Gas chromatographic-mass spectrometric analysis of hexose monophosphate shunt activity in cultured cells.
Gas chromatographic-mass spectrometric analysis of hexose monophosphate shunt activity in cultured cells.
复制标题
培养细胞中己糖单磷酸分流活性的气相色谱-质谱分析。
DOI：
10.1016/s0006-291x(89)80072-5
复制
发表时间：
1989
影响因子：
3.1
通讯作者：
M. Garwood
中科院分区：
文献类型：
--
作者：
S. L. Mitchell;B. Ross;T. Krick;M. Garwood
本文介绍了一种GC/MS方法，用于监测肿瘤细胞中通过己糖一磷酸分流(HMPS)降解为乳酸的葡萄糖的相对量。C6胶质瘤细胞分别在添加[1-13C]葡萄糖和含[1-13C]葡萄糖+0.001 mM吩嗪甲硫酸盐(PMS)的培养液中孵育。用测定三甲基硅基衍生物的m/z219/220和117/118离子确定的[13C]乳酸与[12C]乳酸的比值来计算HMPS的活性。PMS使C6胶质瘤细胞HMPS活性在孵育2小时和12小时分别增加3.2倍和4.8倍。将GC/MS结果与核磁共振测定的[3-13C]乳酸/[3-12C]乳酸比值进行了比较。与核磁共振法相比，GC/MS法所需的样品量更少，灵敏度更高。
A GC/MS method is described for monitoring the relative amount of glucose degraded to lactate via the hexose monophosphate shunt (HMPS) in neoplastic cells. C6 glioma cells were incubated in medium supplemented with [1-13C]glucose and medium containing [1-13C]glucose with 0.001mM phenazine methosulfate (PMS). The ratio of the [13C]lactate to [12C]lactate determined from the measurement of the m/z 219/220 and 117/118 ions of the trimethyl silyl derivative, was used to calculate HMPS activity. PMS increased HMPS activity in C6 glioma cells by 3.2 and 4.8 fold at 2 and 12 hours of incubation respectively. GC/MS results were compared with1H NMR measurements of the [3-13C]lactate/[3-12C]lactate ratio. The GC/MS method was found to require less sample size and yielded better sensitivity than the NMR method.