Metabolomic Approach Reveals the Nutritional Potential of Brosimum gaudichaudii, an Underexploited Native Brazilian Fruit

Abstract

Brazilian Savanna biodiversity is abundant in native fruits with nutritional and sensory properties with potential technological applications. However, the information available regarding the proprieties of Brazilian native fruits is still limited. This study aimed to analyze the metabolic profile of Brosimum gaudichaudii fruits (BGF) using metabolomics approaches, besides its physicochemical and nutritional proprieties. BGF exhibited protein content at least two times higher than other native fruits. Fifty-five metabolites were identified by the metabolomics approaches, mainly fatty acids (n = 22), carbohydrates (n = 13), organic acids (n = 6), and polyphenols (n = 5). The most abundant constituents were D-fructose (42.97%) for sugars, L-asparagine (6.47%) for amino acids, succinic acid (44.11%) for organic acids, and linolenic acid (3.04%) for fatty acids. Highlighted phytochemicals included chlorogenic acid, α-amyrin, and γ-tocopherol. Additionally, this study revealed that BGF is a good source of vitamin C and dietary fiber and an excellent source of vitamin A. Future studies providing nutritional information and suggesting consumption methods and potential industrial applications are essential to familiarize consumers with these foods. Additionally, clarifying the appropriate planting and harvesting seasons can revive traditional practices, especially for native fruits like BGF.

Keywords:
Brosimum gaudichaudii; unconventional food plant; metabolite profiling; GC MS; LC-MS; HPLC

Introduction

The Savanna biome encompasses approximately 22% of the Brazilian territory and is characterized by a rich biodiversity of native fruits, which possess nutritional and sensory properties with potential for technological applications. Despite this, only 30% of the Savanna biome’s fauna and flora are well-documented. In recent years, there has been a growing interest in studying native fruits and developing new products derived from them. However, research on their physicochemical and nutritional characteristics remains limited.¹,²

In this context, studies on the metabolite profile of native fruits, particularly Brosimum gaudichaudii fruits (BGF), are crucial for regional development, promoting consumption, and reviving traditional food culture.³ BGF, belonging to the Moraceae family, is a native plant of the Savanna biome, commonly known as “mama-cadela”, “arbóreo de cadela”, or “algodãozinho do campo”.⁴ As an unconventional food plant, BGF fruits are not commonly consumed and are often neglected or underutilized by the population.⁵ Various unconventional fruits, such as araça, buriti, cagaita, yellow mombin, mangaba, jatobá, jabuticaba, cambuci, pequi, pitanga, gabiroba, and lobeira, are notable for their nutritional and technological potential. However, BGFs remain underexplored.¹,³,⁶

BGF has a pasty consistency, yellow-orange color, and sweet taste, and is typically consumed in natura or used in jams and ice creams. It is rich in soluble (SDF) and insoluble dietary fiber (IDF), as well as bioactive compounds such as phenolic compounds, carotenoids, and vitamin C, which possess antioxidant properties and have been linked to various health benefits. Additionally, BGF shows potential as an ingredient to enhance the nutritional quality of food products. However, it is crucial to characterize the physicochemical and nutritional properties of this native Brazilian fruit to identify its potential applications in food production.²,⁷

Studies on native or wild plants such as BGF have gained importance, as a deeper understanding of their chemical and metabolic composition can aid in species preservation and offer more precise nutritional recommendations for the local population, particularly within the Brazilian Savanna biome. However, the nutritional and physicochemical properties of BGF remain largely unexplored. Further research is needed to elucidate its potential for both direct consumption as a native fruit and its viability for industrial processing applications. Hence, the present work aimed to analyze the physical, physicochemical characteristics and to apply untargeted and targeted metabolomics approaches in order to detect the main nutrients found in BGFs.

Experimental

Sampling

BGFs (n = 5 kg) at mature stage were collected in standard commercial greenhouses in Uberlândia, Minas Gerais, Brazil (18°54’41”S; 48°15’44”W). Mature fruits were harvested when reached their completely yellow orange coloring peel. All analysis were performed in edible part of BGF (pulp with a thin peel). Samples were frozen in liquid nitrogen and stored at -80 °C for further analysis.

Material and equipment

The standards used in this work were D-glucose, D-fructose, maltose, sucrose, D-galactose, myo-inositol, citric acid, malic acid, tartaric acid, L-alanine, L-serine, L-proline, L-aspartate, and L-glutamate, methyl laurate, methyl tetradecanoate, methyl palmitate, methyl octadecanoate, methyl arachidate, methyl docosanoate, methyl lignocerate, methyl linoleate, (Z)-9-oleyl methyl ester, methyl linolenate, methyl palmitoleate, lycopene, lutein, β-carotene, sodium hydroxide, metaphosphoric acid, phosphoric acid, methanol, Folin-Ciocalteau reagent, sodium nitrite, aluminium chloride, ribitol, chloroform, methoxyamine hydrochloride, pyridine, n-tridecane, sodium sulfate, hexane, toluene, hydrochloric acid, and n-methyl-n-(trimethylsilyl) tri-fluoroacetamide. All standards, reagents and solvents used were from Sigma-Aldrich, Saint Louis, MO, USA.

