Nativa, Sinop, v. 11, n. 2, p. 259-265, 2023.

Pesquisas Agrárias e Ambientais

DOI: https://doi.org/10.31413/nativa.v11i2.15977

ISSN: 2318-7670

Extract from

Bixa orellana

L. seed: development of biodegradable packaging

and biological activities

Leonardo Almeida JATOBÁ1, Antonio Carlos Pereira de MENEZES FILHO2* ,

Marconi Batista TEIXEIRA2, Frederico Antônio Loureiro SOARES2,

Carlos Frederico de Souza CASTRO2, Porshia SHARMA3, Matheus Vinícius Abadia VENTURA1,2

1UniBRAS University Center of Southwest Goiás, Rio Verde, GO, Brazil

2Goiano Federal Institute, Rio Verde, GO, Brazil.

3Institute for Innovative Learning and Research (IILR), Madhya Pradesh, Indore, India.

*E-mail: astronomoamadorgoias@gmail.com

Submitted on 07/19/2023; Accepted on 08/14/2023; Published on 08/17/2023

ABSTRACT: Active biodegradable packaging with Bixa orellana extract has potential for producing new

products that combine natural renewable use and food conservation. B. orellana seeds were collected and their

extract was produced. The packages were produced and incorporated with different concentrations of B. orellana

extract. Phytochemical parameters were tested in several groups of special metabolism; physicochemical for

thickness, moisture, solubility, UV-Vis light transmission, color L*, a* and b*; biodegradability in soil and

antifungal activity on Sclerotinia sclerotiorum, Colletotrichum acutatum and Colletotrichum gloeosporioides. The packaging

showed potential for use as a new product for food, the large number of phytocompounds allows an active

packaging, in addition to physicochemical characteristics of thickness, humidity and ideal solubility and use for

foods that need protection at different wavelengths in the UV-Vis that deteriorate important characteristics of

the product. Moreover, it demonstrated an effect with biodegradability of less than 40 days and antifungal

activity on phytopathogenic fungi that cause deterioration in vegetables of agricultural interest.

Keywords: Bixa genus; bixin; carotenoids; natural Polymer; norbixin; polymer matrix.

Extrato das sementes de

Bixa orellana

L.: desenvolvimento de embalagens

biodegradáveis e atividades biológicas

RESUMO: Embalagens biodegradáveis ativas com extrato de Bixa orellana apresentam potencial para a

produção de novos produtos que aliam o uso renovável natural e a conservação de alimentos. Sementes de B.

orellana foram coletadas e o extrato produzido. As embalagens foram produzidas e incorporadas com diferentes

concentrações do extrato de B. orellana. Foram testados parâmetros fitoquímicos em diversos grupos do

metabolismo especial; físico-químicos para espessura, umidade, solubilidade, transmissão de luz UV-Vis, cor

L*, a* e b*; biodegradabilidade em solo e atividade antifúngica sobre Sclerotinia sclerotiorum, Colletotrichum acutatum

e Colletotrichum gloeosporioides. As embalagens demonstraram potencial para uso como novo produto para

alimentos, o grande número de fitocompostos admite uma embalagem ativa, além de características físico-

químicas de espessura, umidade e solubilidade ideais e uso para alimentos que necessitam proteção em

diferentes comprimentos de ondas no UV-Vis que deterioram características importantes do produto. Além

disso, demonstrou efeito com biodegradabilidade inferior a 40 dias e atividade antifúngica sobre fungos

fitopatogênicos que causam deterioração em vegetais de interesse agrícola.

Palavras-chave: gênero Bixa; bixina; carotenóides; matriz polimérica; norbixina; polímero natural.

1. INTRODUCTION

Bixa orellana is a South and Central American native plant

that can also be grown in Peru, Mexico, Ecuador, Indonesia,

India, Kenya, and East Africa. The seeds contained in it have

natural dyes (bixin and norbixin) that are being used in the

Pharmaceutical and Food industries as they contribute in

several biological activities (HIRKO; GETU, 2022).

