"Dilute-and-shoot" rapid-separation liquid chromatography tandem mass spectrometry method for fast detection of thiodiglycolic acid in urine.

A new sensitive rapid-separation liquid chromatography tandem mass spectrometry approach for the determination of thiodiglycolic acid (TDGA) in urine has been developed. The use of the "dilute-and-shoot" method helps to shorten the sample preparation stage and provides a sensitive and direct approach for TDGA determination in urine. Chromatographic separation of the analyte and other urine compounds was achieved using a reverse-phase liquid chromatography column with mobile phases consisting of 0.1% formic acid in water and acetonitrile in a gradient elution mode. For the identification and quantification of TDGA electrospray ionization-tandem mass spectrometry monitoring, two precursor-to-product ion transitions were used. The method demonstrates good linearity and has a detection limit of 50 ng mL⁻¹ in urine.

Monitoring of hydrolysis products of mustard gas, some sesqui- and oxy-mustards and other chemical warfare agents in a plant material by HPLC-MS/MS.

At present, there is a real threat of chemical warfare agents being used in terrorist acts and military clashes. Sulfur and nitrogen mustards are blister agents with high lethality and rapid disruption of armed forces. These highly poisonous substances are hydrolyzed to the characteristic marker compounds when released into the environment. Analysis of environmental objects allows to establish the fact of alleged use of chemical warfare agents and to reveal their type. However, water and soil samples are not always reliable for retrospective analysis. The resulting chemical warfare agent markers may be washed out from the application site over time by groundwaters or atmospheric condensations. This study shows the potential for using plants as a convenient material for retrospective analysis. Garden cress (Lepidium sativum) was chosen as a model plant for this purpose, since it can be easily and quickly grown hydroponically. The plants were cultivated in the environment of the selected markers to study an accumulation of these compounds by the plants. An effective and fast method of homogenization with subsequent ultrasonic extraction was applied. The extracts were analyzed using a specially developed and validated HPLC-MS/MS approach. Separation of the hydrophilic markers was carried out on a reversed-phase column with a polar endcapping. Sensitive mass spectrometric detection was performed in the multiple reaction monitoring mode. Achieved limits of detection for most markers were in the range of 2-40 ng mL-1. It was discovered from the research that after the removal of markers from the growing medium the plants are able to store and concentrate these markers for at least 5 weeks, ensuring a high retrospectivity of the analysis. The obtained results indicate the perspective of using plants as additional objects of analysis during the investigation of incidents related to the use of chemical warfare agents. However, more complex plants and models should be studied in the future.

Simultaneous determination of organophosphorus nerve agent markers in urine by IC-MS/MS using anion-exchange solid-phase extraction.

In this study a comprehensive approach for determination of low molecular organophosphorus nerve agent markers - highly polar alkylphosphonic acids and much less polar alkyl methylphosphonic acids is presented. Accurate, sensitive and simultaneous determination of the nerve agent markers in human urine was performed by ion chromatography and tandem mass spectrometry using deuterated internal standards. Analysis of the urine extracts was conducted on an anion-exchanger based on poly(styrene-co-divinylbenzene) substrate with a high degree of crosslinking and a covalently-bonded branched functional layer. The use of this type of column allowed achieving high values of retention factors for alkylphosphonic acids and alkyl methylphosphonic acids due to combination of anion-exchange and hydrophobic interactions between the analytes and the stationary phase of the column. Prior to the analysis, the urine samples were purified using anion-exchange cartridges for solid-phase extraction, and high recovery values were achieved for each analyte. The developed IC-MS/MS technique was validated for linearity, limit of detection, limit of quantification, precision and accuracy using two LC-MS/MS instruments. The proposed approach was successfully tested on the urine samples, provided by the Organisation for the Prohibition of Chemical Weapons in the frame of the 4th Biomedical Proficiency Test.

'Dilute-and-shoot' RSLC-MS-MS method for fast detection of nerve and vesicant chemical warfare agent metabolites in urine.

A sensitive screening method based on fast liquid chromatography tandem mass-spectrometry (RSLC-MS-MS) has shown the feasibility of separation and detection of low concentration ß-lyase metabolites of sulfur mustard and of nerve agent phosphonic acids in urine. The analysis of these compounds is of interest because they are specific metabolites of the chemical warfare agents (CWAs), sulfur mustard (HD), sarin (GB), soman (GD), VX and Russian VX (RVX). The 'dilute-and-shoot' RSLC-MS-MS method provides a sensitive and direct approach for determining CWA exposure in non-extracted non-derivatized samples from urine. Chromatographic separation of the metabolites was achieved using a reverse phase column with gradient mobile phases consisting of 0.5% formic acid in water and acetonitrile. Identification and quantification of species were achieved using electrospray ionization-tandem mass-spectrometry monitoring two precursor-to-product ion transitions for each compound. The method demonstrates linearity over at least two orders of magnitude and had detection limits of 0.5 ng/mL in urine.