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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Sequential injection-cation exchange micro-column system for hemoglobin typing to differentiate HbE carriers

Supaporn Kradtap Hartwell1, Worathip Sripaoraya, Somchai Lapanantnoppakhun

  • 1Department of Chemistry and Center for Innovation in Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand. kradtas@yahoo.com

Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry
|March 11, 2010
PubMed

A weak cation exchange micro-column was incorporated into a sequential injection (SI) system to perform automatic hemoglobin (Hb) typing as an alternative way to measure HbE. Separation of HbF, HbA and HbA(2)/HbE was performed using phosphate buffer solutions in the pH range of 6-7 to create pH gradient mobile phase. The resultant chromatogram showed relative amounts of HbE to other types of hemoglobins in more quantitative detail than the conventional techniques such as dichlorophenol indophenol precipitation and micro-column anion exchange. The system is more economical than a commercially available ion-exchange HPLC analyzer for hemoglobin testing, though analysis time per run is longer due to the aspiration operation of the syringe pump of the SI system. It has been demonstrated that the system can differentiate negative (normal) from positive (HbE carriers) subjects.

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