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.
Highlights
Type 2 diabetes mellitus (T2DM) is a metabolic disorder that affects millions of individuals worldwide and it is usually associated with prevalent comorbidities such as hypertension, hyperlipidemia or obesity
The method consisted of a simple liquid–liquid extraction of 300 μL of serum with ethyl acetate
Validation procedures which were performed to evaluate the differences in the recoveries of spiked 2HB between water and serum showed no significant differences in the slope coefficient (α) of 3-point 2HB-spiked curves prepared in water or serum (Figure 1A)
Summary
Type 2 diabetes mellitus (T2DM) is a metabolic disorder that affects millions of individuals worldwide and it is usually associated with prevalent comorbidities such as hypertension, hyperlipidemia or obesity. Serious complications may develop as a result of T2DM, such as retinopathy, nephropathy or cardiovascular disease, among others [1]. A key mechanism related to T2DM is insulin cellular resistance, which leads to hyperglycemia [2]. Insulin resistance is described as a common mechanism for different associated comorbidities such as obesity and metabolic associated fatty liver disease [3,4]. T2DM is a chronic disease with different therapeutic options available, a great endocrinological challenge in recent years has been the identification of subjects who are at risk of it and the prevention of its early development. Prediabetes, which is defined by plasma glucose concentrations higher than normal but below the defined threshold of diabetes, is related to an increased risk of T2DM and cardiovascular disease [5]
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