Service of Biochemistry and Molecular Genetics, Hospital Clinic Universitari, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain.
Department of Biomedicine, University of Barcelona, 08905 Barcelona, Spain.
Molecules. 2022 Mar 15;27(6):1889. doi: 10.3390/molecules27061889.
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 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 .
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 是未来发生糖代谢异常和 2 型糖尿病的早期生物标志物，并且可能与正常个体向空腹血糖受损或胰岛素抵抗的进展有关。在本研究中，我们开发并验证了一种简单、特异和灵敏的气相色谱-质谱（GC-MS）方法，旨在专门定量血清 2HB 水平。采用乙酸乙酯进行液-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度、精密度和回收率，定量限为 5 µM。经过三次冻融循环、室温下和 4°C 下长达 24 小时，血清中 2HB 水平稳定。在微波辐射下衍生的提取物稳定长达 96 小时。在 EDTA 血清或血浆样本中测量的 2HB 浓度无差异。总之，该方法可用于快速、精确和准确地定量评估糖代谢异常和 的血清样本中的 2HB。
