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Comparison of Gd DTPA-BMA (Omniscan) versus Gd HP-DO3A (ProHance) Retention in Human Bone Tissue by Inductively Coupled Plasma Atomic Emission Spectroscopy
Wendell A. Gibby, MD,* Krissa A. Gibby,† and W. Andrew Gibby‡

Rationale and Objectives: Human bone tissue was collected following administration of a clinical dose of gadolinium chelate (0.1 mmol per kg) to patients undergoing hip joint replacement surgery to determine if measurable differences in Gd deposition occur between 2 widely available magnetic resonance contrast agents. Materials and Methods: Gd HP-DO3A (ProHance), Gd DTPABMA (Omniscan), and an age-matched control population without history of gadolinium chelate administration were compared. Bone samples were collected fresh, placed in refrigeration, and subsequently frozen. Tissue digestion was performed using a microwave tissue digester and concentrated nitric acid. A method of tissue analysis was created for gadolinium using inductivity coupled plasma atomic emission spectroscopy (ICP-AES). Results: Tissue retention was 1.18 Ϯ .787 ␮g Gd/g bone (N ϭ 10) for Omniscan and 0.466 Ϯ .387 ␮g Gd/g bone (N ϭ 8) for ProHance measured by ICP-AES. Conclusion: Omniscan (Gd DTPA-BMA) left 2.5 times more Gd behind in bone than did ProHance (Gd HP-DO3A). Key Words: human bone gadolinium retention, gadolinium ICPAES, Gd HP-DO3A, Gd DTPA-BMA (Invest Radiol 2004;39: 138 –142)

adolinium (Gd) chelates are widely used as contrastenhancing agents for magnetic resonance imaging. The structure of the chelate in part determines the kinetic lability of the metal ligand and, in turn, can affect the release of the metal in vivo. Numerous in vitro studies have indicated


Received August 29, 2003 and accepted for publication, after revision, November 23, 2003. From the *Riverwoods Advanced Imaging Center, Provo, Utah; the †Department of Tumor Biology, Georgetown University, Washington, DC; and ‡Magnetic Research Inc., Provo, Utah. Reprints: Wendell A. Gibby, MD, Riverwoods Advanced Imaging Center, 280 W. Riverpark Dr., Ste. 100, Provo, UT 84604. E-mail: Copyright © 2004 by Lippincott Williams & Wilkins ISSN: 0020-9996/04/3903-0138 DOI: 10.1097/01.rli.0000112789.57341.01

differences in Gd chelate affinities and in Gd chelate kinetic labilities. These can be affected by a variety of factors, including the pH, the availability of other metal ions that compete with the Gd on the chelate, and the structure of the chelate. Gd DTPA-BMA is a substituted open-chain chelate which, although having somewhat similar thermodynamic stability1 to the ring compound Gd HP-DO3A, has vastly different kinetic stabilities as a result of the relatively rigid ring structure of HP-DO3A.2 Animal studies have indicated repeatedly an increase in Gd retention for the more labile Gd DTPA-BMA material both as a chemical entity and as formulated for clinical use. However, there have been no human studies to date that have attempted to demonstrate a significant difference in Gd retention. The retention of Gd could be important clinically, because Gd is not a naturally occurring biologic constituent and, once within the tissues of animals, persists for long periods of time.3 It has significant toxicities, both in in vitro and in vivo experiments.4 – 6 For example, it is the most potent calcium antagonist known.7,8 Gd has the potential of leeching into membranes, bone, and enzymatic structures, causing as-yet undetermined long-term consequences. Therefore, the release of Gd into the human body is of significant clinical interest.3,9,10 This study was undertaken to compare 2 U.S. Food and Drug Administration-cleared, commonly used Gd-based magnetic resonance imaging chelates, Gd DTPA-BMA and Gd HP-DO3A, in their retentive properties in human bone tissue. Human bone tissue was selected for 2 reasons: 1) it is available in certain orthopedic procedures, whereas other tissues such as the liver, spleen, and so on, are not readily acquired; and 2) bone is one of the target organs in which Gd retention occurs.

