a

ISSN 0101-2061 (Print)

Food Science and Technology ISSN 1678-457X (Online)

DDOI: https://doi.org/10.1590/fst.10519

Health benefits of chocolate consumption with high concentration of cocoa

incorporated from triterpenic acids, isolated from Mansoa Hirsuta DC.
Maria Patrícia MILAGRES1, Daniel Melo SILVA1, Ivan de Oliveira PEREIRA2,
Ludimila Mascarenhas SENHORINHO2, Antônio Euzébio GOULART SANT’ANA3, Talita Batista MATOS1* 

Abstract
The objective of the work was to develop a 70% cocoa chocolate containing ursolic and oleanolic acids isolated from Mansoa
hirsuta DC and to evaluate the effects of its consumption on the health of individuals. Physical-chemical, microbiological and
sensorial analyzes were made in the chocolates. In order to evaluate the health of individuals, a clinical trial was conducted with
the developed chocolate, in which 45 volunteers, during 04 weeks, were divided into three groups: a test group, a placebo group
and a control group. The individuals were submitted to laboratory and anthropometric tests on two occasions, before and after
the clinical trial weeks. The analyzes revealed that the developed chocolate presented physical-chemical and microbiological
characteristics within the standards accepted for bitter chocolate and presented good sensorial acceptance. The results of the clinical
trial showed that the volunteers presented reduction in the anthropometric measurements and changes in the laboratory tests.
Keywords: food choices; healthy eating; innovation; chocolate.
Practical Application: Functional chocolate contributes to increasing the quality of health and the well-being of consumers.

1 Introduction
Chronic non-communicable diseases (CNCD) are now the In addition to cocoa, other Brazilian plants have beneficial
leading causes of morbidity and mortality worldwide (Zhou et al., effects, among which the Mansoa hirsuta DC stands out, a plant
2018). Estimates indicate that this number is expected to increase in the semi-arid region of Brazil rich in triterpene acids. The use
significantly over the next few years, reaching 52 million deaths of natural foods containing oleanolic acid or ursolic acid, which
by 2030 (Arokiasamy et al., 2017). are triterpenes widely distributed in the plant kingdom, in folk
medicine is diverse and very old. These are known to possess
Thus, it is important to prevent the risk factors associated anticholesterolemic, hypoglycemic, anti-hepatotoxic, antioxidant,
with these diseases, which includes changes in eating habits anti-inflammatory, antifungal and antibiotic activity. Also,
(Libman et al., 2015; Ding et al., 2016). However, several studies they are said to inhibit the growth of tumors and pathogens
(Gough & Conner, 2006; Cheval et al., 2017) have mentioned the (Pereira et al., 2017; Zhang et al., 2019).
lack of time and sensory pleasure as motivators for unhealthy
eating. The sensory properties of food (intrinsic attributes), Although it is present in numerous plants, obtaining these
which encompasses taste, texture, appearance and odor, are two acids is usually done through its isolation from apple peels,
among the most important predictors of food choice by humans an expensive process (Yin et al., 2018). However, recently, these
(Asioli et al., 2017). same compounds were extracted and isolated from Mansoa hirsuta
DC, a Brazilian semi-arid plant, in a process of extraction and
Thus, chocolate can be a tasty and practical alternative for this isolation carried out under patent deposit nº BR 1020150081804,
issue, besides minimizing the lack of pleasure in healthy eating. which will be used in this work.
The benefits of using bitter chocolate for health are widely known;
Thus, the objective of this work was to develop a 70% cocoa
it is a good source of flavonoids and its consumption is associated
chocolate incorporated with ursolic and oleanolic acids and to
with decreased risk of death from cardiovascular disease, decreased
evaluate the effects of its consumption on the health of individuals.
blood pressure, inhibition of platelet aggregation and antioxidant
activity (Carvalho et al., 2018). In addition to flavonoids, it has
some phytochemicals that have well-being-stimulant effects in 2 Materials and methods
the brain, methylxanthines (Quelal-Vásconez et al., 2019), and The experiments were carried out in the laboratories of the
there is evidence that bitter chocolate consumption produces Instituto Federal Baiano Campus de Uruçuca and in the city of
anti-inflammatory effects (Colombo et al., 2015). Itajuípe, Bahia. The study population consisted of a convenience

