm. The method produces a linear response over the concentration range of 0.0106 to 0.6810 mg/mL with detection limit of 0.0029 mg/mL. Considering the specifications of this method, the system was found to be suitable for rapid, direct routine analysis and stability studies of insulin.