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求教:GC-MS同位素标记定量

作者 cat-liujiang
来源: 小木虫 450 9 举报帖子
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请教一个问题,看到一篇文献,做的同位素标记前体物饲喂,最后用GC-MS检测同位素标记的代谢物含量
这些化合物的分子量与他们标记的 target ion差别很大
比如第一个化合物分子量是89,3个碳都同位素标记后,应该是92啊,但他的target ion却是221
所有化合物似乎都对不上号,不知道是咋回事儿

Supp. Table 3. Metabolites identified in RCH2 bacterial lysates and their corresponding 13C target ion used for recovery comparison, retention time (RT) and RT of authentic standards. Putative metabolite annotations are indicated by italics (retention time and/or spectra could not be confirmed with an authentic standard).                                        
Compound        13C target ion         Chemical Formula                               
alanine                      221                       C3H7NO2
valine                      220                           C5H11NO2
isoleucine              163                       C6H13NO2
proline                      146                       C5H9NO2
glycine                      249                        C2H5NO2
uracil                       245                      C4H4N2O2
serine                       206                      C3H7NO3

用的是安捷伦的气质
Agilent 7890 gas chromatograph (Agilent Technologies, Santa Clara, CA)
Agilent 5977 single quadrupole mass spectrometer (MS)

http://www.sciencedirect.com/sci ... 71714003502#appsec1
3.5. Microbial metabolite adsorption and recovery
To explore the types of microbial metabolites that adsorb to soil and therefore may represent those that are unavailable to microbes in situ, 13C-labeled RCH2 bacterial lysates were incubated with soil and extracted with water (Fig. 3). K2SO4 (10 mM) was compared to water to determine if this classical extractant was more efficient at metabolite recovery. The most intense peaks from the unlabeled RCH2 lysates (grown with 12C-acetate) were selected for further analysis in labeled samples (compound identification, 13C incorporation and recovery calculations). Spectra from unlabeled RCH2 were compared to labeled cultures, which resulted in the identification of 45 fully-labeled metabolites (Fig. 4, Supp. Table 3). The 13C target ion for each metabolite was quantified for all samples and recovery was calculated. Only 27% of the quantified metabolites showed >50% recovery in the WEOM, with the majority of those being neutral metabolites. All of the polyamine cations identified in this experiment (putrescine, cadaverine, lysine, spermine and spermidine) and most of the anionic metabolites were among those that adsorbed rapidly to soil (Fig. 4). The inclusion of 10 mM K2SO4 as a recovery extractant did not drastically improve or reduce recovery compared to water alone. 返回小木虫查看更多

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  • yjjc2012

    代谢产物又不是喂食进去的前提,结构完全不一样的

  • cat-liujiang

    引用回帖:
    2楼: Originally posted by yjjc2012 at 2017-08-14 11:04:19
    代谢产物又不是喂食进去的前提,结构完全不一样的

    可以不管是不是前体饲喂什么的,就看最后检测到的产物,也是不对的啊
    比如第一个化合物alanine分子量是89,他的target ion却是221,无法理解

  • yjjc2012

    引用回帖:
    4楼: Originally posted by cat-liujiang at 2017-08-14 11:53:41
    可以不管是不是前体饲喂什么的,就看最后检测到的产物,也是不对的啊
    比如第一个化合物alanine分子量是89,他的target ion却是221,无法理解...

    转变成其他物质了。All of the polyamine cations identified in this experiment (putrescine, cadaverine, lysine, spermine and spermidine) and most of the anionic metabolites

  • 圆圈爱圈圆

    您好,我是做微生物13C标记的,请问13C标记的产物可以直接用GC-MS检测吗?GC-MS不是不能检测出同位素的吗??谢谢啦

  • lisiqi190356

    您好,我是做微生物13C标记的,请问13C标记的产物可以直接用GC-MS检测吗?GC-MS不是不能检测出同位素的吗??谢谢啦

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