A simple CO₂ equilibration method for measuring blood oxygen isotope compositions

Abstract

Rationale: Blood water oxygen isotope compositions can provide valuable insights into physiological processes and ecological patterns. While blood samples are commonly drawn for medical or scientific purposes, blood fractions are infrequently measured for oxygen isotopic compositions (δ 18 O) because such measurements are time consuming and expensive. Methods:We sampled blood from sheep, goats, and iguanas raised in field and animal laboratories into serum, EDTA, heparin, and uncoated plastic vials commonly used in medical and scientific research, then separated red blood cell (RBC) and plasma or serum blood fractions. These were injected into helium-flushed Exetainer tubes where they naturally outgassed endogenous CO 2 (goat blood), or into He-and CO 2 -flushed tubes (iguana blood). The CO 2 gas was sampled on a GasBench II system, and δ 18 O was measured by an isotope ratio mass spectrometer (IRMS).Results: Repeated δ 18 O measurements were stable over multiple days. The addition of desiccated blood solids to water standards had little impact on their δ 18 O measurements, suggesting that organic molecular constituents within blood serum and plasma do not interfere with blood water δ 18 O values. We observed slight but statistically significant δ 18 O offsets between plasma, serum and RBC fractions. Massdependent body water turnover times for iguanas were derived from the data. Conclusions:We demonstrate that a simple blood-CO 2 equilibration method using the GasBench can quickly, reliably and accurately characterize water δ 18 O in the plasma, RBC, and whole blood fractions of mammalian and reptilian blood samples (precision ≤ 0.1‰). This method will expand the application of blood stable isotope analysis in physiological and medical research. | INTRODUCTIONStable isotope analyses of body fluids are a potent tool for evaluating human and animal water budgets, metabolism and energy use (e.g., 1 ).The utility of isotopes in biological and health research was recognized almost immediately after their discovery. In a conversation about the demonstration and publication of oxygen and hydrogen isotopes, 2,3 George de Hevesy, Erich Hofer, and Henry Mosely wondered if isotopes could trace the path of a cup of tea through the human body. 4 Hevesy and Hofer decided to investigate the possibility by providing isotopically heavy drinking water to human subjects while measuring the density of their urine over time, recognizing that