Off-line liquid chromatography-MALDI by with various matrices and tandem mass spectrometry for analysis of glycated human serum albumin tryptic peptides

2007 ◽  
Vol 51 (4) ◽  
pp. 456-461 ◽  
Author(s):  
Annunziata Lapolla ◽  
Francesco L. Brancia ◽  
Jessica Bereszczak ◽  
Domenico Fedele ◽  
Lorenzo Baccarin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Human Serum Albumin ◽  
Tandem Mass Spectrometry ◽  
Serum Albumin ◽  
Human Serum ◽  
Tandem Mass ◽  
Tryptic Peptides

scholarly journals Adduct of the blistering warfare agent sesquimustard with human serum albumin and its mass spectrometric identification for biomedical verification of exposure

2020 ◽  
Vol 412 (28) ◽  
pp. 7723-7737
Author(s):  
Marc-Michael Blum ◽  
Annika Richter ◽  
Markus Siegert ◽  
Horst Thiermann ◽  
Harald John
Keyword(s):  
Mass Spectrometry ◽  
Human Serum Albumin ◽  
Tandem Mass Spectrometry ◽  
Serum Albumin ◽  
Human Serum ◽  
Proteinase K ◽  
Selected Reaction Monitoring ◽  
Hydroxyl Group ◽  
Extensive Literature ◽  
Tandem Mass

Abstract Apart from the well-known sulfur mustard (SM), additional sulfur-containing blistering chemical warfare agents exist. Sesquimustard (Q) is one of them and five times more blistering than SM. It is a common impurity in mustard mixtures and regularly found in old munitions but can also be used in pure form. Compared to the extensive literature on SM, very little experimental data is available on Q and no protein biomarkers of exposure have been reported. We herein report for the first time the adduct of Q with the nucleophilic Cys34 residue of human serum albumin (HSA) formed in vitro and introduce two novel bioanalytical procedures for detection. After proteolysis of this HSA adduct catalyzed either by pronase or by proteinase K, two biomarkers were identified by high-resolution tandem mass spectrometry (MS/HR MS), namely a dipeptide and a tripeptide, both alkylated at their Cys residue, which we refer to as HETETE-CP and HETETE-CPF. HETETE represents the Q-derived thio-alkyl moiety bearing a terminal hydroxyl group: “hydroxyethylthioethylthioethyl.” Targeting both peptide markers from plasma, a micro liquid chromatography–electrospray ionization tandem mass spectrometry method working in the selected reaction monitoring mode (μLC-ESI MS/MS SRM) was developed and validated as well suited for the verification of exposure to Q. Fulfilling the quality criteria defined by the Organisation for the Prohibition of Chemical Weapons, the novel methods enable the detection of exposure to Q alone or in mixtures with SM. We further report on the relative reactivity of Q compared to SM. Based on experiments making use of partially deuterated Q as the alkylating agent, we rule out a major role for six-membered ring sulfonium ions as relevant reactive species in the alkylation of Cys34. Furthermore, the results of molecular dynamics simulations are indicative that the protein environment around Cys34 allows adduct formation with elongated but not bulky molecules such as Q, and identify important hydrogen bonding interactions and hydrophobic contacts.

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