N R Anderson1, J Nicholas, M R Holland, R Gama. 1. Department of Clinical Chemistry, New Cross Hospital, Wolverhampton, WV10 0PQ, UK. neil.anderson@rwh-tr.nhs.uk
Abstract
BACKGROUND: We investigated whether increased protease activity explains the increased in vitro degradation of intact parathyroid hormone (iPTH) observed in serum when compared to EDTA plasma. METHODS: Pre-dialysis blood samples for iPTH were taken from 11 patients with chronic renal failure and collected into plain glass tubes, tubes containing 200 KIU/mL aprotinin (a protease inhibitor) and EDTA tubes. All sample aliquots were separated at 20 min, 1 h, 2 h, 4 h, 8 h and 24 h post collection. RESULTS: Over 24 h, iPTH concentrations remained unchanged in EDTA tubes. iPTH concentrations were significantly lower in both plain tubes (P < 0.01) and aprotinin tubes (P < 0.001) at 24 h when compared to the baseline sample (20 min). At 24 h, iPTH concentrations in EDTA tubes were higher than in plain tubes (P < 0.001) and aprotinin tubes (P < 0.01). The addition of aprotinin to plain tubes significantly reduced the degradation of iPTH (P < 0.05) at 24 h. CONCLUSION: Aprotinin significantly reduces the in vitro degradation of iPTH in plain tubes at 24 h from 24.7% to 9.6%. We suggest that increased protease activity contributes to the decline in serum iPTH over time. As this is observed in serum and not plasma it suggests that the increased protease activity may be due to the clotting process.
BACKGROUND: We investigated whether increased protease activity explains the increased in vitro degradation of intact parathyroid hormone (iPTH) observed in serum when compared to EDTA plasma. METHODS: Pre-dialysis blood samples for iPTH were taken from 11 patients with chronic renal failure and collected into plain glass tubes, tubes containing 200 KIU/mL aprotinin (a protease inhibitor) and EDTA tubes. All sample aliquots were separated at 20 min, 1 h, 2 h, 4 h, 8 h and 24 h post collection. RESULTS: Over 24 h, iPTH concentrations remained unchanged in EDTA tubes. iPTH concentrations were significantly lower in both plain tubes (P < 0.01) and aprotinin tubes (P < 0.001) at 24 h when compared to the baseline sample (20 min). At 24 h, iPTH concentrations in EDTA tubes were higher than in plain tubes (P < 0.001) and aprotinin tubes (P < 0.01). The addition of aprotinin to plain tubes significantly reduced the degradation of iPTH (P < 0.05) at 24 h. CONCLUSION: Aprotinin significantly reduces the in vitro degradation of iPTH in plain tubes at 24 h from 24.7% to 9.6%. We suggest that increased protease activity contributes to the decline in serum iPTH over time. As this is observed in serum and not plasma it suggests that the increased protease activity may be due to the clotting process.