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. 2024 May 29;25(11):5919.
doi: 10.3390/ijms25115919.

Development of a New Binary Matrix for the Comprehensive Analysis of Lipids and Pigments in Micro- and Macroalgae Using MALDI-ToF/ToF Mass Spectrometry

Mariachiara Bianco  1 , Giovanni Ventura  1   2 , Davide Coniglio  1 , Antonio Monopoli  1 , Ilario Losito  1   2 , Tommaso R I Cataldi  1   2 , Cosima D Calvano  1   2
Affiliations

Development of a New Binary Matrix for the Comprehensive Analysis of Lipids and Pigments in Micro- and Macroalgae Using MALDI-ToF/ToF Mass Spectrometry

Mariachiara Bianco et al. Int J Mol Sci. .

Abstract

While edible algae might seem low in fat, the lipids they contain are crucial for good health and preventing chronic diseases. This study introduces a binary matrix to analyze all the polar lipids in both macroalgae (Wakame-Undaria pinnatifida, Dulse-Palmaria palmata, and Nori-Porphyra spp.) and microalgae (Spirulina-Arthrospira platensis, and Chlorella-Chlorella vulgaris) using matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The key lies in a new dual matrix made by combining equimolar amounts of 1,5-diaminonaphthalene (DAN) and 9-aminoacridine (9AA). This combination solves the limitations of single matrices: 9AA is suitable for sulfur-containing lipids and acidic phospholipids, while DAN excels as an electron-transfer secondary reaction matrix for intact chlorophylls and their derivatives. By employing the equimolar binary matrix, a wider range of algal lipids, including free fatty acids, phospholipids, glycolipids, pigments, and even rare arsenosugarphospholipids were successfully detected, overcoming drawbacks related to ion suppression from readily ionizable lipids. The resulting mass spectra exhibited a good signal-to-noise ratio at a lower laser fluence and minimized background noise. This improvement stems from the binary matrix's ability to mitigate in-source decay effects, a phenomenon often encountered for certain matrices. Consequently, the data obtained are more reliable, facilitating a faster and more comprehensive exploration of algal lipidomes using high-throughput MALDI-MS/MS analysis.

Keywords: MALDI MS/MS; alga; binary matrix; chlorophylls; lipids.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
MALDI-MS of an EquiSPLASH™ LIPIDOMIX® standard mixture of 13 deuterated standard lipids containing 15:0 and 18:1(d7) fatty acyl chains using DAN (A), 9AA (B), and the binary mixture of DAN/9AA (C) as matrices. The matrix signals are denoted by an asterisk.
Figure 2
Figure 2
MALDI-MS of Wakame lipid extract using DAN (plot A), 9AA (plot B), and a binary DAN/9AA mixture (plot C) as matrices. The matrix signals are denoted by an asterisk.
Figure 3
Figure 3
MALDI-MS/MS of Pheophytin a at m/z 870.54 (plot A), sulfoquinovosyldiacylglycerol at m/z 819.57 (plot B), phosphatidylinositol at m/z 835.53 (plot C), and sulfoquinovosyldiacylglycerol/phosphatidylglycerol at m/z 765.48 (plot D). DAN/9AA was used as a matrix.
Figure 4
Figure 4
MALDI-MS/MS of phosphatidylglycerol at m/z 719.49 (plot A), phosphatidylethanolamine at m/z 688.50 (plot B), and arsenosugar PL at m/z 957.50 (plot C). Binary DAN/9AA was used as a matrix.
Figure 5
Figure 5
MALDI-MS in the m/z range of 200–500 of microalgae Spirulina (plot A), Chlorella (plot B), and macroalgae Dulse (plot C), and Nori (plot D) using the binary mixture of DAN/9AA. Matrix signals are indicated by *.
Figure 6
Figure 6
MALDI-MS in the m/z range of 500–1000 of microalgae Spirulina (plot A), Chlorella (plot B), macroalgae Dulse (plot C), and Nori (plot D) using the binary mixture of DAN/9AA.
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