The hepatotoxic alkaloids known to occur in tansy ragwort (Senecio jacobaea L.) are also present in honey produced from the nectar of this species. These alkaloids, which inclued senecionine, seneciphylline, jacoline, jaconine, jacobine, and jacozine, are potentially carcinogenic, mutagenic, and teratogenic and may pose health hazards to the human consumer.

Site specific amide hydrogen/deuterium content of oxidized and reduced Escherichia colithioredoxin, and alkylated derivatives, Cys-32-ethylglutathionylated and Cys-32-ethylcysteinylated thioredoxins are measured, after exposure for 20 s to D(2)O/phosphate buffer (pH 5.7), by electrospray mass spectrometry. The degree of deuteration of Oxi-TRX and Red-TRX correlated with the rates of H/D exchange measured previously by NMR. The ethylcysteinyl modification was shown to minimally perturb the active site of the reduced protein, but showed more global effects on structures of alpha-helices and beta-strands distant from the site of modification. In contrast, the larger ethylglutathionyl group had little effect on the protein's overall conformation, but significantly affected the structure of loops close to the active site. A molecular model of GS-ethyl-TRX derived from molecular simulation allowed the H/D exchange results to be interpreted in terms of specific interactions between the alkyl chain and the protein surface. The specific conformation of the ethylglutathione modification was predicted to be fixed by salt bridges between the carboxylates of the gamma-Glu and Gly of glutathione and the guanidinium of Arg-73 and epsilon-amino group of Lys-90 of the protein. Specific hydrogen bonding interactions between the glutathione carbonyl oxygens and the amide protons of thioredoxin residues Ile-75 and Ala-93 were predicted. The H/D exchange studies showed low levels of deuterium incorporation at backbone nitrogens of these residues. The data also provided evidence for an unusual amide proton-amide nitrogen hydrogen bond within the ethylglutathionylated chain. These same sets of electrostatic and hydrogen bonding interactions were not predicted or observed for the smaller alkyl modification in Cys-ethyl-TRX.

Fast-atom bombardment mass spectrometry was used to study disulfide bonding patterns in heat-denatured human recombinant macrophage colony stimulating factor (rhM-CSF). The heat-denaturated protein was studied by analysis of the pattern of peptides in the proteolytic digests. Native rhM-CSF is a homodimer with intramolecular disulfide linkages between Cys7-Cys90, Cys48-CysI39, and Cysl02-Cysl46 and intermolecular linkages between Cys31-Cys31, and the pairs Cys157 and Cys159. Brief heating for 1 min leads to partial disulfide bond scrambling. In addition to the native disulfide bonds between Cys7-Cys90, Cys48-Cys139, and Cys31-Cys31, nonnative disulfide bonds were detected between Cys48-Cys90 and Cys48-Cysl02. When heated for 5 min the disulfide bonds of rhM-CSF are completely scrambled and lead to nonnative intramolecular disulfide bonds between Cys48-Cysl02 and Cys90-Cysl02 and one intermolecular disulfide bond between Cys102-Cysl02.

Tansy ragwort (Senecio jacobaea ) was evaluated for animal and human health hazard using the Salmonella/mammalian-microsome mutagenicity test. An acetone extract of tansy ragwort (TR) produced a negative mutagenic response for bacterial tester strains TA1535 and TA100 and a toxic response in tester strains TA1537 and TA98. Assay of this extract in the presence of mammalian liver microsomes (S-9) resulted in positive mutagenic responses in tester strains TA1535, TA1537, TA98 and TA100. Species differences were evaluated by use of liver microsome preparations from induced rat and uninduced sheep, beef, hamster, trout and rat. Only a slight species difference was demonstrated. A mixture of pyrrolizidine alkaloids (PA), extracted from TR flowers, produced a negative response in tester strains TA1535, TA1537, TA98 and TA100. A negative response was also demonstrated when the TR flower PA mixture was assayed with the Salmonella tester strains and induced rat liver microsomes ( IRLM ). A mixture of PA extracted from Senecio longilobus also produced a negative response. The major PA present in TR, jacobine , produced a negative response without and with IRLM exposure in tester strains TA1535, TA1537, TA98 and TA100. Another similar PA, monocrotaline, found in various Crotalaria species also gave a negative response. Milk from TR-fed goats was evaluated for mutagenic response. Milk from goats not receiving TR and from goats receiving TR at a level of 1% of their body weight/day via rumen cannula produced a negative response without liver microsomes present. Milk from TR-fed goats, however, yielded both negative and marginally positive responses for different combinations of tester strains and liver microsome preparations.

A prototype gas chromatograph (GC) electron monochromator (EM) reflectron time-of-flight (TOF) mass spectrometer has been constructed and demonstrated to simultaneously record four-dimensional resonant electron capture (REC) mass spectra (m/z, ion-intensity, electron-energy, and retention time) of electron-capturing compounds in real time. Specifically, complete REC mass spectra of all of the components in a mixture of perfluorocarboxylic acids and in a sample of pentafluorobenzyl alcohol were recorded in the GC mode. For each compound, the data enable one to distinguish different electronic states of the molecular ion and different possible decomposition pathways for each state. This new instrument can be used to obtain analytical information unrecognizable by any other mass spectrometric technique from the isomeric species of a variety of electron-capturing structures.