Physicochemical characterization and antioxidant potential of plant extracts for use in foods

Abstract

The challenge of producing safe, high-quality food has led the food industry to look for natural products. Plants are excellent sources of nutritional compounds and phytochemicals, which can add functional value to products. This research aimed to evaluate plant extracts (açaí, cinnamon, guaraná, hibiscus, jambu, and mate) for their physicochemical properties and to assess their antioxidant potential, thus allowing application in food products. The açaí extract had the highest total carbohydrate content (90.1%), while the cinnamon extract exhibited the highest L* value (67.8%). In the comparative analysis, the guaraná extract had the highest protein content (20.9%), while the jambu extract had the highest lipid content (19.7%) and, consequently, the highest energy value (474 kcal.100 g^-1). The highest amounts of the elements C and N were reported in the guaraná extract (73.3% and 4.4%, respectively), while the elements Ca and P were more pronounced in the hibiscus extract (0.5% and 0.2%). Regarding the color parameters, as expected, the hibiscus extract exhibited more red color (a* = 17.5) in contrast, the jambu extract had a lower luminosity and b* value (28.3 and -1.4, respectively), indicating green and blue tones. The mate extract showed greater antioxidant potential in the three assays, 1574.3 µmol TE g^-1 (ABTS+), 1048.9 µmol TE g^-1 (DPPH) and 2539.9 µmol TE g^-1 (FRAP) and higher content of TPC (776 mg GAE g^-1). The plant extracts have great potential as natural colorants and ingredients in food formulations due to their physicochemical characteristics, such as color, proteins, lipids, minerals, and antioxidant potential.

Keywords:
Acmella oleracea (L.); Cinnamomum zeylanicum Blume; Euterpe oleracea Martius; Hibiscus sabdariffa L.; Ilex paraguariensis A. St. - Hil.; Paullinia cupana Kunth; Spilanthes oleracea L.; Plant extract

Highlights

The dried mate showed the highest content of phenolic compounds and antioxidant potential

Guaraná extract had the highest protein content, while jambu extract had the most lipids

Plant extracts offer potential as natural colorants and functional food ingredients

1 Introduction

The food industry has faced daily challenges with the great demand to develop and produce safe and high-quality food. To satisfy this demand when working with natural inputs, the physicochemical characteristics of these materials and their composition of bioactive compounds should be investigated (Santana & Macedo, 2018; Roggia et al., 2020). Medicinal plants have been known since antiquity and are widely consumed around the world and recognized as excellent sources of nutritional and phytochemical compounds (Chen & Wei, 2017; Franzen et al., 2019, 2020, 2021; Stefaniak & Grzeszczuk, 2019). These phytochemical compounds can be used to add functional value to food products, in addition to improving health and providing clean labels.

Among a broad universe of phytochemicals, the phenolic compounds have been the most investigated due to their inflammatory, antioxidant, and even anti-aging capacity, allowing the production of plant extracts with high economic value. The phenolic profile varies according to the plant species and the extract preparation process (Boscariol Rasera et al., 2019; Magro & Castro, 2020; Magalhães & Santos, 2021; Santos et al., 2022).

These natural antioxidant substances can be used to increase the shelf life of foods, in addition to promoting health benefits, acting in the prevention of various diseases related to oxidative stress, such as cancer, cardiovascular and neurodegenerative diseases, hypertension, and diabetes (Cardozo Junior & Morand, 2016; Kaulika & Febriansah, 2019; Magalhães & Santos, 2021).

In recent years, there has been an increase in the demand and interest of researchers in açaí pulp (Euterpe oleracea Martius), due to its high antioxidant capacity provided the presence of polyphenols and tocopherols, which explains its classification as a functional food for the prevention of several degenerative diseases (Yamaguchi et al., 2015; Monge-Fuentes et al., 2017; Oliveira et al., 2019; Jesus et al., 2020).

Cinnamomum zeylanicum Blume, known as “cinnamon”, is a plant native to regions of South Asia. The parts (barks, leaves, and flowers) of C. zeylanicum are sources of bioactive compounds which have analgesic, antiseptic, anticancer, coagulant, neuroprotective, hepatoprotective, gastroprotective, cardioprotective, antimicrobial and antioxidant potential, in addition to controlling serum and lipid levels and reducing blood cholesterol concentration (Nabavi et al., 2015; Chuesiang et al., 2019; Gomes et al., 2020; Silva et al., 2020).

Guaraná (Paullinia cupana Kunth, Sapindaceae) is a Brazilian plant originally from the Amazon region and its roasted seeds are used for their stimulant, aphrodisiac, and healing properties. The guaraná plant is associated with a wide variety of pharmacological effects, including anticarcinogenic, antiproliferative, antimicrobial, antioxidant, energetic cytoprotective, thermogenic, antidepressant, anxiolytic, and reducing oxidative and metabolic disorders (Hertz et al., 2015; Machado et al., 2015; Matsuura et al., 2015; Kober et al., 2016; Marques et al., 2019).

