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An Electron Paramagnetic Resonance (EPR) spectroscopy study on the γ-irradiation sterilization of the pharmaceutical excipient l-histidine: Regeneration of the radicals in solution

Vallotto, C., Williams, H.E., Murphy, D.M., Ayres, Z.J., Edge, R., Newton, M.E. and Wedge, C.J. (2017) An Electron Paramagnetic Resonance (EPR) spectroscopy study on the γ-irradiation sterilization of the pharmaceutical excipient l-histidine: Regeneration of the radicals in solution. International Journal of Pharmaceutics, 533 (1). pp. 315-319. ISSN 0378-5173

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The effects of γ-radiation sterilization on the parenteral excipient L-histidine were analysed by means of EPR spectroscopy. The irradiation process was found to induce the formation of a deamination radical which was persistent in the solid state. The nature and reactivity of the radicals following dissolution in water was evaluated using spin-trapping EPR experiments. The deamination radical was found to regenerate in solution in the presence of trace metals, potentially leading to radical induced degradation reactions occurring up to an hour after the dissolution process. Understanding this process is significant for the improved design of parental pharmaceutical formulations in which unwanted radical reactions after γ radiation sterilization could lead to degradation of active ingredients.

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Item Type: Article
Uncontrolled Keywords: Electron Paramagnetic Resonance (EPR) Spin-trapping Sterilization Irradiation Dissolution Fenton reaction
Subjects: Q Science > QD Chemistry
Schools: School of Applied Sciences
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References: Abuhanoğlu, G., Özer, A.Y., 2010. Radiation Effects on Pharmaceuticals. FABAD J. Pharm. Sci. 35, 203–217. Ambroż, H.B., Kornacka, E.M., Marciniec, B., Ogrodowczyk, M., Przybytniak, G.K., 2000. EPR study of free radicals in some drugs γ-irradiated in the solid state. Radiat. Phys. Chem. 58, 357–366. doi: 10.1016/S0969-806X(99)00489-2 Aydin, M., 2010. EPR study of free radicals in amino acid derivatives gamma-irradiated at 300 K, Indian J. Pure Appl. Phys. 48, 611–614. Barr, D., Jiang, J., Weber, R.T., 2001. How to quantitate nitroxide spin adducts using TEMPOL. Bruker Biospin Tech. Note. Davies, M.J., 2016. Detection and characterisation of radicals using electron paramagnetic resonance (EPR) spin trapping and related methods. Methods 109, 21–30. doi: 10.1016/j.ymeth.2016.05.013 Davies, M.J., Fu, S., Dean, R.T., 1995. Protein hydroperoxides can give rise to reactive free radicals. Biochem. J. 305, 643–649. doi: 10.1042/bj3050643 Dicle, I.Y., Osmanğolu, Ş., İpek, N., 2015. Electron paramagnetic resonance study of free radicals in γ-irradiated L-glutamine and L-glutamine-t-butyl ester hydrochloride. Radiat. Eff. Defects Solids 170, 65–71. doi: 10.1080/10420150.2014.996879 Food and Drug Administration, 2011. Food And Drug Administration Compliance Program Guidance Manual. US. Foote, C.S., Clennan, E.L., 1995. Properties and reactions of singlet dioxygen, in: Foote, C.S., Valentins, J.S., Greenberg, A., Liebman, J.F. (Eds.), Active Oxygen in Chemistry. Springer, Netherlands, pp. 105–140. doi: 10.1007/978-94-007-0874-7_4 García-Arieta, A., 2014. Interactions between active pharmaceutical ingredients and excipients affecting bioavailability: impact on bioequivalence. Eur. J. Pharm. Sci. 65, 89–97. doi: 10.1016/j.ejps.2014.09.004 Gebicki, S., Gebicki, J.M., 1993. Formation of peroxides in amino acids and proteins exposed to oxygen free radicals. Biochem J. 289, 743–749. doi: 10.1042/bj2890743 Gibella, M., Crucq, A-S., Tilquin, B., Stocker, P., Lesgards, G., Raffi, J., 2000. Electron spin resonance studies of some irradiated pharmaceuticals. Radiat. Phys. Chem. 58, 69–76. doi: 10.1016/S0969-806X(99)00353-9 Gopal, N.G.S., 1978. Radiation sterilization of pharmaceuticals and polymers. Radiat. Phys. Chem. 12, 35–50. doi: 10.1016/0146-5724(78)90072-9 Hasanain, F., Guenther, K., Mullett, W.M., Craven, E., 2014. Gamma Sterilization of Pharmaceuticals—A Review of the Irradiation of Excipients, Active Pharmaceutical Ingredients, and Final Drug Product Formulations. PDA J. Pharm. Sci. and Tech. 68, 113–137. doi: 10.5731/pdajpst.2014.00955 Iravani, S., 2017. ESR of Irradiated Drugs and Excipients for Drug Control and Safety, in: Shukla, A.K. (Ed.), Electron Spin Resonance in Food Science. Academic Press, London, pp. 111-122. doi: 