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Table 4 Examples of conventional chromatographic/spectrometric methods for lipid quantification and profiling of oleaginous microorganisms

From: Lipids detection and quantification in oleaginous microorganisms: an overview of the current state of the art

Method Microorganisms Sample preparation Compound detection and quantification Correlation with method Outcome Reference
FABMS Chlorella vulgaris, Scenedesmus obliquus Algal cells, intact Semi-quantitative analysis of fatty acids, TAGs More flexible and versatile than GC [85]
GC-MS Chlorella kessleri, C. vulgaris, Scenedesmus acuminatus, Spirulina platensis Solvent extraction of dried cells
Saturated and unsaturated straight-chain methyl fatty acids, iso- and anteisoacids branched on other carbon atoms and cyclopropane fatty acids Fatty acid profiling [88]
GC-MS Chlamidomonas nivalis In situ transesterification Fatty acid profiling Fatty acid biomarkers correspond to response and adaptation to NaCl stress [89]
Pyrolysis GC-MS Chlamydomonas reinhardtii, B. braunii Lyophilized algae culture, Immersion into isopropanol C8 to C40 pyrolysis products Automated manner without sample preparation
30 min run times, sequences up to 48 samples
Pyrolysis GC-MS 26 microalgae and cyanobacteria Whole cells, freeze-dried Biochemical composition (lipids, protein, carbohydrate) Gravimetric Fast analysis (1 h), minimal sample amount (< 0.1 mg), no sample preparation [91]
GC-MS sub-microscale assay Mallomonas splendens, Nannochloropsis oculata, Chrysochromulina sp., Emiliania huxleyi, Rhodomonas sp., Prorocentrum micans In situ transesterification Saturated and unsaturated methyl fatty acids BODIPY, flow cytometry Minimal size requirement (250 μg dry biomass per sample), fast, less solvent than AOAC and AOCS [44]
MALDI-TOF-MS, ESI-LTQ-Orbitrap MS 7 algal species Solvent extraction TAG profile Direct analysis [92]
DTD-GC x GC-TOF-MS Limnothrix sp., Scenedesmus acutus, Chironomous plumosus, Daphnia galatea Thermally assisted hydrolysis and methylation FAMEs Analysis of lipid markers for food-web, ecological and clinical studies [93]
MS/MS-CID employing paper spray ionization Kyo-Chlorella, Nannochloropsis Whole cells, lyophilized Polar lipid profiling (glycolipids, phospholipids), double bond position determination Structural determination of polar lipids, minimal sample preparation, low sample amount (0.5 μg) [94]
UPLC-MS, ESI-MS/MS, ESI-MS 4 Dunaliella species Solvent extraction TAGs, free fatty acids Low sample concentration, small-scale experiments [95]
HPLC-ELSD C. reinhardtii, Chlorella strains Solvent extraction from dried pellets TAGs, carotenoids, DAGs, free fatty acids, phospholipids, galactolipids Comparison with TLC/GC, gravimetric, GC/MS 15 min run, detectable TAG levels > 0.5 μg/g dry weight [96]
HPLC-ELSD, preparative Scripsiella sp. Solvent extraction by modified Bligh & Dyer, transesterification and FAME were purified with TLC Polyunsaturated fatty acids Polyunsaturated fatty acids purification, enrichment and purification of uncommon VLC polyunsaturated fatty acids [97]
HPLC-ELSD/MS Chlorella Solvent extraction, Folch method TAGs, hydrocarbons No derivatization needed for determining lipid molecules in one peak [98]