Determination of 125I-labelled thyroxine glucuronide by reversed-phase high-performance liquid chromatography using on-line radiochemical detection to determine UDPglucuronosyltransferase activity.
A convenient, fast and highly sensitive high-performance liquid chromatographic method, using on-line radiochemical detection, is described for the determination of [125I]thyroxine glucuronide. The method involves direct injection of the supernatant, a total analysis time of 30 min and a detection limit of 1 pmol. The results demonstrate that the method is suitable for the determination of UDPglucuronosyltransferase activity with thyroxine as substrate in native hepatic microsomes. The rate of thyroxine glucuronidation in microsomes from rats treated with Arodor 1254 was ten times higher than in control microsomes, indicating that with this method, increases of UDPglucuronosyltransferase thyroxine activities, often associated with hepatic induction process involved in thyroid hypertrophy, can be easily detected. This method could also be applied to all experimental biological systems that involve the separation and quantification of [125I]thyroxine and [125I]thyroxine glucuronide.
Département Analyse, Rhône-Poulenc Rorer, Centre de Recherche de Vitry-Alfortville, Vitry-sur-Seine, France.
G
G
Ducrotoy
L
L
Richert
V
V
De Sandro
D
D
Lurier
E
E
Pacaud
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005965:Glucuronates / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000378:metabolism*; D007457:Iodine Radioisotopes; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D011854:Radiochemistry; D051381:Rats; D011919:Rats, Inbred Strains; D013974:Thyroxine / Q000031:analogs & derivatives* / Q000378:metabolism
D005965:Glucuronates; D007457:Iodine Radioisotopes; C054785:thyroxine glucuronide; D014453:Glucuronosyltransferase; D013974:Thyroxine