Additionally the sperm fertility and the sperm ATP
levels were measured in those experiments, which had a positive
effect on the viability of spermatozoa (sperm incubation in SMIS
containing 0.25 mmol/l palmitic acid, 0.005 mmol/l arachidonic acid,
or 0.25 mmol/l linoleic acid). Sperm incubation in pure SMIS served as
control. Washed spermatozoa from 8 semen samples were used and
incubated in the described solutions for 72 h at 4 °C and at a cell
density of 5–7×107 cells ml−1. Immediately after dilution and after
72 h incubation, subsamples were taken and ATP was extracted with
3 mol/l perchloric acid. Using the UV-spectrophotometric assay of
Bergmeyer (1985) the samples were centrifuged to remove precipitated
protein, neutralized and ATP levels were analyzed.