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Bone turnover markers are differentially affected by pre-analytical handling.

G L Christensen1, J R Halgreen1, M Milenkovski1

  • 1Department of Clinical Biochemistry, Rigshospitalet, Copenhagen, Denmark.

Osteoporosis International : a Journal Established As Result of Cooperation Between the European Foundation for Osteoporosis and the National Osteoporosis Foundation of the USA
|January 26, 2019
PubMed
Summary

Bone turnover marker stability varies by sample type and storage. N-terminal pro-collagen type 1 extension pro-peptide (P1NP) and bone-specific alkaline phosphatase (BAP) are stable in serum and plasma for 72 hours. Osteocalcin (OC) degrades quickly, requiring prompt analysis or freezing.

Keywords:
BAPBone turnover markersCTXOsteocalcinP1NPPre-analyticalStability

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Area of Science:

  • Biochemistry
  • Clinical Chemistry
  • Laboratory Medicine

Background:

  • Bone turnover markers (BTMs) are crucial for diagnosing and monitoring bone diseases.
  • BTMs are often analyzed in central laboratories, necessitating an understanding of their stability in various sample types and storage conditions.
  • Accurate BTM analysis depends on sample integrity during pre-analytical phases.

Purpose of the Study:

  • To evaluate the stability of four key bone turnover markers: N-terminal pro-collagen type 1 extension pro-peptide (P1NP), bone-specific alkaline phosphatase (BAP), C-terminal teleopeptide cross links of collagen type 1 (CTX), and osteocalcin (OC).
  • To determine the impact of sample type (serum, EDTA plasma, LiHep plasma), time before separation, and time after separation on BTM stability.
  • To provide recommendations for optimal sample handling and storage to ensure reliable BTM measurements.

Main Methods:

  • Serum, EDTA plasma, and LiHep plasma samples were collected from osteoporosis patients and healthy controls.
  • Samples were stored at room temperature for up to 72 hours before separation.
  • Post-separation, samples were stored at room temperature for up to 72 hours before analysis using the automated iSYS IDS platform.

Main Results:

  • P1NP and BAP demonstrated stability in both serum and plasma for up to 72 hours pre-centrifugation.
  • CTX levels were consistently higher in EDTA plasma compared to LiHep plasma and serum, with EDTA plasma extending CTX stability.
  • Osteocalcin (OC) exhibited significant degradation across all sample types, with concentrations dropping substantially after 24 hours.

Conclusions:

  • P1NP and BAP are stable in both serum and plasma, making either suitable for analysis.
  • EDTA plasma is recommended for CTX analysis to ensure optimal stability.
  • Osteocalcin (OC) requires prompt analysis (within 3 hours) or storage at -18°C due to its rapid degradation.