NDLI: Rapid acetoacetate analysis on the LKB 8600 Reaction Rate Analyser.

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Rapid acetoacetate analysis on the LKB 8600 Reaction Rate Analyser.

Content Provider	Semantic Scholar
Author	Davidson, D. Fraser
Copyright Year	1978
Abstract	Automation of acetoacetate analysis has previously been by continuous flow methods using colorimetric techniques (Klein and Oklander, 1966; Dickie and Gibson, 1976). The most specific methods appear to be enzymatic using the fl-hydroxybutyrate dehydrogenase (fi HBDH, E.C.1.1.1.30)/NADH system (Williamson et al., 1962). These end-point methods, however, do not readily permit the analysis of large numbers of samples, and the incubation time of an enzymatic end-point assay can take as long as 50 minutes (Gibbard and Watkins, 1968). A kinetic technique would reduce this time considerably. The LKB 8600 Reaction Rate Analyser has many advantages including precise temperature control, ease of use, reliability (Smith et al., 1970), and accuracy (Davidson, 1976). The following method uses trichloroacetic acid as protein precipitant (100 g/l), a 50 ,IL sample volume, a reaction rate mode of measurement, and a one-minute measuring time. D(-)-+l-Hydroxybutyrate dehydrogenase catalyses the reaction: acetoacetate + NADH + H+ = D(-)P-hydroxybutyrate + NAD+. Acetoacetate is quantitated by the initial rate of decrease in absorbance of NADH at 340 nm.
Starting Page	83
Ending Page	99
Page Count	17
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://jcp.bmj.com/content/jclinpath/31/6/602.full.pdf
PubMed reference number	670415v1
Volume Number	31
Issue Number	6
Journal	Journal of clinical pathology
Language	English
Access Restriction	Open
Subject Keyword	3-Hydroxybutyrate Dehydrogenase Acetoacetates Catalysis Deficiency of glucose-6-phosphate dehydrogenase Kilogram per Cubic Meter Kinetics NADH Nicotinamide adenine dinucleotide (NAD) Trichloroacetic Acid
Content Type	Text
Resource Type	Article

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