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Department of Animal Health, Welfare and Nutrition, The Danish Institute of Agricultural Sciences, P.O. Box 50, DK-8830 Tjele, Denmark
Corresponding author: Torben Larsen; e-mail: Torben.Larsen{at}agrsci.dk.
Determination of ß-hydroxybutyrate (BHBA) in blood and milk samples is an important tool in the diagnosis of ketosis in dairy cattle. Apart from semiquantitative cow-side tests, well-established laboratory methods exist for measurements in blood serum or plasma. These spectrophotometric methods are, however, neither convenient nor reliable when transferred to analyses of milk. Due to its nontransparent nature, milk needs extensive pretreatment if traditional analyses are to be used. This paper describes a fluorometric determination of BHBA that is useful without pretreatment in opaque matrices such as milk and in blood plasma. The method is easy to automate, saves labor expenses, and is inexpensive. The analytical accuracy and precision are reliable for intensive as well as large-scale analysis; for example, in-line sampling from automatic milking systems. Analysis of 2500 random milk samples showed a BHBA content ranging from 10 to 631 µM (mean 49 µM). Furthermore, selected samples (n = 295) from diagnosed ketotic animals taken on d –35 to +35 from peak level ranged from 10 to 684 µM (median 79 µM, mean 141 µM). Using the same 1240 blood plasma samples, the fluorometric method was closely correlated with a traditional spectrophotometric method (r = 0.987). Hemolysis of samples does not appear to affect the fluorometric determination of BHBA.
Key Words: ß-hydroxybutyrate • ketosis • milk • blood
Abbreviation key: H1 and H2 = high internal BHBA controls, L1 and L2 = low internal BHBA controls.
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