Received: 10 March 2016 | Revised: 23 May 2016 | Accepted: 11 June 2016

DOI 10.1111/jfpp.13147

ORIGINAL ARTICLE

Brazil nut powdered milk properties

Ariane Kluczkovski | Natacha Lima | Maria Katherine Oliveira

Faculty of pharmaceutical Sciences, Av. Gen.

Rodrigo Ot
avio, 6200, Manaus 69067005,
Brazil
Correspondence
Ariane Kluczkovski, Faculty of
pharmaceutical Sciences, Av. Gen. Rodrigo
Ot
avio, 6200, Manaus, 69067005, Brazil.
Email: mendonca-ariane@hotmail.com
Abstract
The Brazil nut powdered milk was featured in order to evaluate its processing yield and the toxico-
logical aspects concerning selenium (Se) and aflatoxin (AFL). The tests followed a type 22 factorial
design, using the drying adjuvant types (maltodextrin and Arabic gum) as independent variables.
The dependent variables were the extracts technological features and the adjuvant ratios. Among
the tested products, the selected one had 30% Arabic gum and showed 26.83% yield. Its mean
protein content was 30.12% and physicochemical features helped preventing microbial deteriora-
tion throughout 60 storage days at room temperature. However, the Se and AFL contents should
be monitored because the spray dryer atomization appears to concentrate these variables content,
when they are found in the raw material, thus affecting the finished product safety.

Practical applications
The research revealed the properties of the Brazil nut powdered milk as a source of non-dairy pro-
tein, especially to vegan and to healthier consumer’s diet. This approach provides an alternative
product instead soymilk to consumers allergic to bovine milk. The spray drying method adds value
to the raw product (native seed) in order to provide a new ingredient for culinary and industrial
formulations.

KEYWORDS
aflatoxin, Bertholletia excelsa, protein, selenium, spray drier

1 | INTRODUCTION
Several pharmacological studies on Brazil nut (Bertholletia excelsa H.B.
K.) have demonstrated its antioxidant activity and clinical benefits
(Huguenigen et al., 2015; Vadivel, Kunyanga, & Biesalski, 2012). Thus,
the development of new products based on this raw material has
increased. Consequently, there was increase in consumer’s interest on
food or biologically active food components known as functional foods
(Mour~ao, Pontes, Rodrigues, Brasil, & Cavalcante, 2012). Such foods
may be used to prevent and/or treat diseases such as cancer, diabetes,
cardiovascular disease, hypertension, and neurological disorder (Yang,
2009). The health-related properties attributed to some functional
foods may derive from their natural constituents or from the addition
of ingredients that change the food original properties. Therefore,
industries have been seeking different production methods to meet the
consumers’ demands for healthier and natural products. Using the
spray drying method has allowed developing new foods (Gharsallaoui,
Roudaut, Chambin, Voilley, & Saurel, 2007). Studying the shelf life
time, as well as evaluating nutritional values and sensory attributes
such as color, flavor and texture or consistency, are essential to intro-
duce new products into the market. Thus, several raw materials from
the Amazon region have been tested to produce new products. Brazil
nut is one of the most valuable seeds among tropical tree nuts. It is cur-
rently the most important extraction plant product (collected in the for-
est) in the Amazon when it comes to ecological, social, economic and
dietary values. It is recognized for its composition containing high
biological value lipids (60–70%) and proteins (15–20%), vitamins B and
E, and considerable amounts of minerals (Kluczkovski et al., 2015).
Selenium (Se) is the most prominent mineral in it. This mineral is essen-
tial to health, due to its antioxidant action in metabolic processes
(Cardoso et al., 2015). Thomson, Chisholm, Mclachlan, and Campbell
(2006) suggest that consuming two (02) nuts/day (approximately 60
mg/Se) would be enough to trigger the antioxidant effect. The tolerable
upper nutrient intake level (UL) for adult individuals is 400 lg/day
(Institute of Medicine, 2000; World Health Organization - Food and
Agric. Organ. of the United Nations, 2004). The contamination by afla-
toxins (AFL) which are metabolites produced by fungi that are hepato-
toxic and carcinogenic for humans and animals—has also been
frequently reported in Brazil nuts and it should be evaluated (Massi
et al., 2014). Therefore, in addition to the nutritional aspects, new nut

