Development of Method for Serum Preparation and Lipidomics Analysis Based on Rapid Resolution Liquid Chromatography-Mass Spectrometry
A rapid resolution liquid chromatography-mass spectrometric(RRLC-MS) method was developed and validated for the determination of endogenous lipid metabolites in human serum.The samples were pretreated by lipid extraction with methylene chloride-methanol(2 ∶ 1,V/V) mixture solvent and introduced to RRLC-MS under intensive optimization of chromatography conditions and mass spectrometry parameters.The chromatographic separation was performed on an ACQUITY UPLC CSH C18 column(100 mm×2.1 mm,1.7 μm) with mobile phase containing 10 mmol/L ammonium formate(0.1% formic acid)(A) and acetonitrile-isopropyl alcohol(1 ∶ 1,V/V) mixture solution(B) in a linear gradient elution.The MS detection was carried out by the integrated ionization approach using electrospray ionization(ESI) in both positive and negative ion modes and atmosphere pressure chemical ionization(APCI) in positive ion mode.Typical lipid molecules such as free fatty acids(FFAs),phospholipids(PLs),sphingophosphalipids(SPs),glycerolipids(GLs),cholesterol(Chol.) and cholesterol esters(CEs) had been detected with high sensitivity,good precision and reliable stability.Furthermore,the principal component analysis(PCA) results of the quality control(QC) samples among the real sample queue indicated the data reliability of the method in lipidomics applications.