The equipment used in this work was Metrohm pH meter (827 pH Lab, Metrohm, Switzerland), refractometer (Krüss-Optronic GmbH, Germany), high-performance liquid chromatography (HPLC) with diode array detector (DAD) (Thermo Fisher Scientific, Dionex DX 500, USA), liquid chromatography (Prominence model, Shimadzu, Japan)-electrospray ionization-ion trap-mass spectrometry/mass spectrometry (Esquire HCT model, Bruker Daltonics, Germany) (LC-ESI-IT-MS/MS), and gas chromatography-mass spectrometry (GC-MS) (Agilent GC-MS 5977 instrument, Agilent Technologies, USA).

Methods

Physical characterization

BGFs (n = 40) were randomly picked up and the physical parameters were analyzed: total mass (g), seed (g), edible fraction (g), fruit yield (%), transverse diameter (cm) and longitudinal diameter (cm). Fruit yield was calculated as described by Meza et al.,⁸ using the equation 1.

(1) $$\text{Fruit yield}(\mathrm{\%})=[\text{pulp weight}(\mathrm{g})/\text{total mass}(\mathrm{g})]\times 100$$

Physicochemical characterization

For the physicochemical parameters, pH, titratable acidity, and total soluble solids (TSS) content were analysed.⁸ For pH analysis, 10 g of fresh sample were weighted, diluted in 100 mL distilled water and analyzed by a Metrohm pH meter. Titratable acidity was determined by titration with 0.1 M sodium hydroxide and the result expressed in grams of citric acid per 100 g of fresh weight (%). For TSS, fruit sample was filtered to retain solid particles and measured with a refractometer, and the result was expressed in Brix at 20 ºC.

Chemical composition and energy value

Contents of moisture, proteins, lipids, ash, and dietary fiber (DF) of fruits were determined in triplicate according to the standard methods.⁹ Available carbohydrates were calculated by difference: 100 - (moisture + proteins + lipids + ash + DF). Results were expressed as g 100 g^-1 on fresh and dry weights. To calculate energy, 4 kcal g^-1 of available carbohydrates, 4 kcal g^-1 of proteins and 9 kcal g^-1 of lipids were considered.

Soluble sugars by HPLC analysis

Soluble sugars were extracted with 80% ethanol (v/v) at 80 ºC. Extract was vapored at 45 ºC and the volume suspended with water. They were determined by HPLC coupled with a DAD, with Carbopac PA1 column (4 × 250 mm, 5 µm particle size), isocratic flow of 1 mL min^-1 of 18 mmol L^-1 sodium hydroxide for 25 min.¹⁰ The calibration curves were prepared with D-glucose, D-fructose, and sucrose external standards.

Organic acids by HPLC analysis

Organic acids were extracted with 3% metaphosphoric acid in the ratio of 1:8 (m/v). The sample was homogenized, centrifuged and the supernatants were filtered on 0.45 μm membranes. They were determined by HPLC coupled with a DAD, equipped with a μBondpack C18 (300 mm × 3.6 mm internal diameter (i.d.), Waters, USA) and elution (flow rate of 0.5 mL min^-1) was carried out in isocratic conditions with 0.1% phosphoric acid, monitored at 210 nm.¹⁰ The calibration curves were prepared with malic, citric and tartaric acids external standards.

Carotenoids by HPLC analysis

Carotenoids were quantified as described in detail by Meza et al.¹¹ Lycopene, lutein, and β-carotene were used as external standards. The results were expressed as mg of carotenoid per 100 g of fruit in fresh and dry weight.

Total phenolic compounds by spectrophotometric analysis

Extracts were prepared using 2 g of fruits mixed with 70% methanol and determined by Folin-Ciocalteau colorimetric method.¹² The extracts were mixed with Folin Ciocalteau reagent and 0.5 mol L^-1 sodium hydroxide. The absorbance was measured at 760 nm. The results were expressed in mg gallic acid equivalent (GAE) per 100 g of fruit in fresh and dry weight.

