OPA derivatization and neutralization buffer

[dragosb]

Hi again,

Can someone plese tell me why there is a neutralization buffer used in the OPA derivatization procedure. Can it be done without the neutralization? What are the risks, effects?

Thank you

[ACrawshaw]

Hi,

The neutralisation step can be considered for two reasons:

The first is to simply stop the derivatisation reaction. From a quantitative perspective this is important to be done at the same time for each sample to remove any possible differences in the results.

The second reason is that the pH of the derivatised solution is quite high (ca pH 10), the neutralisation step therefore brings the pH back down to neutral conditions. Depending upon your column chemistry, this may also prove to be advantageous for column lifetime.

Hope this helps..