Extraction and estimation of ATP from human spermatozoa

This study attempts to provide guidelines for the extraction and estimation of ATP from spermatozoa. The conditions of extraction of ATP from spermatozoa using trichloroacetic acid were optimized. The efficiency of this extraction procedure was compared to that using boiling Tris buffer. Three commercially available kits which use the firefly luciferase assay for ATP and two luminometers were evaluated for the measurement of the spermatozoa extracts. Elevated levels of Mg2+ and/or Ca2+ did not interfere with the extraction of ATP by trichloroacetic acid or with the subsequent estimation of the nucleotide.