Simultaneous analysis of (13)C-glutathione as its dimeric form GSSG and its precursor [1-(13)C]glycine using liquid chromatography/isotope ratio mass spectrometry.
Simultaneous analysis of (13)C-glutathione as its dimeric form GSSG and its precursor [1-(13)C]glycine using liquid chromatography/isotope ratio mass spectrometry.
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使用液相色谱/同位素比质谱法同时分析 (13)C-谷胱甘肽二聚体 GSSG 及其前体 [1-(13)C]甘氨酸。
DOI：
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发表时间：
2009
影响因子：
2
通讯作者：
J. V. van Goudoever
中科院分区：
文献类型：
--
作者：
H. Schierbeek;D. Rook;F. T. te Braake;K. Dorst;G. Voortman;J. Godin;L. Fay;J. V. van Goudoever
使用稳定同位素测定谷胱甘肽动力学需要精确测量示踪剂和示踪物。以前，前体和合成的产物用两种单独的技术测量，液相色谱/同位素比质谱法（LC/IRMS）和气相色谱/燃烧/同位素比质谱法（GC/C/IRMS）。为了减少样品体积和最大限度地减少分析工作，我们开发了一种方法，在一次分析中同时测定小体积红细胞中的（13）C-谷胱甘肽二聚体形式（GSSG）及其前体[1-（13）C]甘氨酸。在将（13）C-谷胱甘肽转化成其二聚体形式GSSG后，我们使用液相色谱法结合LC/IRMS测定了150 μ L全血中GSSG和甘氨酸的红细胞内浓度和（13）C-同位素富集。结果表明，使用环亮氨酸作为内标物可以可靠地测量浓度（μ mol/mL范围），即精度优于0.1 μ mol/mL。GSSG和甘氨酸的（13）C-同位素富集在同一次运行中测量，给出了可靠的值，具有极好的精密度（标准偏差（sd）<0.3 ‰）和准确度（在0和5 APE之间测量）。这种新的方法开辟了各种动力学研究与相对低剂量的示踪剂管理，降低了研究设计的总成本。此外，仅需要最小的样品体积，即使在非常小的受试者（如早产儿）中也可以进行研究。
Determination of glutathione kinetics using stable isotopes requires accurate measurement of the tracers and tracees. Previously, the precursor and synthesized product were measured with two separate techniques, liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) and gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). In order to reduce sample volume and minimize analytical effort we developed a method to simultaneously determine (13)C-glutathione as its dimeric form (GSSG) and its precursor [1-(13)C]glycine in a small volume of erythrocytes in one single analysis. After having transformed (13)C-glutathione into its dimeric form GSSG, we determined both the intra-erythrocytic concentrations and the (13)C-isotopic enrichment of GSSG and glycine in 150 microL of whole blood using liquid chromatography coupled to LC/IRMS. The results show that the concentration (range of micromol/mL) was reliably measured using cycloleucine as internal standard, i.e. with a precision better than 0.1 micromol/mL. The (13)C-isotopic enrichment of GSSG and glycine measured in the same run gave reliable values with excellent precision (standard deviation (sd) <0.3 per thousand) and accuracy (measured between 0 and 5 APE). This novel method opens up a variety of kinetic studies with relatively low dose administration of tracers, reducing the total cost of the study design. In addition, only a minimal sample volume is required, enabling studies even in very small subjects, such as preterm infants.
缺乏蛋白质的猪在受到炎症应激时无法维持还原型谷胱甘肽的稳态。
DOI：
10.1093/jn/125.6.1462
发表时间：
1995
期刊：
The Journal of nutrition
影响因子：
--
作者：
Jahoor,F;Wykes,LJ;Reeds,PJ;Henry,JF;delRosario,MP;Frazer,ME
通讯作者：
Frazer,ME