Using FTIR spectroscopy for rapid determination of lipid accumulation in response to nitrogen limitation in freshwater microalgae
Abstract
In this study Fourier transform infrared micro-spectroscopy (FTIR) was used to determine lipid and carbohydrate content over time in the freshwater microalgae Chlamydomonas reinhardtii and Scenedesmus subspicatus grown in batch culture in limiting concentrations of nitrogen (N). Both algae exhibited restricted cell division and increased cell size following N-limitation. FTIR spectra of cells in N-limited media showed increasing lipid:amide I and carbohydrate:amide I ratios over time. The use of lipid- and starch-staining dyes confirmed that the observed ratio changes were due to increased lipid and carbohydrate synthesis. These results demonstrate rapid metabolic responses of C. reinhardtii and S. subspicatus to changing nutrient availability, and indicate the efficiency of FTIR as a reliable method for high-throughput determination of lipid induction.
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Nitrogen and phosphorus limitation causes changes in photosynthetic activity, lipid synthesis, and biomass composition in freshwater and marine microalgae.
“Studies have indicated changes in photosynthetic activity, lipid synthesis, and biomass composition in freshwater and marine microalgae as a result of nitrogen and phosphorus limitation [8 -11].”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
Chlamydomonas sp. exhibited an increase in CH intensities (3000-2800 cm-1) with nutrient depletion (NP1) on the 10th and 20th days of growth.
“Chlamydomonas sp. also exhibited an increase in CH intensities (3000-2800 cm -1 ) with nutrient depletion (NP1) on the 10 th (Fig.3) and 20 th (Fig.4) days of growth, in accordance with previous studies [9,13,58].”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The two species M. contortum and Chlamydomonas sp. have produced more neutral lipids (TAGs) with nutritional depletion.
“This indicates that the two species M.contortum and Chlamydomonas sp. have produced more neutral lipids (TAGs) with nutritional depletion [9,20,61 -63] and it is a positive indication that these microalgae species can be tapped for biodiesel production.”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The microalgae cells experienced nutritional stress as a result of low nitrate and phosphate contents, which increased the content of hydrocarbons.
“This implies that the microalgae cells have experienced nutritional stress as a result of the low nitrate and phosphate contents, which have increased the content of hydrocarbons [9,20].”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
Biomolecular groups in Chlamydomonas sp. were identified and variation in peak intensity between NP treatments and days of growth was assessed.
“Similarly, for Chlamydomonas sp ., (Fig.3 and 4) their biomolecular groups were identified and the variation in the intensity of the peaks between NP treatments and the days of growth was assessed and is tabulated in Table 3 [9,20,23,52 -54].”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
PC3 loadings in the amide regions (~1654, 1540 cm-1) are all in the positive direction, indicating that these are likely to be higher on the 20th day compared to the 10th day, where the protein content would be higher in the cells on the 20th day compared to the 10th day.
“PC3 loadings in the amide regions (~1654, 1540cm -1 ) are all in the positive direction indicating that these are likely to be higher on the 20 th day compared to the 10 th day, where the protein content would be higher in the cells on the 20 th day compared to the 10 th day [9,31].”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
Nearly a fivefold increase on the 29th day of sampling has been reported under nitrogen deprivation.
“and nearly a fivefold increase on the 29 th day of sampling, under nitrogen deprivation [9].”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~1740 cm−1 IR band is assigned to ν(C=O) stretches of esters from lipids - Triacylglycerides (TAGs).
“~ 1740 | | m + | | m + | | m + | | s + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~1658 cm−1 Raman band is assigned to cis ν(C=C) from unsaturated lipids.
“~ 1654 | ~ 1658 | s + | w + | s + | vw + | s + | vw + | m + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~2924 cm−1 IR band is assigned to (CH2) asymmetric stretches from hydrocarbons.
“~ 2924 | ~ 2912 | s + | vw + | m + | vw + | m + | vw + | s + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~1654 cm−1 IR band is assigned to ν(C=O), ν(C-N) stretches and δ(N-H) deformations of Amide I from proteins.
“~ 1654 | ~ 1658 | s + | w + | s + | vw + | s + | vw + | m + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~2850 cm−1 IR band is assigned to νs(CH2) symmetric stretches.
