Hammond P W, Wiese W A, Waldrop A A, Nelson N C, Arnold L J
Gen-Probe Inc., San Diego, CA 92121.
J Biolumin Chemilumin. 1991 Jan-Mar;6(1):35-43. doi: 10.1002/bio.1170060108.
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 to the 9 position of an acridinium ester to form the carbinol adduct has also been well documented. We have observed a similar addition of other nucleophiles to the acridinium ring to 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 H2O2) has been demonstrated to 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 DNA probe-based assays. The adduct is highly resistant to hydrolysis under the conditions often desired for DNA probe-based assays (high temperature, elevated pH, extended storage).
吖啶酯（AE）的化学发光反应需要在吖啶环的9位添加过氧化物。向吖啶酯的9位添加氢氧根离子以形成甲醇加合物也有充分的文献记载。我们观察到其他亲核试剂也会类似地加成到吖啶环上形成吖啶加合物。与亚硫酸氢盐形成的加合物在形成速率、逆转速率和反应平衡方面得到了特别充分的表征。已证明形成加合物（除H2O2外）会显著降低相邻酯键对碱性水解的反应活性。当吖啶酯用作基于DNA探针的检测中的标记物时，所得的更稳定的加合物非常有用。在基于DNA探针的检测通常所需的条件下（高温、高pH值、延长储存时间），该加合物对水解具有高度抗性。
