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| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Serum, plasma, urine etc |
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Overview
For quantitative determination of ketone body (acetoacetate and 3-hydroxybutyrate) and evaluation of drug effects on its metabolism. The assay uses OD340nm for signal readout. Compatible sample input includes Serum, plasma, urine etc. Typical stated assay timing is 30 min.
Key elements and design rationale
- Readout format: OD340nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Serum, plasma, urine etc, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.12 mM for interpreting low-signal samples.
- Feature emphasis: Sensitive and accurate. Uses a 10 µL sample. Linear detection range of 0.12 to 8 mM for each ketone body in 96-well plate assay.
Additional feature notes highlight Convenient. The procedure involves adding a single working reagent and reading the optical density at room temperature; High-throughput. Can be automated as a high-throughput 96-well plate assay for many samples per day. Available format information for this listing includes 200 Tests.
Biological background
This product is centered on measurement of ketone body within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
Ketone bodies (acetoacetic acid and 3-hydroxybutyric acid) are produced in the liver mainly from oxidation of fatty acids and are normally present at low concentrations in urine and blood. Increased ketone concentrations in the blood may lead to metabolic acidosis, which has been associated with diabetes, childhood hypo-glycemia, growth hormone deficiency, alcohol or salicylate intoxication, and inborn errors of metabolism. Simple, direct, and automation-ready procedures for measuring acetoacetic acid (AcAc) and 3-hydroxybutyric acid (BOH) are very desirable. BioAssay Systems EnzyChrom™ ketone body assay is based on 3-hydroxybutyrate dehydrogenase catalyzed reactions, in which the change in NADH absorbance, measured at 340nm, is directly related to the AcAc and BOH concentrations,
Detection method
Colorimetric (OD 340 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.12 mM.
Procedures and timing
Stated procedure or timing information: 30 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify ketone body in serum, plasma, urine by OD340 nm readout.
- Compare treatment or phenotype groups using matched serum, plasma, urine handling.
- Monitor time-course or pre/post changes in serum, plasma, urine across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
Can serum and plasma samples be stored frozen?
β-hydroxybutyrate is stable and can be detected in samples that had been stored frozen for more than a month. Acetoacetate (AcAc), on the other hand, is an unstable compound which becomes rapidly decarboxylated. We recommend to use fresh serum or freeze serum samples at -80°C for up to one month.
I seem to be getting very low levels of BOH and AcAc in freshly drawn blood. Perhaps this is because they are breaking down within the hour it takes me to process the plasma out.
In healthy individuals, normal blood values for ketone bodies are actually very low or not detectable. Elevated blood levels of ketone bodies (ketosis) can be caused for example by diabetes, alcohol intoxication, starvation, or physical exercise. You can check for the stability of your analyte by setting aside an aliquot of your sample and add the standard as a positive control.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Vulnerability of the endangered Maugean Skate population to degraded environmental conditions in Macquarie Harbour
Moreno, D et al. (2020). Vulnerability of the endangered Maugean Skate population to degraded environmental conditions in Macquarie Harbour. Assay: Ketone body in maugean skate plasma.
Effects of dietary anaplerotic and ketogenic energy sources on renal fatty acid oxidation induced by clofibrate in suckling neonatal pigs
Lin, X et al. (2020). Effects of dietary anaplerotic and ketogenic energy sources on renal fatty acid oxidation induced by clofibrate in suckling neonatal pigs. International Journal of Molecular Sciences, 21(3), 726. Assay: Ketone body in porcine kidney.
Suppression of enteroendocrine cell glucagon-like peptide (Glp)-1 release by fat-induced small intestinal ketogenesis: A mechanism targeted by Roux-en-Y gastric bypass surgery but not by preoperative very-low-calorie diet
Wallenius, V et al. (2020). Suppression of enteroendocrine cell glucagon-like peptide (Glp)-1 release by fat-induced small intestinal ketogenesis: A mechanism targeted by Roux-en-Y gastric bypass surgery but not by preoperative very-low-calorie diet. Gut, 69(8), 1423-1431. Assay: Ketone body in human serum.
