Polymerization of L-Aspartic Acid to Polysuccinimide and Copoly(Succinimide-Aspartate) in Supercritical Carbon Dioxide

Home , Polyaspartic acid, Polysuccinimide

9259 Letters in Organic Chemistry, 2005, 2, 687-689 687

Kenneth M. Doll, 1 , Randal L. Shogren t2, Ronald A. Holser 1 , J.L. Willett2 and Graham Swift3

1 Food and Industrial Oil and 2Plant Polymer Research Units, National Center for Agricultural Utilization Research, United States Department of Agriculture, Agricultural Research Service, 1815 N. University St., Peoria, IL 61604, USA, 3Folia. Inc., 2800 Milan Court, Birmingham AL 35211 Received June 21, 2005: Revised August 21, 2005: Accepted September 05, 2005 Abstract: We have prepared two polymeric materials from L-aspartic acid in supercritical carbon dioxide, polysuccinimide and copoly(succinimide-aspartate). The polysuccinimide was characterized by JR spectroscopy and GPC analysis. The copoly(succinimide-aspartate) product was also characterized by titrometric analysis. These natural materials may prove useful in reducing our dependence on petroleum products. Keywords: Polysuccinimide, copoly(succinimide-aspartate), polyaspartic acid, supercritical carbon dioxide, natural polymer, amino acid polymer.

The synthesis of polysuccinimide from L-aspartic acid, with or without acid catalysis (7-11]. It can also be Scheme (1) has a long history [1,2] L-aspartic acid or salts polymerized enzymatically [12]. The physical properties of of L-aspartate can be polymerized thermochemically [3-6], polysuccinimide have been studied, and found to be

0 OH )I NH No— OH HO (n+2) acid c at \\ 10 H2Nol L.,..,r:o 0 L OH Scheme 1. The synthesis of polysuccinimide from aspartic acid.

0 NH, )ç2 C N 4O +d Na 0 OrNa OH 0 -H2O -NH, H2N OH OH OH .iL o" idr Scheme 2. The synthesis of the water soluble copoly(succinimide-aspartate).

favorable for applications as absorbents or viscosity Address correspondence to these authors at the Food and Industrial Oil modifiers. Additionally, its amino acid structure makes it and 2 PIant Polymer Research Units, National Center for Agricultural inherently more bio-degradable than carbon chain polymers Utilization Research, United States Department of Agriculture, [13]. Even the highly branched form of polysuccinimide Agricultural Research Service, 1815 N. University St., Peoria, IL 61604, formed from maleic acid and ammonia has been shown to be USA; E-mail: [email protected]; [email protected] —70% bio-degradable in wastewater [14]t. tlhe use of trade, firm, or corporation names in this publication is for the Polysuccinimide can also be reacted with nucleophiles to information and convenience of the reader. Such use does not Constitute form useful derivatives for a variety of applications, such as an official endorsement or approval by the United States Department of paper processing, ion exchange, and biomedical release [15]. Agriculture or the Agricultural Research Service of any product or service to the exclusion of others that may be suitable. Recently, a synthesis to yield an alternative form of

polysuccinimide has been developed [6]. This form of In order to test whether our synthesis in supercritical polysuccinim ide, copoly(succinimide-aspartate; Scheme 2), carbon dioxide was more general, we performed a maintains its water solubility facilitating other polymerization of another natural amino acid, Glutamic developments. Herein we report our patented [16] method for Acid. This polymer is often synthesized enzymatically [17], the synthesis of these two polymeric materials in and has found use as a bio-degradable water absorbtion resin supercritical carbon dioxide solvent. Our synthesis allows and in drug delivery systems [18]. Using the same these materials to be made in good yield without the use of conditions as in the polysuccinimide synthesis, we were able vacuum system, or toxic organic solvents and catalysts. to synthesize poly glutamic acid. Both the IR spectra (Fig. Furthermore the pressures that are required for the reaction (3)) and GPC analysis show that the reaction has occured can be achieved by simply heating the reactor. forming a polymeric material with a MW between 192 and 900 Daltons. Finally, we wanted to test our reaction system on a two It / component system. We synthesized a polymer of the di-acid, adipic acid, with the multifunctional amine, triethylenetetraamine (TETA). In this system, the reactants were mixed well, and polymerized using supercritical CO2 as a solvent. As in the other cases, polymerization and could be confirmed by IR spectroscopy (Fig. (4)). Fig. (1). The IR spectra of of polysuccinimide synthesized in supercritical CO2 at 150 °C (bottom), and 200 °C, (top). Only the higher temperature spectra has a significant imide peak at 1714 cm- 1 indicating conversion to product. The syntheses were monitored by isolation of the product and study by IR spectroscopy. The main feature in the IR spectra (Fig. (1)) is the imide peak with a frequency of 1714 cm The spectra of the product from the syntheses performed at two different temperatures show that at 150 °C, product is not produced to any significant extent, whereas at 200 °C, the product formation is nearly quantitative. GPC analysis (Fig. (2)) shows a similar trend, where the higher temperature synthesis primarily has material with a MW of >4000, and the lower temperature synthesis shows little Fig. (4). The IR spectra of a copolymer of adipic acid and TETA molecular weight building. (Triethylenetetramine) product synthesized in supercritical CO2 at 200 °C (top), and the adipic acid starting material (bottom). Our synthesis of polysuccinimide using our CO2 system follows several of the twelve principles of green chemistry [19,20]. The only byproduct formed in our synthesis is water, which possess no toxicity. The rest of the reagent L- aspartic acid, is incorporated into the product yielding an Fig. (2). GPC analysis of a hydrolyzed sample of overall 86% atom efficiency. The raw material used is polysuccinimde synthesized in supercritical CO 2. The peak at natural and there is no required derivitization or catalyst. 7.918 min corresponds to a MW of -.4200 daltons when Furthermore, the the substances used are not hazardous compared to sodium polyacrylate standards. The hydrolysis minimizing the impact of chemical accidents. The procedure necessary for solubility of the GPC sample may have reduced the observed MW.