Total flavonoids by spectrophotometric analysis

Extracts were prepared using 2 g of fruits mixed with 70% methanol. Methanolic extract was mixed with distilled water, 5% sodium nitrite, 10% aluminium chloride, and 1 mol L^-1 sodium hydroxide.¹² The absorbance was measured at 510 nm. The results were expressed in mg catechin equivalents (CE) per 100 g of fruit in fresh and dry weight.

Vitamin C by spectrophotometric analysis

Vitamin C was determined as described by Menezes Filho et al.⁴ Sample was mixed with distilled water, 20% sulfuric acid, 10% potassium iodide (KI) and 1% starch solution. The solution was titrated with 0.002 mol L^-1 KI. The result was expressed as mg of vitamin C per 100 g of fruit in fresh and dry weight.

Polyphenols by LC-ESI-IT-MS/MS analysis

Flavonoids and phenolic acids were extracted as described by Engelbrecht et al.² The identification of flavonoids and phenolic acids was performed by LC ESI IT MS/MS, with a C18 column (300 mm × 3.6 mm i.d., Waters, Milford, MA). The ESI was maintained in negative mode and mass analyzer was programmed for full scan between m/z 100 1000. The collision energy for negative mode was 3000-3500 V. Polyphenol identification was performed using Data Analysis 4.0 software, comparing mass spectra with available databases: Mass Bank of North America and Mass Bank Europe.

Polar and non-polar metabolite profiling by GC-MS analysis

The analyses of polar and non-polar metabolites were conducted as described by Meza et al.¹³ For polar metabolites extraction, fruits were mixed with methanol, ribitol, chloroform, and Milli-Q water. The upper hydrophilic phase was collected and dried under nitrogen gas. Methoxyamine hydrochloride (20 mg mL^-1) dissolved in pyridine was added to derivatize the samples. For non-polar metabolites extraction, sample was mixed with chloroform, methanol, and n-tridecane. Chloroform and 1.5% sodium sulfate were added to the mixture and centrifuged at 1000 rpm for 5 min at 4 °C, and dried with nitrogen gas. Hexane, toluene, methanol and 8% hydrochloric acid were used to reconstitute the samples. The hexane phase was collected and dried with nitrogen gas. Samples was reconstituted with hexane and pyridine. Previously to GC-MS analysis, N-methyl-N-(trimethylsilyl) tri-fluoroacetamide was added to the derivatized polar and non-polar metabolites samples. Derivatized samples were analyzed using an GC-MS equipped with HP5ms column (30 m × 0.25 m × 0.25 μm) under optimized conditions.¹⁴ A pool of polar metabolite external standards was used: D-glucose, D-fructose, maltose, sucrose, D-galactose, myo-inositol, citric acid, L-alanine, L-serine, L-proline, L-aspartate, and L-glutamate. While, a pool of fatty acid methyl ester external standards was used: methyl laurate, methyl tetradecanoate, methyl palmitate, methyl octadecanoate, methyl arachidate, methyl docosanoate, methyl lignocerate, methyl linoleate, (Z)-9-oleyl methyl ester, methyl linolenate, and methyl palmitoleate. Data acquisition and deconvolution were conducted using MassHunter software (Agilent Technologies). NIST20 and Wiley 12^th Edition mass spectral libraries were used for metabolite identification.

Contribution of BGF to dietary intake recommendations

The Dietary Guidelines for the Brazilian Population¹⁵ was used as a criterion for calculating the portion of BGF (1 portion of fruit = 70 kcal = 68.14 g). BGF was classified as a “source”, “good source” or “excellent source” of a given nutrient when it satisfied 5 to 10%; 10 to 20%; or over 20% of the dietary reference intake (DRI) for adults (both sexes) in the portion, respectively.¹⁵

Statistical analysis

This work was completely randomized with three biological replicates (each replicate was composed by 40 fruits) for all analysis. The results are presented as mean ± standard deviation and expressed as a descriptive statistic (mean, standard deviation and percentage of the abundance for metabolites) by using Graph Pad Prism 3.0 software.¹⁶

Results and Discussion

Physical and physicochemical characterization of BGF

The BGF is characterized by its yellow-orange color, round shape, thin skin, soft flesh, and a single seed.⁴ The physical characterization of BGF is detailed in Table 1. The whole fruit had a total mass of 11.18 g, with the seed massing 3.63 g, and an edible fraction (pulp and peel) of 7.60 g, yielding 69.09% of the fruit. The transverse and longitudinal diameters of the fruits were 2.68 and 2.37 cm, respectively. As BGF is a wild fruit without standardized production and management, significant variation exists in these parameters. Edaphoclimatic differences, such as soil composition, air humidity, and climate, likely account for the observed physical variations in the fruit.¹⁷

Table 1
Physical, physicochemical, and nutritional parameters of Brosimum gaudichaudii fruits

