Low Level Exposure to Sulfur Mustard: Development of a SOP for Analysis of Albumin Adducts and of a System for Non-Invasive Diagnosis on Skin
Abstract
In the third year of the grand period, the fluorescence derivatization of the tripeptide S-(2-hydroxylethylthioethyl)-Cys-Pro-Phe (S-HETE)Cys-Pro-Phe was investigated; the 5/6-carboxyfluoresceine (FAM) derivative of the tripeptide (S-HETE)Cys-Pro-Phe has been prepared. When spiked pronase digests were subjected to derivatization with FAM succinimidyl ester, the FAM-tripeptide could be detected in the digest, albeit after prior clean-up by reversed-phase HPLC. Unfortunately, the FAM-tripeptide could not be determined in a pronase digest of albumin isolated from a highly exposed blood sample. With regard to the most abundant adduct, i.e., the histidine adduct, attempts to isolate the histidine adduct by means of reversed-phase HPLC were not successful yet. A definitive SOP for the tripeptide assay was drafted. The inter-individual and intra-individual variation of the in vitro sensitivity of human blood to sulfur mustard was determined, following the albumin-tripeptide (HETE-Cys-Pro-Phe) SOP. Furthermore, the day-to-day variability was determined. The SOP for the tripeptide adduct has been demonstrated to a scientist of USAMRICD, in an independent institute. The method could be set up within one day and the scientist of USAMRICD was able to perform the entire assay on his own after 2 days.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 01, 2004
- Accession Number
- ADA434336
Entities
People
- Daan Noort
- Govert P. Van Der Schans
Organizations
- Prins Maurits Laboratorium TNO