Application of Design of Experiment and Simulation Methods to Liquid Chromatography Analysis of Topical HIV Microbicides Stampidine and HI443Vivek Agrahari, Jianing Meng, Tao Zhang and Bi-Botti C Youan*
Laboratory of Future Nanomedicines and Theoretical Chronopharmaceutics, Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, USA
- *Corresponding Author:
- Bi-Botti C Youan
Laboratory of Future Nanomedicines and Theoretical Chronopharmaceutics
Division of Pharmaceutical Sciences, School of Pharmacy
University of Missouri-Kansas City, 2464 Charlotte Street
690.71 Kansas City, MO 64108, USA
Tel: +1 816-235-2410
Fax: +1 816-235-5779
E-mail: [email protected]
Received date: December 19, 2013; Accepted date: January 22, 2014; Published date: January 24, 2014
Citation: Agrahari V, Meng J, Zhang T, Youan BB (2014) Application of Design of Experiment and Simulation Methods to Liquid Chromatography Analysis of Topical HIV Microbicides Stampidine and HI443. J Anal Bioanal Tech 5:180. doi: 10.4172/2155-9872.1000180
Copyright: © 2014 Agrahari V, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
This study is intended to determine if experimental design and Monte Carlo simulation methods can be utilized to optimize the liquid chromatography (LC) analysis of active molecules. The method was applied for the simultaneous analysis of two vaginal microbicides, stampidine (STP) and HI443 in bulk and nanoformulations. The Plackett-Burman design was used for screening; whereas, Box-Behnken design was used to evaluate the main and interaction effects of the selected factors on the responses, namely peak area of STP (Y1), HI443 (Y2), tailing of STP (Y3), and HI443 (Y4). The Monte Carlo simulation was applied to get the minimum defect rate of the process. The optimized LC conditions were found to be X1; flow rate: 0.6 mL/min, X2; injection volume: 18 μL, and X3; initial gradient acetonitrile ratio: 92% v/v with a minimal defect rate of 0.077%. The optimized method was applied to determine the percent encapsulation efficiency (%EE) and in vitro release profile of STP and HI443 from solid lipid nanoparticles (SLNs). The % EE of STP and HI443 in SLNs was found to be 30.56 ± 9.44 and 94.80 ± 21.90% w/w, respectively, (n=3). It was observed that the release kinetics of STP followed the first order, whereas, HI443 followed the Peppas kinetic model in SLNs. The LC method was also applied for the estimation of molar extinction coefficients (ε270) of both drugs for the first time. The values were observed as 7,569.03 ± 217.96 and 17,823.67 ± 88.12 L/mol/cm for STP and HI443, respectively, (n=3). The results suggest that experimental design and Monte Carlo simulation can be effectively used to reduce the defect rate and to optimize the chromatographic conditions for the analysis of active drug molecules as applied in this study.