Parameter	Fresh weight	Dry weight
Physical parameters
Fruit (total mass) / g	11.18 ± 1.41	NA
Seed / g	3.63 ± 0.77	NA
Edible fraction (pulp + peel) / g	7.60 ± 1.01	NA
Fruit yield / %	69.09 ± 12.59	NA
Transverse diameter / cm	2.68 ± 0.17	NA
Longitudinal diameter / cm	2.37 ± 0.13	NA
Physicochemical parameters
pH	6.07 ± 0.10	NA
Titratable acidity / (% citric acid)	5.14 ± 0.18	NA
Soluble solids / ºBrix	20.07 ± 0.29	NA
Chemical composition and energy
Moisture / (g 100 g-1)	69.69 ± 0.43	NA
Proteins / (g 100 g-1)	3.25 ± 0.18	10.47 ± 0.70
Lipids / (g 100 g-1)	2.03 ± 0.02	6.71 ± 0.05
Ash / (g 100 g-1)	0.45 ± 0.09	1.49 ± 0.29
Soluble dietary fiber / (g 100 g-1)	1.23 ± 0.55	4.06 ± 1.82
Insoluble dietary fiber / (g 100 g-1)	5.71 ± 1.15	18.82 ± 3.80
Total dietary fibera / (g 100 g-1)	6.94 ± 0.60	22.88 ± 1.98
Available carbohydratesb / (g 100 g-1)	18.02 ± 0.23	59.44 ± 0.76
Energy / (kcal 100 g-1)	102.73 ± 0.38	338.93 ± 1.24
Primary metabolites / (mg 100 g-1)
Glucose	309.42 ± 27.39	1020.84 ± 90.38
Fructose	499.07 ± 20.39	1646.57 ± 67.27
Sucrose	212.68 ± 11.67	701.68 ± 38.51
Malic acid	1119.21 ± 73.62	3692.55 ± 242.91
Citric acid	378.71 ± 22.43	1249.46 ± 73.99
Tartaric acid	48.19 ± 2.49	158.99 ± 25.33
Secondary metabolites / (mg 100 g-1)
Total phenolic compounds	74.04 ± 2.49	244.27 ± 8.22
Total flavonoids	1.15 ± 0.10	3.79 ± 0.32
Lutein	1.25 ± 0.17	4.14 ± 0.56
Lycopene	6.81 ± 3.93	22.48 ± 12.98
β-Carotene	10.15 ± 1.85	33.48 ± 6.09
Vitamin C	20.02 ± 0.97	66.06 ± 3.19

• ^a
  Total dietary fiber calculation: soluble dietary fiber + insoluble dietary fiber.
• ^b
  Available carbohydrates calculation: 100 - (moisture + proteins + lipids + ash + total dietary fiber). Results expressed as mean ± standard deviation. n = 3 (each replicate was composed by 40 fruits). NA: not applicable.

Moreover, the physicochemical parameters of BGF were analyzed and reported in Table 1. BGF showed medium pH of 6.07, acidity of 5.14% and TSS of 20.07 ºBrix. Similar results of pH (5.96) and acidity (5.85%) were reported by Land et al.¹⁸ The main soluble solids found in BGF are D-fructose (48.87%), D-glucose (30.30%) and sucrose (20.83%), which are parameters related to desired sensory characteristics such as taste and flavor, and consequent consumer acceptance.⁸ Determining the physicochemical parameters of native fruits, particularly BGF, is crucial for the food industry. The balance between soluble solids and acidity levels directly impacts the quality of the fruit and its suitability for processing.⁴

Nutritional characterization of BGF

Chemical composition

The chemical composition of BGF was presented in both fresh and dry weight (Table 1). BGF had a high moisture content (69.69 g 100 g^-1), which demonstrated the potential for the development of various products, such as jams, ice creams, and beverages as previously demonstrated Land et al.¹⁸ The content of protein in BGF was 3.25 g 100 g^-1, while in dry weight was 10.47 g 100 g^-1. The protein recommendation for adults (both sexes) is 0.80 g of good quality protein per kg body weight per day, value obtained from available nitrogen balance studies. BGF had 2.21 g of protein per portion, so it is not considered a source of protein as it would only provide 3.94% of the daily recommendations for a 70 kg adult.¹⁵ This result was expected, since fruits are not generally considered sources of protein. The lipid contents of these native fruits were 2.03 and 6.71 g 100 g^-1 in fresh weight (f.w.) and dry weight (d.w.), respectively. The ash contents of BGF were 0.45 and 1.49 g 100 g^-1 in f.w. and d.w., respectively.