The usage of substances extracted from special

metabolism of plants have been gaining popularity due to

their various qualities that help development of active and

intelligent packaging. It is observed that bixin and norbixin

(carotenoids) are potentially able to work as antioxidant,

antifungal, anticarcinogenic and among other biological

activities. Bixin is a linear apocarotenoid having double

bonds and costitutes up to 80% of the total carotenoid

content present in B. orellana (annatto or urucum) seeds

(LUIGGI et al., 2020).

The incorporation of plant extracts in natural polymeric

matrices is beneficial as it provides; reduction of synthetic

packaging, easily degradable on discarding in atmosphere, has

little to no pollution rate and also serves the purpuse of

preservation of food from harmful micro-organisms and

oxidation (Shaikh et al., 2021). The usage of biodegradable

materials in food packaging counts as a promising alternative

as it reduces investment and also the treatment cost during

waste management.

Extract from Bixa orellana L. seed: development of biodegradable packaging and biological activities

Nativa, Sinop, v. 11, n. 2, p. 259-265, 2023.

260

Some studies demonstrate the incorporation of bixin into

a polymeric matrix; this results in formation of great barrier

against oxygen, water vapour, thermal stability and to

different wavelengths of light such as ultraviolet (UV) and

visible (Vis) (STOLL et al., 2021a). Although, there is no

experimental evidence of using polymeric matrix of starch

obtained from Maranta arundinacea L. (arrowroot or araruta)

with extract of B. orellana. Thus, there is no known proof of

interaction between this polymer and bixin extract obtained

from the seeds. Thus, this study aimed to evaluate the

addition of ethanolic extract of Bixa orellana seeds regarding

phytochemical constitution, formation of biodegradable

packaging (BP), physicochemical characteristics,

biodegradability and antifungal activity at different

concentrations.

2. MATERIAL AND METHODS

2.1. Reagents, equipments and glasswares

Ethyl alcohol P.A - ACS (Neon, Brazil), glycerin P.A –

ACS (Dinâmica, Brazil), fungicide Frowncide 500 SC (ISK,

Brazil) Fluazinam concentration (500 g/L-1), Tween 80 P.A

– ACS (Dinâmica, Brazil), PDA culture medium (Himedia,

India).

Digital analytical balance (Shimadzu, Mod. AY220,

Brazil), digital colorimeter (HunterLab, Mod. ColorFlex EZ,

U.S.A), digital caliper (Mitutoyo, Mod. 500-196-30, Japan),

digital thermometer (Bmax, Mod. TP-101, Brazil), electric

greenhouse with forced air circulation (Thoth, Mod. th-510-

480, Brazil), lyophilizer (Terroni, Mod. LS6000 B, Brazil),

magnetic stirrer with heating (Solab, Mod. SL 91, Brazil), UV-

Vis spectrophotometer (Bel, Mod. M-51, Italy).

Beaker 500 mL (Deltex, Brazil), glass rod (Uniglas,

Brazil), precipitation cup 250 and 500 mL (Uniglas, Brazil),

polystyrene Petri dish (FirstLab, Mod. FL3-9015RI, China),

qualitative filter paper (Unifil, Brazil), universal tip 1 mL

(Olen, U.S.A), variable micropipette 10-1000 µL (Peguepet,

Brazil).

2.2. Species collection and identification

Bixa orellana seeds were collected in July 2022 in the

municipality of Rio Verde, Goiás State, Brazil, with the

following geographic coordinate (17º48’00.0’’S and

50º54’41.5’’W). The species was identified, exsiccate

herborized and deposited in the Herbarium of the

Laboratório de Sistemática Vegetal at Instituto Federal

Goiano, Rio Verde, Goiás State, Brazil. Voucher (HRV:

17.786).

2.3. Seed processing

The seeds were grounded in a food processor for 3 min.

After this, the obtained seed powder was stored in a plastic

food packaging and was kept at -12 °C until further analysis.

2.4. Production of extract

Extraction was carried out with 50 g sample and 100 mL

ethanol (m/v); it was kept under constant magnetic agitation

for 4 h. Contact between solvent and raw material was kept

for seven days at room temperature (25 °C), in the dark. It

was manually agitated on a daily basis. The mixture that

resulted from the extraction was separated through filtration

(Whatman qualitative filter), followed by solvent evaporation

which was carried out by a vacuum rotary evaporator at 70

°C. The lyophilized extract was kept under refrigeration at -

12 °C until further analysis. The extracts were lyophilized.

