Simultaneous quantification of labeled (2)H5-glycerol, (13)C6-glucose, and endogenous D-glucose in mouse plasma using liquid chromatography tandem mass spectrometry.
Simultaneous quantification of labeled (2)H5-glycerol, (13)C6-glucose, and endogenous D-glucose in mouse plasma using liquid chromatography tandem mass spectrometry.
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使用液相色谱串联质谱法同时定量小鼠血浆中标记的 (2)H5-甘油、(13)C6-葡萄糖和内源性 D-葡萄糖。
DOI：
10.1007/s00216-015-9015-5
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发表时间：
2015
影响因子：
4.3
通讯作者：
Wang,Rong
中科院分区：
文献类型：
--
作者：
Jahouh,Farid;Wang,Rong
监测生物液体中葡萄糖和甘油或其标记衍生物的水平以进行动力学研究一直是具有挑战性的，特别是在小鼠中，因为体积有限，加之血浆的复杂性。为此，我们开发了一种简单、快速、灵敏的方法，用于同时测量标记的2h5 -甘油和13c6 -葡萄糖以及内源性d -葡萄糖的绝对浓度，使用液相色谱-串联质谱（LC-MS/MS）。在我们的研究中，15.0 μL的小鼠血浆经过一步蛋白沉淀，然后进行LC-MS/MS分析。分析物的定量是通过监测其相应的去质子化分子离子的产物离子扫描，并通过为每个分析物选择特定的产物离子（相当于前体离子到产物离子的转变）构建提取的离子片段图来进行的。稀释小鼠血浆中th2h5 -甘油和13c6 -葡萄糖的检出限（LOD）为1.0 μM，定量限（LOQ）为5.0 μM，对应血浆中50 μM或血浆中甘油和葡萄糖分别为4.60和9.01 mg/dL。2h5 -甘油提取率为81.9% (CV = 8.1%)， 13c6 -葡萄糖提取率为26.2% （CV = 13.6%）。
Monitoring the level of glucose and glycerol or their labeled derivatives in biological fluid for kinetic studies has always been challenging, especially in mice, because of the limited volume in addition to the complexity of plasma. For such application, we developed a simple, fast, and sensitive method for the simultaneous measurement of absolute concentrations of labeled2H5-glycerol and13C6-glucose as well as endogenous D-glucose using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). In our study, 15.0 μL of mouse plasma was processed by a one-step protein precipitation, followed by LC-MS/MS analysis. The quantification of the analytes was carried out by monitoring the product ion scan of their corresponding deprotonated molecular ions and constructing the extracted ion fragmentogram by choosing a specific product ion for each analyte (equivalent to precursor ion to product ion transitions). The limit of detection (LOD) was evaluated to be 1.0 μM for both2H5-glycerol and13C6-glucose, and the limit of quantitation (LOQ) was observed to be 5.0 μM for both2H5-glycerol and13C6-glucose in diluted mice plasma that corresponds to 50 μM in plasma or 4.60 and 9.01 mg/dL of glycerol and glucose in plasma, respectively. The extraction recoveries are 81.9 % (CV = 8.1 %) for2H5-glycerol and 26.2 % (CV = 13.6 %) for13C6-glucose.