Simultaneous determination of polyamines and catecholamines in PC-12 tumor cell extracts by capillary electrophoresis with laser-induced fluorescence detection.
A method to detect polyamines and catecholamines in PC-12 tumor cell extracts by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) is described for the first time. Both derivatization conditions and buffer concentrations and pH were optimized. Under optimized conditions the polyamines (putresine, spermine, spermidine) and catecholamines (dopamine, norepinephrine, epinephrine, serotonin) were derivatized with fluorescein isothiocyanate and separated at 25 kV in a fused-silica capillary (50 microm ID x 40 cm) with 0.1 M borate, pH 9.0, in less than 18 min. The influence of running buffer conditions, such as buffer pH and concentrations, were also investigated. Linearity of the analytes ranged from 0.05 to 1.0 micromol/L, and the detection limit (S/N = 3 ) ranged from 0.03 to 2.50 nmol/L. The concentrations of polyamines and catecholamines in PC-12 tumor cell extracts were determined with this method.
Du
Ming
M
Department of Chemistry and Center for Environmental Science & Technology, University of Missouri-Rolla, 1870 Miner Circle, Rolla, MO 65409, USA.
Flanigan
Virgil
V
Ma
Yinfa
Y
D000818:Animals; D002395:Catecholamines / Q000032:analysis*; D002457:Cell Extracts / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods; D006863:Hydrogen-Ion Concentration; D016716:PC12 Cells; D011073:Polyamines / Q000032:analysis*; D051381:Rats
D002395:Catecholamines; D002457:Cell Extracts; D011073:Polyamines