Accession Number:

ADA434336

Title:

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

Descriptive Note:

Annual rept. 1 Dec 2003-30 Nov 2004

Corporate Author:

PRINS MAURITS LABORATORIUM TNO RIJSWIJK(NETHERLANDS)

Report Date:

2004-12-01

Pagination or Media Count:

62.0

Abstract:

In the third year of the grand period, the fluorescence derivatization of the tripeptide S-2-hydroxylethylthioethyl-Cys-Pro-Phe S-HETECys-Pro-Phe was investigated the 56-carboxyfluoresceine FAM derivative of the tripeptide S-HETECys-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.

Subject Categories:

  • Biochemistry
  • Anatomy and Physiology
  • Chemical, Biological and Radiological Warfare

Distribution Statement:

APPROVED FOR PUBLIC RELEASE