Fig. (3). The IR spectra of polyglutamate product synthesized in supercritical CO2 at 200 °C (top), and glutamic acid starting material (bottom). The polymerization is shown by the product peak at 1724 cm-1. Fig. (5). The reactor utilized in these syntheses.

Polymerization of L-Aspartic Acid to Polysuccinimide Letters iii Organic Chemistry, 2005, VoL 2, No. 8 689

supercritical carbon dioxide serves two purposes. First, it is synthesis. A solid, light brown material was isolated and a good media for dispersing the reactants. Second, it ground into a tine powder with a mortar and pestle (2.8 g; effectively removes the water produced by the condensation 68% yield after grinding). Titration by first acidifying with reaction allowing the reaction to proceed without the HCI, then titrating with NaOH showed 0.45 equivalents of phosphoric acid catalyst often used in other syntheses. It is carboxylate per 100 g of polymer, similar to the expected also important to note that the only high pressure equipment 0.43 demonstrated for a 1:1 copolymer of succinimide and required for our reaction is a reasonably inexpensive pressure aspartate. reactor (Fig. (5)). In conclusion, we have demonstrated a Poly glutamic acid (Sigma, 99%) and the Adipic acid synthesis of the useful products, polysuccinimide and (Aldrich, 99%), tri ethyl en etetraam ine (TETA, Aldrigh, Tech copoly(succinimide-aspartate), and we have also 60%) copolymer were synthesized by the same method as demonstrated the generality of our system as well. the polysuccinimide. A slightly larger 600 mL parr reactor was used without a reactor liner. EXPERIMENTAL The reactor employed in this reaction was a 450 mL Parr ACKNOWLEDGEMENTS Series 4560 Bench Top Mini Stirred Reactor, equipped with This research was funded in part by the USDA-ARS a standard impeller stirrer (Fig. (5)). Teflon or glass reactor CRIS# 3620-41000-088, and by a cooperative research and liners were used to facilitate reactor cleaning. development agreement, CR.ADA# 58-3K95-3-0971, with First, 1.58 g (0.01 mol) l-aspartic acid (Aldrich, 98+%), Folia Inc. was added to the reactor liner which was placed in the reactor and flushed with nitrogen for I mm, then pressurized with nitrogen to -.0.69 MPa(lOO PSI) in order to test for leaks in REFERENCES the system. The nitrogen was vented to —0.07MPa (10 PSI). [I] Dessaignes, M. Coniptes, Rendus, Chime. 1850, 31, 432434. The reactor was pressurized to 5.38 MPa (780 PSI) from a 121 Schiff, H. Chem, Ber. 1898, 30, 2449-2459. CO2 tank (Air Products) equipped with a syphon tube. The [ 3 1 Roweton, S.; Huang, S. J., Swift, G. J. Environ. Polym. Degrad. 1997, 5, 175-I81. reactor cooling water was started, and the temperature set to [4] Matsubara. K.; Nakato, R.; Tomida, M. Macromolecules 1998, 3/, 70 C. The pressure was vented to 7.63 MPa (1106 PSI) at 1466-1472. 70 °C, which gives a CO 2 density of 0.16 g /mL. The [5] Matsubara, K.; Nakato, R.; Tomida, M. Macromolecules 1997, 30, reactor temperature was set to 205 °C, the stirring set to 400 2305-2312. RPM. The reaction was run for 4 hrs. The pressure was [6] Sikes, C. S. Imide-free and Mixed Amice/Imide Thermal Synthesis ofPolyaspartate. US 5981691, Nov. 9, 1999. slowly vented to 0.07 MPa (10 P51), the reactor pressurized [7] Chang, C. J.; Swift, G. J. M. S. Pure and App. Chem. 1999, A36, with nitrogen to 0.80 MPa (116 PSI), and cooled overnight. 