کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1199843 1493566 2014 14 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Chiral chromatography–tandem mass spectrometry applied to the determination of pro-resolving lipid mediators
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Chiral chromatography–tandem mass spectrometry applied to the determination of pro-resolving lipid mediators
چکیده انگلیسی


• A very sensitive and selective method to analyze lipid mediators was developed.
• Separation of epimers using a combination of chromatographic stationary phases.
• Chiral chromatography and determination of pro-resolving mediators.
• Validation according to the FDA guidelines was performed.

Pro-resolving lipid mediators are a class of endogenously synthesized molecules derived from different fatty acids, such as arachidonic, docosahexaenoic or eicosapentaenoic acid, which are derived into four different product families: lipoxins, resolvins, maresins and protectins. For quantitation of these compounds, a sensitive, selective and robust liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantitation of lipoxin A4, 6-epi-lipoxin A4, lipoxin B4 and lipoxin A5, the D-series resolvins D1 and D2 as well as aspirin-triggered lipoxin A4 and resolvin D1, maresin and protectin and the pathway markers 17(S)-hydroxy-docosahexaenoic acid and 17(R)-hydroxy-docosahexaenoic acid in cell culture supernatants. For this purpose, a chiral column was connected in series with a reversed-phase column to achieve efficient analyte separation and high sensitivity. Sample pre-treatment included a fast and simple liquid–liquid extraction procedure. Limits of quantitation in the range of 0.1–0.5 ng/mL cell culture media, absolute recoveries between 90 and 115%, intra- and interday precision of less than 13% and an accuracy of less than 11% were obtained. Stability of the samples after 60 days storage at −80 °C, three freeze/thaw cycles and 4 h at room temperature has been demonstrated for all analytes. Sample extracts can be stored at 7 °C for 24 h without degradation of the analytes. Deviations of less than 13% in the accuracy, evaluated in terms of relative error, were obtained. The suitability of the method has been demonstrated in cell culture supernatants of human polymorphonuclear leukocytes, stimulated with 15R-hydroxy-eicosatetraenoic acid and in cell culture media of human polymorphonuclear leukocytes co-incubated with human platelets. From all studied analytes, lipoxin A4 and 6-epi-lipoxin A4 were found in cell culture media under both incubation conditions, while 15-epi-lipoxin A4 was additionally detected in cell culture supernatants of polymorphonuclear leukocytes stimulated with 15R-hydroxy-eicosatetraenoic acid.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1360, 19 September 2014, Pages 150–163
نویسندگان
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