Separation of polyethylene glycols and their fluorescein-labeled compounds depending on the hydrophobic interaction by high-performance liquid chromatography.
The separation and characterization of fluorescein-labeled polyethylene glycols (PEG) is described. Firstly, the polyethylene glycols labeled with fluorescein isothiocyanate (FITC) were synthesized and separated using Sephadextrade mark LH-20 medium by a step gradient. Secondly, a TSK GEL G4000 PW XL column was developed for determining the FITC derivatives of PEG. The retention mechanism is based on the hydrophobic interaction between the FITC derivatives of PEG and the packing material of the TSK GEL G4000PW XL column. The retention time of the PEG compounds increased by adding of salts in the mobile phase and decreased by adding of organic modifier. In addition, various salts in the eluent can also change the chromatographic behavior of these compounds. Finally, the pH of the mobile phase can have an impact on the retention time of the PEG compounds.
Liu
Min
M
Fudan-Pharmco Targeting Drug Research Center, Department of Pharmaceutics, School of Pharmacy, University of Fudan, Shanghai 200032, China.
Xie
Cao
C
Pan
Hong
H
Pan
Jun
J
Lu
Weiyue
W
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019793:Fluorescein / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D015394:Molecular Structure; D011092:Polyethylene Glycols / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results
D011092:Polyethylene Glycols; D019793:Fluorescein