Disposable
test strips and a
wet chemistry method for measuring each of beta-hydroxybutyrate alone, combined beta-hydroxybutyrate and acetoacetate or total
ketone bodies (i.e., beta-hydroxybutyrate, acetoacetate and
acetone) in human bodily fluid samples, including but not limited to
urine,
saliva or sweat are described. The
test strips need only be dipped in the sample and can be used by anyone in almost any milieu. Measurement can be made electrochemically, spectrophotometrically, fluorometrically or by comparision to a color standard. Combined acetoacetate and beta-hydroxybutyrate which account for 97-98% of total
ketone bodies and may be measured in a cyclic reaction that occurs at pH about 7.0 to about 8.3 with beta-
hydroxybutyrate dehydrogenase, (beta-HBD),
nicotinamide adenine dinucleotide, a tetrazolium dye precursor and an
electron mediator. Using this reaction, false positive results obtained from
urine samples taken from patients on sulfhydryl drugs are avoided. beta-HBD from some sources was found to cause false negative results in samples (e.g.
urine) containing high
chloride content due to
chloride inhibition of beta-HBD. Using a simple test for
chloride inhibition, it was found that beta-HBD from
Alcaligenes is not so inhibited. Using either beta-HBD that is not inhibited by chloride or using 10-20 times the
normal concentration of this
enzyme eliminates false negatives in samples having substantial chloride content, such as urine, both in the reaction described above and in other reactions disclosed for measuring each of beta-hydroxybutyrate alone, combined beta-hydroxybutyrate and acetoacetate and total
ketone bodies, all of which reactions occur in the
pH range of about 8.6 to about 9.5.