Assay of fatty acid omega-hydroxylation using high-performance liquid chromatography with fluorescence labeling reagent, 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one (BrMB).
A sensitive nonradioisotopic method is reported for measuring microsomal lauric acid omega-hydroxylation activity. The assay is based upon separation and detection of 12-hydroxylauric acid formed by means of high-performance liquid chromatography following fluorescence labeling of the carboxyl group with 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one (BrMB). The use of 10-hydroxycapric acid as an internal standard affords the accurate and reproducible assay. The differential effect of dehydroepiandrosterone, a peroxisome proliferator, on the omega-hydroxylation activity in the liver and kidney of rats is also reported.
Yamada
J
J
Department of Clinical Biochemistry, Tokyo College of Pharmacy, Japan.
Sakuma
M
M
Suga
T
T
D000818:Animals; D048588:Benzoxazines; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003687:Dehydroepiandrosterone / Q000494:pharmacology; D005456:Fluorescent Dyes; D006880:Hydroxy Acids / Q000032:analysis* / Q000378:metabolism; D006900:Hydroxylation; D007672:Kidney Cortex / Q000187:drug effects / Q000201:enzymology*; D007850:Lauric Acids / Q000032:analysis* / Q000378:metabolism; D008297:Male; D008861:Microsomes / Q000187:drug effects / Q000201:enzymology*; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D010078:Oxazines; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values; D013050:Spectrometry, Fluorescence
D048588:Benzoxazines; D005456:Fluorescent Dyes; D006880:Hydroxy Acids; D007850:Lauric Acids; D010078:Oxazines; C030358:lauric acid; C073982:3-bromomethy-7-methoxy-1,4-benzoxazin-2-one; D003687:Dehydroepiandrosterone