J Food Process Preserv 2016; 1-6 wileyonlinelibrary.com/journal/jfpp V

C 2016 Wiley Periodicals, Inc. | 1
2 |
products should be subjected to toxicity studies. A thick and white
extract may be obtained from fresh Brazil nut or from the “cake.” This
extract is used to replace cow’s milk in some areas of the Amazon
region. Popularly known as “Brazil nut milk,” it is a protein-rich aqueous
extract from this nut and is used as an ingredient in the Amazon
regional cuisine, presenting good market potential (Felberg, Antoniassi,
Deliza, Freitas, & Modesta, 2009). However, the product shelf life
needs to be extended, and a technological process such as spray drying
the liquid milk into dry extract (powder) could be an alternative. The
soy milk powder, for instance, is a vegetable-origin product with nutri-
tional properties, well known in the market of some consumer groups,
and it may be obtained by spray-drying process (Osthoff, Hugo, Wyk,
Wit, & Meyer, 2010). In this context, the Brazil nut powder milk could
be an alternative to soybean milk powder and a study was carried out
to evaluate the effect of processing Brazil nut water-soluble extract
into milk powder by spray drying, as well as to feature the new prod-
uct’s nutritional and technological aspects.
2 | MATERIAL AND METHODS
2.1 | Raw material featuring
The current study used 05 different lots of Brazil nuts coming from
processing plants in the Brazilian Northern region. The nuts were dehy-
drated and shelled. They had hull and were vacuum packed in flexible
laminate packages. The nuts were visually selected, and those looking
injured or showing stains were removed from the lot. Each lot was ana-
lyzed for acid index, moisture content (mc), and for total AFL content,
according to the AOAC International (2005), in order to evaluate raw
material quality.
2.2 | Obtaining the Brazil nut milk
The milk was obtained by the Brazil nut “cake.” At first, Brazil nuts
were dehulled according to Ferberg, Cabral, Gonçalves, and Deliza
(2002). The Brazil nuts were previously dehulled by soaking in 2%
sodium hydroxide (NaOH) at the ratio 2:1 (NaOH: Brazil nut). They
were boiled for 1 min, dried at 708C in an oven and placed in cold
water for 10 min, and then manually dehulled under running water and
immersed in cold water for 10 min to remove NaOH residues. NaOH
absence was checked using 0.1% phenolphthalein solution drops. After
dehulling, the nuts were pressed (mechanical pressing in TecnalV), to
R
separate the oil and the Brazil nut “cake.” The residue (cake) was
packed in aluminum foil and in plastic bags (300 g each) and it was
stored under refrigeration (2188C). As for extract obtainment, the
“cake” was homogenized in a blender with filtered water at 758C, at
the ratio 2:1 (water:cake) for 3 min. until it got homogeneous consis-
tency. It was then filtered and centrifuged to obtain the water-soluble
extract. The extracts were added up with: 500 ppm citric acid and
0.2% w/v potassium sorbate. They were packed in 100 mL Nylon-
polyethylene thermally sealed bags, pasteurized at 72 6 28C for 20
min, and cooled by immersion in ice. They were stored under refrigera-
tion after reaching room temperature (208C). The methodology used to
KLUCZKOVSKI ET AL.
T A B LE 1 Brazil nut dry-extracts type 22 qualitative factorial design
Factors Level (%)a
(A) Maltodextrin (2) 0
(1)10
(B) Arabic Gum (2) 0
(1) 30
a
In dry base.
obtain the extract was adapted from Souza, Holanda, Maia, Junior, and
Figueiredo (1987) and Cardarelli and Oliveira (2000).
2.3 | Obtaining the dry extract
This process used a Mini Spray Dryer (Model MSD 1.0) by Labmaq
BrasilV
R with 1.0 mm vacuum diameter. The drying operating conditions
were (a) Temperature: 1408C inlet and 90 6 38C outlet; and (b) 6.5 mL/
min feeding flow. The influence of different adjuvant types on the fea-
tures of dry products produced from water-soluble extract was eval-
uated using the type 22 qualitative factorial design. The herein studied
qualitative factors were drying adjuvants such as maltodextrin and ara-
bic gum in the ratios described in Table 1. The adjuvants were added,
in calculated weight ratio, to the extraction solution dry residue. Their
proportions had been chosen according to good yields obtained by pre-
liminary tests in spray drying by varying these additives and their
ratios.
The lots corresponding to 1 L extraction solution drying were pre-
pared for each experiment, as described in Table 2.
When all the studied factors were at the lower level (experiment 1),
an adjuvant-free dry product was obtained and it was conventionally
called spray-dried extract (ESA). The other adjuvant-containing products
were called spray-dried products (PSA) and they presented different
adjuvant type and concentration numberings. The material to be dried
was stirred in magnetic stirrer throughout the entire procedure to gen-
erate a product with homogeneous features. ESA and PSA were placed
in opaque bottles to be protected from light. A sample was removed
from the treatment showing the best yield as well as the best organo-
leptic features to be characterized according to the analysis mentioned
below.
2.4 | Physicochemical features of the product
The following variables were analyzed according to AOAC International
(2005): pH, titratable acidity, water activity (Aw) and composition (mc,
lipids, raw protein, carbohydrates, and total fiber). Density (g/cm3at
258C) was determined by gently pouring the brazil nut powder into a
10 cm3 graduated cylinder and was calculated as the ratio of the
weight (g) of the sample contained in the cilinder to the volume occu-
pied (i.e., 10 cm3) (Gallo, Llabot, Allemandi, Bucata, & Pinna, 2011). The
Se content was set according to Olson, Palmer, and Cary (1975).
2.5 | Product yield
The product yield was calculated as the ratio of the amount of powder
collected after every spray-drying experiment to the initial amount of
KLUCZKOVSKI ET AL. | 3

