Analysis of intact heparin by capillary electrophoresis using short end injection configuration.
A capillary electrophoresis method for the analysis of intact heparin was developed using a phosphate buffer and a fused silica capillary. Operational parameters such as pH and concentration of the running buffer were investigated. The short end injection configuration permitted a gain on peak efficiency, on the analysis time and on the repeatability of both migration times and peak areas, through a reduction of the migration distance. Moreover, the beneficial effect of the presence of sodium chloride in the heparin sample on the peak efficiency was demonstrated and the influence of the salts on the conformation of the heparin was discussed. The optimized method (short end injection configuration, 50mM phosphate buffer pH 3, heparin sample prepared in 10 g/L NaCl solution) was validated in terms of linearity, reproducibility and specificity according to ICH requirements.
Duchemin
V
V
ROCHE, Service de développement analytique, 52 rue M. et J. Gaucher, 94120 Fontenay-sous-Bois, France.
Le Potier
I
I
Troubat
C
C
Ferrier
D
D
Taverna
M
M
D002021:Buffers; D019075:Electrophoresis, Capillary / Q000379:methods*; D006493:Heparin / Q000032:analysis* / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D002021:Buffers; D006493:Heparin