The Hibiscus sabdariffa L. plant is a species of the Hibiscus genus and belongs to the Malvaceae family, originating in Asia and Africa. In traditional medicine, hibiscus is used as a diuretic and for the treatment of gastrointestinal disorders, liver infections, fever, and hypertension. Hibiscus is considered a functional food in Asian countries, it is a rich source of flavonoids and the economic interest is in dehydrated calyxes, used worldwide for the production of teas, foods, conservatives, and antioxidants (Nazratun Nafizah et al., 2017; Su et al., 2018; Vargas-León et al., 2018).

Acmella oleracea (L.), also known as Spilanthes oleracea L., from the Asteraceae family, was discovered in Peru and is found in tropical and subtropical regions, where it is known as jambu. It is an important medicinal plant, traditionally used for its analgesic and anti-inflammatory properties, but also for antipyretic, anticonvulsant, antidiarrheal, antidiuretic, antiseptic, antifungal, antiprotozoal and insecticidal properties (Lalthanpuii et al., 2020; Maggini et al., 2021; Uthpala & Navaratne, 2021).

Ilex paraguariensis A. St. - Hil., popularly known as yerba mate, is a species native to temperate and subtropical regions of South America, belonging to the Aquifoliaceae family. The compounds present in I. paraguariensis, have several therapeutic properties and are recommended as hypocholesterolemic, hepatoprotective, diuretic, digestive, nervous system stimulant, antioxidant, anti-inflammatory, antirheumatic, and lipolytic agent. It is also indicated for the treatment of occasional asthenia and as an adjunct in the treatment of overweight (Fagundes et al., 2015; Gambero & Ribeiro, 2015; Antunes et al., 2018).

Many in vitro antioxidant assays are available in the literature to assess the bioactivity of extracts, and the most common methodologies include 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and Ferric Reducing Antioxidant Power (FRAP) assays. These methods differ in the principles of the chemical reaction, the target molecule, the way of expressing the results, the pH of the environment, the reaction time, and the plant matrix, among others. Therefore, methodologies using different mechanisms and chemical reagents are required to evaluate the in vitro antioxidant capacity of natural extracts. It is also important to take into account the polarity of the antioxidant compounds present in the plant matrix (Oliveira et al., 2015; Granato et al., 2018).

Thus, this study aimed to determine the total phenolic compounds (Folin-Ciocalteu), the antioxidant potential (ABTS⁺, DPPH, and FRAP assays) and to investigate the physicochemical characterization of the plant extracts of E. oleraceae (Açaí), C. zeylanicum (Cinnamon), P. cupana (Guaraná), H. sabdariffa (Hibiscus), S. oleraceae (Jambu) and I. paraguariensis (Dried Mate).

2 Materials and methods

2.1 Material

The plant extracts were kindly donated by Grupo Centroflora/Givaudan® (Rod. Eduardo Zuccari, km 21,5, Botucatu, SP, Brazil), and registered in the National System for the Management of Genetic Heritage and Associated Traditional Knowledge (SisGen) under registration number A69B2CF.

The extracts of the species E. oleraceae (Açaí) batch number 160620.02807, product code number 100343, C zeylanicum (Cinnamon) batch number 251018.00885, product code number 200064, P. cupana (Guaraná) batch number 200521.03897, product code number 100350, H. sabdariffa (Hibiscus) batch number 020221.03470, product code number 100101 and I. paraguariensis (Dried Mate) batch number 041120.03238, product code number 100383, were presented in the form of a fine soluble powder and the extract of S. oleraceae (Jambu) batch number 010921.04255, product code number 100400, was presented in the form of a viscous liquid. All extracts of this study are used as food ingredients and additives by the supplier company, which attests to its safety by the ABNT-NBR 14725-2: 2009 – corrected version 2:2010.

The extracts were obtained from plant parts such as leaves, flowers, fruits and seeds, specific to each species through a standardized extraction, concentration and drying process, as described by the manufacturer. For the jambu extract, specifically, the oleo-resin extraction and concentration processes were carried out. The solvents used in the extraction processes were water and ethanol, as described by the manufacturer.

The dry extract obtained represents the soluble fraction of the plant parts in the solvent used. This form allows for better standardization and greater stability for analysis and application. The dry extract maintains most of the soluble bioactive compounds present in the plant matrix, as confirmed by the antioxidant analyses performed.

2.2 Reagents for the analysis of phenolic compounds and antioxidant potential

The ABTS, potassium persulphate, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), DPPH, Ferric Chloride Hexahydrate (FeCl3·6H2O), (TPTZ), Folin and Ciocalteau’s phenol reagent, and sodium carbonate were purchased from Sigma-Aldrich (Steinheim, Germany). All other chemicals were purchased in the grade commercially available.

2.3 Preparation of extracts for the analysis of phenolic compounds and antioxidant potential

To prepare the extracts for analysis, 0.25 g of each plant material was placed in test tubes with 10 mL of distilled water and homogenized for 1 hour. After homogenization, sample dilutions (extract:distilled water) of 1:50, 1:100, 1:300, and 1:500 were made for the analysis.

