468 Magnetic Resonance Imaging 0 Volume 8, Number 4, 1990
and then lowered to pH 1 using 12 N HCl. The result-ing precipitate was removed by suction filtration,washed with absolute ethanol until the washings werecolorless, and recrystallized from 95% ethanol. ThisHCl salt was then dissolved in 500 mL of water andneutralized with 5 N NaOH, then raised to pH 12.5,and the free base extracted with methylene chloride(4 x 500 mL). The methylene chloride solution wasdried to give 136 g (65%) of a pale yellow oil. NMR(D,O): 6 2.46 (s, 8H), 3.55 (s, 4H), 7.10 (m, 4H), 7.55(t, J= 12.5 Hz, 2H), 8.25 (d, J= 10 Hz, 2H), 7.50 (t,J = 10 Hz, 2H), 8.40 (d, J = 10 Hz, 2H).
Synthesis of N,N”-2-Bis(2-pyridylmethyl)-N,N,W-Tris(t-Butylcarboxymethyl)Diethylenetriamine
To a solution of N, N”-2-bis(2_pyridyl)diethylenetri-amine (23.6 g, 82.6 mmol) and diisopropylethylamine(53.4 g, 0.4 mol) in 1.2 L of methylene chloride atroom temperature was added dropwise t-butylbromo-acetate (50 g, 0.2 mol) in 300 mL of methylene chloride.After stirring for 24 hr, the solution was evaporatedto dryness and placed under vacuum for 2 hr to re-move excess diisopropylethylamine. The crude solidwas taken up in 1.5 L of methylene chloride, washedwith 0.2 N NaOH, water (2 x 250 mL), brine (200mL) and dried (MgS04). The methylene chloride wasremoved, 200 mL of ethyl acetate added and this so-lution passed through 300 g of silica gel in a buchnerfunnel using EtOAc to elute the product. The purefractions (TLC: MeOH/CH2C12: 3/7) were combinedto give 40.6 g (79.5%) of the ester, NMR (CDC&) 61.26 (s, 9H), x 1.31 (s, 18H), 2.62 (s, 8H), 3.12 (s,2H), 3.17 (s, 4H), 3.78 (s, 4H), 7.02
10 Hz,2H), 7.35
(d, J = 10
Synthesis of N,N”-2-Bis(2-Pyridylmethyl)Diethylenetriamine-N,W,N”-Triacetic Acid
The tris(t-butylcarboxymethyl)ester (24.8, 0.1 mol)was dissolved in a solution of 600 mL of methylenechloride containing 380 mL of trifluoroacetic acid.The solution was stirred for 48 hr, evaporated underreduced pressure and diluted with 50 mL of water.This solution was applied to 200 mL of AG50-X8, H+form, 100-200 mesh and after washing with water un-til neutral, the product was eluted with 1 N NH40H.After removal of NH40H solution, the product wastaken up in 24 mL of water, adjusted to pH 10 andthe solution applied to AGl-X8, acetate, 100-200mesh. The column was washed with three bed volumesof water and the product eluted with 2 N HOAc togive 12.0 g (69%) of product after several lyophiliza-tions. NMR (D,O) 6 3.02
Hz, 4H), 3.08
J = 6
Hz, 4H), 3.14 (s, 4H), 3.41 (s, 2H), 4.08 (s, 4H),
7.52 (m, 4H), 8.05 (t, J=
10 Hz, 4H), 8.40
10 Hz, 2H).
Potentiometric titrations, to determine the acid pro-tonation constants as well as the metal ion stabilityconstants of DTPA-BMA and DTPA-BP, were car-ried out with an automatic titrator system.3 The sys-tem was controlled by a BASIC computer programwhich displays the data in tabular form concurrentwith a high resolution plot. Components of the au-totitrating system included a Fisher digital pH meter,Corning glass and AgCl reference combination elec-trode, and a Metrohm digital autoburette. In each ex-periment, temperature was maintained at 25.O”C + 0.1and ionic strength was kept constant at 0.10 M withNaCl. The concentration of metal ions and ligand wasmaintained between 3 x lop3 M and 5
10m3M, and0.1000 M HCl was used as the titrant to minimize ionicstrength changes during the course of a titration. Ad-dition of a small amount of NaOH prior to the titra-tions was used to raise the solutions to pH 11.Stability constants for Gd3+, Zn2+, Cu2+, andCa2+ complexes of DTPA-BP and the Zn2+, Cu2+and Ca2+ complexes of DTPA-BMA were determinedby direct titration. For these systems the complexeswere found to be greater than 25% dissociated at pH2, while the data for the GdDTPA-BMA system re-vealed the complex to be dissociated less than 1% atpH 2. Thus GdDTPA-BMA was studied using a li-gand-ligand competition titration. In this experimenta 1: 1: 1 molar ratio of Gd3+, DTPA-BMA, andEDTA was titrated. EDTA forms a complex withGd3+ whose stability constant is accurately known.At high pH, Gd3+ is primarily bound to EDTA. TheGd3+ ion was readily transferred to DTPA-BMA asthe pH was lowered to 2. The rate of metal transferwas sufficiently fast that equilibrium pH measure-ments could be made in a reasonable time period af-ter each addition of acid.
Proton association constants for DTPA-BMA werecalculated using a BASIC computer program writtenfor polyprotic weak acid equilibrium calculations. Thestability constants of the metal ion complexes werecalculated using a BASIC program designed for metalion, ligand and proton systems containing a variety ofspecies. Both programs employed a modified Newton-Raphson algorithm which solved the simultaneousnonlinear mass balance equations, yielding -log[H+]values. The evaluated equilibrium constants were variedby a combined Simplex/Marquardt nonlinear regres-