DOI: 10.3303/CET24109055 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Paper Received: 4 December 2023; Revised: 21 February 2024; Accepted: 28 March 2024 
Please cite this article as: Sandoval J., Naranjo K., Casas L., 2024, Sunscreen Production from Chlorella Vulgaris, Chemical Engineering 
Transactions, 109, 325-330  DOI:10.3303/CET24109055 
  

 CHEMICAL ENGINEERING TRANSACTIONS  
 

VOL. 109, 2024 

A publication of 

 

The Italian Association 
of Chemical Engineering 
Online at www.cetjournal.it 

Guest Editors: Leonardo Tognotti, Rubens Maciel Filho, Viatcheslav Kafarov 

Copyright © 2024, AIDIC Servizi S.r.l. 

ISBN 979-12-81206-09-0; ISSN 2283-9216 

Sunscreen Production from Chlorella Vulgaris 

Juan Sandoval*, Kelly Naranjo, Laura Casas 

Universidad de América, Bogotá, Colombia  

juan.sandoval@profesores.uamerica.edu.co  

Sunscreen use has increased due to climate change and greater widespread awareness of the risk of acquiring 

skin diseases, including cancer, due to ultraviolet radiation from the sun (Hiatt & Beyeler, 2020; Alli et al., 2023). 

This has had a negative impact on water due to the increased release of components from these cosmetic 

products (Couselo-Rodríguez et al., 2022). In this sense, product formulation should consider aspects of 

environmental and social sustainability. Microalgae capture CO2 from the atmosphere, remove contaminants 

from wastewater, including traces of emerging pollutants (Ávila-Cújar et al., 2021), and give an alternative for 

environmentally friendly sunscreens. The initiative becomes a first step towards the possible development of a 

business based on sunscreen production. Microalgal biomass production was carried out under laboratory 

conditions. Subsequently, the biomass was separated from the culture medium, and the components 

responsible for the protective function, such as beta-carotenes and mycosporines, were quantified by 

spectrophotometry. Next, the sunscreen formulation was developed, and finally, the sun protection factor (SPF) 

was determined, in addition to verifying some organoleptic properties related to its application. The product 

obtained achieved an SPF of 8 with in vitro tests, and the use of environmentally friendly components in its 

formulation makes it an attractive alternative from an environmental and social perspective. 

1. Introduction 

In the global context, cosmetic industry generates around $262.21 billion, with skin care products representing 

38% of the market value. Sunscreens, categorized as part of the skincare product range, have increased their 

market share, serving as preventive measures against aging, sunburns, and the development of keratinocyte 

cancers and melanoma due to prolonged exposure to UV radiation (Grand View Research, 2020; Narla & Lim, 

2020). Consequently, these products are considered pharmaceuticals and emerge as an alternative for skin 

cancer prevention, because 1.5 million new cases of skin cancer were reported worldwide in 2020, caused for 

the increased ultraviolet radiation on Earth (World Health Organization, 2022). 

Currently, among various cosmetic products, sunscreens give the most effective protection against UV radiation 

damage. Following Ambruso et al. (2023), sunscreens are "any cosmetic product containing UV filters in its 

formulation with the aim of protecting the skin from harmful solar ultraviolet light, preventing or minimizing the 

damage this radiation can cause to human health". Their formulation includes organic compounds 

(aminobenzoate, benzophenones, and dibenzylmethanes) and inorganic compounds (titanium dioxide and zinc 

oxide) with absorption and reflection properties for solar radiation. However, the environmental impact caused 

by the organic UV filters used in the process is alarming, as their low solubility and high lipophilic properties 

hinder their elimination in wastewater, disrupting aquatic ecosystems and biodiversity. According to the 

environmental organization Green Cross, 25,000 tons of sun creams reach the oceans annually (Crespo, 2022). 

Microalgae offer advantages in terms of biomass production, rapid growth times, and tolerance to wide ranges 

of pH, salinity, and temperature. Besides, they can convert solar energy into chemical energy, producing high-

value-added natural bioactive compounds (Ariede et al., 2017). As a result, they have gained attention in the 

cosmetic industry, for the development of pharmaceutical and cosmetic products. Thus, microalgae and their 

secondary metabolites are a more sustainable alternative for manufacturing ecological ingredients and products.  