“~ 2846 | ~ 2820 | s + | m + | s + | w + | s + | m + | vs + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~3010 cm−1 IR band is assigned to (=CH) stretches from unsaturated lipids.
“~ 3010 | ~ 3014 | w + | w + | w + | vw + | w + | vw + | w + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~1380 cm−1 IR band is assigned to δs(CH3) symmetric bend from lipids and proteins.
“~ 1380 | | vs - | | s - | | m - | | w - |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~1440 cm−1 Raman band is assigned to δ(CH2) scissoring from saturated lipids and hydrocarbons.
“| ~ 1440 | | s + | | s + | | s + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
The ~1266 cm−1 IR band is assigned to δ(=CH) deformations from unsaturated lipids.
“~ 1266 | ~ 1264 | vw + | w + | vw + | w + | vw + | w + | vw + |”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [9]
Previous studies on Chlamydomonas reinhardtii and Scenedesmus subspicatus showed 5- to 6-fold increases in lipid accumulation.
“Previous studies on green algal species like Chlamydomonas reinhardtii and Scenedesmus subspicatus have exhibited 5 to 6 fold increase in lipid accumulation [8]”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [8]
Nile Red staining is affected by uneven dye uptake due to cell wall differences, is not quantitative as chromatographic methods, and cannot distinguish neutral from polar lipids.
“However, it is affected by uneven dye uptake due to differences in cell wall composition among species.Nile red staining is not quantitative as chromatographic methods and cannot distinguish between neutral and polar lipids [5,8].”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [8]
Mid-infra-red has been in use since the 1940s, with single point, point mapping and imaging applied to study microalgae composition, and inferences are drawn from qualitative observations or quantitative integration.
“Mid-infra-red has been in use since the 1940s, with different techniques viz. single point, point mapping and imaging applied to study microalgae composition with mid-infrared peaks associated with proteins, lipids and carbohydrates. Inferences are drawn from qualitative observations from infra-red bands or quantitative integration and calculation of peak heights and ratios [73,74].”
From Factors Affecting Microalgae Production for Biofuels and the Potentials of Chemometric Methods in Assessing and Optimizing Productivity (2019) · cites this paper as [73]
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Moraes, G. P. D., & Vieira, A. A. H. (2014) · PLoS ONE · doi:10.1371/journal.pone.0114458 · Cited by 5 · see citing works
Methods for quantitative determination of microalgal lipid and fatty acids content
Morshchinin, I. V. (2025) · Food systems · doi:10.21323/2618-9771-2025-8-2-267-275 · Cited by 3 · see citing works
- Authors
- Dean, A. P., Sigee, D. C., Estrada, B., & Pittman, J. K.
- Affiliations
- University of Manchester (GB)
- Corresponding author
- Pittman, Jon K.
- Venue
- Bioresource Technology
- Publisher
- Elsevier BV
- Published
- 2010-06
- Volume
- 101
- Issue
- 12
- Pages
- 4499-4507
- ISSN
- 0960-8524 (ISSN-L)
- OpenAlex
- W2163635466
- Cited by
- 523
- References
- 35
- Citations by year
- 2026: 13 · 2025: 17 · 2024: 25 · 2023: 25 · 2022: 41 · 2021: 39 · 2020: 45 · 2019: 46 · 2018: 44 · 2017: 41 · 2016: 33 · 2015: 43 · 2014: 30 · 2013: 35 · 2012: 26
- FWCI
- 12.08
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Bioresource Technology (publishedVersion)
- PubMed (publishedVersion)
- Research Explorer (The University of Manchester) (publishedVersion)
- Funders
- The Leverhulme Trust (F/00 120/AO)
- D.C.S.
- Topics
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
- Physical Sciences › Chemistry › Analytical Chemistry › Spectroscopy and Chemometric Analyses
- Keywords
- lipid accumulation, nitrogen limitation, freshwater microalgae, Chlamydomonas reinhardtii, Scenedesmus subspicatus, FTIR spectroscopy, algal metabolism, cell division
- MeSH terms
- Amides, Carbohydrates, Cell Count, Chlorophyll, Culture Media, Fluorescence, Fresh Water, Lipids, Nitrogen, Phosphorus, Chlamydomonas reinhardtii, Spectroscopy, Fourier Transform Infrared, Biomass, Principal Component Analysis, Scenedesmus, Cell Size, Eukaryota
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