通过反相液相色谱串联质谱同时定量L(+)和D(-)乳酸测定,2型糖尿病患者血浆和尿液样品中的L(+)和D(-)乳酸增加。

Experimental Diabetes Research Pub Date : 2012-01-01 Epub Date: 2012-03-08 DOI:10.1155/2012/234812
Jean L J M Scheijen, Nordin M J Hanssen, Marjo P H van de Waarenburg, Daisy M A E Jonkers, Coen D A Stehouwer, Casper G Schalkwijk
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引用次数: 71

摘要

背景:血浆和尿中d -乳酸水平与糖尿病有关。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。方法:采用内标超高效液相色谱串联质谱法定量测定D-乳酸和l -乳酸。样品用二乙酰- l-酒石酸酐衍生,在C(18)反相柱上分离。分析了对照组、炎症性肠病(IBD)患者和2型糖尿病(T2DM)患者血浆和尿液中的D-和l -乳酸。结果:D-和l -乳酸的定量分析成功。校正曲线在生理和病理生理范围内呈线性关系(r(2) > 0.99)。尿液和血浆的回收率在96% ~ 113%之间。测定间和测定内的变异在2%到9%之间。血浆中d -乳酸和L-乳酸的检出限分别为0.7 μmol/L和0.2 μmol/L。尿中d -乳酸和l -乳酸的检出限分别为8.1 nmol/mmol肌酐和4.4 nmol/mmol肌酐。与对照组相比,IBD和T2DM患者血浆和尿中D-和l -乳酸水平升高。结论:该方法适用于D-乳酸和l -乳酸的定量测定,为D-乳酸作为生物标志物的应用开辟了可能性。
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L(+) and D(-) lactate are increased in plasma and urine samples of type 2 diabetes as measured by a simultaneous quantification of L(+) and D(-) lactate by reversed-phase liquid chromatography tandem mass spectrometry.

Background: 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.

Methods: 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).

Results: 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.

Conclusion: 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.

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Experimental Diabetes Research
Experimental Diabetes Research 医学-内分泌学与代谢
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