The yield seed ethanolic (EEBO) extract from B. orellana

content was determined by equation 1.

Yield (%) = (Em/Sm) * 100 (01)

where: Em = Extract mass; Sm = sample mass.

2.5 Phytochemical analysis

For phytochemical analysis, 50 mL of ethanol solution

containing 5 g (w/v) of extract was prepared. Crude ethanolic

extract of B. orellana seeds was subjected to number of

analysis for phytochemical characterization according to

Bessa et al. (2013) and Shaikh; Patil (2020) with adaptations.

The positivity for each group of special metabolites and

constituents analyzed was achieved from the development of

color and/or precipitate, characteristic for analysis of each

phytoclass. saponins, alkaloids, anthraquinones, catechins,

coumarins, steroids and triterpenoids, phenols, tannins,

flavonoids, quinones, organic acids, carboxylic acids,

polysaccharides, aromatic and aliphatic compounds, oxylates,

bixin and norbixin, reducing sugars, non-reducing sugars and

carbohydrates were evaluated. The detection was based on

visual observations of a color change or precipitate formation

after adding specific reagents.

2.6. Biodegradable packaging preparations

Biodegradable films were obtained by a casting technique,

use of the methodology proposed by Issa et al. (2017),

modified. To produce each film, 5 g arrowroot starch was

dissolved in 100 mL deionized water. The solution was then

moderately agitated at room temperature 25 °C. Afterward, it

was heated at 70 °C, at constant agitation for 30 min.

After starch gelatinization, glycerol was added as a

plasticizer (30% w/w); this dispersion was agitated for 5 more

min. When the filmogenic solutions reached 50 °C, the

control did not show any EEBO incorporated into it, while

EEBO dose of (250 (EESBO), 500 (EESBO), 750 (EESBO)

and 1000 (EESBO) µL mL-1) were added to the others under

constant agitation for 15 min. The seeds extract from B.

orellana was better solubilized as 200 mg EEBO per 1 mL

Tween 80 at 5%. All filmogenic solutions were poured on

polystyrene slabs and dried in an air circulation oven at 30 °C

for about 36 h.

2.7. Characterization of biodegradable packaging

incorporating

2.7.1. Thickness

Biodegradable packaging thickness was measured by a

digital caliper in millimeters (mm). Measurements were

carried out in 10 spots on every packaging and the thickness

mean was calculated.

2.7.2. Moisture content

The moisture content of packaging was weighed and then

dried in an oven at 105 °C for 4 h. Three replicates per

packaging treatment were used, in agreement described by

Santos et al. (2021).

2.7.3. Measurement of water solubility

Biodegradable packaging which measured about (2 x 2

cm2) was dried in oven at 50 °C for 4 h and then weighed so

that initial mass (Mi) could be determined. They were

Jatobá et al.

Nativa, Sinop, v. 11, n. 2, p. 259-265, 2023.

261

immersed in 60 mL distilled water and kept under constant

agitation (170 rpm) at 25 °C for 24 h.

Afterward, solutions with the packaging were filtered

using filter paper (qualitative filter) with packaging had been

previously weighed. Sheets of filter papers with

biodegradable packaging were dried at 105 °C for 24 h and

weighed so that final mass (Mf) could be found, in agreement

with the methodology described by Jahed et al. (2017). Every

treatment was analyzed in quadruplicate. Biodegradable

packaging solubility in percentage (%) was calculated by

equation 2.

Water solubility (%) = Mi – (Mf/Mi) * 100 (02)

2.7.3. Light transmittance (UV-Vis)

Ultraviolet (UV) and visible light (Vis) transmittance of

film was conducted with the help of UV-Vis

spectrophotometer. The BP samples were cut and placed in

quartz cuvettes so that transmittance could be measured over

a wavelenght range between 900 at 200 nm as described by

Hosseini et al. (2015).

2.7.4. Analysis of color

Analysis of BP color was carried out a ColorEZ digital

colorimeter. Parameters under evaluation were L*

(luminosity) (0- lower brightness and 100- higher brightness)

and chromaticity parameters a* (green -180 and red +180)

and b* (yellow +180 and blue -180). The equipment was

calibrated with black crystal, green crystal and white crystal.