2.4 Total phenolic content (TPC)

The total phenolic contents (TPC) were estimated according to the method of Swain & Hillis (1959) with modifications suggested by Pereira et al. (2018). Briefly, 25 µL of diluted extracts were mixed with 25 µL of Folin and Ciocalteau's phenol reagent (50%, v:v). After that, 200 µL of sodium carbonate solution (5%, w:v) was added. The reaction mixtures were allowed to stand for 20 minutes at 40°C. The absorbance was read in a micro plaque reader at 760 nm. TPC were expressed as mg of gallic acid equivalents (GAE) per gram of sample (mg GAE g^-1).

2.5 Antioxidant potential

The antioxidant potentials were measured by different methodologies, as follows: ABTS- and DPPH- radicals scavenging activity (Al-Duais et al., 2009) and FRAP assay (Benzie & Strain, 1996; Wiriyaphan et al., 2012). The detailed procedures were described by Boscariol Rasera et al. (2019). All results were expressed as µmol of Trolox equivalents per g of dried and defatted sample (µmol TE g^-1).

2.6 Physicochemical characterization of plant extracts

The moisture contents of the extracts were determined by the Association of Official Analytical Chemists (2005) method in a Marconi® drying oven, model MA 035/5-PB. The ash contents were determined by incineration in a muffle furnace (Fornitec®, model 1994) at 550 °C (Association of Official Analytical Chemists, 2005). Protein contents were determined by the Kjeldahl method with a conversion factor of 6.25 (Association of Official Analytical Chemists, 2005). Lipid contents were determined according to the method of Bligh and Dyer (1959). Total carbohydrates were estimated by the difference between the total mass and the sum of moisture, ash, protein, and lipids levels. The energy value of the extracts was calculated according to Franzen et al. (2019). The pH was measured using SevenGo™ pH meter SG2 da Mettler Toledo©, and the total soluble solids (TSS) was determined in a digital portable refractometer model PAL-3, Atago® was used. For the elemental quantification of the extracts, a K-alpha XPS spectrometer (Thermo Scientific) was used with a monochromatic Al Kα X-ray source (72 W) as cited by Silva et al. (2022), using 10 different positions of the extract powders. Electron kinetic energy was measured by a 180° hemispherical energy analyzer operated in the analyzer's constant energy mode, and data acquisition and processing were performed using Thermo Advantage software (Thermo Scientific). The color of the extracts was analyzed in a Konica Minolta CR-410 colorimeter, calibrated according to the CIELAB system (L*, a*, b*) with measurement of the luminosity parameters (L*) with a range of 0 = black and 100 = white, a* and b* coordinates responsible for chromaticity (+a* = red, -a* = green and +b* = yellow, -b* = blue), color intensity or chroma (c*) and the hue angle (h°) (illuminant D65). These measurements were made at 10 random points of the samples for a better representation of the color of the extracts.

2.7 Statistical analysis

The results were analyzed according to Tukey’s test, using the software IBM® SPSS Statistics, version 22.0 (USA). The values were expressed as the arithmetic mean and considered statistically different at p-value ≤ 0.05.

3 Results and discussion

The results of the extract characterization are shown in Table 1. The total phenolic contents (TPC) were expressed in mg of gallic acid equivalent per g of extract (mg GAE g^-1) and ranged from 428.46 to 776.09 mg GAE g^-1 for the açaí and dried mate extracts, respectively.

Table 1
Total phenolic contents (TPC) and antioxidant potential measured by ABTS- and DPPH- radical scavenging activities and FRAP assay of the plant extracts.

Samples	TPC	ABTS	DPPH	FRAP
	(mg GAE g-1)	(µmol TE g-1)	(µmol TE g-1)	(µmol TE g-1)
Açaí	428.46d ± 8.01	38.06e ± 6.56	359.03d ± 18.27	63.04e ± 5.44
Cinnamon	432.81cd ± 10.70	253.38c ± 4.86	506.62c ± 31.03	132.04de ± 21.20
Guaraná	652.62b ± 14.55	1140.95b ± 24.52	849.83b ± 63.57	834.79b ± 24.02
Hibiscus	457.95cd ± 7.76	186.58cd ± 5.01	575.78c ± 26.72	240.40cd ± 22.08
Jambu	464.77c ± 7.76	116.39de ± 6.59	473.82cd ± 24.53	294.37c ± 14.14
Dried Mate	776.09a ± 20.09	1574.32a ± 86.87	1048.92a ± 77.58	2539.94a ± 130.51

• Means followed by equal letters (vertical) do not differ statistically from each other by the Tukey’s test at 5% (p-value ≤ 0.05).

It was observed that the dry mate extract presented a statistical difference compared to the other plant extracts analyzed (p < 0.05) for TPC content and antioxidant potential for the three methods applied in this study. Guaraná extract was also highlighted in the comparative results between the four tests.

