Nucleophilic addition to the 9 position of 9-phenylcarboxylate-10-methylacridinium protects against hydrolysis of the ester

✍ Scribed by Hammond, Philip W. ;Wiese, Wendy A. ;Waldrop, Alex A. ;Nelson, Norman C. ;Arnold, Lyle J.

• Publisher: John Wiley and Sons
• Year: 1991
• Weight: 695 KB
• Volume: 6
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170060108

✦ Synopsis

The chemiluminescent reaction of an acridinium ester (AE) requires addition of peroxide to the 9 position of the acridinium ring. The addition of a hydroxide ion t o the 9 position of an acridinium ester t o form the carbinol adduct has also been well documented. We have observed a similar addition of other nucleophiles t o the acridinium ring t o form an acridan adduct. The adduct formed with bisulphite has been particularly well-characterized for rate of formation, rate of reversion, and reaction equilibrium. The formation of an adduct (other than H,O,) has been demonstrated t o decrease significantly the reactivity of the adjacent ester bond to alkaline hydrolysis.

The resulting, more stable adduct is very useful when the acridinium ester is used as a label in D N A probe-based assays. The adduct is highly resistant t o hydrolysis under the conditions often desired for D N A probe-based assays (high temperature, elevated pH, extended storage).