Liquid chromatography electrospray ionization tandem mass spectrometry analysis method for simultaneous detection of trichloroacetic acid, dichloroacetic acid, S-(1,2-dichlorovinyl)glutathione and S-(1,2-dichlorovinyl)-L-cysteine.
Trichloroethylene (TCE, CAS 79-01-6) is a widely used industrial chemical, and a common environmental pollutant. TCE is a well-known carcinogen in rodents and is classified as "probably carcinogenic to humans". Several analytical methods have been proposed for detection of TCE metabolites in biological media utilizing derivatization-free techniques; however, none of them is suitable for simultaneous detection of both oxidative metabolites and glutathione conjugates of TCE in small volume biological samples. Here, we report a new combination of methods for assessment of major TCE metabolites: dichloroacetic acid (DCA), trichloroacetic acid (TCA), S-(1,2-dichlorovinyl)-L-cysteine (DCVC), and S-(1,2-dichlorovinyl) glutathione (DCVG). First, DCA and TCA were extracted with ether. Second, the remaining aqueous fraction underwent solid phase extraction for DCVC and DCVG. Then, DCA and TCA were measured by hydrophilic interaction liquid chromatography ion exchange negative electrospray ionization tandem mass spectrometry, while DCVC and DCVG were measured by reverse phase positive electrospray ionization tandem mass spectrometry. This method was applied successfully to measure all 4 TCE metabolites in as little as 50 microl of serum from mice orally exposed to TCE (2100 mg/kg, 2h). Serum concentrations (mean+/-standard deviation) of the TCE metabolites obtained with this method are comparable or equivalent to those previously reported in the literature: DCA, 0.122+/-0.014 nmol/ml (limit of detection: 0.01 nmol/ml); TCA, 256+/-30 nmol/ml (0.4 nmol/ml); DCVG, 0.037+/-0.015 nmol/ml (0.001 nmol/ml); DCVC, 0.0024+/-0.0009 nmol/ml (0.001 nmol/ml). This method opens new opportunities to increase throughput and decrease number of animals required for mechanistic studies on TCE in rodents.
Kim
Sungkyoon
S
Department of Environmental Sciences and Engineering, School of Public Health, University of North Carolina, CB 7431, Chapel Hill, NC 27599, USA.
Collins
Leonard B
LB
Boysen
Gunnar
G
Swenberg
James A
JA
Gold
Avram
A
Ball
Louise M
LM
Bradford
Blair U
BU
Rusyn
Ivan
I
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003545:Cysteine / Q000031:analogs & derivatives / Q000097:blood; D003999:Dichloroacetic Acid / Q000097:blood; D005978:Glutathione / Q000031:analogs & derivatives / Q000097:blood; D008297:Male; D051379:Mice; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014238:Trichloroacetic Acid / Q000097:blood; D014241:Trichloroethylene / Q000378:metabolism*
D014241:Trichloroethylene; D014238:Trichloroacetic Acid; C039961:S-(1,2-dichlorovinyl)cysteine; C041464:S-(1,2-dichlorovinyl)glutathione; D003999:Dichloroacetic Acid; D005978:Glutathione; D003545:Cysteine