Chromatography

Liquid chromatography tandem mass spectrometry (LC-MS) was carried out with a Shimadzu Prominence HPLC system and SCIEX 6500 mass spectrometer. An aliquot of 20 µl of each sample was injected onto an Synergi 3 µm C18 250 x 2.1 mm column at 50oC for gradient separation. Mobile phases were: A, ddH₂O with 0.1% Formic Acid; and B, Acetonitrile with 0.1% Formic Acid. Gradient schedule starts at 10% B and is linearly increased to 80% B at 11 minutes, then 100% B at 14 minutes, and then returned to 10% B at 16 at minutes and re-equilibration until 20 minutes. Time per sample injection was 20 minutes.

Extraction Protocols:

Extraction protocols for sample processing are dependent on the matrices involved. Please see extraction methods section for more specific instructions.

References

Method was adapted from the following reference:

Singh A, Rahman T, Bartiss R, Arabshahi A, Prasain J, Barnes S, Musteata FM, Sellati TJ. Lipoxin A4, a 5-lipoxygenase pathway metabolite, modulates immune response during acute respiratory tularemia. J Leukoc Biol. 2017 Feb;101(2):531-542. doi: 10.1189/jlb.4A0815-365RR. Epub 2016 Sep 14 [PMID: 27630217]

Resources

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