Received 18 Apr., 2019

Accepted 28 Sept., 2019
1
Universidade Estadual do Sudoeste da Bahia – UESB, Jequié, BA, Brasil
2
Instituto Federal Baiano, Uruçuca, BA, Brasil
3
Universidade Federal de Alagoas – UFAL, Maceió, AL, Brasil
*Corresponding author: [email protected]

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 Dark Chocolate with triterpenic acids

and non-probabilistic sample, composed of men and women hours. The chocolate was then removed from the molds, packed
aged over 18 years, totaling 145 (one hundred and fortyfive) and stored at 25 °C.
volunteers. Of these, 100 volunteers were from the city of Uruçuca
and participated in the sensory analysis (acceptance test) and 2.3 Physical-chemical characterization
45 volunteers were from the city of Itajuípe and participated in
the clinical trial. The analyzes for physical-chemical characterization of
chocolates (test and placebo) were done three times for each
Inclusion criteria were people older than 18 years who wanted repetition, according to the methodology described in the Adolfo
to participate in the whole study process, had availability of time, Lutz Institute’s Manual of Analytical Standards (Instituto Adolfo
liked to eat chocolate and correctly signed the Informed Consent Lutz, 2004). It was also performed analyzes of water content
Form (ICF). Volunteers who presented intolerance/allergy to any (moisture); waste by incineration (ash); pH; total titratable
components of the product were excluded from the study, as well acidity and lipids by Soxhlet with modifications.
as those using medications that interfered with the perception of
odors and flavors and those who had any pathology that could be Protein quantification (nitrogen content) was performed
aggravated by research and/or that impaired sensory perception. by using a modified Micro-Kjeldahlm (Association of Official
Analytical Chemists, 1990) methodology, and the determination
This research was carried out in accordance with the of the total carbohydrates was done by means of the difference in
guidelines and standards of Resolution 466 of December 12, the sum values of moisture, ashes, proteins and lipids. The mean
2012 (Brasil, 2012), sent to the Research Ethics Committee of particle size was determined using a digital micrometer with a
the Universidade Estadual do Sudoeste da Bahia and approved 0-25 μm scale, according to Sampaio (2011).
under No. 46718615.0.0000.0055.
A qualitative analysis was carried out to verify the presence
2.1 Obtaining ursolic and oleanolic acids of the acids in the chocolate samples by using the methodology
of thin layer chromatography, using the ursolic and oleanolic
The ursolic acid and its isomer, the oleanolic acid, were acidspatterns for comparative purposes, under the following
isolated and extracted from the leaves of the Mansoa hirsuta DC conditions: silica gel chromatographic plates, mobile phase
plant, whose patent was registered at the National Institute of chloroform solution: methanol (5% v/v), iodine vapors and
Industrial Property (INPI) under the number BR102015008180 sulfuric vanillin as developing agents.
(Silva et al., 2015). The acids were provided by Laboratory of
Pharmacognosy of the Pharmacy course of the Universidade
2.4 Microbiological analyzes
Estadual do Sudoeste da Bahia after authorization from the
patent’s author. Microbiological analyzes were performed to ensure food safety
for the sensory tests in the two chocolates (test and placebo),
2.2 Chocolate processing following Annex III of Normative Instruction 62 (2003) and in
Petrifilm3M™ plates, according to the recommendations of the
Two chocolate formulations were developed; one containing manufacturer (3M 1997).
the triterpenic acids to be used in the clinical trial of the test
group, and another not containing triterpenic acids, to be used For the preparation of the sample, 25 g of chocolate were
in the clinical trial of the placebo group. used in 225 mL of 0.1% (w/v) sterile peptone saline solution
to obtain a 10-1dilution. This solution was used to carry out all
The chocolate was developed from a formulation containing analyzes performed under a flat surface within the previously
cocoa liquor (64%), cocoa butter (6%), refined sugar (29.54%), prepared flow chamber and forthree times.
liquid soy lecithin (0.4%) and ursolic and oleanolic acids (0.06%),
corresponding to 150 mg of acids/25 g bar of 70%chocolate for the The analysis of mesophilic aerobes was performed in PCA
test group of the clinical trial. While the chocolate formulation medium (Plate Cont Agar). 1mL of the 10-1dilution was inoculated
for the placebo group was cocoa liquor (64%), cocoa butter into sterile Petri dish. 20 mL of the hot (water bath) and molten
(6%), refined sugar (29.60%) and liquid soya lecithin (0.4%). medium was poured into the plate on the diluted sample (10-1).
The test and placebo chocolates were produced by following The plates were inverted and incubated in B.O.D.-type incubators
the same methodology. at 25 °C for 48 hours.
After weighing the ingredients, they were transferred to a For the determination of molds and yeasts from the chocolate
Melanger-type roller mill, in which the steps of refining, mixing samples, it was performed on Petrifilm YM 3M™ (Ref.6407).
of ingredients and conching took place. The ingredients in Then, 1mL of the diluted sample was transferred to Petrifilm.
the roller mill were homogenized and kept under stirring at a After homogenization, the sample was incubated in B.O.D.
temperature of about 54 °C for a 24-hour period until the mass incubators at 25 °C for 5 days.
turned into chocolate.
For the determination of coliforms with differentiation for
Then the chocolate was transferred to a marble table for the E. coli in the samples of chocolate, it was carried out inPetrifilm
tempering step for cooling to 29 °C. After the tempering step, EC 3M™ (Ref.6404). 1mL of the diluted sample was transferred
the chocolate was molded manually into specific 25g molds to Petrifilm. After homogenization, the sample was incubated
(bars) which were then cooled at approximately 5 °C for two in B.O.D. incubators at 35 °C for 48 hours.

