Determination of β-Hydroxybutyrate in Blood and Urine Using Gas Chromatography-Mass Spectrometry
Determination of β-Hydroxybutyrate in Blood and Urine Using Gas Chromatography-Mass Spectrometry
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DOI：
10.1093/jat/33.8.502
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发表时间：
2009-10-01
影响因子：
2.5
通讯作者：
Cooper, Gail A. A.
中科院分区：
文献类型：
--
作者：
Hassan, Huda M. A.;Cooper, Gail A. A.
β-羟丁酸被认为是酒精性酮症酸中毒的潜在生物标志物。建立了一种可靠、灵敏的测定死后血和尿中牛血红蛋白的方法，并以γ-羟基丁酸酯为内标进行了验证。样品经液-液萃取、硅基衍生化后，用气相色谱-质谱仪进行分析。血样和尿样的检出限和定量下限分别为2和7 mg/L和2和6 mg/L。日内精密度和日间精密度用血和尿的变异系数进行测量，质量控制样品在50 mg/L和300 mg/L范围内的日间精密度和日间精密度在1.0~12.4%之间，线性范围为50~500 mg/L，平均相关系数为0.99，血和尿中的平均提取回收率分别为≥82%和≥59%。在冰箱(2-5°C)中保存15天后，血液中添加300毫克/L的BHb保持稳定。对于死者有慢性酗酒史的病例，采用经过验证的方法对死者的死后血液和尿液样本进行分析，以确定BHb作为AKA的潜在标志物的用途。
β-Hydroxybutyrate (BHB) is considered a potential biomarker for alcoholic ketoacidosis (AKA). A robust and sensitive method was developed and validated for the quantitative determination of BHB in postmortem blood and urine using deuterated γ-hydroxybutyrate as an internal standard. Samples were analyzed by gas chromatography-mass spectrometry following liquid-liquid extraction and silyl derivatization. The limits of detection and lower limits of quantification in blood and urine were 2 and 7 mg/L and 2 and 6 mg/L, respectively. The interday and intraday precision was measured by coefficients of variation for blood and urine and ranged from 1.0 to 12.4% for quality control samples spiked at 50 and 300 mg/L. The linear range of 50–500 mg/L resulted in an average correlation ofR2> 0.99, and the average extraction recoveries in blood and urine were ≥ 82% and ≥ 59%, respectively. BHB remains stable in blood spiked at a concentration of 300 mg/L for 15 days when stored within a refrigerator (2–5°C). Postmortem blood and urine samples were analyzed using the validated method for cases where the deceased had a history of chronic alcohol abuse to establish the use of BHB as a potential marker of AKA.