Measurements were conducted on three randomly selected

packaging spots.

2.8. Biodegradable activity

The soil biodegradability test was performed using by the

methodology described by Martucci; Ruseckaite (2009). The

BP samples (2 x 2 cm2) were dried up to constant weight so

that initial mass (Mi) could be determined.

Samples were then placed in open polyethylene packaging

to enable microorganisms and moisture to gain access to

them. Now, these were buried in previously prepared organic

soil (red dystroferric soil type) at constant moisture and room

temperature. Thirty days after the experiment installment, the

biodegradable packages with the samples were removed from

the soil, cleaned with a fine brush and dried up to constant

weight (Mf). Biodegradability in percentage (%) was

calculated by equation 3.

Biodegradability (%) = Mf – (Mf/Mi)*100 (03)

2.9. Antifungal assay

Antifungal activity of BP was analyzed against the

phytopathogenic fungi Sclerotinia sclerotiorum, Colletotrichum

acutatum and C. gloeosporioides by a disk diffusion assay. Petri

dishes, half full with potato, dextrose and agar medium

(PDA), were inoculated with 100 µL suspension with 107

CFU mL-1.

Then, four samples of packaging which had been cut in

circles of about 7 mm were placed on every dish. Petri dishes

were incubated at 25 °C for 15 days. As standard fungicide

Frowncide 500 SC concentration 10 µL mL-1was used.

Finally, diameters of the zone of inhibition were measured

using a digital caliper and expressed in millimeters (mm),

described by Valadares et al. (2020) and Menezes Filho et al.

(2022b) modified.

2.10. Statistical analysis

Analysis was carried out in triplicate and quadruplicate

and the standard deviations (SD) among them were

calculated. The data were collected and represented as the

standard error of the mean. Following ANOVA, a Duncan

test was used to examine the statistical relevance of the mean

differences at P < 0.05, using SigmaPlot statistical software.

3. RESULTS

3.1. Phytochemical screening

A low number of groups of phytomolecules present in

both extract from B. orellana seeds were observed (Table 1).

Some phytomolecules were found in both extracts such as

alkaloids, reducing sugars, flavonoids, phytosterols,

carotenoids, coumarins, organic acids, aromatic compounds,

proteins, hemolytic saponins, bixin and norbixin.

Table 1. Phytochemical prospection of ethanolic extract of Bixa

orellana seeds.

Tabela 1. Prospecção fitoquímica do extrato etanólico das sementes

de Bixa orellana.

Phytogroup

EEBO

Tannins

Blue

Alkaloids

Carbohydrates

Reducing sugars

reducing sugars

Glycosides

Cardiac glycosides

Amino acids

Flavonoids

Phenolics

Phlobatannins

Phytosterols

Cholesterol

Terpinoids

Triterpinoids

Diterpenes

Lignins

Carotenoids

Quinones

Anthraquinones

Anthocyanins

Carboxylic acids

Coumarins

Emodins

Gums &

mucilages

Resins

Fixed oils & Fat

Volatile oils

Organic acids

Aliphatic & aromatic

Red

Polysaccharides

Proteins

Foamy saponin

Hemolytic saponin

Oxylates

Bixin

Norbixin

Note: EEBO: ethanolic extract of Bixa orellana seeds. (-) no detected.

(+) positive. Blue: hydrolyzable or gallic tannins. Red: aromatic

compounds. Source: Authors, 2023.

Nota: EEBO: extrato etanólico das sementes de Bixa orellana. (-) não

detectado. (+) positivo. Azul: taninos hidrolisáveis ou gálico.

Vermelho: compostos aromáticos. Fonte: Autores, 2023.

Extract from Bixa orellana L. seed: development of biodegradable packaging and biological activities

Nativa, Sinop, v. 11, n. 2, p. 259-265, 2023.

262

3.2. Physicochemical characteristics of packaging

The control BP exhibited the lowest values of thickness,

while in the others, the higher the extract doses added to the

BP, the higher their thickness values (Table 2). Moisture

contents of BP depended directly on the amount of extract

that was added to them (Table 2). Control BP had

significantly higher in solubility due to the e -OH group

presence in starch, and thereby affecting the starch BP by its

hydrophilic nature. This consequently leads to higher

interaction with water molecules. Increasing the extract from

250 to 1000 µL has lowered the solubility. Basically, low water

solubility exhibits great water resistance (Table 2).