T A B LE 2 Description of a type 22 qualitative factorial design for Brazil nut dry-extracts production

Combinationsa
Experiment (n. ) Extracts Adjuvants Concentration (%) Process yield (%)
1 ESA Adjuvant-free NA b
4.02 6 0.2
2 PSA1 Maltodextrin 0 20.54 6 0.5
3 PSA2 Maltodextrin 10 11.87 6 0.4
4 PSA3 Arabic Gum 30 26.83 6 0.5
5 PSA4 Arabic Gum 0 19.11 6 0.1
6 PSA5 Maltodextrin1Arabic gum NA 16.34 6 0.2
a
The combinations of factor and level according Table 1; The drying operating conditions for PSA3 were (a) Temperature: 1408C inlet and 90 6 38C
outlet; and (b) 6.5 mL/min feeding flow.
b
Not Applicable.

solids in the feed solution according to Wang, Dufour, and Zhou
(2015).
2.6 | Microbiological analysis
Total coliforms (MPN/g), Salmonella (CFU/g), molds and yeasts (CFU/g)
were evaluated according to American Public Health Association
(2001).
2.7 | Aflatoxin content (AFLs)
The total AFL (B11B21G11G2) was determined by thin layer chroma-
tography (TLC), according to the methodology described by AOAC
International (2005). The method’s limit of detection (LOD) was 2.0 ug/
kg for total AFL.
2.8 | Shelf life
The extracts were subjected to the following analyses: pH, titratable
acidity, and aw at 0, 15, 30, 45, and 60 storage days in controlled envi-
ronment (temperature from 25 to 308C) and 70% relative humidity
(RH) %. All measurements were performed in triplicate.
the analyzed nuts was 2.39% (2.10–2.87). It was similar to that found


by Alvares, Castro, Costa, Lima, and Madruga (2012) who found 2.00–
3.12% for peeled nuts stored at room temperature. According to these
authors, storing the nuts at 28C allows keeping their initial mc levels.
However, the mc content of vacuum-packed peeled nuts was con-
trolled by the packaging typical conditions, since the lots in the current
study were received in vacuum metallized bags. According to Alvares
et al. (2012), the mc content depends, among other factors, on the
packaging type and on the product storage period, which may enable
fungi proliferation, including those producing AFLs. In addition, the
acceptable limit must be achieved during the nuts industrial processing
by means of dehydration stages. Given the high lipid content found in
Brazil nuts, the acidity index evaluation is important as quality parame-
ter. The lots’ mean acidity index was 0.96% (0.94–0.99). Levels below
1.0% indicate that the nuts were suitable for use in the product prepa-
ration, since these values suggest low deterioration levels. The highest
total AFL content in the experiment was 4.0 mg/kg (in the raw material)
and it met the 10 mg/kg maximum limit established for further process-
ing Brazil nuts (Codex Alimentarius Commission (CAC), 2010).
3.2 | Yield and composition