The dried mate extract had the highest TPC content with 776.09 mg GAE g^-1, followed by guaraná extract with 652.62 mg GAE g^-1. The third extract with the highest TPC content was the jambu extract with 464.77 mg GAE g^-1, which did not differ statistically from the hibiscus (457.95 mg GAE g^-1) and cinnamon (432.81 mg GAE g^-1) extracts for this parameter.

The present results are in line with the literature and confirm that yerba mate is an important source of polyphenols, with similar phenolic contents to other polyphenol-rich beverages such as cocoa powder, black tea, and red wine (Blum-Silva et al., 2015; Mateos et al., 2018).

Several studies have pointed out the complexity of the chemical composition of yerba mate. The main active compounds with high concentrations found in the leaves and branches of the plant include polyphenols (chlorogenic acid) and xanthines (caffeine, theobromine, and theophylline), followed by purine alkaloids (caffeic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid) and flavonoids (quercetin, kaempferol, and rutin) (Bojić et al., 2013; Braghini et al., 2014; Cardozo et al., 2021).

Silva et al. (2019b) studied the characterization of solid lipid microparticles with guaraná seed extract (Paullinia cupana) and found the highest content of phenolic compounds of 434.56 mg GAE/g in guaraná seed extract, using pure ethanol (50%) and water (50%) as solvents. Those authors also used spray-drying to produce powdered extracts using 50% ethanol as a solvent and reported a total phenolic content of 437.75 mg GAE/g.

Santana et al. (2019) evaluated the extraction of raw seeds and guaraná (P. cupana) residues using pressurized liquids and supercritical CO₂ combined with enzymes and reported a TPC of 642.45 mg GAE/100 g in the extraction of raw seeds using the enzyme pectinase by the pressurized liquid extraction method. For the supercritical fluid extraction method with the pectinase enzyme, the TPC in the extract was 604.71 mg GAE/100 g. For the guaraná residue extracts, the TPC values were 560.43 mg and 386.20 mg GAE/100 g using the pressurized liquid extraction method and the enzymes cellulase and pectinase, respectively. Concerning the supercritical fluid extraction using the cellulase and pectinase enzymes, the values were 183.97 mg and 188.15 mg GAE/100 g, respectively. Thus, despite the use of different methodologies, the results of this study corroborate the findings of other studies.

The biological properties of jambu are due to its endogenous content of bioactive compounds, such as sterols, coumarins, flavonoids, saponins, terpenoids, polysaccharides, and, mainly, alkylamides (Dallazen et al., 2020).

Among the alkyl amides, spilanthol ((E, E, Z)-2,6,8-decatrienoic acid N-isobutylamide) is considered the most active compound found in S. oleracea. Spilanthol is found mainly in the flowers, leaves, stems, and roots of Spilanthes (Dallazen et al., 2020; Lalthanpuii & Lalchhandama, 2020; Joshi et al., 2020).

Boontha et al. (2020) investigated crude jambu extract and found phenolic contents and antioxidant potential at IC₅₀ of 62.8 and 1.2 mg GAE/mL, respectively. The phenolic content is lower when compared to the result found in the present study (464.77 mg GAE g^-1), with consequent higher antioxidant potential (473.82 µmol TE g^-1), due to the higher concentration of TPC in the jambu extract from the present research.

Swargiary et al. (2019) evaluated the phytochemical properties of S. acmella and reported that the jambu methanolic extract contained a high concentration of TPC (67 µg GAE/mg d.w.) and antioxidant potential by the DPPH, ABTS and superoxide anion scavenging activity, with IC50 values of 730, 57, and 965 µg/mL, respectively. The authors observed a relationship between TPC and antioxidant potential, once a higher TPC content in the plant extract contributes to a higher antioxidant potential. The authors also pointed out that the greater antioxidant potential is due to the ability of phenolic contents to chelate metal ions during the formation of free radicals. Those results corroborate the potential of the jambu extract of this study.

The polyphenols in hibiscus play an important role as an anti-inflammatory agent, improving the antioxidant conditions and regulating the expression of cyclooxygenase-2 (El Bayani et al., 2018). Ojulari et al. (2019) reported that the bioactive compounds from hibiscus were also effective in acting against obesity, with a relevant decline in body weight, suppression of adipogenesis, and inhibition of lipid accumulation.

Plants and their extracts contain compounds that can improve the health state of individuals or reduce the risk of chronic diseases, with an emphasis on the phenolic compounds, which have shown immunoregulatory activity and preventive effects against cancer, diabetes, cardiovascular, and neurodegenerative diseases. These health benefits arise from the antioxidant potential of this compound, which plays a key role in metabolism and against oxidative stress (Shahidi et al., 2019; Borges et al., 2022; Kan et al., 2023; Pasquet et al., 2024).

Phenolic compounds can be consumed by direct absorption from plant-based food products but also extracted from food by-products (plant extracts) and applied to food formulations to produce functional foods (GopikaJayaprakash et al., 2023).

