I have to develop an amino acid method for only one amino acid and I need to derivatize but the HPLC does not have an on-line way to do it. How can I derivatize manually with OPA?
See the water's application note below;
http://www.waters.com/waters/library.htm?cid=511436&lid=1527206&locale=en_US
In that case just request this article.
Article Realities of automating OPA HPLC amino acid analyses
Derivatization occurs rather quickly, about a minute. Much more than that and fluorescence starts to degrade. How would you time that well enough so exactly the same reaction time occurred between each sample? If you feel confident enough in your stopwatch and accurate pipetting skills, perhaps try to do a manual version of an autosampler program?
Also, OPA is sensitive to oxygen.
Do you have a manual injection port? If you do have a sampling needle to inject your samples, there are methods that describe picking up reagents and mixing them in either a well plate or the needle wash port.
Or is this an issue of the programming?
- Badges
- Science method