A novel approach based on solid phase microextraction gas chromatography and mass spectrometry to the determination of highly reactive organic compounds in cells cultures: styrene oxide.
A solid phase microextraction (SPME) gas chromatography/mass spectrometry (GC/MS) method was developed to assess actual doses of highly reactive organic compounds like styrene oxide (SO) in exposed cell cultures. Using SPME, we set up a method to measure accurately extracellular SO concentrations as well as to obtain an approximate assessment of intracellular levels. The SPME-GC/MS method was developed and validated using two different coating materials, carboxen-PDMS and polyacrylate. In cell-free systems, linearity was established over 3 orders of magnitude for both fibers, but carboxen-PDMS showed higher extraction efficiency and a lower limit of detection (0.5 x 10(-7) vs 10(-6) M for polyacrylate). Precision calculated as % RSD was within 4-16% for all intra- and interday determinations. Experiments performed to study SO stability in cell-free medium showed a time-dependent decrease in SO concentration (11% of initial the concentration after 24 h), mostly due to the spontaneous hydrolysis of SO into styrene glycol, which was measured by liquid chromatography/tandem mass spectrometry (LC/MS/MS). When the neuronal cell line (SK-N-MC) was exposed to a nominal concentration of 0.3 x 10(-4) M SO, the actual concentration measured in the supernatant was considerably lower and was found to decrease during incubation. Intracellular SO was estimated indirectly, by difference between the amount measured in the medium without cells and in the supernatant of the cell-containing medium.
Poli
Diana
D
ISPESL Research Center, and Laboratory of Industrial Toxicology, Department of Clinical Medicine, Nephrology and Health Sciences, University of Parma, Italy.
Vettori
Maria Vittoria
MV
Manini
Paola
P
Andreoli
Roberta
R
Alinovi
Rossella
R
Ceccatelli
Sandra
S
Mutti
Antonio
A
D000179:Acrylates; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media / Q000032:analysis; D004121:Dimethyl Sulfoxide; D004129:Dimethylpolysiloxanes; D004852:Epoxy Compounds / Q000032:analysis* / Q000633:toxicity; D005026:Ethylene Glycols / Q000032:analysis / Q000633:toxicity; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D009447:Neuroblastoma; D011108:Polymers; D015203:Reproducibility of Results; D012828:Silicones; D012995:Solubility
D000179:Acrylates; D003470:Culture Media; D004129:Dimethylpolysiloxanes; D004852:Epoxy Compounds; D005026:Ethylene Glycols; D011108:Polymers; D012828:Silicones; C031474:styrene glycol; C013830:baysilon; C013690:styrene oxide; D004121:Dimethyl Sulfoxide