title: Quantitation of Monophosphorylated Lipid A in the Oil-in-Water Adjuvant Delivery Systems Using Transesterification and GC-MS.
abstract: Vaccine delivery systems play pivotal role in effective antigen delivery. These systems often contain adjuvants that stimulate specific immune response and are important for vaccines' efficacy and safety. Oil-in-water vaccine delivery lipid emulsion systems containing monophosphoryl lipid A (MPLA) as immune modulator have been extensively investigated in vaccine trials. Herein, we describe a simple orthogonal method, for quantitative measurement of MPLA in an oil-in-water lipid delivery system using direct transesterification reaction followed by gas-chromatography-mass spectrometry analysis. In this protocol, the transesterification reaction results in the release of fatty acid methyl esters followed by gas-chromatography-mass spectrometry-based targeted quantification of the specific 3-hydroxytetradecanoate fatty acid methyl ester to measure the concentration of MPLA in an oil-in-water lipid emulsion system.
authors.affiliation: Laboratory of Proteomics and Analytical Technologies, Advanced Technology Program, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702-1201. Electronic address: athar.masood@fda.hhs.gov.
authors.forename: M Athar
authors.initials: MA
authors.lastname: Masood
authors.affiliation: Laboratory of Proteomics and Analytical Technologies, Advanced Technology Program, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702-1201.
authors.forename: Josip
authors.initials: J
authors.lastname: Blonder
authors.affiliation: Laboratory of Proteomics and Analytical Technologies, Advanced Technology Program, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702-1201.
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Veenstra
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000032:analysis*; D016503:Drug Delivery Systems; D004655:Emulsions / Q000737:chemistry*; D004951:Esterification; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D008050:Lipid A / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000032:analysis; D009821:Oils / Q000737:chemistry*; D014677:Pharmaceutical Vehicles / Q000737:chemistry*; D014612:Vaccines / Q000008:administration & dosage / Q000032:analysis*; D014867:Water / Q000737:chemistry
chemical_list: D000276:Adjuvants, Immunologic; D004655:Emulsions; D008050:Lipid A; D009821:Oils; D014677:Pharmaceutical Vehicles; D014612:Vaccines; D014867:Water; C048436:monophosphoryl lipid A
keywords: GC-MS quantitation method; fatty acid methyl esters (FAMEs); glucopyranosyl lipid A (GLA); monophosphorylated lipid A (MPLA); oil-in-water emulsion; transesterification; vaccine adjuvants