Concerning the antioxidant potential of the extracts, the dried mate extract showed the highest activity by the FRAP assay, with 2539.94 µmol TE g^-1. Also, this extract exhibited the highest averages compared to the three methods used for assessing the antioxidant potential, with values of 1574.32 and 1048.92 µmol TE g^-1 by ABTS and DPPH, respectively. Mate was the herb with the highest phenolic content, which can explain its higher antioxidant potential.

The guaraná extract showed the second highest antioxidant potential among the extracts studied for the three methods applied in this study, with values of 1140.95 µmol TE g^-1, 849.83 µmol TE g^-1, and 834.79 µmol TE g^-1 for ABTS, DPPH, and FRAP assays, respectively.

Plants produce secondary metabolites called phenolic compounds that act in plant defense processes against pathogens, environmental stress and radiation, providing physical and chemical barriers, in addition to contributing to the color, aroma and flavor of these raw materials. The structure of phenolic compounds can vary widely, but they all share the presence of the phenol group, which consists of a benzene ring linked to a hydroxyl group (OH^-). This hydroxyl group is responsible for giving unique properties to phenolic compounds, such as antioxidant activity and the ability to form bonds with other molecules (Acosta-Estrada et al., 2014; Vuolo et al., 2019).

The structure of phenolic compounds can include several substituents linked to the benzene ring, which is related to their radical scavenging and/or metal chelating activity, making their antioxidant action quite dependent on their structure. With free hydroxyl groups (OH^-) in their structures, phenolic compounds tend to exhibit strong antioxidant activity due to the ability of these groups to donate electrons. Therefore, the number of hydroxyl groups, their position in relation to the carboxyl functional group, glycosylation and the presence of substituents in the rings directly influence the antioxidant potential of phenolic compounds (Oliveira et al., 2015; Franzen et al., 2021; Boscariol Rasera et al., 2023).

Investigating the antioxidant potential and phenolic content of plant extracts is important for correlating the extract composition with health benefits (Magalhães & Santos, 2021). From a nutritional and pharmaceutical point of view, it is important to evaluate the antioxidant potential of phytochemicals in plant materials and their extracts (Schauss, 2016).

Significant differences were observed for the moisture contents of the extracts, with the highest and lowest contents observed for hibiscus and jambu extracts (Table 2), with values of 8.2 and 5.9%, respectively. Moisture content is a critical parameter in plant material since a plant extract with low moisture is essential for maintaining chemical and microbiological stability (Gallo et al., 2015; Simen et al., 2016).

Table 2
Results of the physicochemical characterization, color parameters, and particle size of the plant extracts.

Constituents	Açaí	Cinnamon	Guaraná	Hibiscus	Jambu3	Dried Mate
Moisture1	6.67d ± 0.089	7.12c ± 0.087	7.50b ± 0.040	8.22a ± 0.099	5.90e ± 0.127	7.45b ± 0.128
Ash1	1.33d ± 0.074	0.61e ± 0.063	3.57c ± 0.118	10.41a ± 0.065	0.04f ± 0.022	8.12b ± 0.086
Proteins1	1.34c ± 0.008	3.74c ± 1.633	20.97a ± 1.064	10.57b ± 1.316	9.37b ± 1.569	11.51b ± 0.800
Lipids1	0.47d ± 0.135	0.02e ± 0.016	1.89b ± 0.116	0.19e ± 0.020	19.72a ± 0.136	0.88c ± 0.051
Total Carbohydrates1	90.19a ± 0.204	88.51a ± 1.647	66.07c ± 1.033	70.61b ± 1.330	64.97c ± 1.680	72.04b ± 0.740
Energy Value2	370.34b ± 0.575	369.19b ± 0.142	365.17c ± 0.977	326.44e ± 0.168	474.84a ± 1.048	342.14d ± 0.794
pH	4.36ab ± 0.036	4.40a ± 0.065	4.50a ± 0.104	3.87c ± 0.093	4.24b ± 0.031	4.39ab ± 0.030
Total soluble solids (°Brix)	8.15e ± 0.058	8.70c ± 0.000	6.25f ± 0.058	8.55d ± 0.058	9.05b ± 0.058	9.35a ± 0.058
L*	45.39d ± 0.414	67.88a ± 0.163	52.89c ± 1.693	37.14e ± 1.402	28.39f ± 0.224	58.61b ± 0.713
a*	15.64b ± 0.066	14.04c ± 0.106	15.93b ± 1.317	17.54a ± 0.364	1.26e ± 0.046	6.58d ± 0.326
b*	3.53d ± 0.141	20.44b ± 0.098	19.48c ± 1.252	2.37e ± 0.588	-1.48f ± 0.023	22.61a ± 0.481
c*	16.03d ± 0.083	24.80a ± 0.138	25.21a ± 0.692	17.71c ± 0.424	1.94e ± 0.046	23.55b ± 0.545
h°	12.71e ± 0.475	55.51c ± 0.090	50.71d ± 3.843	7.66f ± 1.758	310.27a ± 0.710	73.78b ± 0.481
Particle size diameter (µm)	57b ± 3.304	89a ± 4.646	51b ± 4.573	44bc ± 2.449	ND	43c ± 1.708