The available carbohydrate contents (calculated by difference) of the BGF were 18.02 and 59.44 g 100 g^-1 in f.w. and d.w., respectively (Table 1). The BGF can be considered a source of available carbohydrates as it provided 9.44% of the daily recommendation for adults in one portion of fruit (Table 2). In addition, the total dietary fiber (TDF) found in BGF was 6.97 and 22.88 g 100 g^-1 in f.w. and d.w., respectively. From this total, IDF (5.71 and 18.82 g 100 g^-1 in f.w. and d.w., respectively) was the fraction that contributed most when compared to SDF (1.23 and 4.06 g 100 g^-1 in f.w. and d.w., respectively) (Table 1). The BGF were considered a good source of DF, as it provided between 12.44-15.73% and 18.88-22.48% of the daily recommendation for man and female adults in one portion of fruit, respectively (Table 2). In terms of energy, BGF had an energy density of 102.73 and 338.93 kcal 100 g^-1 in f.w. and d.w., respectively (Table 1). In this context, BGF showed low energy value, since fresh fruits with 70 to 150 kcal 100 g^-1 are considered low energy density food.¹⁵

Table 2
Contribution of Brosimum gaudichaudii fruit to meet the dietary reference intakes of available carbohydrates, dietary fibre, and vitamins A and C for adults

Age group / years		Available carbohydratesa / (g per day)	Dietary fibreb / (g per day)	Vitamin A / (μg RAE per day)	Vitamin C / (mg per day)
Male
19-50	DRI	130	38	900	90
	daily value / %	9.44	12.44	64.03	15.14
	classification	source	good source	excellent source	good source
51- >70	DRI	130	30	900	90
	daily value / %	9.44	15.73	64.03	15.14
	classification	source	good source	excellent source	good source
Female
19-50	DRI	130	25	700	75
	daily value / %	9.44	18.88	82.34	18.17
	classification	source	good source	excellent source	good source
51- >70	DRI	130	21	700	75
	daily value / %	9.44	22.48	82.34	18.17
	classification	source	excellent source	excellent source	good source

B. gaudichaudiipe
• ^a
  Available carbohydrates calculation: 100 - (moisture + proteins + lipids + ash + total dietary fibre).
• ^b
  Dietary fiber calculation: soluble dietary fiber + insoluble dietary fiber. DRI: dietary reference intakes considered RDA (Recommended Dietary Allowances) values previously described by the literature;¹⁵¹⁹ RAE: retinol activity equivalents.

In the context of the biodiversity of the Brazilian Savanna, BGF can be compared with other native fruits such as araçá, buriti, cagaita, yellow mombin, mangaba, and marolo. Buriti (68.86 g 100 g^-1) and marolo (70.56 g 100 g^-1) have moisture contents similar to BGF. However, araçá, cagaita, yellow mombin, and mangaba have higher moisture values, ranging from 80.41 to 92.8 g 100 g^-1. The lipid content of yellow mombin and buriti is 0.48 and 7.72 g 100 g^-1, respectively, while the protein content of araçá and buriti is 0.42 and 1.73 g 100 g^-1. The ash content of cagaita and buriti is 0.30 and 1.01 g 100 g^-1, respectively. BGF stands out for its protein content, which is at least twice as high as that of other native fruits, while its lipid and ash contents fall within the ranges reported in the literature for other native fruits.³

Primary metabolites in BGF

Primary metabolites such as carbohydrates (n = 13), organic acids (n = 6) and fatty acids (n = 22) were identified by untargeted metabolomics approach (Table 3). The most abundant sugar identified was D-fructose (42.97%) and sucrose (16.31%), representing 80.83% of total carbohydrates. While succinic acid (4.57%) and 2-thiobarbituric acid (4.52%) accounted for most of the organic acid content identified, representing 87.74% of the total organic acids. Moreover, sugars such as D-glucose, D-fructose and sucrose, and organic acids such as malic, citric, and tartaric acids were quantified in the native fruits (Table 1). D-Fructose (1646.57 g 100 g^-1 d.w.) had the highest content among the soluble sugars, followed by D-glucose (1020.84 g 100 g^-1 d.w.) and sucrose (701.68 g 100 g^-1 d.w.). Among the organic acids quantified, malic acid (3692.55 g 100 g^-1 d.w.) had the highest content, followed by citric acid (1249.46 g 100 g^-1 d.w.) and tartaric acid (158.99 g 100 g^-1 d.w.). The composition of soluble sugars and organic acids in the fruit, along with the ratio between them, plays a crucial role in the sensory properties of BGF. This balance between sweetness and acidity is likely responsible for the desirable flavor of the fruit.⁸