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In all analyzes the colonies were counted in colony counts After all the instructions, an anthropometric evaluation
and the results were expressed by the number of Colony-Forming was performed by measuring weight (kg), height (m) and waist
Units per gram of the sample (CFU/g). circumference (cm). Based on these measurements, the Body
Mass Index (BMI=kg/m2) was obtained. Weight measurements
2.5 Sensory analysis were carried out in scales having a maximum capacity of 150 kg
and a minimum of 2 kg, and a 200-cm tape was used to obtain
In order to verify the opinion of the people regarding the height and waist circumference.
sensorial characteristics of the chocolate developed for the test
group (containing acids), an evaluation was carried out, using In addition to the nutritional and anthropometric evaluation,
a sensorial acceptance analysis of the product. For this purpose, clinical tests were also performed to evaluate the lipid profile - total
a 9-pointhedonic scale of was used, according to Amaral et al. cholesterol and its fractions (high density lipoprotein - HDL
(2018) apud Belsito et al. (2017), (9 = extremely liked; 5 = neither and low density lipoprotein - LDL) and triglycerides and fasting
liked nor disliked, and 1 = disgusted extremely). glycemia. For that, participants’ venous blood was collected after
a 12-hour fasting period, in an outpatient clinical laboratory
The chocolate samples were evaluated as for overall impression (LL Laboratório - Itajuípe, Bahia). For the analysis of the
by 100 untrained evaluators. Thus, containers encoded with three laboratory tests the methods according to Lima et al. (2001).
random digits containing 10g of chocolate at room temperature
were served to the evaluators in individual booths using white The participants were submitted to laboratory and
light and individually. anthropometric tests on two occasions: before the first week of
the experiment and immediately after the fourth week of the
Together with chocolate, the evaluator received a hedonic‑scale experiment, in order to evaluate whether there were changes
sensory acceptance form, where he/she was asked to mark his/her in the indicators mentioned above.
overall impression on chocolate according to the proposed scale
In order to compare whether there were differences in the
(Belsito et al., 2017).
variables (weight, BMI, waist circumference, glucose, HDL, LDL,
The results of the acceptance test were submitted to descriptive triglycerides, total cholesterol) before and after the chocolate
statistical analysis by using the Statistical Packpage for the Social consumption, a Kolmogorov-Smirnov test was initially performed
Sciences® software (SPSS, version 21.0). to test the normality of data, and after that definition, we used the
average test for comparison, using Wilcoxon test in the variables
2.6 Clinical trial to evaluate chocolate consumption in the that did not have a normal distribution, and the t test for paired
health of individuals samples in the variables that followed the normal distribution.

