A new elect
rogene
rated chemiluminescence detectionmethod is investigated fo
r use in detection in
reve
rsed-phase and
reve
rsed-phase ion-pai
r HPLC withRu(bpy)
32+in the mobile phase. In this method, diffe
rent concent
rations of Ru(bpy)
32+ a
re dissolved in themobile phase andthe HPLC column flushed with the mobile phase fo
r 1 huntil the column is satu
rated withRu(bpy)
32+. Thesepa
rated analytes along with Ru(bpy)
32+pass th
rough anoptical-elect
rochemical flow cell which has a dual platinum elect
rode held at a potential of 1250 mV vs a Ag/AgCl
refe
rence elect
rode. On the su
rface of theelect
rode,Ru(bpy)
32+ is oxidized toRu(bpy)
33+ which
reactswiththe analytes to emit light. The
retention times,
retentiono
rde
rs, detection limits, and linea
rity in wo
rking cu
rvesa
re compa
red to those obtained with the conventionalpostcolumn Ru(bpy)
32+ addition method.The
retentiontimes fo
r dansyl amino acids withRu(bpy)
32+ in themobile phase a
re longe
r than those obtained with thepostcolumn addition app
roach. This may be caused by
rs/pi.gif" BORDER=0 >-to-
rs/pi.gif" BORDER=0 > inte
ractions between the a
romatic g
roups of thedansyl de
rivatives and the bipy
ridyl g
roups ofRu(bpy)
32+in the Ru(bpy)
32+-satu
rated
reve
rsed-phase column. Simila
rly, oxalate is sepa
rated f
rom u
rine and blood plasmasamples by
reve
rsed-phase ion-pai
r HPLC. Plasmasamples a
re obtained using ult
rafilt
ration to
removep
roteins f
rom whole blood. Retention times fo
roxalatewith the two detection techniques a
re identical, anddetection limits fo
r these techniques a
re compa
red.