Light-scattering detection of phospholipids resolved by HPLC.
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 microg with detection limits below 1 microg 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.
Instituto Tecnología de Alimentos, Centro de Investigación de Agroindustria, Instituto Nacional de Tecnología Agropecuaria Castelar, (B1708WAB) Morón, Província de Buenos Aires, Argentina. adescalzo@cnia.inta.gov.ar
A M
AM
Descalzo
E M
EM
Insani
N A
NA
Pensel
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D010743:Phospholipids / Q000032:analysis* / Q000737:chemistry; D013997:Time Factors
D010743:Phospholipids