Spectrophotometric analysis of pigments

a critical assessment of a high-throughput method for analysis of algal pigment mixtures by spectral deconvolution

Jan Erik Thrane, Marcia Kyle, Maren Striebel, Sigrid Haande, Merete Grung, Thomas Rohrlack, Tom Andersen

Research output: Contribution to journalArticle

24 Citations (Scopus)
4 Downloads (Pure)

Abstract

The Gauss-peak spectra (GPS) method represents individual pigment spectra as weighted sums of Gaussian functions, and uses these to model absorbance spectra of phytoplanktonpigment mixtures. We here present several improvements for this type of methodology, including adaptation to plate reader technology and efficient model fitting by open source software. We use a one-step modeling of both pigment absorption and background attenuationwith non-negative least squares, following a one-time instrument-specific calibration. The fitted background is shown to be higher than a solvent blank, with features reflecting contributions from both scatter and non-pigment absorption. We assessed pigment aliasing due to absorption spectra similarity by Monte Carlo simulation, and used this information to select a robust set of identifiable pigments that are also expected to be common in natural samples. To test the method's performance, we analyzed absorbance spectra of pigmentextracts from sediment cores, 75 natural lake samples, and four phytoplankton cultures, and compared the estimated pigment concentrations with concentrations obtained using high performance liquid chromatography (HPLC). The deviance between observed and fittedspectra was generally very low, indicating that measured spectra could successfully be reconstructed as weighted sums of pigment and background components. Concentrations of total chlorophylls and total carotenoids could accurately be estimated for both sediment and lake samples, but individual pigment concentrations (especially carotenoids) proved difficult to resolve due to similarity between their absorbance spectra. In general, our modified-GPSmethod provides an improvement of the GPS method that is a fast, inexpensive, and high-throughput alternative for screening of pigment composition in samples of phytoplanktonmaterial.

Original languageEnglish
Article numbere0137645
Pages (from-to)1-24
Number of pages24
JournalPLoS ONE
Volume10
Issue number9
DOIs
Publication statusPublished - 11 Sep 2015
Externally publishedYes

Bibliographical note

Copyright the Author(s) 2015. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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