• ND: not determined due to the liquid nature of the extract. Data expressed as mean ± standard deviation. Means within the same line with different letters are significantly different at the 5% level, by the Tukey’s test (p-value < 0.05). ¹(g/100 g). ²(kcal/100 g). ³for the extract of the Jambu species, the averages of the proximal composition are expressed as mL/100 mL of liquid extract, and the energy value is expressed as kcal/100 mL of liquid extract. L*: 0 = black and 100 = white; a*: +a = red and -a = green; b*: +b = yellow and –b = blue; c*: chroma; h°: hue angle. Source: The authors.

A well-dried extract decreases the risk of microbiological contamination and reduces the occurrence of redox reactions. Anthocyanins are sensitive to factors such as temperature, light, pH, oxygen, and others (Lopes et al., 2020). Thus, liquid extracts should be subjected to a freeze-drying process to preserve their chemical and pharmacological properties as well as their physicochemical and microbiological stability (Silva et al., 2019a).

Significant differences were observed for the ash content of the extracts, which ranged from 0.04% to 10.41%. The hibiscus extract had the highest ash content, followed by dried mate and the guaraná extracts, with 10.41%, 8.12%, and 3.57%, respectively. Mineral-rich plant extracts can be used to supplement diets lacking in essential nutrients. Minerals such as iron, zinc, calcium, magnesium, and potassium are vital for the proper functioning of the body and the prevention of nutritional deficiencies, including osteoporosis, anemia, and hypertension, among others (Bhardwaj et al., 2023; Sudha & Saral, 2023).

Some mineral elements are essential cofactors for isoenzymes such as superoxide dismutase, which may contribute to the in vivo antioxidant potential of the extracts (Huehne et al., 2020).

The guaraná extract exhibited the highest protein content, with 20.97%. Protein is an essential nutrient for human growth and well-being. It is necessary for the body's regular biological functions and development, growth, maintenance, maturation, lactation, and reproduction, as well as protecting against cancers and diseases, and its shortage has negative impacts on the human diet (Wang et al., 2022; Zhao et al., 2023; Javed et al., 2024).

Concerning the jambu extract, it presented the highest lipid content with 19.72%. Lipids are the essential components of cell membranes and serve as the building blocks of biomolecules that enable cell function and health. This nutrient is part of the construction of cell membranes and the membranes of numerous essential organelles, such as mitochondria, the Golgi apparatus, and the endoplasmic reticulum (Gunay et al., 2021).

Lipids are mainly responsible for the juiciness, tenderness, smoothness, flavor, nutritional value, and storage stability of food products. In addition, plant-derived lipids have a completely different chemical composition and physicochemical properties from animal fats. Most vegetable oils are in a liquid state at room temperature, while animal fats are in a solid state (Chen et al., 2023).

Carbohydrates are an important source of energy in the human diet. The current global importance of healthy eating of these nutrients has focused on increasing the consumption of carbohydrates from different plant sources (Stylianopoulou, 2023). The açaí and cinnamon extracts had the highest carbohydrate contents, with 90.19 and 88.51% respectively, with significant differences when compared to the other extracts. They are a significant source of micronutrients (vitamins and minerals) and phytochemicals. Consuming only a single food source of carbohydrates in diets can lead to nutritional deficiencies. Therefore, the intake of different carbohydrate sources is very important for nutrient sufficiency and health benefits (Stylianopoulou, 2023).

Vegetables are characterized by their low energy values. The jambu extract had the highest energy value with 474.84 kcal 100 g^-1, probably due to the high lipids content of this extract, which was used to calculate the energy value, along with the macronutrients (proteins, lipids, and carbohydrates). According to the North American National Research Council (1989), the recommended daily energy requirements for men and women aged from 19 and 24 years are 2,900 and 2,200 kcal, respectively. Thus, the results of the present study showed that consuming 100 g of jambu extract can provide 6.1 and 4.6% of the recommended daily energy intake for adult men and women, respectively.

The pH is a parameter that indicates the acidity or alkalinity of a substance or product. In foods and their by-products, pH plays a crucial role in various aspects, influencing both the chemical and sensory characteristics, as well as product safety and stability (Pasquet et al., 2024). The hibiscus and jambu extracts had the lowest pH values of 3.87 and 4.24, respectively, while the guaraná, cinnamon, dried mate, and açaí extracts presented the most alkaline pH, with values of 4.50, 4.40, 4.39, and 4.36, respectively.

The alkaline environment in phenolic-enriched foods leads to the formation of quinones through auto-oxidation process due to the phenol/phenolate ion balance, with a significant reduction of phenolic compounds in the food products (Liu et al., 2022; Pasquet et al., 2024). In turn, quinones are reactive species prone to polymerization, leading to the undesirable browning of foods and a decrease in product quality (Ali et al., 2018; Geng et al., 2023).

