Derivatization of isolated endogenous butyrobetaine with 4'-bromophenacyl trifluoromethanesulfonate followed by high-performance liquid chromatography.
A method for the isolation and chromatography of butyrobetaine from plasma, urine, and liver is described. The recovery of [3H-methyl]butyrobetaine from spiked biological samples was from 76-80%. Spiked samples then were derivatized with 4'-bromophenacyl trifluoromethanesulfonate and the butyrobetaine 4'-bromophenacyl ester was isolated by high-performance liquid chromatography (HPLC). Radioactivity eluted in a single peak which co-chromatographed with authentic butyrobetaine 4'-bromophenacyl ester. Two identical liver specimens were treated according to this isolation procedure. Prior to derivatization, one specimen was treated with butyrobetaine hydroxylase. After derivatization, there was no butyrobetaine 4'-bromophenacyl ester peak in the specimen treated with butyrobetaine hydroxylase. The HPLC detection sensitivity to butyrobetaine 4'-bromophenacyl ester was 1 pmol injected with a signal-to-noise greater than 2:1.
Krahenbuhl
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Department of Veterans Affairs Medical Center, Cleveland, OH 44106.
Minkler
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Hoppel
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D000098:Acetophenones; D000818:Animals; D001622:Betaine / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000493:pharmacokinetics; D001711:Biotransformation; D002331:Carnitine / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D051381:Rats
D000098:Acetophenones; D007202:Indicators and Reagents; D001622:Betaine; C002889:gamma-butyrobetaine; C043597:4'-bromophenacyl triflate; D002331:Carnitine