Development of stable film test for rapid estimation of blood or plasma 3-hydroxybutyrate.
A film test for the rapid detection of plasma/serum 3-hydroxybutyrate (3-OHB) has been developed. The film contains NAD, nitro blue tetrazolium, 3-OHB dehydrogenase, and diaphorase, and the surface is coated with modified biomembrane and can detect 50-1500 microM 3-OHB within 2-3 min. One drop or 50 microliters of plasma/serum or blood is applied to the film, and the violet color is read via reflectance meter after 2 min. Plasma/serum samples greater than 1500 microM 3-OHB can be measured by dilution with saline. In blood with 40% hematocrit, the color developed is 50% less than with plasma/serum, and this was adjusted in the reflectance meter. A good correlation (r = 0.99) was observed between results with automated and film methods and between visual methods and reflectance meter. In insulin-dependent diabetes mellitus, all 3 subjects with positive ketonuria (+ +), 8 of 12 subjects with mild ketonuria (+), and 7 of 25 subjects without ketonuria exhibited elevation of 3-OHB in blood greater than 200 microM. The results indicate that 3-OHB film is valuable not only in the emergency room for the differential diagnosis between ketoacidotic and nonketotic hypersomolar coma but also as a marker for insulin dependency, energy dependency on fatty acid compared with glucose, and metabolic control of diabetes.
Harano
Y
Y
Third Department of Medicine, Shiga University of Medical Science, Japan.
Aoki
T
T
Nakajima
Y
Y
Kojima
H
H
Kosugi
K
K
Harano
A
A
Hidaka
H
H
Yamada
S
S
Uno
S
S
Shigeta
Y
Y
D020155:3-Hydroxybutyric Acid; D000293:Adolescent; D001322:Autoanalysis; D002648:Child; D004355:Drug Stability; D006801:Humans; D006885:Hydroxybutyrates / Q000097:blood*; D008832:Microchemistry; D011934:Reagent Strips
D006885:Hydroxybutyrates; D011934:Reagent Strips; D020155:3-Hydroxybutyric Acid