@article{oai:hirosaki.repo.nii.ac.jp:00003925,
 author = {Seya, Kazuhiko and Motomura, Shigeru and Imaizumi, Tadaatsu and Furukawa, Ken-Ichi},
 issue = {1},
 journal = {弘前医学},
 month = {Oct},
 note = {Two sensitive enzymatic fluorometric assays have been developed for adenosine 3’,5’-cyclic monophosphate
（cAMP） by Sugiyama et al. （Anal. Biochem. 1990） and for guanosine 3’,5’-cyclic monophosphate （cGMP） by Seya
et al. （Anal. Biochem. 1998）. However, to make a new simultaneous comparison of two cyclic nucleotides, distinct
measurement methods cause less reliability and longer measurement time. To overcome these problems, we
developed a simultaneous measurement method for them. All adenosine nucleotides and GMP were enzymatically
degraded using alkaline phosphatase and apyrase. The remaining GDP was converted to GTP by creatine kinase.
Cyclic GMP and cAMP, absorbed into a Sep-Pak amino propyl cartridge, were eluted to separate from GTP, and
then were simultaneously quantified using improved enzymatic fluorometric assay. The detection limits for cAMP
and cGMP were 1 and 5 fmol, respectively. The total measurement time was about 10 h. Using this method, the
basal cAMP and cGMP levels in rat aortic smooth muscle cells were confirmed to 4.1 and 0.042 pmol/mg protein,
respectively. An adrenergic agonist, isoproterenol （1 μM） increased cAMP approximately 6 fold, while nitric oxide
donor, S-nitroso-N-acetylpenicillamine （100 μM） increased cGMP approximately 230 fold. These results suggest that
this simultaneous measurement of cGMP and cAMP can provide a more convenient assessment of them in biological samples., 弘前医学. 68, p.52-61. 2017},
 pages = {52--61},
 title = {Simultaneous Measurement of Adenosine 3’,5’-Cyclic Monophosphate and Guanosine 3’,5’-Cyclic Monophosphate in Biological Samples},
 volume = {68},
 year = {2017}
}