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Light‐scattering detection of phospholipids resolved by HPLC
Author(s) -
Descalzo A. M.,
Insani E. M.,
Pensel N. A.
Publication year - 2003
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-003-1154-1
Subject(s) - chromatography , high performance liquid chromatography , chromatography detector , chemistry , calibration curve , detection limit , solid phase extraction , hexane , elution , quantitative analysis (chemistry) , extraction (chemistry) , methanol , pulmonary surfactant , acetonitrile , reversed phase chromatography , organic chemistry , biochemistry
An improved method for the analysis of phospholipids by normal‐phase HPLC is described. Addition of methanol and acetonitrile to a gradient based on 2‐propanol/hexane/water promoted a rapid separation of major classes of bovine surfactant phospholipids (PL) by using a conventional silica column. The use of an ELSD permitted an accurate analysis of a mixture of PL. Calibration curves were linear within the range of 5–40 μg with detection limits below 1 μg for PE and PC, and CV ranged from 0.6 to 9.6%. PL present in surfactant homogenates were separated by a solid‐phase extraction (SPE) procedure before HPLC analysis. This methodology gave a recovery of 95% and combined SPE‐HPLC and quantification of biological PL within a 30‐min run. The use of ELSD detection of the eluted compounds was precise, linear, and sensitive.