10.1016/B978-0-12-805428-4.00007-6 Jacobs, G.P., 1995. A Review of the Effects of Gamma Radiation on Pharmaceutical Materials. J. Biomater. Appl. 10, 59–96. doi: 10.1177/088532829501000104 Jacobs, G.P., 1985. A Review: Radiation sterilization of pharmaceuticals. Radiat. Phys. Chem. 26, 133–142. doi: 10.1016/0146-5724(85)90177-3 Kaisheva, E.A., Flores-Nate, A., Gupta, S., 2003. Stable lyophilized pharmaceutical formulation of IgG antibodies. US Pat., 2003/0113316 A1. Kamerzell, T.J., Esfandiary, R., Joshi, S.B., Middaugh, C.R., Volkin, D.B., 2011. Protein – excipient interactions : Mechanisms and biophysical characterization applied to protein formulation development. Adv. Drug Deliv. Rev. 63, 1118–1159. doi: 10.1016/j.addr.2011.07.006 Karabulut, B., Yıldırım, İ, 2015. EPR study of gamma irradiated DL-methionine sulfone single crystals. J. Mol. Struct. 1102, 186–189. doi: 10.1016/j.molstruc.2015.08.063 Kuwabara, M., Lion, Y., Riesz, P., 1981. E.S.R. of Spin-trapped Radicals from Sugars. Reactions of Hydroxyl Radicals in Aqueous Solutions and γ-radiolysis in the Polycrystalline State. Int. J. Radiat. Biol. Relat. Stud. Physics, Chem. Med. 39, 451–455. doi: 10.1080/09553008114550561 Lagercrantz, C., Forschult, S., 1968. Trapping of Free Radicals formed by γ-Irradiation of Organic Compounds. Nature 218, 1247–1248. doi: 10.1038/2181247a0 Lehmann, M.S., Koetzle, T.F., Hamilton, W.C., 1972. Precision neutron diffraction structure determination of protein and nucleic acid components. IV. The crystal and molecular structure of the Amino Acid L-Histidine. Chem. Biol. Drug Des. 4, 229–239. doi: 10.1111/j.1399-3011.1972.tb03424.x Makino, K., Riesz, P., 1982. E.S.R. of Spin-trapped Radicals in y-irradiated Polycrystalline Amino Acids. Chromatographic Separation of Radicals. Int. J. Radiat. Biol. 41, 615–624. doi: 10.1080/09553008214550711 Mangion, I., Liu, Y., Reibarkh, M., Williamson, R.T., Welch, C.J., 2016. Using Electron Paramagnetic Resonance Spectroscopy To Facilitate Problem Solving in Pharmaceutical Research and Development. J. Org. Chem. 81, 6937−6944. doi: 10.1021/acs.joc.6b00937 Minegishi, A., Bergene, R., Riesz, P., 1980. E.S.R. of Spin-trapped Radicals in y-irradiated Polycrystalline Amino Acids, N-acetyl Amino Acids and Dipeptides. Int. J. Radiat. Biol. 38, 627–650. doi: 10.1080/09553008014551461 Neyens, E., Baeyens, J., 2003. A review of classic Fenton’s peroxidation as an advanced oxidation technique. J. Hazard. Mater. 98, 33–50. doi: 10.1016/S0304-3894(02)00282-0 Pazos, M., Andersen, M.L., Skibsted, L.H., 2006. Amino Acid and Protein Scavenging of Radicals Generated by Iron/Hydroperoxide System: An Electron Spin Resonance Spin Trapping Study. J. Agric. Food Chem. 54, 10215−10221. doi: 10.1021/jf062134n Pikal, M.J., Costantino, H.R., 2004. Lyophilization of Biopharmaceuticals, Biotechnology: Pharmaceutical Aspects. AAPS Press, Arlington, VA. Rustgi, S., Joshi, A., Riesz, P., Friedberg, F., 1977. E.S.R. of Spin-trapped Radicals in Aqueous Solutions of Amino Acids. Reactions of the hydrated electron. Int. J. Radiat. Biol. 32, 533–552. doi: 10.1080/09553007714551321 Schulman, S.G., Achey, P.M., 2007. Encyclopedia of Pharmaceutical Technology. Marcel Dekker, New York. Stoll, S., Schweiger, A., 2006. EasySpin, a comprehensive software package for spectral simulation and analysis in EPR. J. Magn. Reson. 178, 42–55. doi: 10.1016/j.jmr.2005.08.013 Symons, M.C.R., 1995. Electron spin resonance studies of radiation damage to DNA and to proteins. Rad. Phys. Chem 45, 837–845. doi: 10.1016/0969-806X(94)E0035-H Talbi, S., Raffi, J., Aréna, S., Colombani, J., Piccerelle, P., Prinderre, P., Dolo, J.-M., 2004. EPR study of gamma induced radicals in amino acid powders. Spectrochim. Acta, Part A 60, 1335–1341. doi: 10.1016/j.saa.2003.10.031 Wade, A.M., Tucker, H.N., 1998. Antioxidant characteristics of L-histidine. J. Nutr. Biochem. 9, 308–315. doi: 10.1016/S0955-2863(98)00022-9 Westhof, E., Flossmann, W., Ludemann, H.-D., Muller, A., 1974. ESR and INDO study of radical conformations in irradiated single crystals of L-histidine free base. J. Chem. Phys. 61, 3376–3381. doi: 10.1063/1.1682500 Zs.-Nagy, I., Floyd, R.A., 1984. Hydroxyl free radical reactions with amino acids and proteins studied by electron spin resonance spectroscopy and spin-trapping. Biochim. Biophys. Acta 790, 238–250. doi: 10.1016/0167-4838(84)90028-1
Depositing User: Christopher Wedge
Date Deposited: 02 Oct 2017 08:03
Last Modified: 28 Aug 2021 15:31


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