Table 3
Metabolic profiling of Brosimum gaudichaudii fruits by GC-MS approach

Metabolite	Molecular formula	CAS	Retention time / min	Mass-to-charge ratio (m/z)	Relative abundance / %
Carbohydrates (n = 13)
D-Fructose	C6H12O6	7776-48-9	22; 27; 42	73; 307; 217	42.97
Sucrose	C12H22O11	25702-74-3	50; 40	217; 361	16.31
D-Galactose	C6H12O6	10257-28-0	23; 27; 29; 31; 34	73; 319; 204; 319; 73	3.71
Inositol	C6H12O6	173524-45-3	29; 30; 31; 36	73; 73; 73; 73	3.10
Galactaric acid	C6H10O8	526-99-8	28	333	2.82
D-Glucose	C6H12O6	2280-44-6	28	204	1.70
Maltose	C12H22O11	4482-75-1	37; 38; 39; 41	204; 204; 191; 217	0.66
D-Cellobiose	C12H22O11	16462-44-5	38; 42; 43	204; 204; 204	0.62
Arabinofuranose	C5H10O5	41546-26-3	24	217	0.50
2-α-Mannobiose	C12H22O11	15548-39-7	41	361	0.40
β-D-Glucopyranuronic acid	C6H10O7	23018-83-9	35	73	0.33
D-Mannose	C6H12O6	530-26-7	29; 35; 41	204; 387; 204	0.17
Sedoheptulose	C7H14O7	3019-74-7	32	73	0.04
Total carbohydrates	-	-	-	-	73.34
Amino acids (n = 6)
L-Asparagine	C4H8N2O3	3130-87-8	20; 22; 26	159; 73; 73	6.47
L-Threonine	C4H9NO3	13095-55-1	13; 15	73; 73	0.42
L-Proline	C5H9NO2	1150316-19-0	18; 26	156; 216	0.35
L-Alanine	C3H7NO2	115967-49-2	12; 15; 27; 33	73; 160; 160; 116	0.12
L-Serine	C3H7NO3	302-84-1	12; 14	132; 204	0.09
L-Leucine	C6H13NO2	61-90-5	16; 32	172; 204	0.03
Total amino acids	-	-	-	-	7.47
Organic acids (n = 6)
Succinic acid	C4H6O4	110-15-6	13; 14; 18	147; 147; 73	4.57
2-Thiobarbituric acid	C4H4N2O2S	504-17-6	26	345	4.52
D-Gluconic acid	C6H12O7	133-42-6	29	73	0.73
Shikimic acid	C7H10O5	138-59-0	25	204	0.36
Glucaric acid	C6H10O8	25525-21-7	30	73	0.13
Benzoic acid	C7H6O2	1079-02-3	11; 28	179; 281	0.06
Total organic acids	-	-	-	-	10.36
Saturated fatty acids (n = 15)
Palmitic acid	C16H32O2	408-35-5	29; 30.9; 31.3; 33; 36; 41	74; 31; 31; 299; 311; 371	1.02
Eicosanoic acid	C20H40O2	506-30-9	36; 38	74; 369	0.72
Stearic acid	C18H36O2	57-11-4	33	74	0.57
Myristic acid	C14H28O2	32112-52-0	27	285	0.002
Tetracosanoic acid	C24H48O2	557-59-5	45; 45.2	411; 425	0.07
Tricosanoic acid	C23H46O2	2433-96-7	41; 44	74; 397	0.04
Docosanoic acid	C22H44O2	112-85-6	41; 42	397; 383	0.04
Heneicosanoic acid	C21H42O2	2363-71-5	38; 40	74; 74	1.87
Lauric acid	C12H24O2	143-07-7	39	179	0.02
Pentacosanoic acid	C25H50O2	506-38-7	44	74	0.02
13-Methyltetradecanoic acid	C15H30O2	2485-71-4	27	74	0.02
Undecanoic acid	C11H22O2	112-37-8	20	74	0.02
Nonadecanoic acid	C19H38O2	12707-74-3	35	74	0.01
Hexacosanoic acid	C26H52O2	506-46-7	45; 48	74; 439	0.01
Octanoic acid	C8H16O2	124-07-2	33	165	0.004
Total saturated fatty acids	-	-	-	-	4.44
Unsaturated fatty acids (n = 7)
Linolenic acid	C18H30O2	463-40-1	33	79	3.04
cis-13,16-Docosadienoic acid	C22H40O2	7370-49-2	39; 50	55; 393	0.04
Linoleic acid	C18H32O2	121250-47-3	34	73	0.03
Methyl 13-eicosenoate	C21H40O2	69120-02-1	36	55	0.02
Oleic acid	C18H34O2	112-80-1	34	129	0.01
Myristoleic acid	C14H26O2	544-64-9	27	74	0.01
cis-11,14-Eicosadienoic acid	C20H36O2	5598-38-9	36	67	0.01
Total unsaturated fatty acids	-	-	-	-	3.16
Phytosterols / tocopherols (n = 5 )
α-Amyrin	C30H50O	638-95-9	51	218	0.66
Stigmasterol	C29H48O	83-48-7	50	129	0.06
γ-Tocopherol	C28H48O2	54-28-4	46; 47	488; 416	0.09
δ-Tocopherol	C28H48O2	119-13-1	45	474	0.01
β-Tocopherol	C28H48O2	148-03-8	46	488	0.003
Total phytosterols / tocopherols	-	-	-	-	0.82
Other metabolites (n = 6)
1,2,3-Propanetricarboxylic acid	C6H8O6	850848-65-6	25	273	0.10
Dodecanal	C12H24O	112-54-9	30	73	0.01
2-Piperidinecarboxylic acid	C6H11NO2	119678-10-3	9; 18; 20	84; 156; 73	0.01
2-Hydroxyundecanoic acid	C11H22O3	19790-86-4	21	229	0.01
Tetracosan-1-ol	C24H50O	506-51-4	43	411	0.06
Adenosine	C10H13N5O4	30143-02-3	39	73	0.002
Total other metabolites	-	-	-	-	0.19
Total metabolites	-	-	-	-	100

