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Effect of storage conditions on lactate dehydrogenase released from perfused hearts
Insights
Storing lactate dehydrogenase (LDH) samples at 4 degrees C for 24 hours maintains enzyme activity. Avoid frozen storage to prevent significant loss of LDH marker integrity.
Area of Science:
- Biochemistry
- Physiology
Background:
- Lactate dehydrogenase (LDH) is a critical biomarker for assessing tissue damage.
- Current research practices exhibit variability in storing coronary effluents before LDH activity measurement.
- This study investigates the impact of diverse storage conditions on LDH activity.
Discussion:
- Storage at 4 degrees C for 24 hours showed no significant decline in LDH activity.
- Room temperature (23 degrees C) storage led to a 42% LDH decline within 24 hours and 98% within 48 hours.
- Frozen storage at -20 degrees C and -80 degrees C resulted in substantial LDH activity loss (79% and 40%, respectively).
Key Insights:
- Short-term (24h) storage of coronary effluents at 4 degrees C is reliable for preserving LDH activity.
- Refrigerated storage at 4 degrees C is superior to room temperature or frozen conditions for LDH sample stability.
- Freezing samples significantly compromises the integrity of LDH as a tissue damage marker.
Outlook:
- Standardizing short-term refrigerated storage protocols can enhance the reliability of LDH measurements in research.
- Further studies could explore optimal long-term storage solutions for LDH samples.
- These findings are crucial for consistent and accurate assessment of tissue injury in cardiovascular research.
Background:
Release of lactate dehydrogenase (LDH) is a widely used marker of tissue damage. A review of the literature indicates that after collection of coronary effluents from hearts, a variety of storage conditions have been utilized prior to measurement of LDH activity. The purpose of this study was to determine whether storage conditions reported in the literature affect LDH activity.
Methods:
Isolated rat hearts perfused with Krebs-Henseleit buffer (KHB) were subjected to 30 min of global ischemia followed by normoxic reperfusion to generate tissue damage and elevate LDH release. Coronary effluents were collected and stored at 4 degrees C, 23 degrees C, -20 degrees C, and -80 degrees C prior to measurement of LDH activity.
Results:
After 24 h no decline in LDH occurred at 4 degrees C storage (P>0.05), but a 42% decline occurred at 23 degrees C (P<0.05). After 48 h, activity declined (P<0.05) 11% at 4 degrees C and 98% at 23 degrees C. Frozen storage resulted in a 40% loss at -80 degrees C and a 79% loss at -20 degrees C (P<0.05).
Conclusions:
Short-term storage in Krebs-Henseleit buffer for 24 h at 4 degrees C does not affect LDH activity and frozen storage should be avoided.
