Hi Ashrafi,
Your mobile phase strikes me as rather completed (i.e. too many solvents mixed together). If there is any chance, I would try to make it a little bit less complicated which may help with your reproducibility issues. Also, could you maybe give some more info regarding the "lut" and "zeax" please? In additoon, to help you with your troubleshooting, it would be great to have the proportions of each solvent/modifier for both, mobile phase A and B along with the gradient.
What solvent have you used to make up your beta carotene standard? And at which concentration?
Considering you complicated mobile phase, have you tried to dissolve your standard in mobile phase A?
From the information you supplied, I suspect that your beta carotene keeps crashing out when injected which would give rather irreproducible results. The extend of the "crashing" out would be determined by the concentration. Do you observe any pressure fluctuations/problems?
Good luck!
Whether check the following links are useful.
https://tools.thermofisher.com/content/sfs/brochures/AU-187-HPLC-C30-Beta-Carotene-Iodine-Isomers-LPN3057-EN.pdf
http://www.waters.com/webassets/cms/library/docs/720004550en.pdf
https://www.aup.edu.pk/sj_pdf/DETERMINATION%20OF%20BETA%20CAROTENE%20CONTENT.pdf
http://www.revagrois.ro/PDF/2007_54.pdf
I suggest you use C30 Carotenoid YMC instead of C18, because C18 could not separate their isomers. That make your result strange.
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