Determination of plasma ketone body following oximation-trimethylsily| derivatization using gas chromatography-mass spectrometry selected ion monitoring
혈장 중 케톤체의 옥심-TMS 유도체화 후 GC-MS/SIM을 이용한 분석

Hye-Ran Yoon
2016 Analytical Science and Technology  
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
more » ... ased 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 o C for 30 min for oximation. Then it was trimetyl-silylated with BSTFA at 80 o C 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. 에서 30 분간 반응시켜 oximation 후 BSTFA를 가한 후 trimethylsilylation 유도체화하여 혈장 중에서 케 톤체를 신속하게 정량할 수 있는 GC-MS/SIM 분석법을 개발하였다. 케톤체의 직선성의 범위는 0.001-250 µg/mL 이었고, 혈장에서의 검출한계는 0.1 pg 이었다. 직선성을 가지는 범위에서의 상관계수(R2)는 0.998-0.999이었고, 회수율은 1 µg/mL의 표준품을 첨가하였을 때 88.2-92.3 %, 10 µg/mL의 농도를 첨가 하였을 때 89.5-94.8 % 였으며 RSD는 6.3-9.4 %였다. 이 분석법을 정상인과 케토시스 환자의 검체에 적 용하여 벨리데이션 하였으며, 본 분석방법은 어린이나 성인의 당뇨성 케토시스나 여러 유전성대사질환 환자 중 케토시스를 보이는 환자의 혈장 중 β-hydroxybutyric acid/acetoacetic acid의 비를 계산하여 케톤 체를 신속하고 효율적으로 임상검체 분석에 응용할 수 있음을 보여주었다.
doi:10.5806/ast.2016.29.1.49 fatcat:ujy3323os5cn3j64faamzquwcu