Gas chromatographic-mass spectrometric determination of isotopic enrichment of 6-15NH2 in adenine nucleotides.
Gas chromatographic-mass spectrometric determination of isotopic enrichment of 6-15NH2 in adenine nucleotides.
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气相色谱-质谱法测定腺嘌呤核苷酸中 6-15NH2 的同位素富集。
DOI：
10.1016/0003-2697(85)90374-4
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发表时间：
1985
影响因子：
2.9
通讯作者：
Yudkoff,M
中科院分区：
文献类型：
--
作者：
Lewis,S;Yudkoff,M
本文介绍了一种测定[6- 15 NH_2]腺嘌呤核苷酸同位素丰度的气相色谱-质谱法。该方法涉及腺嘌呤的二叔丁基二甲基甲硅烷基（TBDMS）衍生物的形成，随后用固相离子交换色谱分离核苷酸级分，随后将核苷酸酸水解为游离碱。获得腺嘌呤-diTBDMS和[6- 15 NH 2]腺嘌呤-diTBDMS的质谱以鉴定含有6-NH 2部分的那些离子。基峰（m z 306）由C4 H9（57）的损失形成，约占总离子电流的三分之一。使用m/z 306/m/z 307比率的选择离子监测，通过注射10-20 pmol的从分离的大鼠肝细胞获得的腺嘌呤-diTBDMS衍生物，可重复地测量1.0-50.0 mol%过量的同位素丰度水平。通过腺嘌呤氧化为次黄嘌呤并测定次黄嘌呤-ditbDMS衍生物的富集度，证实了测量的同位素丰度与6- 15 NH 2基团的标记一致。该方法被用来研究[6- 15NH_2]腺嘌呤核苷酸的形成过程中的离体大鼠肝细胞与[15 N]丙氨酸孵育。在60 min孵育时观察到约6.0 mol%的过量水平。
A gas chromatographic-mass spectrometric method for the determination of isotopic abundance in [6-15NH2]adenine nucleotides is described. The method involves formation of the di-t-butyldimethylsilyl (TBDMS) derivative of adenine following isolation of the nucleotide fraction with solid-phase ion-exchange chromatography and subsequent acid hydrolysis of nucleotides to free base. Mass spectra for both adenine-diTBDMS and [6-15NH2]adenine-diTBDMS were obtained to identify those ions containing the 6-NH2moiety. The base peak (m z 306) was formed by loss of C4H9(57) and constitutes approximately one-third of the total ion current. Using selected ion monitoring of the m z 306/m/z 307 ratio, levels of isotopic abundance of 1.0–50.0 mol% excess could be measured reproducibly with the injection of 10–20 pmol of the adenine-diTBDMS derivative obtained from isolated rat hepatocytes. Confirmation that measured isotopic abundance was referable to labeling of the 6-15NH2group was obtained by oxidation of adenine to hypoxanthine and determination of enrichment in the hypoxanthine-diTBDMS derivative. The method was used to study the formation of [6-15NH2]adenine nucleotides during the incubation of isolated rat hepatocytes with [15N]alanine. A level of approximately 6.0 mol% excess was observed at 60 min incubation.
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影响因子：
4.1
作者：
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通讯作者：
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DOI：
10.1002/bms.1200080309
发表时间：
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期刊：
Biomedical mass spectrometry
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DOI：
10.1016/s0378-4347(00)80277-7
发表时间：
1981
期刊：
Journal of Chromatography B: Biomedical Sciences and Applications
影响因子：
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影响因子：
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