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Determination of plasma lactic acid concentration and specific activity using high-performance liquid chromatography
B Bleiberg1, J J Steinberg, S D Katz
1Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461.
Journal of Chromatography
|August 23, 1991
Summary
This study introduces a new analytical method using radioactive tracers and HPLC to measure plasma lactate concentrations and specific activities. This technique accurately determines lactate turnover, uptake, and release rates, crucial for understanding exercise and disease states.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Physiology
Background:
- Lactate metabolism is critical in physiological states like exercise and pathological conditions such as diabetes and shock.
- Accurate assessment of lactate kinetics is essential for understanding metabolic disturbances.
Purpose of the Study:
- To develop and validate a novel analytical method for simultaneous determination of plasma lactate concentrations and specific activities (SAs).
- To enable precise quantification of lactate turnover, organ uptake, and release rates.
Main Methods:
- Utilized radio-active tracers (L-[U-14C]lactate) and high-performance liquid chromatography (HPLC) with a reversed-phase column.
- Employed [3H]Acetate as an internal standard, with specific extraction and derivatization steps for lactate detection.
- Validated the method using control plasma samples, assessing precision, analytical recovery, and linearity.
Main Results:
- Achieved linear calibration curves (r = 0.92) in the range of 0.18-6.0 mM, with 3% precision and 87% analytical recovery.
- Established a detection limit of 36 pmol for lactate.
- Demonstrated the method's utility by measuring systemic lactate turnover at 25.65 mumol/kg per min in a patient with congestive heart failure.
Conclusions:
- The developed isocratic HPLC method accurately measures plasma lactate concentrations and specific activities simultaneously.
- This analytical approach provides a reliable tool for determining systemic lactate turnover, organ uptake, and release rates.
- The method is valuable for research in exercise physiology and clinical conditions involving altered lactate metabolism.