Pyridine Hydrochloride by HPLC

[kalidassa]

Hi Friends can any one have any method or idea to analyse Pyridine Hydrochloride by HPLC. With out making free base is it possible to analyze by HPLC.

[carls]

This is a straight forward analysis using RPLC. What detection system are you using?

ACN/buffer at pH 7 will work fine with C18

[kalidassa]

Hi we are using UV Detector, We tryed with ACN/water. i not used any buffer, Column we used C18. WE NOT GORT ANY RESPONCE.

[carls]

Use 25mM (NH4)2HPO4 at pH 7.5, ACN and 254nm. Make a 1mg/mL standard of pyridine in H2O and inject 1uL. Run a gradient from 5-50% ACN and you should definately see a peak elute. The buffer is necessary ensure pyridine is not ionized which helps in 2 ways - increases hydrophobicity/retention and decreases interaction with silica/tailing