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1.
Article in English | MEDLINE | ID: mdl-28215784

ABSTRACT

The performance of two derivatization and ionization techniques for the quantitative reversed phase liquid chromatography (LC)- mass spectrometry (MS) analysis of hydroxy fatty acids (OH-PUFA) in plasma was evaluated: One used AMPP (N-(4-aminomethylphenyl)pyridinium chloride) leading to a positive charged amid-derivate which can be detected by electrospray ionization (ESI)-MS. Second yielded penta fluorobenzyl bromide (PFB) ester derivates allowing detection in electron capture atmospheric pressure chemical ionization (ecAPCI)-MS. The sensitivity of detection of a comprehensive set of hydroxy fatty acids of n6- and n3- poly unsaturated fatty acids was investigated. On the SCIEX3200 MS the applied PFB derivatization led to poor limits of detection (LOD) of 10-100nM (0.1-1pmol/0.03-0.3ng on column). By contrast, AMPP derivatization led to a similar sensitivity compared to the standard ESI(-) of non derivatized analytes (LOD about 1nM (10fmol/3pg on column)). For several analytes, including 9-HETE, 11-HETE and 17-HDHA the AMPP derivatization improved sensitivity enabling their detection in human plasma. However, precision was reduced by AMPP derivatization and variation in IS recovery indicated a strong matrix influence on the MS-signal. In sum, with the instrumentation used, neither of these derivatization methods improves in our hands the LC-MS based quantification of oxylipins.


Subject(s)
Chromatography, Liquid/methods , Oxylipins/antagonists & inhibitors , Oxylipins/chemistry , Tandem Mass Spectrometry/methods , Blood Chemical Analysis , Humans , Oxylipins/blood , Oxylipins/isolation & purification
2.
J Agric Food Chem ; 64(47): 8973-8976, 2016 Nov 30.
Article in English | MEDLINE | ID: mdl-27933871

ABSTRACT

The product of cytochrome P450 monooxygenase (P450) ω-hydroxylation of arachidonic acid (AA), 20- hydroxyeicosatetraenoic acid (HETE), is a potent vasoconstrictor. Utilizing microsomes as well as individual CYP4 isoforms we demonstrate here that flavonoids can block 20-HETE formation. Apigenin inhibits CYP4F2 with an IC50 value of 4.6 µM and 20-HETE formation in human liver and kidney microsomes at 2.4-9.8 µM. Interestingly, the structurally similar naringenin shows no relevant effect on the formation of 20-HETE. Based on these in vitro data, it is impossible to evaluate if a relevant blockade of 20-HETE formation can result in humans from intake of polyphenols with the diet. However, the potency of apigenin is comparable to those of P450 inhibitors such as ketoconazole. Moreover, an IC50 value in the micromolar range is also described for the inhibition of CYP-mediated drug metabolism leading to food-drug interactions. The modulation of the arachidonic acid cascade by food polyphenols therefore warrants further investigation.


Subject(s)
Apigenin/pharmacology , Arachidonic Acid/metabolism , Cytochrome P450 Family 4/metabolism , Polyphenols/pharmacology , Vasoconstrictor Agents/pharmacology , Cytochrome P450 Family 4/antagonists & inhibitors , Food-Drug Interactions , Humans , Hydroxyeicosatetraenoic Acids/metabolism , Hydroxylation , Inhibitory Concentration 50 , Kidney/drug effects , Kidney/metabolism , Liver/drug effects , Liver/metabolism , Microsomes/drug effects , Microsomes/metabolism
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