High-performance liquid chromatographic separation of biotinylamide analogues used as substrates in biotinidase radioassays.
A simple, one-step protocol for synthesizing biotinylmono[125I]iodotyramine and biotinyldi[125I]iodotyramine, which are used as tracer substrates in biotinidase radioassays, is presented. This synthetic protocol uses reversed-phase HPLC for the isolation of the biotinylamide analogues from the reaction mixture. The HPLC method developed can potentially be applied to a scaled-up synthetic protocol for non-radioactive biotinylmono- and diiodotyramine. In addition, it can be used for the identification of the above-mentioned compounds.
Livaniou
E
E
Radioimmunochemistry Laboratory, Institute of Radioisotopes/Radiodiagnostic Products, NCSR Demokritos, Athens, Greece.
Roboti
A
A
Kakabakos
S E
SE
Nyalala
J
J
Evangelatos
G P
GP
Ithakissios
D S
DS
D000581:Amidohydrolases / Q000378:metabolism*; D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification*; D043523:Biotinidase; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D007457:Iodine Radioisotopes; D013056:Spectrophotometry, Ultraviolet
D007457:Iodine Radioisotopes; C053607:N-(4-hydroxy-3-iodophenylethyl)biotin amide; C053608:N-(4-hydroxy-3,5-diiodophenylethyl)biotin amide; D001710:Biotin; D000581:Amidohydrolases; D043523:Biotinidase