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BHB is significant in the diagnosis and treatment of diabetic ketoacidosis and early diagnosis of diabetes. BHB is catalyzed by β-hydroxybutyrate dehydrogenase to form acetoacetate. NAD+ is reduced to NADH, which then reacts with WST-8 to form a yellow substance with an absorption peak at 450nm. The increase in this yellow substance at 450nm is measured to calculate the BHB content in the sample.