A prospective, randomized, blinded, placebo-controlled The Kolmogorov-Smirnov test was used to verify the normality
clinical trial was conducted to evaluate the effect of 70% cocoa of the dataand to verify whether there were differences in the
consumption added with ursolic and oleanolic acids on individuals’ variables (weight, BMI, waist circumference, glucose, HDL,
health. The population was composed of volunteers who were LDL, triglycerides, total cholesterol), followed by analysis of
registered in a basic health unit of the city of Itajuípe-Bahia, and variance at 5% probability.
the study was conducted from July to August 2016. All average tests were performed at 5% probability using
Of these volunteers, only the ones who were found at their Statistical Packaged for Social Sciences (SPSS, version 21.0).
homes during the study period and who met the inclusion
criteria were included in the clinical trial, which made up a total 3 Results and discussion
of 45 people. All the volunteers were visited by the researcher in
their homes to explain all phases of the research. The volunteers 3.1 Physico-chemical characterization of chocolates
were divided into three groups (a test group, a placebo group and Table 1 shows the physico-chemical analysis of 70% cocoa
a control group) through a drawing, aiming at the randomization chocolate with addition of the mixture of ursolic acids and
of the group. oleanolic acid.
The test group consisted of 15 people who received in daily
visits at their homes a 25 g bar of 70% cocoa chocolate added Table 1. Water content, ashes, pH, acidity, lipids, proteins and average
with ursolic and oleanolic acid, individually packaged, for particle size of the chocolate.
four weeks. On the other hand, the placebo group consisted of
15 people who received in daily visits at their homes a 25 g bar Value ± Standard deviation
of 70% cocoa chocolate individually packaged, for four weeks. Water content (%) 1.72 ± 0.16
The control group consisted of 15 people who were instructed not Ashes (%) 2.30 ± 0.05
to consume any form of chocolate during four weeks of the study. pH 5.58 ± 0.04
All participants were instructed to maintain their daily activities Titratable acidity (%) 8.20 ± 0.40
and continue their usual diet, not inserting new medications or Lipids (%) 29.43 ± 0.65
new physical activities into their routine. The 24‑hour Recall Proteins (%) 8.79 ± 0.48
(Buzzard, 1998) was applied in all visits to establish the daily Total Carbohydrate (%) 57.76 ± 0.00
consumption of each individual, as well as to evaluate if the Averageparticlesize (µm) 19.00± 0.01
usual diet was not modified. Source: Directsurvey.

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 Dark Chocolate with triterpenic acids