Total soluble solids (°Brix) include components such as sugars, vitamins, and minerals. As shown in Table 2, the dried mate extract had the highest solids content, with 9.35 °Brix, while the guaraná extract had the lowest solids value, with 6.25 °Brix. It is known that the extraction of plant extracts involves starch degradation and breakdown of the cell wall matrix, which is associated with an increase in total soluble solids (Shamili, 2019).

Particle size in food affects several parameters, including texture and sensory acceptance, as well as nutrient absorption in the digestive tract. This parameter is fundamental in the food industry to guarantee the quality and stability of products (Yao et al., 2023). The cinnamon extract had the largest particle size, with 89 µm, differing statistically from the other extracts, while the dried mate extract had the smallest particle size, with 43 µm.

A wide variety of powdered particulate raw materials is used in the food industry. To develop ingredients and products with specific desirable characteristics, it is necessary to understand the fundamental properties of particles and particulate food systems (López-Córdoba & Goyanes, 2017; Bazán-Colque et al., 2023).

Concerning the color of the extracts, the cinnamon extract showed greater luminosity (+L), close to white, with an L-value of 67.88. In contrast, the jambu extract had a lower L-value of 28.39, close to a black tone. It is known that the color of plant extracts is influenced by the presence or absence of pigments from plants.

Table 2 presents the results of the physical-chemical analyses of the plant extracts researched in this study.

When the plant extract is included in food formulations, the resulting color has an impact on the sensory acceptance of the food product. In this study, three extracts showed high luminosity, including cinnamon, dried mate, and guaraná extracts, while açaí, hibiscus, and jambu extracts showed low luminosity. All extracts differed statistically for the color parameter L.

Saikia et al. (2015) reported high luminosity of dried carambola pomace extract, which is rich in polyphenols. In the present study, positive a* and b* values were observed for all the powders except for the jambu extract, which showed values close to zero and negative values, corresponding to a darker green and bluish color.

The addition of this extract to food formulations can improve the attractiveness of the product. Color is one of the most important sensory attributes of food and has a significant influence on its attractiveness and acceptance by consumers. The visual appearance of food is the consumer's first contact, and the initial perception is based on color (Lan et al., 2022; Ghosh et al., 2023).

Plant extracts can also be used as natural colorings in foods. These colorants are categorized based on their origin, hue, or chemical structure. Sigurdson et al. (2017) demonstrated the classification of natural colorants as isoprenoid derivatives (carotenoids), flavonoid derivatives (anthocyanins), heterocyclic nitrogen derivatives (betalains), and pyrrole derivatives (chlorophyll). In addition, there are other natural colorants such as phycobiliproteins, alizarin, curcumin, etc (Parlak et al., 2024).

The jambu extract used in the study is characterized as an oleo-resin obtained from hydroethanolic extraction. This final product is a viscous liquid with low humidity and a high content of fat-soluble compounds, mainly vegetable oils and lipophilic bioactive substances (Silva et al., 2017).

The jambu extract had a moisture content of approximately 5.9%, which is due to the small amount of residual water present in the final product, possibly due to the hygroscopicity of the material or the minimal retention of water during processing. However, since the extract is predominantly composed of oils and resins (viscous liquid), the term “oil-resin” more accurately describes its physical and chemical nature (Jerônimo et al., 2024).

Concerning the elemental composition, no significant amounts of minerals were observed for the açaí, and cinnamon extracts when compared to the guaraná, hibiscus, and dried mate extracts. The jambu extract had a liquid nature, thus it was not analyzed for the elemental composition once the technique was suitable only for powdered or solid materials.

Minerals represent 1% to 2% of fresh foods. Plants are characterized by low sodium and high potassium contents. They also contribute to the daily intake of calcium, phosphorus, magnesium, iron, and zinc. Although there is a low bioavailability of iron in this food group, vitamin C can contribute to an increase in its absorption. The most representative anions are phosphate, chloride, and carbonate (Ruiz-López & Ruiz, 2023).

The high-resolution XPS scans of carbon (C 1s), oxygen (O 1s), and nitrogen (N 1s) ranged from 282 to 291, 529 to 537, and 392 to 410 eV, respectively. The XPS methodology provides information on the atomic composition, expressed in percentage of atomic weight, and the type of chemical bond (Lawrie et al., 2007; Silva et al., 2022).

The extracts showed element fractions characteristic of plant raw materials, as can be seen in Table 3.

Table 3
Quantification by X-ray photoelectron spectroscopy (XPS) of the elemental composition of commercially obtained dry extracts of guaraná, hibiscus and mate.

Elements (w%)	Guaraná	Hibiscus	Mate
C 1s	73.38	61.10	63.88
O 1s	22.19	33.65	33.90
N 1s	4.44	4.40	2.22
Ca 2p	-	0.59	-
P 2p	-	0.26	-

• w% = atomic weight %. C = carbon. O = oxygen. N = nitrogen. Ca = calcium. P = phosphorus. Source: The authors.

