Chromatography Forum

I am looking for an HPLC method that would test for the concentration of Tricine in a buffer sample. Realizing that UV will not see this analyte, does anyone know of the best way to set up this analysis?

Actually, you can use UV at 210nm if you don't mind poor detection limits and lots of potential interferences.

Also ion-exchange chromatography with conductivity detection (non-suppressed). Since it is a weak zwitterion you could do anion exchange a high pH or cation exchange at low pH.

It rather depends. Do you want to find tricine where is does not belong in your buffer? Or do you want to assay the tricine content of a tricine buffer?

I'm guessing you don't have an LC-MS? Otherwise, you could probably convert it to a benzyl ester if you want to do reverse phase. That would also increase your UV sensitivity.

Mark,
I am looking for tricine amounts in a buffer that also contains Mg acetate, NaOH, polyvinylpyrrolidinone, Tween. What detection limit could be reached on UV210? Would these other components mask the tricine?

Your best approach for the detection is probably ELSD. It will be hard to determine tricine with UV as it will come close to avoid (unless you use ion pairing reagent). NaOH is not UV active as well as Mg.

We have a method for tricine:

http://www.sielc.com/compound_136.html

With the same column and mobile phase you will be able to determine NaOH (as sodium), magnesium acetate (as magnesium) etc.
http://www.sielc.com/compound_058.html (Primesep A column is about 50% stronger then Primesep 100)

Contact us if you have questions.

Regards,

Vlad

Yes, evaporative light scattering detection would work if you have access to one. Polyvinylpyrrolidone and tween are both complex mixtures, both are detectable by low wavelength UV or ELSD but not by conductivity.