An SPME-GC-MS method using an octadecyl silica fibre for the determination of the potential angiogenesis modulators 17beta-estradiol and 2-methoxyestradiol in culture media.
A simple and easily automable method based on solid-phase microextraction followed by gas chromatographic-mass spectrometric analysis was developed for the determination of two potential angiogenesis modulators 17beta-estradiol (17-BE) and 2-methoxyestradiol (2-MEOE) in culture media. Trifluoroacetic anhydride was used as the derivatising agent. A homemade octadecyl silica coating, characterised by a coating thickness of 72 +/- 10 microm and a good thermal stability until 250 degrees C, was prepared. Experimental design was used to optimise the extraction conditions in terms of derivatisation time, derivatisation temperature and time of extraction. As for method validation, lower limits of quantification of 0.17 and 0.015 microg/l for 17beta-estradiol and 2-methoxyestradiol, respectively, were obtained. Finally, the capabilities of the developed fibres were evaluated for the analysis of the investigated analytes developed by granulosa cells in culture media maintained under normoxic, hypoxic and anoxic conditions, in order to better elucidate their possible role in the angiogenic process. An increase of the production of both 17-BE and 2-MEOE in hypoxic and anoxic conditions seems to be related to the effect of oxygen deprivation.
Bianchi
Federica
F
Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica, Università degli Studi di Parma, Viale Usberti 17/A, 43100 Parma, Italy. federica.bianchi@unipr.it
Mattarozzi
Monica
M
Careri
Maria
M
Mangia
Alessandro
A
Musci
Marilena
M
Grasselli
Francesca
F
Bussolati
Simona
S
Basini
Giuseppina
G
D000077584:2-Methoxyestradiol; D043924:Angiogenesis Modulating Agents / Q000032:analysis*; D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D004958:Estradiol / Q000031:analogs & derivatives / Q000032:analysis*; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006107:Granulosa Cells / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide / Q000737:chemistry*; D052616:Solid Phase Extraction / Q000379:methods*; D013552:Swine
D043924:Angiogenesis Modulating Agents; D003470:Culture Media; D004958:Estradiol; D000077584:2-Methoxyestradiol; D012822:Silicon Dioxide