The ursolic and oleanolic acids were inserted into the chocolate 3.3 Sensory analysis
together. This is due to the difficulty of separating them because
According to the evaluators’ answers, the chocolate sample
they are isomers So, the similarity of physical and chemical
obtained a mean score of 7.29 ± 1.43, ranging from the hedonic
properties between these pairs makes their differentiation and
terms “I liked it a little” and “I liked it a lot”, corresponding to
separation very difficult. Howeverthey share many properties the acceptance region of the hedonic scale. The chocolate was
(Ding et al., 2018), which add value to developed chocolate. well-accepted, presenting 83% of the answers between the scales
The developed chocolate has low water content (1.72%) and 07 and 09.
a 5.58 pH due to these characteristics; it is not necessary to use In addition, difficulty in maintaining a balanced diet is often
additives or preservatives, besides being more durable, which is linked to deprivation of tasty foods (Justo & Ferreira, 2019).
a beneficial factor for health (Pandey et al., 2014). The pH value That is why the food industry needs innovation to meet this
is close to the pH found in other bitter chocolates (Leite et al., demand of tasty, practical and healthy foods. The 70% chocolate
2013), as a larger amount of cocoa liquor decreases the pH added with ursolic and oleanolic acids can return the pleasure
in relation to milk chocolates. A lower pH was also expected of eating and increase adherence rates to healthy diets because
because two acids were added to the formulation. of its practicality and good sensory acceptance.
The developed chocolate has an 8.8% protein content.
Foods rich in good quality proteins are important because they 3.4 Clinical trial
provide essential amino acids to promote growth, especially for
The study population consisted of 45 volunteers, 24 male
the maintenance and development of lean body mass (Devries
(53.3%) and 21 female (46.7%). The mean age of the population
& Phillips, 2015). The lipid concentration is the 29.43% and for was 52 ± 15.01 years. There was no significant difference between
example, regarding total carbohydrates, the developed chocolate the groups in the demographic variables. The intervention took
has 57.76% of carbohydrates. place during four weeks, from July to August 2016. During this
Samples of the analyzed chocolates presented value of fixed period, volunteers maintained their usual activities throughout
mineral residue (2.30 ± 0.05) allowed by the legislation for the research process, including feeding and practicing/not
chocolate, which is a maximum of 2.5% w/w of fixed mineral practicing physical activity.
residue. An amount of ash greater than that allowed by the The trial was characterized by the daily consumption of
legislation may suggest adulteration or contamination by some 25 g of 70% cocoa chocolate plus 150 mg of the mixture of
mineral residue in chocolate (RESOLUTION-CNNPA No.12, ursolic and oleanolic acids for the test group; 25 g of 70% cocoa
1978). chocolate without addition of acids for the placebo group and no
The size of the particles of chocolate may interfere with its consumption of chocolate for participants in the control group.
palatability, directly influencing taste and texture (Saputro et al., The recommended daily dose of ursolic and oleanolic acids
2019). Ideally, this size should be between 20-25 μm. The analyzed ranges from 150-300 mg. We chose to analyze the effect of the
chocolates obtained a 20 μm particle size. lowest recommended dose (150 mg/day), although the acids are
The presence of ursolic and oleanolic acids in the chocolate non-toxic (National Center for Biotechnology Information, 2016).
after processing was confirmed by the thin layer chromatography After the four weeks of experiment it was possible to observe
method using iodine and vanillin vapors as developing agents changes in the health of the volunteers according to Table 2.
and the acid standards for comparison. There was presence of
acids in allfractions. This technique was chosen because it is In the placebo group, it was noted that this food inserted
daily in the diet caused significant changes in the variables weight
easy to apply, simple, low cost and efficient.
(ρ = 0.014), BMI (ρ = 0.015) and waist circumference (ρ = 0.02),
by the t-test for paired samples, a 5% probability, when comparing
3.2 Microbiological analyzes before and after the intervention. These modifications occurred
Microbiological analyzes were performed with the purpose in a negative way for the health of the individuals, that is, the
of providing safety for sensory analysis and for the clinical trial. consumption of 70% cocoa chocolate in the amount used and
The dilution used was at 101. The other smaller dilutions were in the intervention period increased the weight, BMI, and waist
not able to indicate the presence of any microorganism studied. circumference of the people.
The values of colony forming unitsof mesophilic bacteria per This negative modification in the placebo group may have
gram of chocolate (7x101) and molds and yeasts (2.5x101) are been caused by the increased amount of sugar in the formulation,
within the limits allowed by the legislation,which are 104 and since the sugar was used to replace the concentrations of the
103 UFC/g, respectively (Resolution CNNPA No.12, 1978). triterpenic acids of the test chocolate, so the acid chocolate had
The analysis for total coliforms and thermotolerant coliforms 29.54 g/100 g sugar while the placebo chocolate had 29.60 g of
resulted in the absence of microorganisms at a 101dilution, sugar.
being within the quality standard required by the legislation When analyzing the same variables in the test group (weight,
(CNNPA Resolution No. 12, 1978). Thus, the result of these BMI and WC) before and after the consumption of 70% cocoa
analyzes revealed that the chocolate bars were safe for human chocolate incorporated with ursolic and oleanolic acids through
consumption, either for sensory analysis or for the clinical trial. t-test for paired samples at 5% probability, significant changes

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 Milagres et al.

