Design of a stable isotope dilution gas chromatography/mass spectrometric assay for cAMP: comparison with standard protein-binding and radioimmunoassay methods.
Design of a stable isotope dilution gas chromatography/mass spectrometric assay for cAMP: comparison with standard protein-binding and radioimmunoassay methods.
复制标题
cAMP 稳定同位素稀释气相色谱/质谱测定的设计：与标准蛋白质结合和放射免疫测定方法的比较。
DOI：
10.1016/0003-2697(80)90150-5
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发表时间：
1980
影响因子：
2.9
通讯作者：
N. Hunt
中科院分区：
文献类型：
--
作者：
L. P. Johnson;J. Macleod;R. Summons;N. Hunt
一种稳定的同位素稀释测定法，其中皮摩尔量的cAMP可以确定高精度和选择性，使用气相色谱和电子轰击质谱与多离子检测技术。使用合成的[2，8 - 2 H2，6- 15 N]-cAMP作为内标物，通过监测cAMP的三甲基甲硅烷基衍生物和内标物的（M = CH 3）+碎片离子，分别在m z 530和m z 533处获得适当的专属性。从质谱仪的检测下限判断的测定灵敏度为3.0 pmol。使用气相色谱/质谱法测定大鼠肝脏和人尿cAMP水平，并与相同样品的蛋白结合试验和放射免疫测定法测定的水平进行比较。大鼠肝脏样品（cAMP水平为832 pmol/g）和尿液样品（cAMP水平为2.50 μmol/L）的气相色谱/质谱分析的批内变异系数分别为5.3%和6.0%。通过三种测定方法测定的cAMP水平的比较显示相关性在10%变化内。
A stable isotope dilution assay is presented in which picomole quantities of cAMP can be determined with high precision and selectivity using gas chromatography and electron impact mass spectrometry with multiple ion detection techniques. Using synthetic [2,8-2H2,6-15N]-cAMP as the internal standard, suitable specificity was obtained by monitoring the (MCH3)+fragment ions of the trimethylsilyl derivatives of cAMP and the internal standard at m z 530 and m z 533, respectively. The sensitivity of the assay as judged from the lower limit of detection of the mass spectrometer was 3.0 pmol. Rat liver and human urine cAMP levels were assayed using gas chromatography/mass spectrometry and were compared with levels determined by protein-binding assays and radioimmunoassays for the same samples. The intraassay coefficients of variation of the gas chromatography/mass spectrometry assay were 5.3% for the rat liver sample (cAMP level 832 pmol/g) and 6.0% for the urine sample (cAMP level 2.50 μmol/liter). Comparison of the levels of cAMP determined by the three assay methods showed correlation to within 10% variation.