G1311c quaternary pump

Mass spectra were obtained using a Bruker micro-TOFQ mass spectrometer (Bruker Daltonics, Bremen, Germany). Nuclear magnetic resonance spectra were acquired with a Bruker AV-500 FT-NMR spectrometer operating at 500.1 MHz for ¹H and at 125.8 MHz for ¹³C at 25 °C; chemical shifts are expressed in δ referring to the residual solvent signals δ_H 2.50 and δ_C 39.5 for DMSO-d₆, coupling constants, J., are in hertz. All chemical shifts are given in ppm. Column chromatography was performed over a RP-18 reversed-phase silica gel (S-50 μm; YMC, Kyoto, Japan). Analytical HPLC (Agilent technologies, Santa Clara, CA, USA) was performed on an Agilent 1260 system equipped with a G1311C quaternary pump, a G1329B autosampler, a G1316A thermostated column compartment, and a G1314F variable wavelength detector coupled with an analytical workstation. Semipreparative HPLC was performed on an Agilent ProStar SD-1 pump connected with an Agilent ProStar 320 UV–Vis detector (at 360 nm), utilizing a Shim-Pack PREP-ODS column (250 mm × 21.2 mm, i.d., 10 μm, Shimadzu, Kyoto, Japan). HPLC-grade water was purified using a Milli-Q system (Millipore, Boston, MA, USA). All solvents used for the chromatographic separations were distilled before use.

Analysis of phospholipids by liquid chromatography coupled to mass spectrometry was carried out as described elsewhere (34 (link)–41 (link)), using an Agilent 1260 Infinity high-performance liquid chromatograph equipped with an Agilent G1311C quaternary pump and an Agilent G1329B Autosampler, coupled to an API2000 triple quadrupole mass spectrometer (Applied Biosystems). Phospholipid molecular species were identified by comparison with previously published data (34 (link)–41 (link)). Lipid analysis by gas chromatography coupled to mass spectrometry was carried out exactly as described elsewhere (59 (link)–63 (link)), using an Agilent 7890A gas chromatograph coupled to an Agilent 5975C mass selective detector operated in electron impact mode, equipped with an Agilent 7693 Autosampler and an Agilent DB23 column (60 m length × 0.25 mm internal diameter × 0.15 µm film thickness).

Analysis of prostaglandins by LC/MS was carried out exactly as described elsewhere [15 (link),33 (link)], using an Agilent 1260 Infinity high-performance liquid chromatograph equipped with an Agilent G1311C quaternary pump and an Agilent G1329B Autosampler, coupled to an API2000 triple quadrupole mass spectrometer (Applied Biosystems, Carlsbad, CA, USA). Quantification was carried out by integrating the chromatographic peaks of each species and comparing with a calibration curve made with analytical standards.