Enzymatic detection of α-hydroxybutyrate, an important marker of insulin resistance, and comparison with LC-MS/MS detection

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY
Beate Steiner , Christian Leitner , David Stadler , Eva-Maria Prugger , Christoph Magnes , Peter L. Herzog
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引用次数: 0

Abstract

Aim

The metabolite α-hydroxybutyrate (α-HB) is an important marker of insulin resistance and impaired glucose tolerance allowing to identify patients at risk of developing diabetes and related metabolic disorders before any symptoms become apparent. At present, its exact quantification requires mass spectrometry (LC-MS), which is not compatible with routine laboratory use. Accordingly, a simple enzymatic-based method was assessed and its applicability and measuring accuracy compared with LC-MS was investigated.

Methods

Standards, serum, and plasma samples containing α-HB were prepared with routine procedures and their α-HB contents measured with the XpressGT® enzymatic test kit photometrically or with LC-MS and multiple reaction monitoring.

Results

α-HB detection with XpressGT® yielded highly linear calibration curves and 102 % recovery of stocks added to commercial samples. Stability of the analyte in serum and plasma samples prepared with various anti-coagulants was >90 % after 46 h for several widely used preparations and recovery after 3 freeze-thaw cycles was ≥95 % with these anti-coagulants. A direct comparison of 75 samples indicated very good agreement of α-HB levels determined by both methods, 86 % of XpressGT® samples being within ±20 % of LC-MS values and even 93 % within ±20 % considering only samples above 30 μM concentration.

Conclusion

XpressGT®-based detection of α-HB is an easily applicable method which can be used for accurate and reliable quantification of the metabolite in clinical practice. Routine α-HB determination in patients at risk of developing diabetes would allow early establishment of preventive measures or pharmacological intervention reducing the risk for the onset of serious diabetes-related health problems.

酶法检测胰岛素抵抗的重要标志物--α-羟丁酸,并与 LC-MS/MS 检测进行比较
目的代谢物α-羟丁酸(α-HB)是胰岛素抵抗和葡萄糖耐量受损的重要标志物,可在症状明显之前识别出有患糖尿病和相关代谢紊乱风险的患者。目前,α-HB 的精确定量需要使用质谱法(LC-MS),这不符合实验室的常规使用要求。方法用常规程序制备含 α-HB 的标准、血清和血浆样品,用 XpressGT® 酶法检测试剂盒光度法或 LC-MS 和多反应监测法测定其 α-HB 含量。在使用各种抗凝剂制备的血清和血浆样品中,分析物在几种广泛使用的制剂中的稳定性在 46 小时后达到 90%,在使用这些抗凝剂进行 3 次冻融循环后的回收率≥95%。对 75 份样本进行的直接比较表明,两种方法测定的 α-HB 含量非常一致,86% 的 XpressGT® 样品在 LC-MS 值的±20%范围内,甚至 93% 在±20%范围内(仅考虑浓度高于 30 μM 的样本)。对有患糖尿病风险的患者进行常规的 α-HB 检测,可以及早制定预防措施或进行药物干预,降低出现严重糖尿病相关健康问题的风险。
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来源期刊
Practical Laboratory Medicine
Practical Laboratory Medicine Health Professions-Radiological and Ultrasound Technology
CiteScore
3.50
自引率
0.00%
发文量
40
审稿时长
7 weeks
期刊介绍: Practical Laboratory Medicine is a high-quality, peer-reviewed, international open-access journal publishing original research, new methods and critical evaluations, case reports and short papers in the fields of clinical chemistry and laboratory medicine. The objective of the journal is to provide practical information of immediate relevance to workers in clinical laboratories. The primary scope of the journal covers clinical chemistry, hematology, molecular biology and genetics relevant to laboratory medicine, microbiology, immunology, therapeutic drug monitoring and toxicology, laboratory management and informatics. We welcome papers which describe critical evaluations of biomarkers and their role in the diagnosis and treatment of clinically significant disease, validation of commercial and in-house IVD methods, method comparisons, interference reports, the development of new reagents and reference materials, reference range studies and regulatory compliance reports. Manuscripts describing the development of new methods applicable to laboratory medicine (including point-of-care testing) are particularly encouraged, even if preliminary or small scale.
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