Updated publication reference for PubMed record(s): 34609160. The incidence of candidiasis, caused mainly by the opportunistic fungal Candida albicans, remains a major concern in public health due to the emerging antifungal resistance. C. albicans modulates their proteome under different conditions trying to cope with several stressors in host niches. In this study we developed a C. albicans spectral library by analysing a total of 52 samples corresponding to cells treated with 5 and 10 mM hydrogen peroxide, 40 and 60 mM acetic acid and in the interaction with human macrophages (THP-1) at 3, 6 and 9 h (4 biological replicates). This samples represented several morphologies of C. albicans (yeast and hypha) and different culture conditions (temperature, media…) that contributed to the incorporation of different proteins into the library. The analysis of these conditions by DDA mode allowed the construction of a C. albicans spectral library comprising information of 2879 proteins and 25645 peptides which entailed a 46.5% of the total proteome. The C. albicans spectral library created constitutes a powerful proteomic tool available for the scientific community and could be useful for global (DIA) or targeted (SRM and PRM) studies.