Chlorella vulgaris is a spherical microalga, 2-10 μm in diameter, non-motile cell, with a green coloration, which 

reproduces through binary fission via asexual spores, growing rapidly and generating an average of one cell 

every 24 hours (Raiesie et al., 2021). The main components of its algal biomass are proteins, lipids, and 

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carbohydrates, constituting up to 90% of the total dry weight. The remaining 10% includes pigments, vitamins, 

and other minor components (Ahmad et al., 2021). These secondary metabolites result from amphibolic or 

anabolic pathways when supplied with nitrogen and phosphorus, as well as atmospheric CO2. Among these 

compounds are β-carotenoids and mycosporines-like amino acids (MAAs), known for their photoprotective 

activity in absorbing UV radiation at short wavelengths, making them effective protective agents against UV 

radiation (Amador-Castro et al., 2020; Yarkent et al., 2020). In this context, the overall objective of this study 

was to produce a sunscreen from the microalga Chlorella vulgaris on a laboratory scale. 

2. Methodology 

This section is divided in five parts: Microalgae cultivation, microalgae growth, biomass separation, metabolites 

extraction and quantification; and sunscreen formulation.  

2.1 Microalgae cultivation 

Chlorella vulgaris was supplied by microbiology laboratory at the Francisco de Paula Santander University in 

Cúcuta, Colombia. It was cultivated in two glass photobioreactors, one under white, and the other under red 

LED light lamps to study the effect of exposure to different light spectra on cell growth and the production of 

secondary metabolites. In both photobioreactors, Bold Basal Medium (BBM) was used as culture medium 

(Bertoloni, 2022). It was sterilized at 120 °C in a DENTOMAT brand autoclave, model Automat 3000, for 15 

minutes. Both photobioreactors, sealed with Parafilm to avoid contamination, had the same total volume of 150 

mL, comprising 77 mL of BBM, 5 mL of microalgae, and 68 mL of deionized water. Continuous aeration and 

agitation, to ensure effective light and nutrients dispersion, was given by a fun and hoses connected to each 

photobioreactor. The first photobioreactor (culture 1) was placed inside a polystyrene incubator with red light 

tape on its walls to provide illumination to the culture. And the second photobioreactor (culture 2) was outside 

the incubator and illuminated by a white LED light lamp, with an average light intensity of 125.85 Lux to the front 

of the culture. Culture 1 was maintained at a temperature above ambient, specifically at 19 °C, while culture 2 

at 17 °C, the average ambient temperature of Bogotá. pH measurements were taken weekly, staying within the 

range of 8-9 for both experiments (Arredondo & Voltolina, 2007). 

2.2 Microalgae growth 

Microalgae growth was monitored by measuring the optical density of each culture at a wavelength of 640 nm 

using a UV/VIS spectrophotometer, THERMOSPECTRONIC model Genesis 20. This wavelength was chosen 

for its proximity to the chlorophyll absorption peak (675 nm), allowing measurements even at minimal cellular 

concentrations (Zhang et al., 2023). These measurements were taken, by triplicate, every two days. The specific 

growth rate in the exponential growth phase was calculated by equation (1) (Metsoviti et al., 2019): 

𝜇𝑒𝑥𝑝 =
ln(𝛼2 𝛼1⁄ )

(𝑡2 − 𝑡1)
 (1) 

𝛼1 and 𝛼3 are the optical densities at the beginning and at the end of the exponential growth phase. 

2.3 Biomass separation 

Solid biomass was separated from the liquid medium, in a TECNAL centrifuge model 206-baby I for 15 minutes 

at 3000 rpm. Supernatant was discarded, and the precipitate was collected. A further drying process was 

necessary. The samples were placed in an oven BINDER model IP20 at 65 °C for 2 hours. The recovered dry 

biomass was milled and stored at -15 °C (Outon, 2019) and divided into different quantities for use in dry weight 

determination, extraction, quantification of target metabolites, and sunscreen production. Dry weight (in g L-1) 

and productivity (in g L-1 d-1) were calculated using the following equations (Corzo et al., 2019) where 𝑚1 is the 

weight of filled container, 𝑚0 is the weight of empty container, 𝑉𝑐𝑢𝑙𝑡𝑢𝑟𝑒 is the total volume of microalgae culture, 

including medium, at the end of the cultivation; and, t is the cultivation time: 

𝐷𝑟𝑦 𝑤𝑒𝑖𝑔ℎ𝑡 (𝑔 𝐿−1) =
𝑚1 − 𝑚0

𝑉𝑐𝑢𝑙𝑡𝑢𝑟𝑒
 (2) 

𝑃𝑟𝑜𝑑𝑢𝑐𝑡𝑖𝑣𝑖𝑡𝑦 (𝑔 𝐿−1𝑑−1) =
𝐷𝑟𝑦 𝑤𝑒𝑖𝑔ℎ𝑡

𝑉𝑐𝑢𝑙𝑡𝑢𝑟𝑒 ∗ 𝑡
 (3) 

