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- E-ISSN 2288-8985
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Determination of plasma ketone body following oximation-trimethylsilyl derivatization using gas chromatography-mass spectrometry selected ion monitoring
Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
2016, v.29 no.1, pp.49-55
https://doi.org/10.5806/AST.2016.29.1.49
(2016). Determination of plasma ketone body following oximation-trimethylsilyl derivatization using gas chromatography-mass spectrometry selected ion monitoring. Analytical Science and Technology, 29(1), 49-55, https://doi.org/10.5806/AST.2016.29.1.49
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Abstract
A ketone body (acetoacetic acid, β-hydroxybutyric acid, and acetone) increases from blood or urine when bio-energy dependence pays more fatty acid than glucose. However, in case oxidation of fat is greater than the capacity of the citric acid cycle the fatty acid oxidation is made from acetoacetyl CoA to acetoacetate then, again form β-hydroxyburytic acid to acetone, the diffusion take place into the blood. Enzymes that oxidize ketone body in the brain and nerve tissue blood ketone dody is increased during prolonged fasting, brain used it as energy. In this study, we developed the rapid two step derivatization method for sensitive detection of the ketone body by GC-MS/SIM. The plasma was deproteinized and then the hydroxy and carboxyl groups of ketone body are subjected to extraction and drying then, keto-group were derivatized with hydoxylamine at 60 oC for 30 min for oximation. Then it was trimetyl-silylated with BSTFA at 80oC for 30 min and analyzed using a GC-MS. The linear ranges were in between 0.001 μg/mL and 250 μg/mL for β-hydroxy butyrate, and acetoacetate. The method detection limits were below 0.1 pg over each target compound determined. The mean recoveries (%) of target compounds were ranged from 88.2 % to 92.3 % at 1 μg/mL, from 89.5 % to 94.8 % at 10 μg/mL, with RSD of 6.3-9.4 %. This method could be applied to quantification of ketone bodies which are seen in the keto-acidosis in children and adults from a variety of diseases that cause ketones in the blood and urine.
- keywords
- ketosis, ketoacidosis, β-hydroxybutyric acid, acetoacetic acid, oximation-trimethylsilylation, GC-MS, selected ion monitoring
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