Fourier-transform infrared spectroscopic analysis of rabbit lung surfactant : subfraction-associated phospholipid and protein profiles

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DOIResolve DOI: http://doi.org/10.1016/0009-3084(95)02471-T
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TypeArticle
Journal titleChemistry and Physics of Lipids
ISSN0009-3084
Volume77
Issue2
Pages193201; # of pages: 9
SubjectSurfactant; OM; Infrared spectroscopy; Lung
AbstractSurfactant obtained from bronchoalveolar lavage (BAL) can be separated into subfractions based on sedimentation characteristics. It has been suggested that the 10 000 × g, 60 000 × g and 100 000 × g subfractions isolated by this approach represent stages of surfactant extracellular processing. These three subfractions have been reported to differ in their morphology, composition and ability to lower surface tension. We wished to determine if infrared spectroscopy, which may be applied as a non-invasive technique could potentially prove useful for characterization and quantification of bronchoalveolar lavage (BAL) protein and phospholipid, and if this approach could detect differences in intermediate surfactant processing stages. Subfractions were collected from adult rabbit lungs by BAL and differential centrifugation and analyzed by Fourier transform infrared (FT-IR) spectroscopy. Biochemical assay of phospholipid and protein showed differences between subfractions that correlated well with the phospholipid/ protein ratios obtained from FT-IR spectra (r = 0.939; r2 = 0.882). The subfraction sedimenting at 100 000 × g (P100) exhibited spectral shifts in the Amide I band, suggesting that the protein secondary structure was different compared to other fractions. Spectra obtained after separation of lipids and protein components showed an apparent disordering of protein secondary structure but little or no effect on the structure or mobility of phospholipids. These results support the idea that subfractions represent various processing stages of surfactant. In addition, they show that results from FT-IR analyses correlate significantly with traditional biochemical assay methods which may prove of clinical use. They also show that each fraction displays characteristic FT-IR spectra suggesting that this technique may potentially provide a non-invasive method for further detailed analysis of BAL.
Publication date
PublisherElsevier Science Ireland Ltd.
LanguageEnglish
AffiliationNational Research Council Canada; NRC Institute for Biodiagnostics
Peer reviewedYes
NRC number342
NPARC number9742235
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Record identifier2de26009-28dc-446b-9f9d-0defb43b342b
Record created2009-07-17
Record modified2016-11-28
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