Concerning colors of BP under development, (Figure 1)

shows decrease in light transmittance rates of BP

incorporating different doses of extract in the visible and

ultraviolet region (from 200 to 850nm).

Table 2. Thickness, humidity and solubility of biodegradable

packages incorporated with ethanolic extract of Bixa orellana seeds.

Tabela 2. Espessura, umidade e solubilidade das embalagens

biodegradáveis incorporadas com extrato etanólico das sementes de

Bixa orellana.

Film

Thickness

(mm)

Moisture (%)

Solubility (%)

Control

0.24±0.01d

10.40±0.73c

50.43±3.30a

EESBO 250

0.26±0.01d

8,28±0.31d

40.88±3.82b

EESBO 500

0.32±0.01c

9.26±0.45cd

29.99±1.52c

EESBO 750

0.36±0.01b

12.92±1.02b

24.94±2.21c

EESBO 1000

0.43±0.01a

14.65±0.31a

22.95±4.48d

Note: Different letters in the column show statistical difference by the

Duncan’s test (P < 0.05) significance.

Nota: Letras diferentes na coluna apresentam diferença estatística pelo teste

de Duncan com (P < 0,05) de significância.

Figure 1. Transmission of UV-Vis light in biodegradable packaging

incorporated with ethanolic extract from Bixa orellana seeds. Source:

Authors, 2023.

Figura 1. Transmissão de luz UV-Vis em embalagens

biodegradáveis incorporadas com extrato etanólico das sementes de

Bixa orellana. Fonte: Autores, 2023.

Results showed that the use of extract increased BP

luminosity. The highest extract concentrations showed the

best luminosity results, being superior to the standard BP.

The control BP exhibited negative values of a* and b*

chromaticity and was green/blue. Biodegradable packaging

incorporating from 250 to 1000 µL extract exhibited positive

values of a* and b* chromaticity and were red and yellow

(Table 2).

3.3. Biodegradable activity

The lowest concentrations 500 and 250 µL mL-1 showed

the highest biodegradation capacity, showing no statistical

difference with the control (Figure 2). Although they have

shown potential, higher concentrations verified suggest that

they are toxic because they reduce biodegradability and

statistically longer time in the soil.

Table 2. Color parameters of biodegradable packages incorporated

with ethanolic extract from Bixa orellana seeds.

Tabela 2. Parâmetros de cor das embalagens biodegradáveis

incorporadas com extrato etanólico das sementes de Bixa orellana.

Film

Control

14.59±0.24e

0.71±0.09e

0.91±0.05e

EESBO 250

17.41±0.11c

0.34±0.09d

9.03±0.13c

EESBO 500

15.48±0.02d

0.94±0.01c

10.19±0.05b

EESBO 750

19.67±0.03a

1.65±0.03b

8.29±0.03d

EESBO 1000

19.11±0.04b

3.53±0.01a

11.95±0.09a

Note: Different letters in the column show statistical difference by the

Duncan’s test with 5% significance. Source: Authors, 2023.

Nota: Letras diferentes na coluna apresentam diferença estatística pelo teste

de Duncan com 5% de significância. Fonte: Autores, 2023.

Figure 2. Biodegradability test in soil in natura in packages

incorporated with ethanolic extract of Bixa orellana seeds. Note:

Different letters in the column show statistical difference by the

Duncan’s test (P < 0.05) significance.

Figura 2. Ensaio de biodegradabilidade em solo in natura em

embalagens incorporadas com extrato etanólico das sementes de

Bixa orellana. Nota: Letras diferentes na coluna apresentam diferença

pelo teste de Duncan (P < 0,05) de significância.

3.4. Antifungal activity

The B. orellana seed extract incorporated into the BP

exhibited good antifungal potential on the three evaluated

strains. For S. sclerotiorum, no significant difference (P < 0.05)

was observed between the 1000 and 750 µL concentrations

with 58% inhibition. The same was observed for C.

gloeosporioides with inhibition above 25%. For C. acutatum

growth inhibition activity was above 30%.