Patients undergoing a total hip arthroplasty with removal of the femoral head were enrolled after informed consent. Gd DTPA-BMA or Gd HP-DO3A was injected
Investigative Radiology • Volume 39, Number 3, March 2004


3 Ϯ 12. laboratory. and the volume of the sample is then adjusted to 10 mL with distilled water. Table 1 indicates the age distribution and timing of the Gd injection between the 2 groups. Replicates: 3. Target temperature was 95°C and the time at temperature was 15 minutes. 0. Plasma 96.2 parts per million. which contained marrow. Instrument stabilization delay: 5 seconds. 1 mL of the diluted Yttrium solution (10 ␮g/g H20) is added. Standard solutions also contained 100 mL of 70% nitric acid and 36. creating a 10. Power: 1 kW.Investigative Radiology • Volume 39. along with 36.03). The other half was sent to Bracco Research for independent analysis.75 L/min.12 nm. Grating order: 1. March 2004 Comparison of Gd DTPA-BMA vs Gd HP-DO3A Retention intravenously at a dose of . Yttrium and Gd standards were weighed with an OHAUS Explorer analytical balance capable of measuring to 0. significant charring occurs and the sample must be discarded. The parameters for Yttrium were 1-second integration time with automatic background correction and an analog wavelength of 371.68 g CaCl2 per 500 mL H2O.000 ␮g Yt/g H20 solution. 1). The 4-Gd standards were also run at the beginning and end of each run.5 L/min. Gd linearity and limit of detection was measured for standard solutions linear and reproducible to . Software version 1.7 4. Viewing height: 10 mm. Wavelength calibration and torch alignment were performed before each run.036. Peak tracking window: 0. Auxiliary flow: 0. Introduction settings: Sample uptake delay: 10 seconds.0 to 3.9 *There was no significant difference between groups. Gd standard calibration solutions were prepared at: 10.12. Each bone assay was performed in triplicate.080 nm.2 N/A Omniscan (N ‫ ؍‬10) 67. General settings: Scan window first order 0.1 and 1. and moisture less than 3 parts per million. Experimental ICP-AES Equipment and Materials Argon from Praxair: 5. 1. Half of the tissue sample was sent to Magnetic Research Incorporated. 70% (vol/vol): with heavy metals less than 0. Yttrium standard for ICP Y2O3 in 2% HNO3. Ramp time was 5 minutes. Nitric acid. Science. An age-matched control population undergoing hip replacement was also obtained. gadolinium standard for ICP both from E.051. Tissue Digestion Bone tissue was digested by removing 1 g (Ϯ 0.0 ␮g Gd/gm bone (Fig. ICP AES Instrument: Varian Liberty Series 2. Manual sample tube aspiration connected to a short piece of Teflon tubing. 1000 ␮g/mL.4 ProHance (N ‫ ؍‬8) 63.) Reagents The following reagents were used: micron-filtered distilled water. Internal diameter: 0. Rinse time: 300 seconds. Age Distribution and Gadolinium Timing Control (N ‫ ؍‬7) Average patient age* (years) Injection to bone harvest* 55. Composition: hydrocarbons less than 1 part per million. The study was performed under the auspices of the Institutional Review Board. Operating conditions: Nebulizer set at 180 psi. The slope error TABLE 1. The bone samples were watched visually until nearly all the nitric acid boiled away. and . Elemental analysis parameters for Gd were set at 1-second integration time with a polynomial-plotted background correction and an analog wavelength of 342.030 nm. Yttrium stock solution was prepared by diluting 1:100 with 5 mL of the Yttrium stock at 10 ␮g/g H20 solution added to 495 mL of distilled water. for analysis.8 4. This was placed in a Pyrex Star System CEM Digestion Flask. When approximately 2.5000 g Yttrium standard solution (1000 ␮g/mL) and sufficient water to constitute 500 mL of stock solution. Twenty milliliters of the nitric acid reagent was automatically pipetted by the CEM instrument (software version 86005/1. the sample is removed. Frozen. Photo multiplier tube voltage: 640. M. oxygen less than 2 parts per million. Integration time: 1 second. © 2004 Lippincott Williams & Wilkins 139 . unprocessed human bone tissue cut with a standard hacksaw in the pathology Recovery of Gd From Spiked Bone The recovery analysis and limit detection of human bone tissue was performed using bone tissue spiked with varying concentrations of Gd from a single control bone between . Plasma flow: 10. Pump rate: 15 RPMs.0001 g weighed in small plastic Dixie cups. 2).68 g CaCl2 ϫ 2 H2O. Bone samples included a slice of cortex and medullary bone. Fischerbrand-approved pump tubes.2004 ␮g Gd/g H20 in a washed 500 mL Pyrex volumetric flask at 23°C.1 mmol/kg not less than 3 days and not more than 8 days before surgery. 05031.247 nm. Gas inlet pressure set at 100 psi.0 mL of residual acid and dissolved bone are present.0043. Gibbstown. (If the sample is not caught before complete loss of the nitric acid.8 Ϯ 20.001 g) from the hip bone samples. NJ. The femoral heads were cut in half. Provo.5 days Ϯ .0 ultra-high purity. Utah. Number 3.6 Ϯ 9. We avoided the dome of the femoral bone where cartilage and degeneration were present.1 ␮g Gd/g solvent (Fig.4 days Ϯ 1.

