Determination of tetrahydrobiopterin in murine locus coeruleus by HPLC with fluorescence detection.
Tetrahydrobiopterin in the murine locus coeruleus was measured as its fully oxidized form, biopterin, using a HPLC coupled to a fluorescence detector, because tetrahydrobiopterin itself cannot be detected by such means. The differential oxidization method distinguished tetrahydrobiopterin-derived biopterin and dihydrobiopterin-derived biopterin. The protocol reported here is a rapid and sensitive method that facilitates the measurement of tissue and/or cellular tetrahydrobiopterin. Using this assay protocol, we were able to detect and quantify variations in the tetrahydrobiopterin content in the murine locus coeruleus.
Kaneko
Y S
YS
Department of Physiology, Fujita Health University School of Medicine, Toyoake, 470-1192, Aichi, Japan.
Mori
K
K
Nakashima
A
A
Nagatsu
I
I
Nagatsu
T
T
Ota
A
A
D000818:Animals; D001708:Biopterins / Q000031:analogs & derivatives* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005453:Fluorescence; D008125:Locus Coeruleus / Q000378:metabolism*; D051379:Mice; D008809:Mice, Inbred C3H; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D013997:Time Factors
D001708:Biopterins; C017226:7,8-dihydrobiopterin; C003402:sapropterin