2.9 | Statistical analysis
Data related to chemical composition measurements and to dry
extracts yield were statistically analyzed by univariate model. The adju-
vants, the extraction and the double interaction (adjuvant 3 extraction)
were the main variation causes. Tukey’s tests were used to study dif-
ferences between the means.
3 | RESULTS AND DISCUSSION
3.1 | Featuring the raw material
The tests used for raw material physicochemical featuring met the
Brazilian laws (BRAZIL, 1976) that specify the criteria for Brazil nut
standardization, classification and internal commercialization. The eval-
uated parameters confirmed that the present results met the accepta-
ble limits. According to the Brazilian legislation (BRAZIL, 2010), the
nuts mc limit should be reduced to 15% safety limit. The mc content in
Among the herein performed treatments, PSA3 formulation with
26.83 6 0.25% showed the best process yield and used arabic gum in
its formulation. Therefore, the physicochemical and microbiological
evaluations were performed in this product alone and they were com-
pared to those of soybean milk powder previously reported.
The average of titrable acidity (0.96 6 0.5%) and moisture content
(2.39 6 0.20%) of the herein used raw material to the PSA3 showed
the nuts were suitable for being processed.
Table 3 shows data related to it seems that the fat content inter-
feres in the Brazil nut extract spray-drying process, since the dry nut
lipid content ranges from approximately 60–70%, leading to high lipid
content in the extract. Therefore, a defatted extract type is necessary
to the dry extract preparation. Thus, the semi-defatted “cake,” which
showed 44.19% lipid content, was used. In addition, the spray-dried
product (PSA3) showed lipid content decrease by almost 50% (23.57 6
0.83). It may have happened because the adjuvant was well incorpo-
rated, and it encapsulated the fat in the water-soluble extract. The fat
4 | KLUCZKOVSKI ET AL.

T A B LE 3 Composition of the Brazil nut products (raw material (dry seed); defatted “cake”; Brazil nut dry extract; soybean milk powder)

Products
Parametersa Brazil nut seed Dafatted “cake” PSA3b Soybean milk powderc
Ash 3.51 6 0.03 d
3.22 6 0.03 e
3.92 6 0.08 e
7.57
Mc 2.84 6 0.02 2.24 6 0.02 2.06 6 0.20 6.58
Fat 64.76 6 0.35 44.19 6 0,35 23.57 6 0.83 11.57
Total fiber 1.30 6 0.02 1.80 6 0.02 0.22 6 0.15 n.i.
Protein 13.19 6 0.31 14.76 6 0.31 30.12 6 0.27 37.48
Carbohydrates e
12.37 6 0.05 33.79 6 0.05 40.11 6 0.05 26.80
a
g% Dry matter.
b
Dry extract1arabic gum 30%.
c
Reported by Pardeshi et al. (2014).
d
Mean 6 standard deviation.
e
Carbohydrates by difference.

and carbohydrate parameters of PSA3 were superior to those reported
by Pardeshi, Murumkar, and Tayade (2014) in soybean milk powder.
On the other hand, the protein of PSA3 was 7% lower. When it was
compared to the crude protein, PSA3 showed 30.12% mean content
and exceeded the amount of protein in the dried nut and in the semi-
defatted “cake.” Nevertheless, the protein amount of the PSA3 could
not be explained only due to the Brazil nut itself. The Arabic gum 30%
was added as adjuvant and It could increase the protein content, as
some of the protein and carbohydrate were added. Additionally, the
amount of protein in the Brazil nut powdered milk could be affected by
modifications during the spraydrying process. Al-Assaf, Philips, Aoki,
and Sasaki (2007) showed that molecular associations in arabic gum
can lead to an increase in molecular weight in the solid state by matu-
ration under controlled heat. The process does not involves change in
the basic structural components and, while the maturation occurs, the
level of association increases giving way to the adjuvant with higher
molecular weight and protein content. On the other hand, arabic gum
is widely used due to its excellent emulsifying capacity, attributed to
this protein fraction of proteins, capacity for protecting the active sub-
stance and a good flow conferred to the particles (Costa et al., 2015).
In this context, the protein fraction in the product should be further
investigated for its bioactive nature to assess its future use as nutra-
ceutical or functional food. Bioactive peptides from plant-origin protein
hydrolysates, for instance, are currently being studied as functional
foods (Li-Chan, 2015). The density (g/cm3) at 258C mean was 1.02 sim-
ilar to the findings of Oguntunde and Akintoye (1991) for soymilk with
T A B LE 4Effect of spray-drying treatment on Brazil nut extract and
the aflatoxin level
range of 1.01–1.05. Generally speaking, the results of the PSA3 prod-
uct showed that fat, protein and carbohydrates levels are nutritionally
attractive in the Brazil nut product as a lactose free alternative in the
diet. Thus, it is also necessary to further evaluate the antioxidant activ-
ity with clinical application in order to see whether other scientific
studies have already reported any beneficial effect of tree nut con-
sumption on health.
3.3 | Toxicity
3.3.1 | Aflatoxins
The development of new Brazil nut products includes AFLs evaluations
to ensure the new product safety, as the seed is commonly studied
concerning quality and AFL contamination (Freitas-Silva & Ven^ancio,
2011). According to Table 4, there was AFL content increase after the
first processing. The raw material with 4.0 mg/kg met the legal limits
for shelled nuts. However, the AFL content in the finished product
increased to 65 mg/kg at the end of the spray-drying processing. Thus,
it was necessary to put away the initial production and to use new raw
material with AFLs content below the LOD (<2.0 mg/kg) to allow the
development of a safe product.
3.3.2 | Selenium
The PSA3 product showed Se content of 1.200 lg/100 g at the end of
the processing. Thus, when one considers the consumption of a table-
spoon (10 g) of the product diluted in a 100 mL glass, 120 lg/Se is pro-
portionally obtained. Based on the glutathione (GSH-Px) enzyme’s
activity maximization, this value is above the Se recommended dietary
allowance (RDA) (55 lg/day) for an adult human. Thus, the dry product

