Presentation Title

Quantification of Ketone Bodies in Saliva using Gas Chromatography-Mass Spectrometry

Format of Presentation

Poster to be presented the Friday of the conference

Presenter Information

Austin PietramalaFollow

Abstract

Type 1 diabetes mellitus is a growing epidemic in today’s world. A complication associated with this disease is diabetic ketoacidosis, and if left untreated it can result in a coma or death. There are commercial methods to analyze ketone concentrations through urine and blood, but both have their respective flaws. Saliva consists mostly of water, but also contains hormones and other metabolites. Using saliva as a diagnostic model is slowly gaining popularity, but there is very little research on the quantification of ketone bodies in saliva. Therefore, the experiment looks to quantify the amount of acetoacetate and beta-hydroxybutyrate in saliva, urine, and blood in participants who are in ketosis, using gas chromatography-mass spectrometry. Participants were recruited, and samples of their blood, urine, and saliva were received before and after completing a 4-day ketogenic diet. All samples were acidified, and the matrices were evaporated. The samples were spiked with both ketone bodies, derivatized using BSTFA + 1% TMCS at 80°C for 30 minutes, and the headspace was analyzed. The expected results include blood and urine having the highest concentrations of beta-hydroxybutyrate and acetoacetate, respectively, and saliva having more beta-hydroxybutyrate than acetoacetate, but less than the amount seen in blood. This is because saliva and plasma are isotonic. Future work will look at how the concentration of ketone bodies changes over time to see how the ketone bodies are distributed across the body.

Department

Chemistry

Faculty Advisor

Kingsley Donkor

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Quantification of Ketone Bodies in Saliva using Gas Chromatography-Mass Spectrometry

Type 1 diabetes mellitus is a growing epidemic in today’s world. A complication associated with this disease is diabetic ketoacidosis, and if left untreated it can result in a coma or death. There are commercial methods to analyze ketone concentrations through urine and blood, but both have their respective flaws. Saliva consists mostly of water, but also contains hormones and other metabolites. Using saliva as a diagnostic model is slowly gaining popularity, but there is very little research on the quantification of ketone bodies in saliva. Therefore, the experiment looks to quantify the amount of acetoacetate and beta-hydroxybutyrate in saliva, urine, and blood in participants who are in ketosis, using gas chromatography-mass spectrometry. Participants were recruited, and samples of their blood, urine, and saliva were received before and after completing a 4-day ketogenic diet. All samples were acidified, and the matrices were evaporated. The samples were spiked with both ketone bodies, derivatized using BSTFA + 1% TMCS at 80°C for 30 minutes, and the headspace was analyzed. The expected results include blood and urine having the highest concentrations of beta-hydroxybutyrate and acetoacetate, respectively, and saliva having more beta-hydroxybutyrate than acetoacetate, but less than the amount seen in blood. This is because saliva and plasma are isotonic. Future work will look at how the concentration of ketone bodies changes over time to see how the ketone bodies are distributed across the body.