شماره ركورد كنفرانس :
3550
عنوان مقاله :
3rd Official OPCW Biomedical Proficiency Test: Verification of Nerve Agents Exposure in Human Plasma by Gas and Liquid Chromatography-Tandem Mass Spectrometry
پديدآورندگان :
Naseri Mohammd Taghi Defence Chemical Research Laboratory (DCRL), Karaj-Iran , Babri Mehran Babri@dcrl.ir Defence Chemical Research Laboratory (DCRL), Karaj-Iran; , Mirbabaei Fatemeh Defence Chemical Research Laboratory (DCRL), Karaj-Iran , Hakamizadeh Moones Defence Chemical Research Laboratory (DCRL), Karaj-Iran , Mousavi faraz Sajjad Defence Chemical Research Laboratory (DCRL), Karaj-Iran , Ashrafi Davood Defence Chemical Research Laboratory (DCRL), Karaj-Iran , Sarabadani Mansour Defence Chemical Research Laboratory (DCRL), Karaj-Iran , Ebrahimi Leial Defence Chemical Research Laboratory (DCRL), Karaj-Iran
تعداد صفحه :
1
كليدواژه :
Plasma Nerve Agents , protein adduct , Tandem Mass Spectrometry , peptide
سال انتشار :
1397
عنوان كنفرانس :
بيست و پنجمين سمينار ملي شيمي تجزيه انجمن شيمي ايران
زبان مدرك :
انگليسي
چكيده فارسي :
The purpose of this study was to analyse human plasma samples for the protein adducts (or biomarkers) with different analytical methods for the presences or absence of nerve agents exposure. Organophosphorus Nerve agents (OPNAs) including tabun (GA), sarin (GB), soman (GD), cyclosarin (GF), and VX target the enzyme Butyrylcholinesterase (BuChE) and albumin to cause acute toxicity [1-2]. Albumin is abundant protein in blood and is a good target protein to investigate exposure to nerve agents. It can be phosphylated by OPNAs on the tyrosine residue [3]. In this Biomedical Proficiency Test, plasma samples (6 × 5 mL vial with codes P301-P306) receipted 5 March, 2018 from Organisation for the Prohibition of Chemical Weapons (OPCW) Laboratory (Rijiswijk, The Netherlands), were analyzed for verification of a human exposure to nerve agents. The spiking protocol was kept unknown for participating laboratories. Laboratories are encouraged to identify nerve agents aged and/or non-aged adducts of BuChE and/or albumin with two techniques such as GC-MS/MS and LC-MS/MS. When tandem mass spectrometric identification is performed by multiple reaction monitoring (MRM) using GC or LC-MS/MS, the data from at least two diagnostic precursor-product ion transitions corresponding to the chemical of interest must be recorded. The signal-to-noise ratio for each diagnostic ion must be ≥ 5:1. BuChE activity was measured spectrophotometrically using an Ellman assay as a preliminary test to diagnose exposure to OPNAs. Fluoride regeneration method (6 M potassium fluoride) and clean-up with HLB solid phase extraction step identified Fluorotabun (Ethyl N,N-dimethylphosphoramidofluoridate) released from the protein using GC-MS/MS (+CI) in MRM with two ion/transition m/z 156→128 and 110 and GC-HRMS (EI) in selected ion monitoring (SIM) with two ions with exact mass of 126.01140 and m/z 155.05109. Totally protein precipitation, enzymatic digestion with Pronase E, clean-up with 10KD MWCO Identified the Ethyl N,N-dimethylphosphoramido-tyrosine (GA-Tyr) using LC-MS/MS (+ESI) in MRM with two ion/transition m/z 317→226 and 198. BuChE from plasma purified with Sepharose Q fast flow gel (anion exchange)-procainamide affinity gel chromatography and digestion with pepsin, clean-up with C18 solid phase extraction identified the two modified aged nonapeptide adduct FGES(Ethyl phosphate)AGAAS and FGES(Phosphate)AGAAS using LC-MS/MS (+ESI) in MRM with two ion/transition m/z 904→778 and 673 and m/z 876→778 and 673 respectively, that all of results unambiguously verified samples to contaminated with Tabun. Reference chemicals (own synthesis) were used for the comparison to the data obtained with the test samples. The plasma samples with code P301 and P306 identified as blank sample. The Defense chemical research laboratory (DCRL) identified all the spiked chemicals (Tabun) in the concentration range of 10-60 µg L-1 in plasma samples P302, P303, P304 and P305 correctly with sufficient analytical data and obtained maximum score A in the Third Official OPCW Biomedical Proficiency Test.
كشور :
ايران
لينک به اين مدرک :
بازگشت