Serum beta-crosslaps are an established laboratory test to investigate bone metabolism. However, lytic conditions may affect the measurement of serum beta-crosslaps. Therefore, we investigated the effect of cathepsin C and human leukocyte lysate on serum beta-crosslaps in relation to temperature and time. We divided eight serum samples with elevated beta-crosslaps levels into three aliquots and stored them at 4, 21 and 37 degrees C. Another five serum samples were divided into three aliquots and adjusted to contain different cathepsin C concentrations (50, 250, 500 IU/l). These aliquots were divided again and stored at 4, 21 and 37 degrees C. Finally, three aliquots from three additional serum samples were treated with human leukocyte lysate (100, 300, 500 microl), divided again and stored at 4, 21 and 37 degrees C. Measurements of serum beta-crosslaps were then carried out before and immediately after manipulation, and after 2 and 5 days of storage. When stored at 21 degrees C, serum beta-crosslaps diminished significantly (25% after 5 days), but no significant change was detectable when they were stored at 4 degrees C. Cathepsin C induced up to a 14% increase in beta-crosslaps while human leukocyte lysate caused up to a 17% decrease. This study demonstrates that the influence of proteolytic conditions on the serum concentration of beta-crosslaps is not uniform. Leukocyte lysate decreased serum beta-crosslaps while the addition of cathepsin C increased their concentration. Therefore, serum should be separated from the whole blood immediately after coagulation and stored until analysis in a deep freezer.