ADITYA, Fitra Ari (2026) Enhancement of Lipid Production in Chlorella vulgaris as a Biofuel Feedstock through Hematite Nanoparticle (α Fe₂O₃) Supplementation with In Vivo Nile Red Staining Analysis. Undergraduate thesis, Fakultas Sains dan Matematika.

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Abstract

Microalgae Chlorella vulgaris is a promising candidate for third-generation
biofuel feedstock due to its rapid growth, high biomass productivity, and
substantial lipid accumulation. This study aimed to evaluate the effects of
hematite nanoparticles (α-Fe₂O₃) on the growth, lipid accumulation, and biofuel
productivity of C. vulgaris, using Nile Red (NR) staining as a rapid in vivo lipid
quantification method. Five α-Fe₂O₃ concentrations (0, 10, 25, 50, and 75 mg/L)
were tested with two replicates. Parameters measured included optical density
(OD₆₈₀), dry biomass, lipid content (%), lipid yield (g/L), lipid productivity
(g/L/day), fluorescence intensity (RFU), and cell morphology. Results indicated a
significant dose-dependent increase (p < 0.05) in both growth and lipid
accumulation, with the highest biomass observed at 75 mg/L (44.705 g/L),
representing a 430% increase compared to the control (8.417 g/L). Lipid content
rose from 15.2% in the control to 37.2% at 75 mg/L, while the highest lipid yield
and productivity were 16.63 g/L and 1.66 g/L/day, respectively, accompanied by
an estimated biofuel potential of 14.97 mL/L/day. Fluorescence analysis
confirmed enhanced intracellular lipid accumulation, with RFU values peaking at
4395 ± 105 under 75 mg/L treatment, alongside an increased mean cell diameter
(1.336 µm). NR staining revealed strong, uniform yellow-orange fluorescence in
treated cells, indicating broad lipid distribution without impairing photosynthetic
functionality. These findings demonstrate that α-Fe₂O₃ supplementation
significantly enhances both growth and lipid biosynthesis in C. vulgaris,
highlighting its potential as an effective strategy for scalable biofuel production.
Keywords: Chlorella vulgaris, α-Fe₂O₃, lipid, Nile Red, biofuel.

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