2.4 Metabolites extraction and quantification 

Beta-carotenoids were extracted and quantified 45 days after the initiation of the culture and following the 

acquisition of dry microalgal biomass, following the standardized protocol by Ibanez (2018). The dry biomass 

326



used for this process was obtained from the culture exposed to white LED light, as the quantity obtained from 

the red LED culture was insufficient for analysis. Initially, using an analytical balance (RADWAG model AS-

220/C/2), 0.1 g of dry microalgal biomass was weighed and added to a 15 mL Falcon tube containing 7 mL of 

the extraction solution (90% acetone). The mixture underwent manual agitation for 30 seconds followed by 

ultrasonic bath treatment for 10 minutes (Chronis et al., 2021). After this, it was centrifuged using a TECNAL 

centrifuge model 206-baby I for 20 minutes at 3200 rpm. The supernatant was collected in a 25 mL volumetric 

flask, and the process was repeated twice until the matrix decolorization was achieved (Ibanez, 2018). The total 

concentration of beta-carotene was determined by spectrophotometry using acetone as a blank. In addition, the 

25 mL volumetric flask containing the supernatant was levelled with acetone. Following this, the absorbance of 

the supernatant was measured at a wavelength of 450 nm to calculate the concentration of beta-carotene in μg 

mL-1 using the following equation (Ibanez, 2018) 

𝐶 =
𝐴𝑀𝐴𝑋 ∗ 𝑀 ∗ 1000

휀 ∗ 𝛿
 (4) 

Where 𝐴𝑀𝐴𝑋 is the absorbance at 450 nm; M is the molecular weight of carotene (537 g mol-1); 휀 is the molar 

extinction coefficient of the β - carotenoid in acetone (140663 L mol-1 cm-1) and δ is the optical path in cm. 

For mycosporines (MAAs) extraction methanol and acetone were used as polar extraction solvents to evaluate 

their effectiveness. Acetone was used as extraction solvent for four samples of which two came from the Culture 

1 (white light) and the other two from the Culture 2 (red light). Each of the samples were crushed, after mixed 

with silica gel, in a mortar for 3 minutes and added to test tubes containing 4 mL of each solvent. Following this, 

all samples were stored at a temperature of -20 °C for 22 h in a dark place (Outon, 2019). Finally, the samples 

were centrifuged for 20 minutes at 4200 rpm, precipitate was separated and the supernatant went to another 

test tube (Outon, 2019). Optical density of the supernatant at a wavelength between 326-704 nm were 

measured. General equation to quantify mycosporines was 

𝑇𝑜𝑡𝑎𝑙 𝑀𝐴𝐴𝑠 𝐶𝑜𝑛𝑡𝑒𝑛𝑡 =
𝐶1 ∗ 𝐴𝜆1

− 𝐶2 ∗ 𝐴𝜆2

𝐷𝑟𝑦 𝑤𝑒𝑖𝑔ℎ𝑡
∗

𝐴𝑀𝐴𝐴𝑆

𝐴𝑐ℎ𝑙𝑎

∗ 1000 (5) 

𝐴𝑀𝐴𝐴𝑠 is the area under the curve at wavelengths from 326 to 356 nm; 𝐴𝑐ℎ𝑙𝑎 is the area under the curve at 

wavelengths from 621 to 704 nm, corresponding to chlorophyll – a. The exact parameters 𝐶1, 𝐶2 and the 

wavelengths 𝜆1, 𝜆2depend on which solvent was used: acetone or methanol (Huang et al., 2016; Outon, 2019).  

2.5 Sunscreen formulation 

A semi-solid formulation was prepared by means of an oil-in-water (O/W) emulsion, intended for topical 

application with a protective effect (Rios, 2020). The compounds used were separated in PHASE A (oily) and 

PHASE B (aqueous). All compounds were added according to limits allowed by the FDA (U.S. Food & Drug, 

2023). Finally, protection factor (SPF) was evaluated by an in vitro method, specifically applying UV 

spectrophotometry to samples onto a 1X2 cm strip of a tape in a wavelength range from 290 to 320 nm, with 

intervals of 5 nm, where the determination of the SPF, which is the correlation between the erythema effect (EE) 

and the radiation intensity () at each wavelength () adapts to the following equation (Widsten et al., 2020) 

where CF is the correction factor which values 10, and 𝐴𝑏𝑠 is the optical absorbance at each wavelength in the 

range (Martínez-Inda et al., 2023): 

𝐹𝑃𝑆 = 𝐶𝐹 ∗ ∑ 𝐸𝐸(𝜆) ∗ 𝐼(𝜆) ∗ 𝐴𝑏𝑠(𝜆)

320

290

 (6) 

3. Results 

3.1 Effect of light on Microalgae growth 

Red light LED, used in Culture 1, led to a specific growth rate in the exponential phase of 0.22 d-1, a productivity 

of 0.52 g L-1 d-1, and a dry weight of 14.47 g L-1. As for Culture 2, the white LED light intensity led to a specific 

growth rate in the exponential phase of 0.31 d-1, a productivity of 1.24 g L-1 d-1, and a dry weight of 34.61 g L-1. 

