Abstract: TH-PO0997
Effect of Sample Preservation Method on Kidney Adenylate Levels: Perchloric Acid vs. Flash Freezing
Session Information
- Transplantation: Basic - Immune Biology, Tissue Injury, and Emerging Technologies
October 22, 2026 | Location: Exhibit Hall A, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Transplantation
- 2001 Transplantation: Basic
Authors
- Cuadrao, Sebastian, University of South Florida, Tampa, Florida, United States
- Tian, Zhi, University of South Florida, Tampa, Florida, United States
- Liu, Ruisheng, University of South Florida, Tampa, Florida, United States
Background
Snap freezing has been the most standard method of long-term organ preservation, due to its efficiency and simplicity. Although this method proves beneficial for multiple types of measurements, Adenosine Triphosphate (ATP) preserved in perchloric acid show improved results for ATP preservation due to the acid stopping all enzymatic activity within the cell. This study suggests utilizing perchloric acid in organ sample preservation as an alternate and improved method for researchers to measure ATP values, specifically during investigation of organ transplant viability.
Methods
Mice were anesthetized in a box with 2.5L of Isoflurane and then moved into a surgical table where it stayed anesthetized with oxygen through an external tube throughout the procedure. Lungs, liver, brains, kidney and heart were collected and put in two groups:1) Snap Frozen and 2) 500uL of 0.4M perchloric acid. Furthermore, samples were put in groups of weeks 0,1,2 and 4 weeks after euthanization. Perchloric acid treated samples were homogenized with Radnot homogenizing tubes on ice. Samples were then centrifuged for 10 minutes at 4°C at 11,000g, removing proteins from solutions and isolates the adenylate pool. The supernatant was collected into a new 1.5ml tube and neutralized with 50ul of 2M K2CO3. Solution was kept on ice for 10 minutes and then stored in -80°C freezer for one hour. Samples were centrifuged again for 10 minutes at °4C at 11,000g. Fresh samples were then measured using High Performance Liquid Chromatography (HPLC)
Results
Results in the kidneys show that samples protected with perchloric acid show a small steady decline of ATP across 4 weeks of measurement. Snap frozen treated show a significant falloff of ATP after week 1 and exponential increase of AMP, showing that it does not protect ATP in the long term conventional method of storage.
Conclusion
This method highlights the importance of perchloric acid in significantly reducing the degradation of ATP across various tissue samples across four weeks. Perchloric acid produced more consistent and accurate ATP concentration measurements than compared with snap frozen samples, which most likely was caused by hydrolysis still occurring. Overall, this study demonstrates the need of an optimized ATP preservation method and can be served as a foundation for future transplant research to improve organ viability for the improvement of transplant surgeries.