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Table 10 Comparison between conventional and non-conventional methods for lipids determination

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

Methods Advantages Disadvantages
Conventional
 Gravimetric The easiest method to quantify total lipids in a short time, no need for special equipment or training Requires cell disruption, extraction, not suitable for small amounts of sample
 Chromatographic coupled with detector Well-established procedures for lipid profiling Requires cell disruption, extraction, and (in situ) transesterification, multi-step process; Depends on microalgae strains and cellular extraction method
Non-conventional
 FTIR Identification of lipid classes on the basis of standards, cellular content determination without disruption of cells, small amount of samples FTIR band shifts are species-specific, cannot differentiate different species when a mixture of cultures is used
 Raman spectroscopy Label-free, no sample preparation, in vivo analysis, real-time, non-destructive Fluorescent pigments and carotenoids interfere with analysis; quantification is focused mainly on determining the degree of unsaturation, there is need for calibration based on other methods such as gravimetric analysis; difficult to translate to an industrial setting
 NMR Can be non-destructive, application to whole cells, extracted and transesterified lipids Need for internal standards for quantification; difficult to translate to an industrial setting
 Dielectric spectroscopy Rapid, non-invasive, and label-free method; potential for automated process for biomonotring Total lipid detection, no profiling
 Fluorescence spectrometry High-throughput potential Uptake and intensity depends on dye and cell wall; only detects neutral lipids
 Nile Red Mostly used in literature Photo-bleaching of agent; poor penetration of cells; interference with chlorophyll autofluorescence and green fluorophores; limited specificity to lipids
 BODIPY 505/515 More lipid-specific, narrower emission spectrum, higher sensitivity, and better reproducibility, requires lower concentration of solvent carrier Interference with green fluorophores
 AC-202 More sensitive than BODIPY; low-background signal Not used for quantification yet
 Colorimetry High-throughput potential
Low cost
Total lipid detection only
 SPV Color formation is stable for hours
Fast, low detection limit
Solvent extraction can be omitted depending on the assay
Requires presence of double bond or free hydroxyl groups (unsaturated lipids)
Is dependent on standard
 TAG kit Fast, automated process Detects only TAGs; requires cell disruption and extraction
 Copper extraction Fast, high detection limit, can be adapted to microcentrifuge format Requires solvent addition and cell disruption