Optimized method for analysis of ethanolamines, hydrolysis products of nitrogen mustards, from urine samples using LC-MS/MS

Hamzah, Nurhazlina; Kjellberg, Matti A.; Vanninen, Paula

doi:10.1016/j.jchromb.2021.122762

Cited by 6 publications

(7 citation statements)

References 24 publications

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“…From urine samples, only HN1‐GSH2 (diglutathionyl conjugate) and ethyldiethanolamine (EDEA), hydrolysis product of HN1, were detected by LC‐MS in full‐scan mode. The recoveries for both HN1‐GSH2 and EDEA were >80%, which is at the same level as recoveries of EDEA for our previous validated urine extraction method 6 . The extracted ion chromatograms (EICs) of HN1‐GSH2 extracted from urine sample are shown in Figure S3 (supporting information).…”

Section: Resultssupporting

confidence: 59%

“…The concentration prepared for this reference was 50 μg/mL, and the conjugates were identified using LC‐MS in full‐scan mode. An amount of 50 μL of reference chemical was added into 500 μL of urine sample (5 μg/mL), mixed on vortex for 10 s and left at room temperature for 1 h. The solid‐phase extraction protocol of the validated method for extraction of EAs in urine sample was followed 6 . The final residue was dissolved in mobile phase A (0.1% formic acid in water) before being analysed by LC‐MS in full‐scan mode.…”

Section: Methodsmentioning

confidence: 99%

“…The recoveries for both HN1-GSH2 and EDEA were >80%, which is at the same level as recoveries of EDEA for our previous validated urine extraction method. 6 The extracted ion chromatograms (EICs) of HN1-GSH2 extracted from urine sample are shown in Figure S3 (supporting information).…”

Section: Conjugate Confirmationmentioning

confidence: 99%

“…4 Most of the methods for detecting EAs have been developed for the analysis of urine samples. [5][6][7][8][9][10] However, the methods for analysing protein and DNA adducts of NMs are preferable as they are stable from several weeks to months after exposure. 11,12 For sulphur mustard (SM), several methods were developed to determine β-lyase metabolites [13][14][15][16][17] for urine samples.…”

Section: Introductionmentioning

confidence: 99%

“…Lastly, the validated method for EAs was tested for determination of NMs-conjugates from urine samples. 6 2 | EXPERIMENTAL…”

Section: Introductionmentioning

confidence: 99%

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Glutathione conjugation of nitrogen mustards: In vitro study

Hamzah

Kjellberg

Vanninen

2023

Rapid Comm Mass Spectrometry

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Rationale: This paper describes an in vitro study designed to identify metabolic biomarkers resulting from the conjugation of nitrogen mustards (NMs) with glutathione (GSH). The method developed is essential in providing evidence in the event of NM exposure in biomedical samples.Methods: The mass spectral characterization of the proposed NMs-GSH conjugates was performed with liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS). The final reaction mixtures were analysed in positive electrospray ionisation (ESI) at different incubation times.Results: This study identified three types of conjugates in addition to ethanolamines, the hydrolysis products of NMs. Monoglutathionyl, diglutathionyl and phosphorylated conjugates were produced for each of the NMs, bis(2-chloroethyl) ethylamine (HN1), bis(2-chloroethyl)methylamine (HN2) and tris(2-chloroethyl)amine (HN3). The monoglutathionyl conjugates consisted of HN1-GSH, HN2-GSH and HN3-GSH. The spontaneous and primary conjugates of diglutathionyl were HN1-GSH2, HN2-GSH2 and HN3-GSH2. These included phosphorylated conjugates, namely HN1-GSH-PO 4 , HN2-GSH-PO 4 and HN3-GSH-PO 4 , as might have formed due to hydrolysis in phosphate buffer. Conclusions:The mass spectral data of all conjugates formed in the presence of all NMs and GSH are reported in this study. These GSH metabolites can be used to confirm NMs toxicity in biological samples such as urine.

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Section: Resultssupporting

confidence: 59%

Section: Methodsmentioning

confidence: 99%

Section: Conjugate Confirmationmentioning

confidence: 99%