Proc. Natl. Acad. Sci. USA Vol. 84, pp. 4099-4102, June 1987 Biophysics Deuterium nuclear magnetic resonance measurements of blood flow and tissue perfusion employing 2H20 as a freely diffusible tracer (rat liver/surface coil/spin-lattice relaxation/heavy water) JOSEPH J. H. ACKERMAN*t, COLEEN S. EwY*, NANCY N. BECKER*, AND ROBERT A. SHALWITZtt *Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110; and *Department of Chemistry, Washington University, St. Louis, MO 63130 Communicated by William D. Phillips, January 28, 1987 ABSTRACT The use of deuterium oxide (21120) is pro- We propose herein the analogous use of 2H20 as a posed as a freely diffusible nuclear magnetic resonance (NMR) diffusible blood-flow and tissue-perfusion tracer where deu- blood flow and tissue perfusion tracer of potential clinical terium NMR is used to monitor label intensity as a function utility. Deuterium is a stable, nonradiative isotope commer- of time. Deuterium is a nonradiative, naturally abundant cially available as 2H20 at enrichment levels of essentially (0.0156%), quadrupole nuclide with only 0.01 the sensitivity 100%-i.e., 110 molar equivalent deuterium. This high con- of 'H. Nevertheless, the combination of an efficient centration, together with the short relaxation time of the spin quadrupole relaxation mechanism and the commercial avail- 1 (quadrupole) deuterium nuclide, provides substantial sensi- ability of 2H20, essentially 100% enriched to ca. 110 molar tivity for NMR spectroscopy. As a result, when 2H20 is equivalent deuterium, can provide substantial NMR signal- administered in a bolus fashion to a specific tissue or organ in to-noise in the intact subject. This is readily illustrated herein vivo, the deuterium NMR intensity time course can be ana- through surface-coil NMR (19, 20) examination of rat liver in lyzed, using mathematical models developed by others for vivo employing an animal model developed previously in our radiolabeled tracers, to measure the rate of blood flow and laboratory. Portions of this study have been presented in tissue perfusion. Such an application is demonstrated herein at abbreviated format (22, §). a static magnetic field of 8.5 tesla. Using single-compartment flow modeling, hepatic blood flow and tissue perfusion in fasted METHODS (18 hr) male Sprague-Dawley rats was determined to be 61 ± 17 (mean ± SD) ml/100 g per min (n = 5). Male Sprague-Dawley rats (Sasco, Omaha, NE) were fasted for 18 hr, weighed (ca. 225 g), and anesthetized with The measurement ofregional blood flow and tissue perfusion Halothane (5% for induction, 2% for surgery, 1% for main- provides an important clinical indicator oftissue viability and tenance). Animals remained under anesthesia throughout the vascular competency. Nuclear magnetic resonance (NMR) entire experimental protocol and were sacrificed at its con- flow measurement techniques, including those based on clusion. Since magnetic resonance imaging localization tech- spin-imaging procedures, that employ observation of the niques were unavailable, spatial localization was achieved naturally abundant 1H resonance from H20 have been surgically in our studies. A 4-cm transverse, subcostal developed (for reviews, see refs. 1-3). These techniques incision was made in the abdomen to expose the liver. The generally rely on the spatial evolution of 1H magnetization xiphoid process was removed and the falciform ligament was after the initial preparation of a nonequilibrium spin-state cut. An infusion catheter was secured in a branch of the population distribution. Therefore, magnetic resonance flow superior mesenteric vein and kept patent with heparinized measurements are often limited by the lifetime of the non- saline (0.9% NaCl) infused at 0.013 ml/min. The abdomen equilibrium state and, thus, by the water spin-lattice and was wrapped in plastic wrap to prevent moisture loss and the spin-spin 1H relaxation times. However, such approaches are animal was secured in the cradle of the NMR probe. (Body extremely attractive because oftheir completely noninvasive temperature was maintained at 370C by warm water circula- nature and their development remains an area of very active tion.) The probe and animal were then brought to the vertical position and the surface-coil antenna was placed on a investigation (4, 5). liver. Alternatively, as described in the early pioneering work of 1-mm-thick glass plate positioned over the Kety (6-8), one can introduce an exogenous substance as a Following standard blood-flow measurement protocols (9), a radiolabel. Al- the time evolution of the deuterium NMR integrated signal blood-flow tracer, usually one containing intensity from rat liver in vivo was followed with typically though the administration of the tracer substance introduces ml of an invasive character to the flow measurement, the high 0.51-sec time resolution after bolus injection of0.2 2H20 sensitivity of nuclear decay detection and the use of into a branch of the superior mesenteric vein. radiolabels that decay slowly relative to blood