Human serum albumin (HSA) was successfully bonded to silica with s-triazine
as activator. The coupling reaction by this method was rapid and effective
. The triazine-activated silica is relatively stable and can be installed f
or at least 1 month without obvious loss of reactivity when stored below 30
degreesC, pH below 7. It was observed that the amount of bound HSA reached
120 mg/g silica calculated from the UV absorbance difference of the HSA so
lution. d,l-tryptophan was selected as the probe solute to characterize the
properties of HSA bonded s-triazine chiral stationary phase, and separatio
n factor of 9.4 was obtained for d,l-tryptophan. Furthermore, the amount of
effective HSA on silica was measured by high-performance frontal analysis,
and only 16.8 mg/g silica was responsible for the resolution of d,l-trypto
phan. These results indicate that the amount of both the bound and effectiv
e HSA on silica with triazine as activator was much higher than those by th
e Schiff base coupling method. Different kinds of enantiomers were resolved
successfully on the aminopropylsilica-bonded HSA s-triazine chiral station
ary phase. (C) 2000 Wiley-Liss, Inc.