The highest cellular growth rate, dry weight, and productivity for the Culture 2, with white LED light, is the same 

behaviour found in literature (Martinez et al., 2022). 

327



3.2 Metabolites extraction and quantification 

The amount of biomass obtained in Culture 1 was not sufficient to perform carotenoid extraction, only for 

mycosporines, the first procedure made under four different conditions: with two solvents and evaluating the 

use of silica gel in sample drying. The results are shown in Table 1. 

Table 1: Quantification of mycosporines 

 

Solvent 

Culture 1 Culture 2 

With silica 

gel 

Without silica 

gel 

With silica 

gel 

Without silica 

gel 

Methanol 6.61 μg g-1 6.22 μg g-1 2.98 μg g-1 1.93 μg g-1 

Acetone 5.10 μg g-1 3.96 μg g-1 2.12 μg g-1 1.98 μg g-1 

 

A higher amount of mycosporines was obtained from the microalgal biomass of Culture 1 compared to Culture 

2, which is consistent with Llewellyn et al. (2020) in their work with Chlorogloeopsis fritschii, where they found 

that the impact of exposure to UVB radiation and red light on MAA biosynthesis was greater compared to white 

light. Based on these results, a factorial regression "MAAs concentration versus extraction solvent; desiccant" 

was applied by means of free Minitab 17 ®, resulting in a variance analysis in which it was found that the solvent 

(effect A) and desiccant (Effect B) had an effect of -1.15 and -0.68 on mycosporines extraction, respectively. 

Furthermore, it was found that the effect of both factors is not statistically significant at a significance level of 

0.05 because the P-values are higher, being 0.53 for factor A and 0.70 for factor B. Finally, the concentration of 

β-carotenoids for Culture 2 was 0.094 μg mL-1. 

3.3 General and protective features of the sunscreen 

In Figure 1 the final product is shown. However, the formulation included all the separated biomass of Chlorella 
vulgaris instead of only the metabolites. From the picture, it is evident its green color and viscous appearance. 
Besides, its consistency was creamy-gel, smooth to the touch, and had a light and fluid texture, facilitating its 
application. Additionally, it was free from lumps and particles, indicating that the compounds used in the 
formulation exhibited compatibility with each other, ensuring skin safety when applied. 

 

Figure 1: Sunscreen produced from the microalgae Chlorella vulgaris on a laboratory scale 

Then, pH and sun protection factor, SPF, were determined to assess security and effectiveness of the sunscreen 

formulation, obtaining values of 6 and 7.74, respectively, from which it is considered that the sunscreen is secure 

for skin care and presented a medium level of photoprotection according to the standard of the European 

cosmetic and perfumery association, COLIPA, (Schneider et al., 2020). Finally, it is also important to consider 

that the SPF depends not only on the concentration of the pigments, but also on the quantity used and on the 

specific method used. 

 

328



4. Conclusions 

The white LED light cultivation showed the highest growth rate: 0.31 d-1, dry weight: 34.61g L-1, and productivity: 

1.24 g L-1 d-1 compared to the red LED light cultivation with 0.22 d-1; 14.47 g L-1 and 0.52 g L-1 d-1 respectively. 

However, the mycosporines concentration was higher in the cultivation under red LED light, with a value of 6.61 

μg g-1, while for the cultivation under white LED light, it was 3.38 μg g-1. As a possible strategy to increase 

mycosporines production, microalgae growth should be made with white LED light until stationary phase, and 

then red LED light should be used. In the sunscreen, the dry biomass of Chlorella vulgaris was used as a new 

UV filter, not only mycosporines and carotenoids, resulting in a product with a pH of 6, a characteristic green 

color of the microalga, a creamy-gel consistency, and a light and fluid texture. The SPF of the sunscreen was 

7.74, classifying it as a medium protection photoprotector. The results obtained in this work demonstrate the 

potential of Chlorella vulgaris to be used in the production of biological sunscreens. It is also needed to make 

an economical evaluation in future research to improve the profitability of the process. 

Acknowledgments 

This work was (partially) supported by Programa Iberoamericano de Ciencia y Tecnología para el Desarrollo 

(CYTED) (through RED RENUWAL 320rt0005).  

Universidad de America has also helped to the experimental work of this project.  

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