Trace isotope analysis of Ricinus communis seed core for provenance determination by laser ablation-ICP-MS.

The castor bean plant, Ricinus communis, grows wild throughout many regions of Australia. The seeds of the plant contain the schedule 1 chemical agent ricin, a type II ribosomal inhibiting protein. Currently there are limited analytical techniques that can be applied in analysis of the seeds to establish attribution. In this study, laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was used for the analysis of seeds collected from 68 plants across 38 locations around Australia. Of the 92 elemental isotopes measured, fifteen (24Mg, 27Al, 44Ca, 53Cr, 55Mn, 57Fe, 60Ni, 65Cu, 66Zn, 75As, 85Rb, 88Sr, 98Mo, 138Ba and 202Hg) yielded data that were relevant to all collection sites. Data were further analysed using multivariate statistical analysis which facilitated the potential for the identification of unique provenance isotopes. Furthermore, this analysis indicated that 59Co was present at significant levels in Victorian and Sydney specimens only.

De novo sequencing of RCB-1 to -3: peptide biomarkers from the castor bean plant Ricinus communis.

Ricinus communis (also know as the castor bean plant) whose forbears escaped from suburban gardens or commercial cultivation grow wild in many countries. In temperate and tropical climates seeds will develop to maturity, and plants may be perennial. In Australia these plants have become widespread and are regarded as noxious weeds in many localities. The seeds of R. communis contain ricin, a protein toxin which can easily be extracted into an aqueous solution. Ricin is toxic by ingestion, inhalation, and injection. The history of terrorist and anarchist interest in the use of seeds from R. communis has driven the development of strategies for determination of cultivar and geographic location of the source of an extract of wild-grown castor bean seed. This forensic information is of considerable interest to law enforcement and intelligence organizations. During forensic studies of both the metabolome and proteome of extracts from eight specimens of six different cultivars of R. communis ("zanzibariensis" collected from Kenya and Tanzania, "gibsonii", "impala", "dehradun", "carmencita", and "sanguineus" collected from Spain and Tanzania), three peptide biomarkers (designated Ricinus communis biomarkers, or RCB) were identified in both the MALDI and electrospray LC-MS spectra. Two of these peptides (RCB-1 and RCB-2) were present in varying amounts in all cultivars, while RCB-3 was present only in the "carmencita" cultivar. The amino acid sequences of RCB-1 to -3 were determined using LC-MS(n) fragmentation and de novo sequencing on both the intact and the carbamidomethyl modified peptides. The connectivity of the two disulfide bonds that were present in all three RCB were determined using a strategy of partial reduction and differential alkylation using tris-(2-carboxyethyl)phosphine with N-ethylmaleimide to reduce and alkylate the most accessible disulfide bond, followed by reduction and alkylation of the remaining disulfide bond with dithiolthreitol and iodoacetamide. The possible functional role of RCB-1 to -3 in R. communis seeds is also discussed.