flow result in The surface-coil antenna was a coaxial double-coil arrange- measurement methods of considerable practical utility ment similar to that described for in vivo 13C{'H} experiments that (20). An outer 2-cm-diameter coil tuned to 360 MHz was used (9-11). Importantly, by employing tracer substances to observe 'H (i.e., H20) for optimization of the static field freely diffuse through aqueous spaces (i.e., intracellular, (8.5 tesla) homogeneity while the smaller inner 1-cm-diame- interstitial), a quantification oftissue perfusion is achieved in ter coil was used for deuterium observation at 55 MHz. contrast to a measurement of simply linear or plug flow During the measurement of hepatic blood flow (HBF) and velocity in large vessels. One such tracer substance that has data and were to seen considerable use in combination with positron emission tissue perfusion, collection storage assigned detection is H2150 (12-18). Abbreviation: HBF, hepatic blood flow. tTo whom reprint requests should be addressed. The publication costs of this article were defrayed in part by page charge §Ewy, C. S., Bennett, D. L., Ackerman, J. J. H. & Shalwitz, R. A., payment. This article must therefore be hereby marked "advertisement" Fifth Annual Meeting of the Society of Magnetic Resonance in in accordance with 18 U.S.C. §1734 solely to indicate this fact. Medicine, Aug. 19-22, 1986, Montreal, pp. 86-87 (abstr.). Downloaded by guest on September 30, 2021 4099 4100 Biophysics: Ackerman et al. Proc. Natl. Acad. Sci. USA 84 (1987) computer memory throughout the experiment to avoid disk External access timing delays. A nonlinear least squares fitting routine reference was used to fit the experimental resonance lines to a Lorentz- ian lineshape (23). From this simulated lineshape function, an A analytical (i.e., closed form) determination of the integrated resonance area was obtained. RESULTS AND DISCUSSION The results ofa spin-lattice relaxation time (T1) measurement (24) on the naturally abundant deuterium present in hepatic tissue water (1H2HO) are shown in Fig. 1. Nonlinear least squares analysis in terms of a three-parameter, single- exponential magnetization-recovery time course yields a time constant T1 = 0.25 sec. This relatively short T1 allows rapid signal-to-noise enhancement through pulsed Fourier B transform time-averaging techniques (25) and, in combina- tion with a high isotopic enrichment of 2H20, provides for short time resolution serial monitoring of the deuterium resonance intensity. Fig. 2 illustrates a natural abundance background deuterium signal from rat liver in vivo (160 scans External summed over 82 sec) and the hepatic signal a few seconds reference after bolus injection of 0.2 ml of 2H20 into a branch of the superior mesenteric vein (1 scan). The superior mesenteric vein leads directly into the portal vein through which ca. 70% of the hepatic blood supply passes. Clearly, this model 10 8 6 4 system using 2H20 as a freely diffusible tracer has adequate ppm signal-to-noise and time resolution to demonstrate the mea- FIG. 2. Representative deuterium spectra from rat liver in vivo at surement of blood flow and tissue perfusion in the intact 55.28 MHz. (A) Natural abundance deuterium spectrum consisting of animal. 160 free induction decays accumulated and summed over 82 sec and Fig. 3 shows that immediately following bolus administra- Fourier-transformed after application of a 10-Hz exponential filter. The external reference resonance represents a small capillary, tion, there is a very rapid marked increase in integrated positioned between the lobes of the liver, containing C62H6 plus resonance intensity as the deuterated tracer enters the liver chromium acetyl acetonate as a relaxation (and shift) agent. (B) vascular space and diffuses into the tissue. Concurrent rapid Single transformed free induction decay (0.20-sec accumulation) of proton exchange yields 1H2HO. This is followed by a slow the same liver as in A 4 sec after a bolus injection of 0.2 ml of 2H20 exponential decrease in integrated signal intensity (note the into a branch of the superior mesenteric vein. A 10-Hz exponential logarithmic ordinate in Fig. 3) indicative of hepatic 'H2HO filter was applied prior to Fourier transformation. 80- washout by ongoing blood flow. No overshoot of integrated signal intensity prior to washout is observed showing that HBF is slow enough to ensure accurate approximation of 2H20 as a freely diffusible tracer (13). 60- Given a one-compartment model (rapid tracer diffusion between vascular and extravascular spaces) and assuming a bolus tracer input (i.e., a pseudo delta function), homoge- neous tissue vascularization with no intracompartment tracer 4._m 4) concentration gradients, and negligible recirculation of la- 14 beled material, an exponential decay of tissue tracer concen- tration is expected (9-11). The tracer washout exponential time constant (7) can be related to blood flow per unit volume (or mass) of tissue through the central volume principle of 20- tracer kinetics (10, 11).