AFL (mg/kg) use must be oriented with emphasis in the product dilution and in the

Treatment Raw material Brazil nut product previous Se content analysis, in each produced lot, in order to avoid

1 a
4.0 65.0 the toxicity level to the consumer, since the Se content in the raw

2 b
<2.0 c
<2.0 c material varies. Se is present in sulfur amino acids such as selenome-
thionine and selenocysteine in plants. These plants show different
a
First treatment: raw material (dried Brazil nut) discarded after spray-
capacities to absorb and accumulate the Se from the soil and are classi-
drying processing.
b
Second treatment. fied as accumulating or non-accumulating plants. The Brazil nut tree is
c
Limit of detection (LOD) for total AFL (B11B21G11G2). an example of Se accumulating plant, but it may vary according to the
KLUCZKOVSKI ET AL.
geographic area in the Amazon region (Pacheco & Scussel, 2007). On
the other hand, in the case of non-accumulating plants, such as soy-
beans, this element’s concentration limit is at most 6 mg/100 g, even
when the plants are grown in seleniferous soils (Whanger, 2002).
According to Turakainem, Hartikainen, and Sepanen (2005), Se has the
property to improve plant capacity and to combat the oxidative stress
caused by oxygen free radicals. On the other hand, the presence of
high concentrations of it in the plant may cause toxicity and enable oxi-
dative reactions. A study conducted by Menezes and Souza (Souza &
Menezes, 2004) found amounts of 713 lg/100 g in Brazil nut “cake.” It
helps justifying that the Se content found in nut products may reach
levels above those recommended for beneficial effects on human
health.
3.4 | Microbiological assays and shelf life
The results of the PSA3 microbiological analysis met the standards set
by the current legislation. They also showed that the product is fit for
consumption because PSA3 does not represent health risks, since it
showed no microbiological growth up to 60 days. Small aw values oscil-
lations were observed in the PSA3 product throughout the studied
sixty (60) storage days. According to Tukey’s test, at the end of the
storage, these values were not significantly different from those
observed at the baseline (p > .05). Since preventing microbial growth is
important for food preservation, the aw content is an important vari-
able to prevent food deterioration and to extend its shelf life. The aw
content is considered to be a stability parameter, since the maximum
aw reference value recommended to prevent Brazil nut contamination
by AFLs-producing fungi is 0.70 (CAC, 2010). The PSA3 aw values
remained below the reference value (< .70), making the product stable
for 60 days. By taking into account that the shelf life of reference soy-
bean commercial products is 2 years, PSA3 needs monitoring in future
shelf life studies. The pH and acidity index parameters were also stable
throughout the storage period, despite the product high protein and
lipid.
4 | CONCLUSION
The Brazil nut milk powder showed 26.83% yield and better fat con-
tent than the soybean product. However, new adjuvants should be
tested in future studies in order to improve its yield. The results of
AFLs and Se contents were higher than the values obtained in the
dried nuts. Thus, these contents require special attention in the raw
material analytical evaluation in the product preparation, since the
spraying process seems to increase the content of these variables. The
stability of physicochemical features such as pH, aw and mc appears to
help ensuring the absence of microbiological contamination and
extending the product shelf life. Thus, the Brazil nut milk powder may
be used in the food industry as raw material and alternative ingredient
regarding food developed for special purposes. Therefore, the product
peptides require further studies on their functional aspects. It is also
necessary to define the probable consumer public, such as vegetarians
| 5
or people who need protein supplement as an alternative to cattle
dairy products, such as individuals with lactose intolerance.
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