Determination of recombinant human epidermal growth factor (rhEGF) in a pharmaceutical formulation by high performance liquid chromatography with electrochemical detection.
A novel HPLC method with electrochemical detection has been developed for the determination of recombinant human epidermal growth factor (rhEGF) in pharmaceutical products. rhEGF was separated from other components in formulation on a reversed-phase C18 column with 24% acetonitrile in 0.1 M phosphate buffer (pH 4.75). The optimum electrochemical oxidation of EGF was obtained at 0.85 V vs. Ag/AgCl in a glassy carbon working electrode due to electroactive tyrosine, tryptophan, methionine, and arginine residues. The quantitation range was from 1.0 to 200 ng of rhEGF with the linear correlation coefficient greater than 0.999. The method was successfully applied for the quantitation of rhEGF in a pharmaceutical preparation.
Lee
K W
KW
College of Pharmacy, Chungbuk National University, Cheongju, Korea.
Hwang
K H
KH
Kim
C S
CS
Han
K
K
Chung
Y B
YB
Park
J S
JS
Lee
Y M
YM
Moon
D C
DC
D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004563:Electrochemistry; D004815:Epidermal Growth Factor / Q000032:analysis*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000032:analysis; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D011994:Recombinant Proteins; D004815:Epidermal Growth Factor