GC-MS: gas chromatography-mass spectrometry; CAS: chemical abstracts service registry number.

The most abundant amino acid found in BGF was L-asparagine (6.47%), representing 86.6% of total amino acids. Asparagine contributes to the storage and transport of nitrogen in plants and its accumulation in BGF may be associated with stress conditions, particularly in situations where the plant cannot support the normal level of protein synthesis.²⁰ The seasonal climate of the Savanna biome, characterized by a rainy season in spring and summer and a dry season in fall and winter, along with high temperatures, can create stressful conditions that lead to changes in the metabolism of the fruit. Additionally, other amino acids present in smaller quantities, such as proline (4.68%) and serine (1.20%), contribute to the sweetness of BGF.

Regarding the fatty acid profile, heneicosanoic acid (1.87%) and palmitic acid (1.02%) accounted for 65.09% of the total saturated fatty acids, while linolenic acid (3.04%) contributed 96.20% of the total unsaturated fatty acids. The fatty acid composition is critical for the quality of ripe fruit, as it is associated with the fruit’s taste and the biosynthesis of aroma compounds.¹¹ However, studies on the volatile compound profile of BGF are essential to better understand its aromatic composition.

Secondary metabolites in BGF

Secondary metabolites are crucial nutrients for humans, as they play significant roles in providing health benefits. The bioactive compounds, including total phenolic compounds, total flavonoids, carotenoids, and vitamin C, of BGF were analyzed in both fresh and dry weights (Table 1). The content of total phenolic compounds in BGF was 74.04 mg 100 g^-1 (f.w.), while the total flavonoids was 1.15 mg 100 g^-1 (f.w.) (Table 1). The following phenolic compounds (n = 2) and flavonoids (n = 3) were identified in the BGF samples by LC-ESI-IT-MS/MS approach in this study: chlorogenic acid, 4-O-caffeoylquinic acid, hesperidin, quercetin-4’-O-glucoside (spiraeoside) and quercetin-3-O-glucuronide (Table 4). Polyphenols are pivotal as antioxidant agents, with chlorogenic acid particularly noteworthy for its potential to mitigate health risks, including inflammation, diabetes, and cardiovascular diseases, owing to its antioxidant properties.²¹

Table 4
Identification of polyphenols (phenolic acids and flavonoids) in Brosimum gaudichaudii fruits by LC-ESI-IT-MS/MS approach

Identified compound	Retention time / min	m/z of ion molecular [M - H]	m/z of fragment ions (MS/MS) (relative abundance / %)	Score of similarity / %
Phenolic acids
Chlorogenic acid (3-O-caffeoylquinic acid)	10.6	353.27	191.07 (100); 179.05 (49.5); 192.07 (13.9); 191.47 (12.2); 179.39 (9.9)	98.30
4-O-Caffeoylquinic acid isomer 1	13.4	353.32	173.08 (100); 179.07 (45.4); 191.08 (22.60); 173.45 (13.80); 180.07 (11.7)	86.90
Chlorogenic acid isomer 1	14.1	353.26	191.07 (100); 191.52 (13.2); 192.04 (5.9); 179.08 (3.7); 173.00 (0.9)	99.90
4-O-Caffeoylquinic acid isomer 2	14.8	353.24	173.09 (100); 179.06 (37.6); 191.11 (14.9); 173.53 (10.3); 180.01 (4.9)	88.41
Chlorogenic acid isomer 2	17.2	353.29	191.06 (100); 192.06 (12.8); 191.46 (12.4); 179.04 (5.0); 207.08 (4.5)	99.50
Flavanones
Hesperetin 7-O- Rutinoside (hesperidin)	23.2	609.29	301.15 (100); 300.19 (20.5); 301.57 (12.0); 302.19 (11.9); 343.19 (10.6)	98.10
Flavonols
Quercetin-4’-O-glucoside (spiraeoside) isomer 1	24.6	463.30	301.16 (100); 300.17 (27.2); 302.13 (14.0); 301.58 (10.7); 299.20 (4.4)	95.50
Quercetin-4’-O-glucoside (spiraeoside) isomer 2	24.9	463.29	301.17 (100); 302.16 (23.9); 300.17 (17.6); 301.59 (12.1); 303.19 (6.0)	93.10
Quercetin-3-O-glucuronide	25.2	477.22	301.17 (100); 301.61 (9.6); 302.18 (8.5); 303.14 (5.0); 255.20 (3.7)	99.00

