Scheijen Jean L J M, Hanssen Nordin M J, van de Waarenburg Marjo P H, Jonkers Daisy M A E, Stehouwer Coen D A, Schalkwijk Casper G
Laboratory for Metabolism and Vascular Medicine, Department of Internal Medicine, Maastricht University, P. Debeyelaan 25, P.O. Box 5800, 6202 AZ Maastricht, The Netherlands.
Exp Diabetes Res. 2012;2012:234812. doi: 10.1155/2012/234812. Epub 2012 Mar 8.
Plasma and urinary levels of D-lactate have been linked to the presence of diabetes. Previously developed techniques have shown several limitations to further evaluate D-lactate as a biomarker for this condition.
D- and L-lactate were quantified using ultraperformance liquid chromatography tandem mass spectrometry with labelled internal standard. Samples were derivatized with diacetyl-L-tartaric anhydride and separated on a C(18)-reversed phase column. D- and L-lactate were analysed in plasma and urine of controls, patients with inflammatory bowel disease (IBD), and patients with type 2 diabetes (T2DM).
Quantitative analysis of D- and L-lactate was achieved successfully. Calibration curves were linear (r(2) > 0.99) over the physiological and pathophysiological ranges. Recoveries for urine and plasma were between 96% and 113%. Inter- and intra-assay variations were between 2% and 9%. The limits of detection of D-lactate and L-lactate in plasma were 0.7 μmol/L and 0.2 μmol/L, respectively. The limits of detection of D-lactate and L-lactate in urine were 8.1 nmol/mmol creatinine and 4.4 nmol/mmol creatinine, respectively. Plasma and urinary levels of D- and L-lactate were increased in patients with IBD and T2DM as compared with controls.
The presented method proved to be suitable for the quantification of D- and L-lactate and opens the possibility to explore the use of D-lactate as a biomarker.
D-乳酸的血浆和尿液水平与糖尿病的存在有关。先前开发的技术在进一步评估D-乳酸作为这种疾病的生物标志物方面显示出一些局限性。
使用带有标记内标的超高效液相色谱串联质谱法定量D-乳酸和L-乳酸。样品用二乙酰-L-酒石酸酐衍生化,并在C(18)反相柱上分离。在对照组、炎症性肠病(IBD)患者和2型糖尿病(T2DM)患者的血浆和尿液中分析D-乳酸和L-乳酸。
成功实现了D-乳酸和L-乳酸的定量分析。在生理和病理生理范围内,校准曲线呈线性(r(2)>0.99)。尿液和血浆的回收率在96%至113%之间。批间和批内变异在2%至9%之间。血浆中D-乳酸和L-乳酸的检测限分别为0.7μmol/L和0.2μmol/L。尿液中D-乳酸和L-乳酸的检测限分别为8.1nmol/mmol肌酐和4.4nmol/mmol肌酐。与对照组相比,IBD和T2DM患者的血浆和尿液中D-乳酸和L-乳酸水平升高。
所提出的方法被证明适用于定量D-乳酸和L-乳酸,并为探索将D-乳酸用作生物标志物提供了可能性。