Measurement of ketone bodies in subcellular fractions using a spectrophotometric iron-chelate assay |
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| Authors: | RI Kientsch-Engel EA Siess OH Wieland |
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| Institution: | 1. Forschergruppe Diabetes des Städtischen Krankenhauses München-Schwabing, Kölner Platz 1, D-8000 Munich 40, Federal Republic of Germany;2. Institut für Klinische Chemie des Städtischen Krankenhauses München-Schwabing, Kölner Platz 1, D-8000 Munich 40, Federal Republic of Germany |
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| Abstract: | A sensitive spectrophotometric assay for 3-hydroxybutyrate determination in biological samples is described. Linearity between the amount of 3-hydroxybutyrate and ΔA546 was obtained in the range of 0.3 to 4.0 nmol 3-hydroxybutyrate/assay. The same method is applicable for acetoacetate determination after its enzymatic reduction. The assay proved to be useful for the study of the subcellular distribution of ketone bodies in isolated liver cells. The assay procedure is adequate to measure the concentration of ketone bodies in 5-mg and 20μl samples from liver and blood, respectively. |
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| Keywords: | 3-HOB 3-hydroxybutyrate AcAc acetoacetate BPS bathophenanthroline sulfonate 3-HOB-DH 3-hydroxybutyrate dehydrogenase (EC 1 1 1 30) |
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