times 10 mL of solution per 1 g bone. March 2004 FIGURE 1.1 but less than 0.8845. Number 3.54 ϩ . 140 © 2004 Lippincott Williams & Wilkins . was 16%. FIGURE 2. times the density correction factor of water.0075 RESULTS The detection limit of Gd for this system is greater than 0. The recovery of Gd in the spiked control samples was 54%.Gibby et al Investigative Radiology • Volume 39.2 ␮g/g bone. The ␮g/Gm of Gd in the bone sample is equal to measured ␮g/g of bone divided by the internal Yttrium standard. The coefficient of determination (r2) was . The formula used was: Actual Concentration Gd ϭ Measured Gd . divided by the bone weight. The human data was then normalized for Gd recovery using spiked bone samples of Figure 2.

6 primarily within the liver and bone. monkeys injected with Gd DTPA-BMA demonstrate all of the signs of zinc deficiency. Thermodynamic stability measurements at a pH of 7. being bound to membranes.117 Nϭ7 Standard Deviation .787 t Test Control v Omniscan 0. lipid and carbohydrate metabolism. These were subsequently corrected to a large extent by adding more free ligand to Magnevist’s formulation.227 t Test Control v ProHance 0. However.4.09%] of the total zinc pool in the body).11 Unlike metals with a known biologic function. Thermodynamic stability is measured by titrating the metal to the chelate at a pH of approximately 11.”11 all of the currently used magnetic resonance contrast agents have the potential for transmetallation in vivo. Gd does not have a known pathway for excretion from within the body.3 Gd metal injected intramuscularly induces sarcomas4 and “granulomatous neoplasms. hypotension. the potential of transmetallation should be considered for long-term sequelae. Results of Gadolinium Bone Retention Control Average ␮g/g fresh bone tissue Ϫ0. and we tightly controlled the time from injection to bone harvest.13 The excess chelate scavenges the bioavailable metals. including in vivo transmetallation11. At this pH. The more important aspect of chelate safety is that of kinetic stability. including testicular atrophy. free Gd initially goes to the liver and is then transferred to bone with negligible elimination over the next 3 weeks.22–24 suggests that there is a significant difference between the propensity of these 2 chelates to give up Gd in vivo. or other weak chelates such as citrates and phosphates. We did not attempt to determine any relationship between the time after Gd injection and residual trace metal present. changes in nucleic acid. Studies with human volunteers have shown that a single dose of Gd DTPA-BMA removes approximately 32% of total plasma zinc (albeit a small fraction [. enzymes. Yet a variety of studies.”5 These agents are also known to interfere with coagulation. The thermodynamic equilibrium constant calculated in vitro is simply not germane.7 High concentrations of heavy lanthanide metals in animals are known to be toxic and can cause. it can persist for long periods of time. Number 3. For example. Furthermore.21 and human case reports. bone substrate. One of the primary causes of transmetallation in vitro is the competition by other bioavailable metals such as zinc and copper that attack the chelate complex. Once a Gd ion is released.0004 Gd HP-DO3A Average ␮g/g fresh bone tissue . with repeated high dosages in subacute toxicity studies in animals. Once within the tissues. there are metals that will attack the