Determination of Dansyl Amino Acids and Oxalate by HPLC with Electrogenerated Chemiluminescence Detection Using Tris(2,2'-bipyridyl)ruthenium(II) in the Mobile Phase
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- 作者：David R. Skotty ; Won-Yong Lee ; and Timothy A. Nieman
- 刊名：Analytical Chemistry
- 出版年：1996
- 出版时间：May 1, 1996
- 年：1996
- 卷：68
- 期：9
- 页码：1530 - 1535
- 全文大小：157K
- 年卷期：v.68,no.9(May 1, 1996)
- ISSN：1520-6882
文摘
A new electrogenerated chemiluminescence detectionmethod is investigated for use in detection in reversed-phase and reversed-phase ion-pair HPLC withRu(bpy)32+in the mobile phase. In this method, different concentrations of Ru(bpy)32+ are dissolved in themobile phase andthe HPLC column flushed with the mobile phase for 1 huntil the column is saturated withRu(bpy)32+. Theseparated analytes along with Ru(bpy)32+pass through anoptical-electrochemical flow cell which has a dual platinum electrode held at a potential of 1250 mV vs a Ag/AgCl reference electrode. On the surface of theelectrode,Ru(bpy)32+ is oxidized toRu(bpy)33+ which reactswiththe analytes to emit light. The retention times,retentionorders, detection limits, and linearity in working curvesare compared to those obtained with the conventionalpostcolumn Ru(bpy)32+ addition method.The retentiontimes for dansyl amino acids withRu(bpy)32+ in themobile phase are longer than those obtained with thepostcolumn addition approach. This may be caused by
-to- interactions between the aromatic groups of thedansyl derivatives and the bipyridyl groups ofRu(bpy)32+in the Ru(bpy)32+-saturatedreversed-phase column. Similarly, oxalate is separated from urine and blood plasmasamples by reversed-phase ion-pair HPLC. Plasmasamples are obtained using ultrafiltration to removeproteins from whole blood. Retention times foroxalatewith the two detection techniques are identical, anddetection limits for these techniques are compared.
-to- interactions between the aromatic groups of thedansyl derivatives and the bipyridyl groups ofRu(bpy)32+in the Ru(bpy)32+-saturatedreversed-phase column. Similarly, oxalate is separated from urine and blood plasmasamples by reversed-phase ion-pair HPLC. Plasmasamples are obtained using ultrafiltration to removeproteins from whole blood. Retention times foroxalatewith the two detection techniques are identical, anddetection limits for these techniques are compared.