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 50^oC 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 derived from the following

[1] PLoS Genet. 2013 Jan; 9(1): e1003271.
Published online 2013 Jan 31. doi: 10.1371/journal.pgen.1003271
A Heterogeneous Mixture of F-Series Prostaglandins Promotes Sperm Guidance in the Caenorhabditis elegans Reproductive Tract
Hieu D. Hoang,  Jeevan K. Prasain,  Dixon Dorand,  and Michael A. Miller, *

Resources

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