The variability of control samples is the main problem in clinical proteomics and reliable sources for estimation of variability of normal content of different proteins are not numerous. Most of the investigations of normal human tear proteome include pool of samples as reference materials. But it is impossible to use such experimental approach to establish the range of variability of different tear proteome components. This study is the first attempt to estimate the variability of proteins content in healthy donors tear using high-performance liquid chromatography and tandem mass spectrometry. The protein profiles from 11 individual healthy donors were analyzed. Essential variability of the minor proteins was revealed while the presence of 6 major proteins in all 11 samples was invariable. We found the Lacritin glycosylation in all samples of tear fluid received from healthy donors. It seems that this modification is typical for healthy donors tear. The analysis of the pool samples was also carried out to estimate the availability for this approach for the search of ophthalmologic diseases biomarkers.