Quantitative determination of lysophosphatidic acids (LPAs) in human saliva and gingival crevicular fluid (GCF) by LC-MS/MS.

摘要: Lysophosphatidic acid (LPA) is a phospholipid mediator that plays multiple cellular functions by acting through G protein-coupled LPA receptors.LPAs are known to be key mediators in inflammation,and several lines of evidence suggest role for LPAs inflammatory periodontal diseases. A simple and sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method has been developed validated quantify species (LPA 18:0, 16:0, 18:1 20:4) human saliva gingival crevicular fluid (GCF). 17:0 was used as an internal standard the were extracted from liquid-liquid extraction using butanol. Chromatography performed Macherey-Nagel NUCLEODUR® C8 Gravity Column (125mm × 2.0mm ID) mixture methanol/water: 75/25 (v/v) containing 0.5% formic 5mM ammonium formate (mobile Phase A) methanol/water : 99/0.5 phase B) at flow rate 0.5 mL/min. detected linear ion trap-triple quadrupole spectrometer total run time 8.5 min. The limit quantification (LOQ) 1 ng/mL all validatedover range 1-200 ng/mL. GCF over ranges 10-500 18:0 5-500 20:4. This LC-MS/MS assay successfully applied obtain quantitative data individual levels control subjects patients various All four consistently elevated samples obtained diseases, which supports thepathogenesis