Plasma ADP levels: direct determination with luciferase luminescence using a biometer

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• Accurate quantification of adenosine-5'-diphosphate (ADP) in plasma is essential for understanding platelet function and various physiological processes.
• Existing methods for measuring low plasma ADP levels can be challenging and require sensitive techniques.

Purpose of the Study:

• To develop and validate a sensitive method for determining low plasma concentrations of adenosine-5'-diphosphate (ADP).
• To apply this method for the measurement of ADP in human and rabbit plasma.

Main Methods:

• Utilized the luciferin-luciferase luminescence reaction for ADP detection.
• Employed enzymatic conversion of ADP to ATP using creatine phosphate and creatine kinase after endogenous ATP removal.
• Quantified luminescence using a Dupont Biometer.

Main Results:

• Achieved high precision with mean coefficients of variation of 2.1% and 1.8% for ADP standards at 0.02 and 2.2 micromol/liter, respectively.
• Determined mean human plasma ADP concentrations of 0.13 ± 0.025 micromol/liter (n=10).
• Determined mean rabbit plasma ADP concentrations of 0.07 ± 0.05 micromol/liter (n=5).

Conclusions:

• The described luciferin-luciferase assay provides a reliable and sensitive method for quantifying low plasma ADP levels.
• This method offers potential for broader applications in clinical diagnostics and biomedical research.
• The findings establish baseline ADP levels in human and rabbit plasma using this novel technique.