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cat-liujiang鐵桿木蟲(chóng) (正式寫(xiě)手)
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[求助]
求教：GC-MS同位素標(biāo)記定量 已有1人參與
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請(qǐng)教一個(gè)問(wèn)題，看到一篇文獻(xiàn)，做的同位素標(biāo)記前體物飼喂，最后用GC-MS檢測(cè)同位素標(biāo)記的代謝物含量 這些化合物的分子量與他們標(biāo)記的 target ion差別很大 比如第一個(gè)化合物分子量是89，3個(gè)碳都同位素標(biāo)記后，應(yīng)該是92啊，但他的target ion卻是221 所有化合物似乎都對(duì)不上號(hào)，不知道是咋回事兒 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 用的是安捷倫的氣質(zhì) 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. |
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2樓2017-08-14 11:04:19
3樓2017-08-14 11:04:59
cat-liujiang
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4樓2017-08-14 11:53:41
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6樓2017-09-18 09:25:46
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7樓2017-09-18 09:31:27
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gcms測(cè)量的是分子量，所以當(dāng)然可以測(cè)出c13 標(biāo)記的化合物。實(shí)際上有的醫(yī)院就用質(zhì)譜測(cè)定c13標(biāo)記的二氧化碳(用碳十三標(biāo)記的尿素讓檢測(cè)者服用，可以測(cè)定有無(wú)幽門(mén)螺桿菌細(xì)胞內(nèi)起催化作用的尿素分解酶) 發(fā)自小木蟲(chóng)Android客戶端 |
8樓2017-09-18 14:53:07
lisiqi190356
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9樓2017-09-19 14:55:16
風(fēng)過(guò)疏竹
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，想問(wèn)一下，如果我要定量13c標(biāo)記抗生素的濃度，該怎么做？10樓2017-09-29 15:46:22
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