Present review is to introduce an omasal sampling technique in rumen proteolysis and to consider some information on the omasal sampling technique with particular emphasis on methodological aspects. Use of the omasal sampling technique provides a new opportunity for accurate estimation of rumen metabolism with overcoming limitations of previous in vivo, in vitro and/or in situ methods. The potential advantages of the present technique compared with post-ruminal sampling techniques include following points; 1) only rumen cannulated animals are required, 2) less endogenous nitrogen (N) is contaminated in omasal digesta and 3) omasal digesta are devoid of exposure to acid peptide hydrolysis occurring in the abomasum. Estimates of soluble non-ammonia N (SNAN) in omasal digesta indicate that the assumptions underlying the in situ method that rapidly degradable N fraction can be degraded at an infinite rate and only insoluble dietary N escapes the rumen may be not valid. Quatitatively higher peptide concentration rather than free amino acid and soluble protein in escapable SNAN suggests that hydrolysis of peptide to amino acid may be the rate-limiting step in rumen proteolysis.