TY - JOUR
T1 - Capillary electrophoresis of carboxylated carbohydrates III. Selective precolumn derivatization of glycosaminoglycan disaccharides with 7- aminonaphthalene-1,3-disulfonic acid fluorescing tag for ultrasensitive laser-induced fluorescence detection
AU - El Rassi, Ziad
AU - Postlewait, Jennifer
AU - Mechref, Yehia
AU - Ostrander, Gary K.
N1 - Funding Information:
Z.E.R. acknowledges the partial ®nancial support by the Cooperative State Research Service, U.S. Department of Agriculture, under Agreement 94-37102-0989, and G.K.O. acknowledges the ®nancial support of NIH Grant CA54590.
PY - 1997/1/15
Y1 - 1997/1/15
N2 - Eight different glycosaminoglycan-derived disaccharities were selectively labeled via their carboxylic acid group with 7-aminonaphthalene- 1,3-disulfonic acid (ANDSA) by a condensation reaction between the amino group of ANDSA and the carboxylic acid group of the saccharides in the presence of water-soluble carbodiimide. This derivatization reaction yielded stable derivatives with percentage yields as high as 97%. The ANDSA disaccharide derivatives were readily detected by on-column laser induced fluorescence (LIF) with a He-Cd laser at 325 nm. With LIF, the limit of detection was at the nanomolar level, three orders of magnitude lower than the limit of detection of underivatized disaccharides in the uv at 231 nm. In addition, due to the presence of two strong sulfonic acid groups in the ANDSA tag, the derivatives were readily separated at acidic pH (i.e., pH 4.0-5.0) using 100 mM sodium acetate buffers as the running electrolytes. The addition of polycationic spermine in small amounts to the running electrolytes provided different selectivity with baseline resolution in the pH range 6.0- 7.0, and the excess ANDSA migrated ahead of the ANDSA disaccharides.
AB - Eight different glycosaminoglycan-derived disaccharities were selectively labeled via their carboxylic acid group with 7-aminonaphthalene- 1,3-disulfonic acid (ANDSA) by a condensation reaction between the amino group of ANDSA and the carboxylic acid group of the saccharides in the presence of water-soluble carbodiimide. This derivatization reaction yielded stable derivatives with percentage yields as high as 97%. The ANDSA disaccharide derivatives were readily detected by on-column laser induced fluorescence (LIF) with a He-Cd laser at 325 nm. With LIF, the limit of detection was at the nanomolar level, three orders of magnitude lower than the limit of detection of underivatized disaccharides in the uv at 231 nm. In addition, due to the presence of two strong sulfonic acid groups in the ANDSA tag, the derivatives were readily separated at acidic pH (i.e., pH 4.0-5.0) using 100 mM sodium acetate buffers as the running electrolytes. The addition of polycationic spermine in small amounts to the running electrolytes provided different selectivity with baseline resolution in the pH range 6.0- 7.0, and the excess ANDSA migrated ahead of the ANDSA disaccharides.
UR - http://www.scopus.com/inward/record.url?scp=0031568286&partnerID=8YFLogxK
U2 - 10.1006/abio.1996.9905
DO - 10.1006/abio.1996.9905
M3 - Article
C2 - 9025945
AN - SCOPUS:0031568286
SN - 0003-2697
VL - 244
SP - 283
EP - 290
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -