Peak AAA fatty acid homolog contaminants present in the dietary supplement l-Tryptophan associated with the onset of eosinophilia-myalgia syndrome

Abstract

The eosinophilia-myalgia syndrome (EMS) outbreak that occurred in the USA and elsewhere in 1989 was caused by the ingestion of Showa Denko K.K. (SD) L-tryptophan (L-Trp). "Six compounds" detected in the L-Trp were reported as case-associated contaminants. Recently the final and most statistically significant contaminant, "Peak AAA" was structurally characterized. The "compound" was actually shown to be two structural isomers resulting from condensation reactions of L-Trp with fatty acids derived from the bacterial cell membrane. They were identified as the indole C-2 anteiso (AAA₁-343) and linear (AAA₂-343) aliphatic chain isomers. Based on those findings, we utilized a combination of on-line HPLC-electrospray ionization mass spectrometry (LC-MS), as well as both precursor and product ion tandem mass spectrometry (MS/MS) to facilitate identification of a homologous family of condensation products related to AAA₁-343 and AAA₂-343. We structurally characterized eight new AAA₁-XXX/AAA₂-XXX contaminants, where XXX represents the integer molecular ions of all the related homologs, differing by aliphatic chain length and isomer configuration. The contaminants were derived from the following fatty acids of the bacterial cell membrane, 5-methylheptanoic acid (anteiso-C8:0) for AAA₁-315; n-octanoic acid (n-C8:0) for AAA₂-315; 6-methyloctanoic acid (anteiso-C9:0) for AAA₁-329; n-nonanoic acid (n-C9:0) for AAA₂-329; 10-methyldodecanoic acid (anteiso-C13:0) for AAA₁-385; n-tridecanoic acid (n-C13:0) for AAA₂-385; 11-methyltridecanoic acid (anteiso-C14:0) for AAA₁-399; and n-tetradecanoic acid (n-C14:0) for AAA₂-399. The concentration levels for these contaminants were estimated to be 0.1-7.9 μg / 500 mg of an individual SD L-Trp tablet or capsule The structural similarity of these homologs to case-related contaminants of Spanish Toxic Oil Syndrome (TOS) is discussed.

Keywords: Case-associated contaminants; Dietary supplements; Eosinophilia-myalgia syndrome; L-Tryptophan; Structural homologs; Toxic oil syndrome; peak AAA.