Effect of Temperature, Wavelength, pH, Ion Pair Reagents and Organic Modifi ers' Concentration on the Elution of Cystatin C. Stability of Mobile PhaseManar Khalid Fayyad1, Adel Khalil Misha2and Othman Ibrahem Yousef Al-Musaimi3*
- *Corresponding Author:
- Othman Ibrahem Yousef Al-Musaimi
Department of Chemistry
University of Ha'il
Ha'il –Saudia Arabia
Tel: 962 78 5782259
E-mail: [email protected]
Received date: September 20, 2010; Accepted date: September 29, 2010; Published date: October 01, 2010
Citation: Fayyad MK, Misha AK, Yousef Al-Musaimi OI (2010) Effect of Temperature, Wavelength, pH, Ion Pair Reagents and Organic Modifiers' Concentration on the Elution of Cystatin C. Stability of Mobile Phase. J Anal Bioanal Tech 1:103. doi: 10.4172/2155-9872.1000103
Copyright: © 2010 Fayyad MK, 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.
Robustness of an analytical chromatographic method for separation of cystatin c has been veri fi ed. Changes in many parameters were carried out, such as, wavelength, column oven, mobile phase composition, chromatographic column.
Imperative changes have altered the ef fi ciency of the chromatographic separation; such changes include pH alteration of the mobile phase as well as alkyl sulfonate molarity changing.
All robustness conditions showed no major effect on the chromatographic separation of the analyte except with the changes related to TFA and alkyl sulfonate ion pair reagents. Peak area RSD, asymmetry and No. of theoretical plates were < 0.7%, < 1.2 and > 10,000, respectively. Results obtained using mobile phase after 6 months of storage have proven its stability and possibility of use. Gradient elution mode was utilized to elute cystatin c with a UV detection of 224 nm. Ace and Waters C8 (150 x 4.6 mm i.d., 5 μ m) as chromatographic columns were used.