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Decreased storage stability of creatine kinase in a cardiac reperfusion solution
Insights
Creatine kinase (CK) stability in Krebs-Henseleit solution was assessed. Refrigeration at 5°C maintains CK activity for one week, while albumin (≥25 g/L) stabilizes frozen samples for prolonged storage.
Area of Science:
- Biochemistry
- Cardiology
- Biotechnology
Background:
- Creatine kinase (CK) is a key biomarker for myocardial cellular damage.
- Assessing CK levels in reperfusate aids in evaluating cellular injury after heart preservation.
- Understanding CK stability in preservation solutions is crucial for accurate diagnostics.
Purpose of the Study:
- To determine the stability of creatine kinase (CK) in Krebs-Henseleit (KH) solution.
- To investigate the impact of cold storage temperatures and albumin concentrations on CK stability.
- To identify optimal conditions for preserving CK activity in preserved heart samples.
Main Methods:
- Isolated rat hearts were preserved for 24 hours and reperfused using KH solution.
- CK activity in KH reperfusate was measured after storage at different temperatures (5°C, -10°C, -200°C).
- Albumin (49, 25, 12, 6 g/L) was added to assess its stabilizing effect on CK in frozen samples (-10°C) over three weeks.
Main Results:
- CK activity remained high (93%) after one week of refrigeration at 5°C.
- Freezing at -10°C significantly reduced CK activity (63%), while nitrogen freezing (-200°C) caused drastic loss (1.6%).
- Albumin concentrations of 25 g/L or higher maintained 100% CK activity in samples frozen at -10°C for three weeks.
Conclusions:
- Refrigeration at 5°C for one week is suitable for maintaining CK activity in KH solution.
- For extended frozen storage (-10°C), albumin (≥25 g/L) is essential to stabilize CK and ensure reliable assessment of myocardial damage.
Abstract:
Creatine kinase (CK; EC 2.7.3.2) has been used as an indicator of myocardial cellular damage. In this study we used a Krebs-Henseleit (KH) solution to reperfuse isolated rat hearts after 24 h of cold preservation and collected the KH reperfusate for assay of CK to assess cellular damage. We wanted to determine the stability of CK in the KH solution at different cold-storage temperatures and albumin concentrations. CK activity (mean +/- SEM) after one week of refrigeration (5 degrees C) was 93% +/- 1% of control values, whereas CK activity in nitrogen-frozen (-200 degrees C) samples was only 1.6% +/- 1% of control values, and that in samples frozen at moderately low temperatures (-10 degrees C) was 63% +/- 1% of control values. To enhance stability, we added albumin at several concentrations (49, 25, 12, and 6 g/L) to reperfusion collections in which CK had been previously determined. Specimens were frozen (-10 degrees C), then re-analyzed for CK weekly for three weeks. CK activity was maintained (100% +/- 5%) only in samples containing 25 g/L or more albumin. These data suggest that refrigeration (5 degrees C) for one week maintains normal CK activity in KH solution; however, if prolonged storage is necessary, a stabilizer such as albumin (greater than or equal to 25 g/L) will maintain analyte stability in frozen storage (-10 degrees C) for at least three weeks.