Dissolution of biotite, the main Fe-bearing mineral in granitic bedrock, is of particular importance for the remediation of reducing conditions after the ingress of oxygen, such as after mining activities or the construction of deep repositories for toxic waste. This study investigated the leaching of biotite of size fraction 0.053–0.075 mm under anaerobic conditions at room temperature and pH 4 and 6.5 for a maximum of 160 days. The changes in the concentrations of the major elements in the leaching solutions were monitored. In addition, Fe(II) was analysed separately. pH-independent rate coefficients kH+ were 4.8∙10−10, 6.9∙10−10, 6.3∙10−11, and 1.0∙10−12 mol1-n m−2 s-1, for Fe, Fe(II), Mn, and Si, respectively. The corresponding proton reaction orders nH+ were 0.61, 0.63, 0.33, and 0.09, respectively. The corresponding parameters for Al were not evaluated because of a suspected gibbsite precipitation at pH 6.5. The dissolution of biotite was found to be incongruent (non-stoichiometric) with respect to both the dissolving elements and the pH value. At pH 4, the dissolution was dominated by the octahedral layer element Fe, whereas at pH 6.5, the dissolution of the tetrahedral element Si dominated. There was no evidence of secondary phase formation, and the biotite leaching rates were consistent with those reported in previous studies conducted under aerobic conditions. In addition, the Fe(III)/Fetot ratio of biotite remained essentially unchanged before and after the experiment. This indicates that the anaerobic conditions alone have little effect on the rate and nature of biotite dissolution, although they may influence vermiculite formation. Therefore, biotite dissolution rates previously obtained under aerobic conditions may also be valid under anaerobic conditions.