Table 4. Antifungal activity of biodegradable packaging

incorporated with ethanolic extract of Bixa orellana seeds.

Tabela 4. Atividade antifúngica das embalagens biodegradáveis

incorporadas com extrato etanólico das sementes de Bixa orellana.

Strain

Inhibition Zone (%)

1000

750

500

250

58.75b

55.95b

32.83c

19.82d

31.91b

29.67bc

27.93c

22.57d

26.31b

23.92b

19.54c

15.62d

1 = Sclerotinia sclerotiorum. 2 = Colletotrichum acutatum. 3 = Colletotrichum

gloeosporioides. Standard 100a. Control 0.00e.

Note: Different letters in the line show statistical difference by the Duncan’s

test (P < 0.05) significance.

Nota: Letras diferentes na linha mostram diferença estatística pela

significância do teste de Duncan (P < 0,05).

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263

4. DISCUSSION

The diversity of plants with a formidable source of active

compounds is extremely high (COSTA; FREIRE, 2018;

MENEZES FILHO et al., 2023). Here, our plant B. orellana

exhibit this potential through many of it's group having

important phytomolecules for several sectors such as

pharmaceutical, agricultural and food. Our findings

corroborate the study by Santos et al. (2022) where they

found results similar to ours for the phytochemistry of B.

orellana seed extract, although our results were more complex

for other phytochemical groups. According to Gutierrez-del-

Rio et al. (2021) and Stoll et al. (2023), some phytochemical

groups such as terpenoids (monoterpenes or carotenoids)

such as bixin and norbixin present in B. orellana seeds have

antioxidant action. Carotenoids can eliminate reactive species

of oxygen, nitrogen, superoxide, hydroxyl or peroxyl radicals.

In addition, when the extract is incorporated into the

polymeric matrix or directly into the product, it conserves

meat and other products by preventing lipid oxidation

(PACHECO et al., 2018).

Polymeric matrices based on natural starch have high

specificity, a characteristic of great importance in

incorporating extracts, essential oils, resinous oil, fixed oil,

among others. Among the various characteristics, we can

mention the physicochemical parameters such as the

intramolecular arrangement that undergoes changes, which

are positive mainly in the physicochemical parameters.

Increase in BP thickness were observed, and this may be

related to conformational changes in the starch chain by the

addition of B. orellana seed extract. Package thickness is an

important parameter that affects mechanical properties and

permeability to oxygen, carbon dioxide and water vapour. In

addition, the drying conditions and preparation methods are

factors that also influence the thickness of the packaging and

that must be verified (HOSSEINI et al., 2015).

Furthermore, denser BP were observed in this study due

to a greater distribution of carotenoids in the starch

polymeric matrix, retaining more moisture. It was observed

that B. orellana extract induces a high sensitivity to moisture

absorption, which can also affect the mechanical and thermal

properties of BP (THAKUR et al., 2019).

Although, we observed an increase in moisture content;

the high concentration of carotenoids (bixin or norbixin) in

the extract can form hydrogen bonds within the starch

carbohydrate chain. Thus, this limits the bonding of the

hydrogen group with water. This can result in BP starch with

extract to not completely solubilized due to lack of affinity

for water molecules. It was observed that with increasing

concentration the solubility decreases. In the same sense Nor

Adilah et al. (2018) found lower solubility with the addition

of increasing concentrations of carotenoid-rich mango

extract.

Regarding the colors of the biodegradable packaging

developments, the transmission scan shows a decrease in the

light transmission rates of BP incorporating different doses

of EEBO in the ultraviolet and visible regions (from 200 to

850 nm). Our packages visually showed an increasing reddish

tone as a higher concentration of extract was added.

Additionally, luminosity increased as higher extract

concentrations were added. Chroma a* tended to reddish

tones and chroma b* to yellow. Similar results were observed

by Santos et al. (2020) for packaging incorporated with

Capsicum chinense extract using arrowroot as a polymeric

matrix.

Color is an important parameter to be evaluated, as it

influences product acceptance (ROMANI et al., 2018;

RAMBABU et al., 2019; SANTOS et al., 2022). The

biodegradable packaging used is usually transparent so that

the product can be seen. However, colored and opaque

packaging helps to protect foods exposed to UV-Vis light,

especially in foods with a high fat content, such as meat (beef,

pork, fish), thus preventing them from suffering oxidative

degradation (PRIYADARSHI et al., 2021; SANTOS et al.,

2022).

