A micromethod for quantitative determination of iodoamino acids in thyroglobulin

被引:10
作者
Baudry, N [1 ]
Mallet, B [1 ]
Lejeune, PJ [1 ]
Vinet, L [1 ]
Franc, JL [1 ]
机构
[1] FAC MED, UNITE 38 INSERM, F-13385 MARSEILLE 5, FRANCE
关键词
D O I
10.1677/joe.0.1530099
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
We describe a new method for quantification of iodoamino acids after enzymatic hydrolysis of thyroglobulin. The procedure involves separation of monoiodotyrosine (MIT), diiodotyrosine, tri-iodothyronine and thyroxine by reverse phase HPLC with a Vydac C-18 stationary phase and a mobile phase of water-acetonitrile-acetic acid. The separation is monitored by sensitive spectrophotometric detection through a 96-well microplate system based on the catalytic Sandell-Kolthoff reaction of iodide on the oxidation of arsenic(III) by cerium(IV). This new microassay is particularly convenient because of its high sensitivity and its rapidity (less than 2 h). It can detect 1 pmol MIT and 0.5 pmol of the other three iodoamino acids with a recovery higher than 96%. Moreover, the 96-well microplate system allows many samples to be tested simultaneously and avoids the use of radio-labeled iodine.
引用
收藏
页码:99 / 104
页数:6
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