A LC-MS/MS Method for Quantifying Adenosine, Guanosine and Inosine Nucleotides in Human Cells.
A LC-MS/MS Method for Quantifying Adenosine, Guanosine and Inosine Nucleotides in Human Cells.
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DOI：
10.1007/s11095-016-2040-z
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发表时间：
2017-01
影响因子：
3.7
通讯作者：
Kiser, Jennifer J.
中科院分区：
文献类型：
--
作者：
Jimmerson, Leah C.;Bushman, Lane R.;Ray, Michelle L.;Anderson, Peter L.;Kiser, Jennifer J.
开发并验证一种同时测量外周血单核细胞（PBMCs）、红细胞（rbc）和干血斑（DBS）中单磷酸（MP）和三磷酸（TP）形式的腺苷、鸟苷和肌苷的方法。固相萃取细胞裂解物，然后去磷酸化为摩尔当量核苷，LC-MS/MS定量。对腺苷、鸟苷和肌苷的定量范围分别为10-2000、1.0-200和0.25-50 pmol/样品，该方法均呈线性。所有三种分析物的测定内（n=6）和测定间（n=18）精密度（%CV）在1.7%至16%之间，而准确度（%偏差）在- 11.5%至14.7%之间。单磷酸核苷酸的浓度低于三磷酸核苷酸（肌苷除外），三种核苷酸在外周血中的浓度均高于红细胞（ATP、GTP和ITP分别为10倍、55倍和5.6倍）。与配对的RBC裂解物相比，DBS样品的TP平均（SD）降低了- 26% (22.6%)，MP水平提高了184%(173%)，表明DBS中TP水解。该方法对单磷酸和三磷酸形式的腺苷、鸟苷和肌苷核苷酸的生理相关浓度具有准确性和精确性，为临床研究提供了核苷酸定量的生物分析工具。
To develop and validate a method for the simultaneous measurement of adenosine, guanosine, and inosine derived from mono (MP) and triphosphate (TP) forms in peripheral blood mononuclear cells (PBMCs), red blood cells (RBCs) and dried blood spots (DBS). Solid phase extraction of cell lysates followed by dephosphorylation to molar equivalent nucleoside and LC-MS/MS quantification. The assay was linear for each of the three quantification ranges: 10–2000, 1.0–200 and 0.25–50 pmol/sample for adenosine, guanosine, and inosine, respectively. Intraassay (n=6) and interassay (n=18) precision (%CV) were within 1.7% to 16% while accuracy (%deviation) was within −11.5 % to 14.7 % for all three analytes. Nucleotide monophosphates were less concentrated than triphosphates (except for inosine) and levels in PBMCs were higher than RBCs for all three nucleotides (10, 55, and 5.6 fold for ATP, GTP and ITP, respectively). DBS samples had an average (SD) of −26% (22.6%) lower TP and 184% (173%) higher MP levels compared to paired RBC lysates, suggesting hydrolysis of the TP in DBS. This method was accurate and precise for physiologically relevant concentrations of adenosine, guanosine and inosine nucleotides in mono- and triphosphate forms, providing a bioanalytical tool for quantitation of nucleotides for clinical studies.
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影响因子：
3.7
作者：
Chen, Ping;Liu, Zhongfa;Liu, Shujun;Xie, Zhiliang;Aimiuwu, Josephine;Pang, Jiuxia;Klisovic, Rebecca;Blum, William;Grever, Michael R.;Marcucci, Guido;Chan, Kenneth K.
通讯作者：
Chan, Kenneth K.
影响因子：
29.4
作者：
Hitomi, Yuki;Cirulli, Elizabeth T.;Goldstein, David B.
通讯作者：
Goldstein, David B.
影响因子：
5.8
作者：
SMOLENSKI, RT;MONTERO, C;SIMMONDS, HA
通讯作者：
SIMMONDS, HA
影响因子：
13.5
作者：
De Franceschi, L;Fattovich, G;Corrocher, R
通讯作者：
Corrocher, R
影响因子：
2.2
作者：
Hugo Aragon-Martinez, Othoniel;Galicia, Othir;Martinez-Morales, Flavio
通讯作者：
Martinez-Morales, Flavio