Table 2. Parameters of volunteers before and after the clinical trial.

Test group Placebo group Control group
Variable (mean ± deviation) (mean ± deviation) (mean ± deviation)
Before After Before After Before After
Weight (kg) 70.07±15.99* 68.87±15.04* 73.73±17.70* 75.40±18.23* 68.19±16.02 68.10±15.48
BMI (kg/m2) 25.42 ± 4.62* 24.99±4.28* 26.12±4.87* 26.68±4.89* 24.59±4.15 24.57±4.01
WaistCircumference– WC (cm) 92.13±14.43* 88.73±13.26* 93.80±13.45* 97.13±13.30* 86.60±12.36* 87.07±11.61*
*Significant change in columns p<0.05.

(ρ<0.05) were noted. However, these occurred in a beneficial way group also showed an increase in their WC measurement as
for the group, since the consumption of that chocolate reduced 46.6% (n = 7) of the subjects presented increase of 1 to 3 cm in
the anthropometric measurements. the circumference.
In the control group, the analysis of the variables: weight, BMI Considering that waist circumference measurements reflect
and WC before and after the experimental period through the the visceral fat content and are also strongly associated with
t-test for paired samples at 5% probability showed no significant total body fat and this fact correlates with the risk of CNCD
changes (ρ>0.05). (Hernandez et al., 2017), this reduction was very important for
the health of individuals in the test group.
After consumption of 70% cocoa chocolate incorporated
with ursolic and oleanolic acid, 53.3% (n = 8) of the subjects The increasing incidence of chronic noncommunicable
in the test group decreased weight measurements. Of these, diseases associated with poor diet is increasing and worrying,
50% (n = 4) lost approximately 2 kg. On the other hand, in the especially the increase in overweight, obesity and their associated
placebo group, 86.6% (n = 13) of the individuals gained weight complications (Arokiasamy et al., 2017). Thus, a balanced diet
during the intervention, of which 23.7% (n = 3) acquired 3 kg. and food additions that may aid in weight, BMI and waist
In the control group, 60% (n = 9) remained at the same weight. circumference loss are essential for the maintenance of a body
mass within the parameters of normality. And it was observed in
The association between weight gain, abdominal increase,
this study that the developed chocolate helped in the reduction
physical inactivity and the onset of chronic non-communicable
of these anthropometric variables.
diseases is evident, and eating practices are considered elements
that can be modified in the prevention of comorbidities linked In the statistical evaluation of the clinical trials, no significant
to overweight and obesity (Hernandez et al., 2017). Thus, the difference was observed in the variables glucose (ρ = 0.414) and
weight loss observed in the test group was a very relevant result. LDL (ρ = 147), comparing the values collected before and after the
consumption of 70% cocoa chocolate with ursolic and oleanolic
As a consequence of weight reduction, the decrease in Body
acids in the test group, according to the Wilcoxon test at 5%
Mass Index (BMI) was observed in the same individuals who
probability. No difference was observed in this group between
hadweight reduction in the test group. Thus, 53.3% (n = 8) of
the values of total cholesterol (ρ = 0.580), triglycerides (ρ = 0.124)
the individuals had a BMI loss, of which four had BMI reduction
and HDL (ρ = 0.113) before and after the consumption of 70%
between 0.5 and 1.0 kg/m2. The subjects in the placebo group
cocoa chocolate with ursolic and oleanolic acids by the t-test
gained weight (n = 13) and increase between 0.5 and 1.0 kg/m2
for paired samples at 5% probability.
was observed in 61.54% (n = 8) of the individuals. In the control
group the majority (60%) of the subjects remained with no The results of the clinical trials of the placebo group also
changes in the BMI. did not indicate statistical differences (ρ> 0.05) in the variables
glucose, LDL, HDL, total cholesterol before and after consumption
This reduction in BMI seen in this study is considered
of 70% cocoa chocolate with placebo for ursolic and oleanolic
fundamental for health maintenance, since this is a good indicator
acids by Wilcoxon’s test at 5% probability, and in the variable
of obesity and overweight. Evidence suggests that high values
triglycerides (ρ = 0.803) by the t-test for paired samples at 5%
(> 30 kg/m2) of this variable are associated with the risk of
probability.
chronic non-communicable diseases. This result may be related
to the incorporation of ursolic acids to chocolate, since the use In the control group, no significant difference (ρ>0.05) was
of these is related to the decrease of body mass index in study observed in the variables glucose, LDL, HDL, total cholesterol,
for evaluate the effect of ursolic acid on metabolic syndrome and triglycerides before and after the intervention period without
(Ramírez-Rodríguez et al., 2017). chocolate consumption by the t-test for paired samples at 5%
probability, as shown in Table 3:
The analysis of the waist circumference of the individuals
allowed verifying that 73.3% (n = 11) of the individuals in the test When comparing the test, control and placebo groups, the
group had reduction in measurements. Of these, 27.7% (n = 3) statistical difference (ρ<0.05) after intervention of the values of
had more than 8cm of reduction in waist circumference. In the glucose, LDL, HDL, totalcholesterol and triglycerides (ρ<0.05)
placebo group the result was different, 46.6% (n = 7) of subjects were observed by analysis of variance at 5% probability. In the
had an increase of 1 to 3cm in waist circumference, while only one test group, greater reductions were observed in the clinical
individual presented no changeinhis measurements. The control variables after the consumption of 70% cocoa chocolate with

