Abstract：
Circulating levels of 2-hydroxybutyrate (2HB) are highly related to glycemic status in different metabolomic studies. According to recent evidence, 2HB is an early biomarker of the future development of dysglycemia and type 2 diabetes mellitus and may be causally related to the progression of normal subjects to impaired fasting glucose or insulin resistance. In the present study, we developed and validated a simple, specific and sensitive gas chromatography-mass spectrometry (GC-MS) method specifically intended to quantify serum levels of 2HB. Liquid-liquid extraction with ethyl acetate was followed by 2 min of microwave-assisted derivatization. The method presented acceptable accuracy, precision and recovery, and the limit of quantification was 5 µM. Levels of 2HB were found to be stable in serum after three freeze-thaw cycles, and at ambient temperature and at a temperature of 4 °C for up to 24 h. Extracts derivatized under microwave irradiation were stable for up to 96 h. No differences were found in 2HB concentrations measured in serum or plasma EDTA samples. In summary, the method is useful for a rapid, precise and accurate quantification of 2HB in serum samples assessed for the evaluation of dysglycemia and diabetes mellitus.
摘要：
在不同的代谢组学研究中，2-羟基丁酸（2HB）的循环水平与血糖状态高度相关。根据最近的证据,2HB是血糖异常和2型糖尿病未来发展的早期生物标志物，可能与正常受试者空腹血糖受损或胰岛素抵抗的进展有因果关系。在本研究中,我们开发并验证了一个简单的，特异性和灵敏的气相色谱-质谱（GC-MS）方法专门用于定量2HB的血清水平。用乙酸乙酯液-液萃取，然后进行2分钟的微波辅助衍生化。该方法具有可接受的准确性，精度和回收率，定量限为5µM。在三个冻融循环后，血清中的2HB水平稳定，在环境温度和4°C的温度下长达24小时。在微波辐射下衍生的提取物稳定长达96小时。在血清或血浆EDTA样品中测得的2HB浓度没有差异。总之,该方法是有用的快速，血清样本中2HB的精确和准确定量评估，以评估血糖异常和糖尿病。