963-970. A solid, light red product was isolated and ground with a [8] Kang, H. S.; Shin, M.-S.; Kim, J.-D.; Yang, J.-W. Polymer Bulletin mortar and pestle (1.07 g; 78% yield after grinding). The 2000, 45, 39-43. [9] Kang, H. S.; Yang, S. R.; Kim, J.-D.; Han, S.-H.; Chang, 1. —S. product was characterized by Infrared spectroscopy (IR) (Fig. Langmuir2001, 17, 7501-7506. (1)), which was carried out on a Thermonicolet Avatar 370 [101 Suwa, M.; Hashiczume, A.; Morishima, V.; Nakato, T.; Tomida, spectrophotometer using transmission sample holder and M. Macromolecules 2000, 33, 7884-7892. standard potassium bromide pellets. Gel Permeation [II] Tomida, M.; Nakato, T.; Kuramochi, M.; Shibata, M.; Matsunami, Chromatography (GPC) (Fig. (2)) was performed on a S.; Kakuchi, T. Polymer 1996, 37, 4435-4437. [12] Matsumura, S.; Tsushima, Y.; Otozawa, N.; Murakami, S.; Waters 1525 HPLC system with a Waters 717 plus Toshima, K.; Swift, G. Macromol, Rapid Commun, 1"9,20,7-11. autosampler and a Waters 2996 photodiode array detector [13] Rypacek, F.; Dvorak, M.; Stefko, I.; Machova, L.; Skarda, V.; and analyzed at 218 nm. A Phenomenex Poly-sep-GFC- Kubies, D. Poly(aminoacid)s and Ester-Amide Copolymers: tailor- P2000 column was used. The samples were dissolved in IN Made Biodegradable Polymers, ACS Symposia Series, Chapter 16, Volume 786: American Chemical Society NaOH solution then an aliquot was added to a 10 mM TRIS 2001; 258-275. [14] Swift, G. Degradable Polymers, second edition; Scott, G. Ed. buffer (pH 8), 100 mM NaCl solution and analyzed by the Kulwer Academic Publishers, The Netherlands 2002; 379412. GPC. [IS] Swift, G.; Redlich, G. H. Methods of Synthesis of Polysuccinimide, cnnnivmerc nnivc,,rr,n,mA p ...A •h .. ..,t 0,.,.... of fl-I IYau.T tI,,..IvI.- r flicilt The product, copoly(succinimide-aspartate) (I to I molar application US 2004/0102602 Al, May 17, 2004. ratio) was synthesized by a similar method, although a 116] Swift, G.; Doll, K. M.; Shogren, R. L.; Holser, R. A.; Willett, J. L. mixture of aspartic salts had to be made initially. First, 13.3 Synthesis of polysuccinimide and copoly(succinimide-aspartate) g (0.] mol) 1-aspartic acid was stirred with 5.1 mL 9.83 M in a supercritical fluid. US 6887971 B2, May 3, 2005. Ashiuchi, M.; Shimanouchi, K; Nakamura, H.; Kamei, T; Soda, NaOH solution (0.05 mol NaOH) and 3.25 mL of K.; Park, C; Sung, M-H.; Misono, H App! Environ Microbial. concentrated 15.43 M NH 40H (0.05 mol NH4 0H) in 100 2004, 70, 4249-4255. ml- of Nanopure H 2O. This mixture was stirred for 15 mm [18] Sanda, F.; Fujiyama, T.; Endo, T. Macromol, Chem. Phys. 2002, and dried in a forced air oven for at 80 °C for 14 hrs. After 203, 727-734. [19] Anastas, P. T., Kirchhoff, M, M. Acc. Chem. Res. 2002, 35, 686- drying, the mixture was a solid with a moisture content of 694. 3.5%. A portion of this solid (5.35 g) was ground with a [20] Warner, J. C.; Cannon, A. S.; Dye, K. M. Environ. Impact Assess. mortar and pestle and added to a reactor liner. The reaction Rev. 2004, 24, 775-799. was then carried out as was done in the polysuccinimide Supplied tbi U.S. DSpailmS.it at Agdcultuie, National Center for Agficuttural UtlIf.tI.n Research, Peoria, Illinois