chelate and displace the Gd. fatty degeneration of the liver. This was because the numbers of patients was limited.6.14 However. March 2004 Comparison of Gd DTPA-BMA vs Gd HP-DO3A Retention Table 2 represents the normalized.5 Ϯ .0165 © 2004 Lippincott Williams & Wilkins 141 . it is immediately carried away by other biologic material. there is no equilibrium. the more relevant stability constant for in vivo use is the conditional stability constant.466 Nϭ8 Standard Deviation .4 Ϯ 1.18 N ϭ 10 Standard Deviation .8. in the body.9 days for Gd HPDO3A. skin lesions with ulceration. although it could be the cause of the wellknown elevated serum Fe levels reported initially with the introduction of Gd DTPA. ataxia. and death. and gastritis. However.0021 Gd DTPA-BMA Average ␮g/g fresh bone tissue 1. However. Both chelates have the same number of coordinating sites. Gd is one of the most potent inorganic calcium antagonists known. HP-DO3A is a ring compound that is TABLE 2. measured Gd content in micrograms of gadolinium per gram of bone.4 would indicate that only 1 atom of 1017 (a very small number) of Gd would be loose from the chelate at any given time. The concentration of these metals depends on the microenvironment for the chelate.15 One of the mistakes made in evaluating chelates for in vivo use is to compare thermodynamic stability constants.16 –19 and Gd retention in animals20. there are no competing hydrogen ions for the chelate and a theoretical maximum stability for the chelate is obtained. DISCUSSION Despite claims to the contrary that “there seems to be no dissociation of Gd within the body. In the milieu of the body. Physicians injecting such material into the body should be aware of the potential deleterious effects of free Gd. writhing. which is calculated at a pH of 7. It might never “see” the chelate again. and reasonable efforts should be made to select ligands that reduce this effect. Some chelates such as Gd DTPA-BMA formulate with 5% excess calcium ligand.387 t Test Omniscan v ProHance 0. sedation.4 days in the case of Gd DTPA-BMA and 4. In small quantities. consider 2 chelates of near-equivalent conditional thermodynamic stability: Gd HPDO3A and Gd DTPA.12 In the dosages given clinically. among other things. 4. labored respiration.Investigative Radiology • Volume 39. transmetallation is not known or suspected to be an acute problem.

it is likely that most of the Gd detected 3 to 8 days after administration is released from the original chelate. Stability of gadolinium complexes in vitro and in vivo. Nordenbo AM. 9. Lancet. et al. 3. WIP:1199. et al. Tweedle MF. animal experiments have shown dissociation in vivo. 1992. Acute deterioration of myasthenia gravis after intravenous administration of gadolinium DTPA [Letter]. On the other hand. pregnant patients).8:467– 481. 1994. Invest Radiol. In: Toxicity of Heavy Metals in the Environment. 1980:33–34. 1996. Somnier FE. Stability assessment of gadolinium complexed by P-31 and H-1 relaxometry. Goudemant J-F. J Nucl Med. Weinmann H-J. Tweedle MF. Tweedle MF. 142 © 2004 Lippincott Williams & Wilkins . Talbot RB. Brasch RC. Tien RD.30:372–380. Number 3. White DL. Invest Radiol. et al. Hattner R. et al. 18. del Pozo E. 1992. Puttagunta NR. Thus. Br J Pharmacol. Deposition of gadolinium after high and low doses of gadolinium chloride to mice: organ distribution. 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