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Lee, Jun, Lee, Han, Song, Park, and Song: Evaluation of the T-KB-H and 3-HB Kits for the Measurement of Serum Ketone and β-Hydroxybutyric Acid



Diabetes mellitus and alcohol consumption are the most common causes of ketoacidosis in adults. Recently, β-hydroxybutyric acid (βHBA) was reported to be a potential serum biomarker in the diagnosis and monitoring of ketoacidosis. We evaluated the performance of T-KB-H and 3-HB kits for the measurement of ketone bodies [acetoacetate (AcAc)+βHBA] and βHBA, respectively.


Quantitative enzymatic assays were performed using the T-KB-H and 3-HB kits (Nittobo Medical Co., Japan) and the Architect ci16200 Integrated System (Abbott Laboratories, USA). Simultaneously, the ketone body levels in these serum samples were determined by gas chromatography-mas spectrometry (GC-MS). We evaluated precision and linearity of these kits and correlation with GC-MS, and established reference intervals in children and adults.


The coefficients of variation for the T-KB-H and 3-HB kits were less than 4.0% at analyte levels of 50, 100, and 400 µmol/L. Linearity was observed for AcAc and βHBA over a 0-1,000 µmol/L range (R2<0.99). Results from the T-KB-H and 3-HB kits were in good agreement with those from the GC-MS analysis, with correlation coefficients of 0.94 for AcAc and 0.96 for βHBA. Reference intervals determined for the T-KB-H kit were 9.8-270.1 µmol/L and 18.5-531.8 µmol/L in children and adults, respectively. For the 3-HB kit, the reference intervals were 6.4-234.0 µmol/L and 16.0-437.2 µmol/L in children and adults, respectively.


The T-KB-H and 3-HB kits displayed good precision, clinically acceptable linearity, and reliable correlation with an established assay. This indicates that the kits can be used clinically for measuring serum ketone bodies.

Figures and Tables

Fig. 1
Linearity of concentrations measured by the T-KB-H kit. (A) β-hydroxybutyric acid, (B) acetoacetate.
Fig. 2
Linearity of concentrations measured by the 3-HB kit for β-hydroxybutyric acid.
Fig. 3
Comparison of acetoacetate (AcAc) and β-hydroxybutyric acid (βHBA) measurements obtained with the T-KB-H kit, the 3-HB kit, and gas chromatography-mass spectrometry (GC-MS). (A) and (C), Scatter plots with ordinary linear fit with the Passing-Bablock regression analysis for AcAc and βHBA. (B) and (D), Bland-Altman plots for AcAc and βHBA.
Table 1
Precision of the T-KB-H and 3-HB kits for total ketone and β-hydroxybutyric acid assays

Abbreviation: CV, coefficient of variation.


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