Edible dairy formula fortified with coconut oil for neuroprotection against aluminium chloride-induced Alzheimers disease in rats
Khalil, HMA et al. (2020). Edible dairy formula fortified with coconut oil for neuroprotection against aluminium chloride-induced Alzheimers disease in rats. Journal of Functional Foods, 104296. Assay: Ketone body in rat serum.
Reduced fatty acid uptake aggravates cardiac contractile dysfunction in streptozotocin-induced diabetic cardiomyopathy
Umbarawan, Y et al. (2020). Reduced fatty acid uptake aggravates cardiac contractile dysfunction in streptozotocin-induced diabetic cardiomyopathy. Scientific Reports, 10(1), 20809. Assay: Ketone body in mouse serum.
Variation of body condition and plasma energy substrates with life stage, sex, and season in wild-sampled nurse sharks Ginglymostoma cirratum
Moorhead, SG et al. (2020). Variation of body condition and plasma energy substrates with life stage, sex, and season in wild-sampled nurse sharks Ginglymostoma cirratum. Journal of Fish Biology. Assay: Ketone body in shark plasma.
Dexamethasone-induced Kruppel-like factor 9 expression promotes hepatic gluconeogenesis and hyperglycemia
Cui, A., Fan, H., Zhang, Y., Zhang, Y., Niu, D., Liu, S. & Liu, Y. (2019). Dexamethasone-induced Kruppel-like factor 9 expression promotes hepatic gluconeogenesis and hyperglycemia. The Journal of clinical investigation 130:2266-2278. Assay: Ketone body in mice serum.
Autophagy regulates lipid metabolism through selective turnover of NCoR1
Saito, T., Kuma, A., Sugiura, Y., Ichimura, Y., Obata, M., Kitamura, H. & Saito, I. (2019). Autophagy regulates lipid metabolism through selective turnover of NCoR1. Nature communications 10(1):1567. Assay: Ketone body in mice plasma.
Infusion of butyrate affects plasma glucose, butyrate, and beta-hydroxybutyrate but not plasma insulin in lactating dairy cows
Herrick, K. J., Hippen, A. R., Kalscheur, K. F., Schingoethe, D. J., Ranathunga, S. D., Anderson, J. L. & Van Eys, J. E. (2018). Infusion of butyrate affects plasma glucose, butyrate, and beta-hydroxybutyrate but not plasma insulin in lactating dairy cows. Journal of dairy science, 101(4), 3524-3536. Assay: Ketone body in cow plasma.
CD36 is indispensable for nutrient homeostasis and endurance exercise capacity during prolonged fasting
Iso, T., Haruyama, H., Sunaga, H., Matsui, H., Matsui, M., Tanaka, R. & Hanaoka, H. (2018). CD36 is indispensable for nutrient homeostasis and endurance exercise capacity during prolonged fasting. Physiological reports, 6(19), e13884. Assay: Ketone body in mice serum.
Physiological consequences of environmental contamination in an elasmobranch with matrotrophic histotrophy, the Round Stingray (Urobatis halleri)
Lyons, K. (2018). Physiological consequences of environmental contamination in an elasmobranch with matrotrophic histotrophy, the Round Stingray (Urobatis halleri). Assay: Ketone body in stingrays plasma.
Treatment of NASH with gemcabene
Bisgaier, Charles L., and Daniela Carmen Oniciu (2017). Treatment of NASH with gemcabene. U.S. Patent No. 9,849,104 Assay: Ketone body in mice plasma.
Premorbid obesity, but not nutrition, prevents critical illness-induced muscle wasting and weakness
Goossens, C., Marques, M. B., Derde, S., Vander Perre, S., Dufour, T., Thiessen, S. E. & De Bock, K. (2017). Premorbid obesity, but not nutrition, prevents critical illness-induced muscle wasting and weakness. Journal of cachexia, sarcopenia and muscle, 8(1), 89-101. Assay: Ketone body in mice serum.
Single-dose infusion of sodium butyrate, but not lactose, increases plasma beta-hydroxybutyrate and insulin in lactating dairy cows
Herrick, K. J., Hippen, A. R., Kalscheur, K. F., Schingoethe, D. J., Casper, D. P., Moreland, S. C., & Van Eys, J. E. (2017). Single-dose infusion of sodium butyrate, but not lactose, increases plasma beta-hydroxybutyrate and insulin in lactating dairy cows. Journal of dairy science, 100(1), 757-768. Assay: Ketone body in cow plasma.
