Chromatography Forum

Hellow every Body,[Cyanouric chloride :

Cyanouric chloride : Derivatizing solvent: 5% Aniline in Acetic acid.
Sample Preparation: 0.0050 gm Cyanouric chloride into 25 ml volumetric flask + 5 ml of derivatizing solvent, sonicate the solution up 5 minutes and finally make up the solution with Acetonitrile.
Observation-1 : On HPLC one peak was obtained up to 99 + and other peak was obtained 0.5%.
Observation-2 : Follow the same procedure of sample preparation as described above but two equivalent peaks were observed. One has mass of 298 and other has 354. As per search it was found that two Cl removed with two aniline and all three Cl were removed with aniline [i.e di (m/z = 298)and tri (m/z = 354)aniline substituted triazine were formed.]
Now another experiment was performed and here derivatizing agent was 5% aniline in Acetonitrile. Here we got Mono and di substituted triazine and substitution was performed by Anilin.
The HPLC condition: ACN (0.1% Fromic acid) : Water (0.1% H-COOH) : : 75 : 25
Wavelength: 270 nm
Flow : 1.0ml/min , Column : Inertsil ODS-3 250mm x 4.6mm and 5µm
Oven : 40°C
The question is that how can I got Only one peak??????
Another is if the cyanouric chloride has Cyanouric acid then How can this will predict????
For the formation of cyanouric chloride we prepared the sample into Acetonitrile : Water : : 50 : 50 premix and in 100 % ACN also.
But when we mixed cyanouric acid solution into derivatized solution (Observation-2) no cyanouric peak will found.
SO over all what is the procedure t separate the cyanouric acid with cyanouric chloride.

Best regards

Praveen

Hi,

Do you have to use a C18 column? Can you use a HILIC column?

Cheers,
ES

Hi

Again the same problem - acid (alkaline) and salt. They can not be properly divided into RP-18 as they are in dynamic equilibrium as soon as we change the phase pH - we break balance in one of form.
And when using RP-18 at least one of our solvent (water) is not identiffirenten to the balance of ions in the object of research.

It is impossible to quantitatively derivatized separately. As for the quantitative derivatization, we have to take a large excess of derivatization reagent and lead to severe conditions, which respond both forms of the substance.

There's good anecdote. Since the topic is relevant in Afghanistan - only to bring it to an end.
Captive Sgt in Talib duvane:
"- What a fool I was been! Colonel was righ , was said me: Learn technique .... "[/b]