Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy

Two green microalgae species Monoraphidium contortum (M. contortum) and Chlamydomonas sp. that were identified to accumulate lipids were subjected to four different nutrient treatments (NP1-NP4), ranging in nitrate (0.05–5 mM N) and phosphate (2.8–264 μM P) concentrations, at a fixed N:P ratio of ∼18. The effect of nutrient variation on lipid productivity in the species was investigated using second derivative (SD) FTIR and Raman spectroscopy of algal biomass. SD spectral analysis revealed high production of lipid in the form of hydrocarbons (CH) (3000–2800 cm−1), triacylglycerides (TAGs)(∼1740 cm−1), saturated (SFA)(∼1440 cm−1), and unsaturated fatty acids (UFA)(∼3010 cm−1) for the nutrient deplete condition (NP1) in both species. Changes in signals attributed to lipids in proportion to other biochemical components were consistent with physiological changes expected from nutrient depletion. Relative signal intensities for lipids showed a significant increase in NP1, in particular, CH, TAGs in relation to protein signals (in SD-FTIR), and SFA, UFA in relation to carotenoid signals (in SD-Raman). PCA performed on the negative spectral values of the SD-FTIR and SD-Raman data for the four NP treatments enabled discrimination not only between the species but also between the NP treatments and the timing of harvest. M. contortum was found to contain a relatively higher proportion of CH, TAGs, SFA, and UFA compared to Chlamydomonas sp. Peak areas from the negative SD spectra, informed by PCA analysis, enabled capturing quantifiable changes in a manner that is consistent with known microalgal physiology. SD-FTIR and SD-Raman spectroscopy have been shown to possess superior potential to capture relevant microalgal physiological changes.