A method for the determination of betaine in tissues using high performance liquid chromatography.
Betaine is a major metabolite of choline in liver and kidney and may be an important product of choline metabolism in other tissues. The available methods for assay of betaine, however, are time consuming and relatively insensitive. We describe a modification of published methods that provides a sensitive and specific assay for betaine by derivatization and HPLC separation with UV quantitation. Betaine and other water-soluble choline metabolites are extracted from biological samples and separated by HPLC based on mobility of 14C-labeled internal standards. The betaine fraction is collected and derivatized with 4'-bromo-phenacyl triflate. The betaine-triflate derivative is quantitated by UV absorbance and the result is corrected for possible losses due to incomplete extraction recovery and incomplete derivatization by simultaneous measurement of radioactivity from the derivatized 14C-betaine internal standard. Betaine concentrations determined with this procedure are reported for several adult and fetal rat tissues.
Mar
Mei Heng H.
MH
Department of Nutrition, Schools of Public Health and Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
Ridky
Todd W.
TW
Garner
Sanford C.
SC
Zeisel
Steven H.
SH