The guaraná extract is composed of carbon (C 1s, 282 to 291 eV), oxygen (O 1s, 529 to 536 eV), and nitrogen (N 1s, 392 to 410 eV). The hibiscus extract showed the highest concentration of chemical elements, probably due to its higher ash content when compared to the other extracts studied. In turn, the hibiscus extract is composed of carbon (C 1s, 282 to 294 eV), oxygen (O 1s, 529 to 537 eV), nitrogen (N 1s, 397 to 405 eV), calcium (Ca 2p, 344 to 360 eV) and phosphorus (P 2p, 130 to 138 eV). Finally, the dried mate extract is composed of carbon (C 1s, 282 to 298 eV), oxygen (O 1s, 529 to 537 eV), and nitrogen (N 1s, 392 to 410 eV).

The C 1s and O 1s scans of the extracts showed binding energies characteristic of hydrocarbons (CH- and C-C, ∼284 eV) and carboxyl groups (RO-C=O, ∼533 eV). N 1s scans show binding energies for the amine group (C-NH2, ∼400 eV). For the hibiscus extract, calcium (Ca 2p, ∼347 eV) and phosphorus (P 2p, ∼134 eV) fractions were found, which include chemical compounds such as carbonate, nitrate, and calcium oxide as well as phosphate and phosphide ions (Silva et al., 2022).

The C 1s binding energy distributions (high-resolution scans) showed an increase in peak intensity at ∼285 eV and a decrease at ∼287 eV, corresponding to hydrocarbons. On the other hand, the O 1s high-resolution scans showed a high intensity of peaks near 533 eV, which referred to carboxyl groups. According to Sari & Chellam (2013), the increase in the content of hydrocarbon groups with an N 1s atom may be due to the amino acid side chain, while the increase in carboxyl groups may be due to the deposition of mixtures of bitter compounds and sugars.

The aromas and flavors of food are made up of various chemical elements that bind together to form various compounds, including volatile substances (Morais Souto et al., 2023).

The extracts in this study had significant concentrations of carbon (C), oxygen (O), and nitrogen (N), which belong to various classes of organic compounds. The specific characteristics of aromas and flavors depend on the unique molecular structure of each compound and their relative concentrations in a given substance. Therefore, the plant extracts should be characterized from the chemical point of view with the aim to use them as ingredients in food products to enhance or intensify aromas and flavors, as well as provide color to the product.

4 Conclusion

The phenolic contents and the antioxidant potential of the plant extracts differed greatly depending on the plant species. The dried mate extract showed the highest content of phenolic compounds and antioxidant potential.

The plant extracts studied presented good sources of phenolic compounds, with important antioxidant potential that can be considered beneficial for health, such as guaraná and dried mate extracts. Regular consumption of foods rich in phenolic compounds is associated with several health benefits, such as reducing the risk of cardiovascular disease, cancer, diabetes and other chronic conditions.

The physicochemical characterization of the plant extracts showed a large amount of nutrients with low energy values that are important for the diet. The hibiscus extract had the higher ash content and the higher concentration of chemical elements, as well as a more acidic profile (lower pH) among the extracts studied. The guaraná extract had the highest protein content, while jambu extract had the highest lipid content and energy value. On the other hand, the açaí and cinnamon extracts had the highest carbohydrate contents.

Concerning the color of the extracts, the cinnamon, guaraná, and dried mate extracts exhibited lighter color, while the açaí, hibiscus, and jambu extracts were darker. Regarding the particle size, the cinnamon extract had the higher particle size, while the dried mate extract had the smaller particle size among the extracts studied.

The elemental analysis of the extracts showed chemical elements characteristic of aromas and flavors as a function of the chemical bonds that form the compounds.

The physicochemical and nutritional results showed the effectiveness of using plant extracts as coloring agents and ingredients in food formulations, which may be an interesting strategy in the food industry.

Acknowledgements

The authors are grateful to CAPES with Finance Code 001 for the financial and fellowship support and the Centroflora/Givaudan® Group for preparing and donating the plant extracts used in this research. This research used facilities of the Brazilian Nanotechnology National Laboratory (LNNano), part of the Brazilian Centre for Research in Energy and Materials (CNPEM), under the supervision of the Brazilian Ministry for Science, Technology, and Innovations (MCTI).

• Cite as:
  Franzen, F. L., Boscariol Rasera, G., Silva, K. F. C., Castro, R. J. S., Oliveira, M. S. R., & Bolini, H. M. A. (2025). Physicochemical characterization and antioxidant potential of plant extracts for use in foods. Brazilian Journal of Food Technology, 28, e2024085. https://doi.org/10.1590/1981-6723.08524
• Funding: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Finance Code 001).

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Edited by

• Associate Editor: Maria Teresa B. Pacheco

Publication Dates

• Publication in this collection
  21 Mar 2025
• Date of issue
  2025

History

• Received
  22 Aug 2024
• Accepted
  30 Dec 2024