Metformin does not prevent glucose intolerance but improves renal function and reduces oxidative stress in type 1 diabetes (Doctoral dissertation)
Driver, C. (2016). Metformin does not prevent glucose intolerance but improves renal function and reduces oxidative stress in type 1 diabetes (Doctoral dissertation). Assay: Ketone body in rat serum and urine.
Grapefruit derived flavonoid naringin improves ketoacidosis and lipid peroxidation in type 1 diabetes rat model
Murunga, A. N., Miruka, D. O., Driver, C., Nkomo, F. S., Cobongela, S. Z., & Owira, P. M. (2016). Grapefruit derived flavonoid naringin improves ketoacidosis and lipid peroxidation in type 1 diabetes rat model. PloS one, 11(4), e0153241. Assay: Ketone body in rat serum and urine.
The effects of naringin on glucose tolerance and ketoacidosis in type 1 diabetes (Doctoral dissertation)
Neondo, M. A. (2016). The effects of naringin on glucose tolerance and ketoacidosis in type 1 diabetes (Doctoral dissertation). Assay: Ketone body in rat serum and urine.
Fatty Acid Binding Protein 4 and 5 Play a Crucial Role in Thermogenesis under the Conditions of Fasting and Cold Stress
Syamsunarno, MRAA et al (2014). Fatty Acid Binding Protein 4 and 5 Play a Crucial Role in Thermogenesis under the Conditions of Fasting and Cold Stress. PloS one 9.3: e90825. Assay: Ketone body in mouse serum.
Inflammation and Resolution Are Associated with Upregulation of Fatty Acid beta-Oxidation in Zymosan-Induced Peritonitis
Fujieda, Y et al (2013). Inflammation and Resolution Are Associated with Upregulation of Fatty Acid beta-Oxidation in Zymosan-Induced Peritonitis. PloS one 8.6: e66270. Assay: Ketone body in mouse liver tissue.
Increased susceptibility of natural killer T-cell-deficient mice to acetaminophen-induced liver injury
Martin-Murphy, BV et al (2013). Increased susceptibility of natural killer T-cell-deficient mice to acetaminophen-induced liver injury. Hepatology 57.4: 1575-1584. Assay: Ketone body in mouse serum.
A Critical Role of Fatty Acid Binding Protein 4 and 5 (FABP4/5) in the Systemic Response to Fasting
Syamsunarno, MRAA et al (2013). A Critical Role of Fatty Acid Binding Protein 4 and 5 (FABP4/5) in the Systemic Response to Fasting. PloS one 8.11: e79386. Assay: Ketone body in mouse serum.
Comparison of the metabolic profiling of hepatitis B virus-infected cirrhosis and alcoholic cirrhosis patients by using (1) H NMR-based metabonomics
Qi S et al (2012). Comparison of the metabolic profiling of hepatitis B virus-infected cirrhosis and alcoholic cirrhosis patients by using (1) H NMR-based metabonomics. Hepatol Res 42(7):677-85. Assay: Ketone body in human serum.
Klusonova P et al (2011) Local metabolism of glucocorticoids in Prague hereditary hypertriglyceridemic rats–effect of hy
Klusonova P et al (2011) Local metabolism of glucocorticoids in Prague hereditary hypertriglyceridemic rats–effect of hypertriglyceridemia and gender. Steroids 76(12):1252-9. Assay: Ketone Body in rat Hepatocyte culture.
Metabolic profiling based quantitative evaluation of hepatocellular metabolism in presence of adipocyte derived extracellular matrix
Sharma NS et al (2011). Metabolic profiling based quantitative evaluation of hepatocellular metabolism in presence of adipocyte derived extracellular matrix. PLoS One 6(5):e20137. Assay: Ketone body in rat hepatocyte culture.
Metabolic profiling based quantitative evaluation of hepatocellular metabolism in presence of adipocyte derived extracellular matrix
Sharma NS et al (2011). Metabolic profiling based quantitative evaluation of hepatocellular metabolism in presence of adipocyte derived extracellular matrix. PLoS One 6(5):e20137. Assay: Ketone Body in human serum.
