| Project/Area Number |
04807157
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Physical pharmacy
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
KAI Masaaki Kyushu University, Faculty of Pharmaceutical Sciences, Assistant professor, 薬学部, 助手 (00160953)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Guanine / Nucleic acid-related compound / High-performance liquid chromatography / Fluorescence detection / Chemiluminescence detection / Phenylglyoxal / Selective reagent / Sensitive analysis |
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| Research Abstract |
Pre-column and post-column fluorescence derivatization methods for the high-performance liquid chromatographic (HPLC) determination of guanine-containing compounds have been studied on the basis of the fluorescence reaction with phenylglyoxal (PGO). The detection limits for the compounds tested were 140-720 fmol and 3-10 pmol per 100-ul injection volume in the pre- and post-column derivatization methods, respectively. The PGO reagent was also applied to the measurement of the nuceobase-pair of guanine-cytosine in double-stranded DNAs.
Alkoxyphenylglyoxals as the analogues of PGO were synthesized and found to be more sensitive reagents for the guanine-containing compounds. A novel chemiluminogenic method was developed on the basis of the fluorescence reaction of the guanine-containing compounds with PGO in a phosphate buffer (pH 6.0) at 37 ﾟC for 20 min, followed by a chemiluminescence reaction with N, N-dimethylformamide in a weakly alkaline medium. The chemiluminescence method was approximately 20-fold more sensitive than the fluorimetric one.
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