Packaging must have a short degradation time, thus

avoiding large volumes in landfills. That is why during the

development of a new package the biodegradability time

must be observed. In our study, the biodegradability of the

packages in soil in nature was performed to determine their

degradation time and confirm their potential as alternatives

to replace synthetic polymers, which require a long time for

degradation; greater than 500-1000 years. Biodegradation is

defined as the loss of mechanical properties, fragmentation

or chemical modifications due to the action of

microorganisms and enzymes present in the natural

microbiota of the soil or beach sand. According to Filipini et

al. (2020) biodegradability is influenced by the presence of

gases, humidity, light, temperature and natural biota.

Our biodegradability time is similar to studies that include

plant extracts in starch polymeric matrix or similar (RIAZ et

al., 2020). We noticed that higher concentrations of extract

become toxic to soil microorganisms, as we found that there

was a decrease in the percentage of biodegradability.

Antifungal and antibacterial activity is reported for B. orellana

extract in different extraction solvents, Jena; Bhatnagar

(2021) found inhibition activity on Aspergillus flavus and

Aspergillus niger for B. orellana extracts produced from n-

hexane solvents, chloroform, ethyl acetate and methanol;

Majolo et al. (2013) observed that the hydroethanolic extract

of B. orellana showed intense antibacterial activity on

Salmonella serovar Enteritidis, Escherichia coli, Staphylococcus

aureus, Enterococcus faecalis and Listeria monocytogenes, and on

Candida albicans by the ethanolic extract by Poma-Castillo et

al. (2019).

The antifungal effect is one of the bioactive

characteristics that enhance the use of a new active packaging

capable of inhibiting the development of deteriorating

microorganisms. Our results were encouraging and are

similar to other studies with extracts incorporated into a

natural polymeric matrix (FILIPINI et al., 2020; DEY et al.,

2021).

5. CONCLUSIONS

The phytochemical study showed richness in several

important groups that present biological activities, including

antifungal. The interaction between the Bixa orellana seed

extract and the arrowroot polymeric matrix showed potential

for the development of new biodegradable active packaging,

this statement was observed by the good results on the

physicochemical characteristics, high biodegradability and

antifungal potential on food phytopathogens.

Due to the satisfactory results obtained, new studies can

evaluate mechanical characteristics, in vitro and in vivo, and

Extract from Bixa orellana L. seed: development of biodegradable packaging and biological activities

Nativa, Sinop, v. 11, n. 2, p. 259-265, 2023.

264

crystalline morphology of packages produced with the

ethanolic extract of Bixa orellana seeds.

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Acknowledgments: We thank Goiano Federal Institute, Rio

Verde, Goiás, Brazil; to the Technological Chemistry, Plant

Products Post-Harvest and Water and Effluent Laboratories, and to

the Master's and Doctorate in Agrochemistry.

Author Contributions: L.A.J. - oriented, writing (original draft),

investigation and data collection and final writing; A.C.P.M.F. - co-

advisor, conceptualization, study design, methodology, submission,

final corrections and publication; P.S. - Experimental validation and

translation; C.F.S.C., M.B.T. and F.A.L.S. - acquisition of funding;

M.V.A.V. - advisor, conceptualization, study design, supervision,

statistical analysis. All authors read and agreed to the published

version of the manuscript.

Funding: Goiás Research Support Foundation (FAPEG) for the

doctoral scholarship for the second author; National Council for

Scientific and Technological Development (CNPq); Financier of

Studies and Projects (FINEP), and Higher Education Personnel

Improvement Coordination (CAPES).

Institutional Review Board Statement: Not applicable.

Informed Consent Statement (Ethics Committee of the area):

Not applicable.

Data Availability Statement: Study data can be obtained by

request to the corresponding the second author, via e-mail.

Conflicts of Interest: There are no conflicts of interest between

the authors. Supporting entities had no role in the design of the

study; in the collection, analysis, or interpretation of data; in the

writing of the manuscript, or in the decision to publish the results.