²⁴²⁵

The carotenoids quantified were lutein (1.25 mg 100 g^-1 f.w.), lycopene (6.81 mg 100 g^-1 f.w.) and β-carotene (10.15 mg 100 g^-1 f.w.) (Table 1). Carotenoids are significant phytochemicals for human health, serving as precursors to vitamin A and exhibiting antioxidant activities. Within the plant, carotenoids play a crucial role as aroma precursors and contribute to the color transition during fruit ripening, thereby influencing the flavor and visual perception of ripe fruit.²² The fruits of BGF had 18.21 mg 100 g^-1 of carotenoids or 845 µg RAE 100 g^-1, classified as an excellent source of vitamin A, satisfying 64.03 and 82.34% of the needs for this nutrient for adult men and women, respectively (Table 2). Other study found 25.26 mg of carotenoids per 100 g of BGF from Goiás (Brazil) and identified zeaxanthin, β-cryptoxanthin and β-carotene by HPLC analysis.²³

The level of vitamin C was 20.02 mg 100 g^-1 (f.w.) in BGF (Table 1). Vitamin C is crucial as both an antioxidant and a cofactor in redox reactions. It also plays a regulatory role in controlling cell differentiation, which is linked to the development of certain cancers. Recent studies²⁶ have highlighted the significance of ascorbate in activating epigenetic mechanisms that govern cell differentiation, the dysregulation of which can contribute to the onset of specific cancer types. The BGF were considered a good source of vitamin C as it meets 15.14 and 18.17% of the daily recommendations for male and female adults, respectively, in one portion (Table 2).

Moreover, phytosterols and tocopherols were identified by GC-MS approach (Table 3). The phytosterols were the more abundant in BGF than the tocopherols. The phytosterols identified were α-amyrin (0.66%) and stigmasterol (0.06%), while tocopherols were γ-tocopherol (0.09%), δ-tocopherol (0.01%) and β-tocopherol (0.003%). Phytosterols and tocopherols are vital compounds for health. Phytosterols are linked to a decrease in LDL cholesterol and total cholesterol levels, while tocopherols exhibit antioxidant properties, effectively scavenging lipid peroxyl radicals and reactive oxygen species.¹²,¹³

Conclusions

Brazilian BGF exhibited abundant primary metabolites, including protein, D-fructose, L-asparagine, succinic acid, heneicosanoic acid, and linolenic acid, as well as secondary metabolites such as β-carotene, chlorogenic acid, α-amyrin, and γ-tocopherol. These compounds contribute to the fruit’s quality and its nutritional attributes related to health. BGF has been highlighted as an excellent source of vitamin A and a good source of vitamin C and dietary fiber. This study addressed the important aspect of researching unconventional native Brazilian plants. However, further studies providing nutritional information and suggesting consumption methods and potential industrial applications are crucial to increase consumer acceptance of these foods. Additionally, information regarding the correct planting and harvesting seasons can help revive traditional practices, especially for native fruits like BGF.

Acknowledgments

We would like to thank the Food Research Center (FoRC) for the technical support and the São Paulo Research Foundation (FAPESP, grants 2013/07914 8 and 2015/06336 6), the Coordination for the Improvement of Higher Education Personnel (CAPES, grants 88882.376973/2019-01, 88882.376974/2018 01, and 88881.145838/2017-01), and Minas Gerais State Research Support Foundation (FAPEMIG, Grant IC FAPEMIG20170021) for the financial support.

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Edited by

Editor handled this article: Andrea R. Chaves (Associate)

Publication Dates

• Publication in this collection
  26 Aug 2024
• Date of issue
  2025

History

• Received
  10 May 2024
• Accepted
  05 Aug 2024