A bioanalytical method using liquid chromatography-tandem mass spectrometry was developed for the analysis of fluconazole in Candida albicans after incubation with phenolic compounds, which have been proved possessing antifungal properties and have synergetic activity against C. albicans when in combination with fluconazole. Samples of C. albicans thallus obtained by centrifuging the mixed culture after 24 h incubation were saponified and centrifuged. The supernatant was evaporated to dryness, reconstituted, and injected on a C18 column using an organic-aqueous mobile phase. The chromatographic run time was 3.5 min per injection, with retention times of 2.4 min for fluconazole. The detection was by monitoring fluconazole at m/z 305-->191. The standard curve range was 1.0-100.0 ng ml(-1) with a mean correlation coefficient 0.9992. The precision and accuracy of the quality control (QC) samples were R.S.D < 5.5%, R.E. <3% for intra-day and R.S.D. <6.2%, R.E. <4% for inter-day. The concentration of fluconazole in C. albicans was found to be increased with the increment of the tested compounds concentration when they were in combination.