Capillary gas chromatography of hexadecylphosphocholine in Caco-2T cells and cell culture media.
The gas-chromatographic determination of hexadecylphosphocholine (HePC), an experimental antitumor agent of the alkyllysophospholipid group, in Caco-2T cell culture and cell culture media is described. The Caco-2T cells were treated with HePC at a concentration of 40 micrograms/ml (9.8 microM) and the uptake of the drug into the cells (calculated per milligram protein) was measured after 48 h culture (37 degrees C, 10% CO2). Also, a reversibility test for another 48 h was carried out in which the retention of the drug was measured. The toxicity of HePC on Caco-2T cells in viability assays was determined. Before the capillary gas-chromatographic determination, sample cleanup was performed by solid-phase extraction (SPE) on a weak cation-exchange column of the CBA (carboxylic acid) type. For quantitation, racemic 1-O-octadecyl-2-O-methylglycero-3-phosphorylcholine (ET-18-OMe) was added as internal standard, followed by derivatization with trimethylsilylbromide. The results showed that HePC taken up by the cells during 48 h of treatment was still detectable 48 h after removal of the drug from the medium.
Steelant
W F
WF
Laboratory for Pharmaceutical Biotechnology, University of Ghent, Belgium.
Bruyneel
E A
EA
Mareel
M M
MM
Van den Eeckhout
E G
EG
D000970:Antineoplastic Agents / Q000494:pharmacology; D016923:Cell Death; D002849:Chromatography, Gas; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D006801:Humans; D010742:Phospholipid Ethers / Q000032:analysis*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000032:analysis / Q000494:pharmacology; D014297:Trimethylsilyl Compounds / Q000737:chemistry; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D003470:Culture Media; D010742:Phospholipid Ethers; D014297:Trimethylsilyl Compounds; D010767:Phosphorylcholine; C026659:edelfosine; C068554:trimethylsilyl bromide; C039128:miltefosine