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 Dark Chocolate with triterpenic acids

Table 3. Clinical parameters of the volunteers before and after the clinical trial.
Test group Placebo group Control group
Variable (mean ± deviation) (mean ± deviation) (mean ± deviation)
Before After Before After Before After
Glucose 85.00±6.28 83.43±12.68 83.25±8.23 84.03±13.09 85.13±5.23 84.50±2.54
Total cholesterol 195.54±24.30 198.43±34.76 196.98±23.33 197.97±29.67 195.97±22.22 196.07±32.11
HDL 56.29±7.56 52.00±7.02 55.97±5.67 54.30±2.07 55.31±35.67 55.98±23.78
LDL 100.00±14.31 115.00±35.22 101.10±9.021 109.00±23.56 103.19±13.45 104.56±11.89
Triglycerides 197.14±24.08 155.79±58.72 195.13±24.08 182.89±32.15 186.23±34.56 183.83±36.79
Source: Directsurvey.

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The results of this study show that it was possible to develop Methods of Analysis of the Association of Official Analytical Chemists.
and characterize 70% cocoa chocolate with addition of ursolic Arlington: AOAC.
acid and oleanolic acid, so that the formulation showed good Belsito, P. C., Ferreira, M. V. S., Cappato, L. P., Cavalcanti, R. N., Vidal,
sensory acceptance, which indicates its possible inclusion in the V. A. S., Pimentel, T. C., Esmerino, E. A., Balthazar, C. F., Neto, R.
healthy foods market. It also showed that regular consumption of P. C., Tavares, M. I. B., Zacarchenco, P. B., Freitas, M. Q., Silva, M.
C., Raices, R. S. L., Pastore, G. M., Pollonio, M. A. R., &Cruz, A.
a 25 g bar of 70% cocoa chocolate containing 150 mg of ursolic
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4 Conclusion normas regulamentadoras de pesquisas envolvendo seres humanos
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in the clinical parameters in relation to the other experimental of nutritional and physicochemical proprieties of milk chocolates
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