Abstract: The method for efficiency calibration of tritium sample in liquid scintillation spectrometry was presented. The quenching effects from different chemical quenchers (Acidbase, CH₃NO₂, CCl₄, CH₃COCH₃) and color quencher (Na₂CrO₄) were studied. For each quencher, the methods of sample channel ratio (SCR), spectrum index of the sample (SIS) and spectral quenching parameter of the external standard (SQP(E)) were used for efficiency calibration respectively, and three methods were compared. The results show that the quenching from the various chemical quencher can be unified for one chemical quenching for efficiency calibration. There is great difference in the correction curves of chemical quenching and color quenching, and the fact is independent of the used efficiency calibration method. The SCR method is not advantageous for the tritium sample with low radioactivity or strong quenching. The SQP(E) method is independent of the sample count rate, and it is especially suitable for the efficiency calibration of low radioactivity tritium. The SIS method can be used for samples with high radioactivity. The accurate efficiency calibration for various quenching can be carried out by combining the SIS method and the SQP(E) method.