Analysis of Lysozyme using HPLC

[wongyl4985]

Hi,

I am analyzing lysozyme chloride using C18 column with hplc system. Sadly to say my method is not working because the resolution between my lysozyme and nearest peak is less than 2. May i know do you have any method in analyzing this compound using hplc system? Thanks in advance.

[unmgvar]

can you give more details of the method and column?
RT of the compounds?
mobile phase
gradient
injection volume
detector used

[wongyl4985]

Thanks for the reply. I am currently using a C18 stable bond with 25cm, 4.6mmID column. The mobile phase composition is mixture of pH2.8 Ammonium Acetate: Acetonitrile with UV vis detector. The retention time of the lysozyme is around 7 min. However, there is a peak besides the lysozyme peak and it is difficult to separate this peak with the main peak even i have tried different ratio of mobile phase composition. Looking for you soon reply. Thanks.

[unmgvar]

can you give further details
there are many c-18 columns, there are not all the same
can you say more about the composition of the method?
a mixture of a buffer and ACN makes for most of the applications in the world

what about the retention time of the compounds?
can you say what you are looking for? are we talking of oxidized forms?
something else?
knowing this can also help

[sepscientologist]

If you are running a gradient then just make it more shallow (slower composition change over time)