Identification and quantitative determination of major...

Chemistry: analytical and immunological testing – Including chromatography

Reexamination Certificate

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C436S527000, C436S805000, C422S082050, C422S082090

Reexamination Certificate

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08062899

ABSTRACT:
An improved chromatographic method for the separation, identification and quantification of contaminants associated with Eosinophelia-Myalgia Syndrome (EMS) includes a high pressure liquid chromatography (HPLC) method which utilizes a silica-based column and a gradient mobile phase. The method provides for effective, efficient, and economical quantitative and qualitative identification of 1,1′-Ethylidenebis L-tryptophan (EBT), 2,3-indolymethyl L-tryptophan (Peak 200), and 2-anilino-L-alanine (Peak 1).

REFERENCES:
patent: 5993653 (1999-11-01), Ahmed et al.
Phenomenex Gemini Two-In-One-Technology (Twin Technology), 2004.
Zhao et al., 1999, Carcinogenesis, vol. 20, No. 11, pp. 2101-2108.
Hill, Robert H., Jr. et al., “Contaminants in L-Tryptophan Associated with Eosinophelia Myalgia Syndrome,” Arch. Environ. Contam. Toxicol. 25,134-142 (1993).
Williamson, B.L., et al., “On-line HPLC-tandem mass spectrometry analysis of contaminants of L-tryptophan associated with the onset of eosinophelia-myalgia syndrome,” Toxicology Letters, 92(2), 139-148 (1997)—Abstract only.
ESA Note 5600A, “Tryptophan Contamination,” ESA, Inc.
Trucksess, MW, et al., “High-performance liquid chromatographic determination of 1,1′-ethylidenebis(L-tryptophan) in L-tryptophan preparations,” J. Pharm. Sci., 83(5), 720-722 (May 1994)—Abstract only.
Zangrilli, JG, et al., “1,1′-Ethylidenebis[L-tryptophan], an impurity in L-tryptophan associated with eosinophelia-myalgia syndrome, stimulates type I collagen gene expression in human fibroblasts in vitro,” Biochem. Mol. Biol. Int., 37(5), 925-933 (Nov. 1995)—Abstract only.
Trucksess, MW, “Separation and isolation of trace impurities in L-tryptophan by high-performance liquid chromatography,” J. Chromatogr., 630(1-2), 147-150 (Feb. 5, 1993)—Abstract only.
Smith, M.J., et al., “1,1′-Ethylidenebis(l-tryptophan), Structure Determination of Contaminant “97”—Implicated in the Eosinophelia-Myalgia Syndrome (EMS),” Tetrahedron Letters, 32(8), 991-994 (1991).
Love, L. A., et al., “Pathological and Immunological Effects of Ingesting L-Tryptophan and 1,1′-Ethylidenebis(L-Tryptophan) in Lewis Rats,” J. Clinical Invest., 91, 804-811 (Mar. 1993).

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