ORGANIC
LETTERS
2002
Vol. 4, No. 25
4487-4490
Synthesis of a New Fluorescent Probe
Specific for Catechols
Douglas E. Stack,* Anastacia L. Hill, Clark B. Diffendaffer, and Nicole M. Burns
Department of Chemistry, UniVersity of Nebraska at Omaha, 6001 Dodge Street,
Omaha, Nebraska 68182-0109
Received September 30, 2002
ABSTRACT
The synthesis of a new fluorescent probe, specific for the catechol moiety, has been conducted by preparation of r,r-dibromomalonamides
containing an appropriate fluorophore. N,N′-Bis-anthracen-9-ylmethyl-2,2-dibromomalonamide reacted with various catechols in the presence
of cesium carbonate to generate highly fluorescent derivatives.
The catechol moiety is an important structural unit in many
biomolecules. Analytical assay of various catechol-containing
substances, such as catecholamines and catechol estrogens,
has received considerable attention. High-performance liquid
chromatography (HPLC) using electrochemical detection
(HPLC-ED),1 precolumn HPLC fluorescent derivation,2 and
gas chromatography interfaced to mass spectrometry (GC/
MS)3 have all been employed in detecting catechols. We are
interested in the detection of catechol estrogen-DNA adducts
(CE-DNA) generated by the reaction of catechol estrogen
quinones (CE-Q) with DNA bases, namely guanine and
adenine.4 Recent interest in CE-DNA adducts stems from
their use in studies aimed at understanding the etiology of
hormonal carcinogenesis, and the hope they may be em-
ployed as biomarkers for the early detection of estrogen-
induced cancers.5
Recent publications indicate that production of CE-DNA
adducts in vivo occurs at very low levels, ca. at the low
femtomolar level.5d,e Detection at this level by HPLC-ED
was not possible due to the diminished selectivity of HPLC-
ED at these levels. Fluorescent derivation using 1,2-di-
phenylethylenediamine (DPE) has been used for femtomolar
detection of catecholamines. Precolumn derivation with DPE
requires prior oxidation of the catechol ring system so that
condensation of the DPE amino groups with quinone
carbonyls can occur.2 We tried applying the DPE methodol-
ogy toward CE-DNA adducts with no success. 1-Pyrene-
sulfonyl chloride was used as a precolumn fluorescent probe
for the CE-DNA adduct, 4-OHE2-1-N7Gua (see Figure 1)
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Musso, N. R.; Vergassola, C.; Pende, A.; Lotti, G. J. Liq. Chromatogr.
1990, 13, 1075.
(2) (a) Mitsui, A.; Nohta, H.; Ohkura, Y. J. Chromatogr. Field 1985,
344, 61. (b) Ohkura, Y.; Nohta, H. Trends Anal. Chem. 1992, 11, 74.
(3) Castagnetta, L. A.; Granata, O. M.; Arcuri, F. P.; Polito, L. M.; Rosati,
F.; Cartoni, G. P. Steroids 1992, 57, 437.
(4) (a) Stack, D. E.; Byun, J.; Gross, M. L.; Rogan, E. G.; Cavalieri, E.
L. Chem. Res. Toxicol. 1996, 9, 851. (b) Cavalieri, E. L.; Stack, D. E.;
Devanesan, P. D.; Todorovic, R.; Dwivedy, I.; Higginbotham, S.; Johansson,
S. L.; Patil, K. D.; Gross, M. L.; et al. Proc. Natl. Acad. Sci. U.S.A. 1997,
94, 10937.
(5) (a) Devanesan, P.; Santen, R. J.; Bocchinfuso, W. P.; Korach, K. S.;
Rogan, E. G.; Cavalieri, E. Carcinogenesis 2001, 22, 1573. (b) Devanesan,
P.; Todorovic, R.; Zhao, J.; Gross, M. L.; Rogan, E. G.; Cavalieri, E. L.
Carcinogenesis 2001, 22, 489. (c) Todorovic, R.; Devanesan, P.; Higgin-
botham, S.; Zhao, J.; Gross, M. L.; Rogan, E. G.; Cavalieri, E. L.
Carcinogenesis 2001, 22, 905. (d) Cavalieri, E. L.; Devanesan, P.; Bosland,
M. C.; Badawi, A. F.; Rogan, E. G. Carcinogenesis 2002, 23, 329. (e)
Cavalieri, E. L.; Li, K.-M.; Balu, N.; Saeed, M.; Devanesan, P.; Higgin-
botham, S.; Zhao, J.; Gross, M. L.; Rogan, E. G. Carcinogenesis 2002, 23,
1071. (f) Cavalieri, E. L.; Rogan, E. G.; Chakravarti, D. Cell. Mol. Life
Sci. 2002, 59, 665.
10.1021/ol027000j CCC: $22.00 © 2002 American Chemical Society
Published on Web 11/21/2002