Vitamins quantification

[Lorespain]

Hi everybody! I am trying simultaneous quantification of vitamins A, E and D3. I tested them in spectrometry UV to know the best wavelenght for the LC analysis and I found different wavelengh for each one. Now, I have a doubt: have the wavelengh choosed for run LC an influence on the peak areas of the vitamins? How I chose the best wavelengh for them together? Thank you in advance

[Bryan Evans]

Below is are fat soluble vitamins separated on Cadenza CD-C18 (UV @ 280nm):

http://www.imtaktusa.com/site_media/fil ... TI121E.pdf

[HW Mueller]

Since the question, "have the wavelengh choosed for run LC an influence on the peak areas of the vitamins?" has not been addressed:
Of course, if you choose a wavelength where your analyte does not absorb you don´t get a peak at all.
If your detector allows this you can change the wavelengths to the max lambda of each compound in the region where it elutes. According to Bryan a good compromise is to use 280 nm for all. In any case you have to calibrate.