Amx 400 mhz spectrometer

Platelet-activating factor (PAF; β-acetyl-γ-O-hexadecyl-L-α-phosphatidylcholine), formyl-met-leu-phe (fMLP), and Gue1654 (7-(methylthio)-2-[(2,2-diphenylacetyl)amino]benzo[1,2-d:4,3-d′]bisthiazole) were obtained from Sigma-Aldrich, whereas interkleukin-8 (IL-8) was purchased from R&D Systems. 5-Oxo-ETE³⁵ and LTB₄³⁶ were prepared by chemical synthesis as previously described. All reactions were carried out using dry solvents under argon atmosphere. High-resolution mass spectra were recorded on an AccuTOF mass spectrometer by positive ion ESI mode with DART as an ion source. ¹H NMR and ¹³C NMR spectra were recorded in CDCl₃ using TMS as an internal standard on a BRUKER AMX 400 MHz spectrometer at rt. All compounds were analyzed by TLC, HRMS and NMR. Prior to biological assay, the purity of all final compounds was determined to be >95% by NMR and HPLC. HPLC conditions: Waters 2695 Alliance System with Waters Novapak C18 (150 × 3.9 mm) column and photodiode array detector (Waters Model 2996), gradient mobile phase between H₂O/MeCN/MeOH (56:22:22) to H₂O/MeCN/MeOH (16:42:42) over 30 min, both solvents contained 0.02% acetic acid, and a flow rate was 1 mL/min.

All reagents and solvents used for chemical synthesis were purchased from Sigma-Aldrich, Saint Louis, MO. All reactions were carried out under an argon atmosphere using dried, nitrogen-purged glassware and dry solvents. Reaction progress was monitored using Merck TLC Silica gel 60 F₂₅₄ plates. The plates were visualized using 254 nm UV light, iodine chamber and anisaldehyde dip where appropriate, followed by gentle warming. ¹H NMR and ¹³C NMR spectra were recorded on a BRUKER AMX 400 MHz spectrometer at rt in CDCl₃, using TMS as an internal standard. The following abbreviations are used for the description of NMR spectra: s- singlet; d- doublet; t- triplet; q- quartet; dd- doublet of doublets and m- multiplet. High resolution mass spectrometry (HRMS) was performed using an AccuTOF mass spectrometer with positive ion ESI mode and DART as an ion source. The purity of all tested compounds was determined to be >95% by a combination of HPLC, NMR and HRMS.

All reactions were carried out under an argon atmosphere using dry solvents. DMSO was purchased from Aldrich as anhydrous grade and used without further purification. Potassium tert-butoxide solution (1.0 M in THF), titanium tetrachloride (1.0 M in CH₂Cl₂), tetraethylammonium iodide, tert-Butyl (chloro)diphenylsilane (TBDPS-Cl), and 3,4-Dihydro-2H-pyran (DHP) were purchased from Aldrich. The catalysts, p-toluenesulfonic acid monohydrate and 10% Pd/C were purchased from Aldrich. All compounds were analyzed by TLC, NMR and HRMS. ¹H NMR and ¹³C NMR spectra were recorded at rt on a BRUKER AMX 400 MHz spectrometer in CDCl₃ using TMS as an internal standard. High-resolution mass spectra were recorded using a JEOL DART-AccuTOF mass spectrometer (JEOL USA, Inc, Peabody, MA). Prior to biological assay, the purity of all final compounds was determined to be >95% by a combination of HPLC, NMR and HRMS.