FRUIT AGROINDUSTRIAL

5
WASTES FOR PREPARING
BEVERAGES FOR MEDICINAL
PURPOSES BY SUPERCRITICAL
FLUID EXTRACTION
TECHNOLOGY: ANDES BERRY
(RUBUS GLAUCUS BENTH) CASE
Sonia C. Lizcano⁎, Javier A. Dávila⁎, Valentina Hernández†
⁎
Department of Engineering, Chemical Engineering Program, Universidad
Jorge Tadeo Lozano, Bogotá, Colombia †TecnoParque Nodo Manizales,
Servicio Nacional de Aprendizaje, SENA Regional Caldas, Manizales, Colombia

5.1 Overview
Tropical countries have a wide variety of fruits that contain valu-
able compounds, which could be obtained not only from the pulp but
also from the peel and seed. Fruit processing generates a significant
amounts of residues that correspond to the nonedible parts that are
disposed in an inadequate manner but its valuable compounds have
different potential applications. Since a lot of these valuable com-
pounds contained in fruits could be used for pharmaceutical, cos-
metic, and chemical applications; the valuable biologically active
compounds such as antioxidants, vitamins, phenolic compounds, and
anthocyanins are the most attractive compounds to be extracted, and
thus, could be used for preparing beverages with high functional qual-
ity to be used as health promoters. However, the applications of these
compounds are constrained for the extractions and concentrations
yields because the technologies are not well established yet. Because
for conserving the chemical properties of the valuable compounds
it is an important factor in the extraction process, it is necessary to
improve such processes to increase the yields of the extraction and to

Production and Management of Beverages. https://doi.org/10.1016/B978-0-12-815260-7.00005-5

© 2019 Elsevier Inc. All rights reserved. 151
152 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

obtain solvent-free extracts as well as to enhance the concentration

and stabilization of valuable compounds for allowing medicinal appli-
cations for human health and well-being.
In this sense, Andes berry (Rubus glaucus benth) has antioxidant
properties due to its phenolic compounds such as cafeic, chlorogenic
acids, and quercitin. This fruit also has significant contents of ascorbic
acid and it is an excellent source of anthocyanins, which have appli-
cations in the field of cancer. Owing to this fact Andes berry is a good
source of valuable compounds (phenolic compounds, anthocyanins
among others), which can be obtained as extracts and can be used for
beverage preparation aimed for medicinal applications. In this sense,
this chapter describes the use of Andes berry waste for obtaining extracts
rich in phenolic compounds and anthocyanins using supercritical CO2
extraction. The functional quality of the extracts is assessed according
to the contents of total phenolic compounds (TPCs), anthocyanins con-
tent, and the total antioxidant activities as the main characteristics for
beverages preparation. Pressure and cosolvent (ethanol) to solid matrix
ratio are evaluated for assessing the extraction yield as well as functional
quality. Thus, potential applications of fruit agroindustrial wastes from
Andes berry processing are evaluated for obtaining extracts that can be
aimed to prepare beverage for medicinal purposes.

5.2 Fruits as Source of Valuable Compounds

Because of its geographic location and its wide range of climates,
Colombia is the third-ranked country in the world in terms of biodi-
versity, housing approximately the 10% of the world species, and the
fourth-ranked country in terms of hydrographic resources (Mongabay,
2010). According to Procolombia (2016) 10.9% of the country was de-
voted to fruit production in Colombia in 2013, which corresponded
to a production of 9.5 million tons of fruits. In the country, approxi-
mately 48 fruit species can be obtained, including both perennial
and nonperennial fruits (Alarcón García et al., 2015). Among the pe-
rennial fruits, the top five species cultivated in Colombia are orange
(Citrus sinensis), mango (Mangifera indica), avocado (Persea ameri-
cana), guava (Psidium spp.), and tangerine (Citrus spp.), comprising
44.8% of the total cultivated area devoted to fruit production, with-
out considering domestic plantain and banana production (Alarcón
García et al., 2015). Similarly, the most important nonperennial fruits
(without considering plantain and banana) are pineapple (Ananas
comosus), blackberry (Rubus ulmifolius), tamarillo fruit (Solanum be-
taceum), and lulo (Solanum quitoense), comprising 23.48% of the total
cultivated area (Alarcón García et al., 2015). Table 5.1 summarizes the
crop production per year of some fruits in Colombia and its respective
agroindustrial wastes.
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 153

Table 5.1 Some Fruits and Agroindustrial Wastes

Produced in Colombia
Crop Agroindustrial
Agroindustrial Production Wastes (tons/
Waste (tons/year) year) References
Plantain peel 2.724.888 416.908 de Fonseca (2009), Motato et al. (2006), Castellanos
and Lucas (2011), and Duque et al. (2015)
Rejection bananas 2.034.340 364.420 Motato et al. (2006)
Banana stem 226.037 Motato et al. (2006) and Duque et al. (2015)
Coffee pulp 514.128 216.962 Rodríguez (2009) and Pandey et al. (2000)
Coffee husks 71.493 Rodríguez (2009) and Pandey et al. (2000)
Guava peel and seed 440.102 59.854 Torres (2010), Yepes et al. (2008), and González
(2010)
Pineapple peel 397.824 149.184 Duque et al. (2015), Yepes et al. (2008), Rodríguez
and Hanssen (2007), and Ministerio de Agricultura
y Desarrollo Rural and Corporación Colombia
Internacional (2002)
Pineapple leaf 79.565 Yepes et al. (2008), Ministerio de Agricultura
y Desarrollo Rural and Corporación Colombia
Internacional (2002) and García and Torres (2003)
Orange peels, seeds, 322.989 161.495 de Fonseca (2009), Yepes et al. (2008), Rodríguez and
and bagasse Hanssen (2007), López et al. (2011), and Espinal et al.
(2005)
Papaya peel and 262.914 93.710 Yepes et al. (2008)
seed
Peel, seeds, and 243.375 65.711 Duque et al. (2015), Yepes et al. (2008), Ospina et al.
whole mango (2012), and Mejía et al. (2007)
Tamarillo peel 130.211 11.719 Yepes et al. (2008), Cerón et al. (2010), Revelo (2011),
and Sánchez and Murillo (2010)
Mandarin wastes, 123.641 68.003 Espinal et al. (2005) and Navarrete et al. (2010)
seeds, and peel
Coconut fiber 93.206 27.962 Bonilla-Lavado et al. (2006)
Blackberry seeds 86.176 56.014 Sánchez and Murillo (2010), Franco and Giraldo
(1998), and Díaz (2011)
Passion fruit peel 81.089 42.166 Yepes et al. (2008), Durán and Méndez (2008), and
Ministerio de Agricultura y Desarrollo Rural and
Corporación Colombia Internacional (2003)
Cocoa pulp 39.534 13.178 Cuéllar (2010) and Villegas et al. (2007)
Cocoa seed hulls 3.953 Villegas et al. (2007) and Serra Bonvehı́ and Ventura
Coll (1999)

Continued
154 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Table 5.1 Some Fruits and Agroindustrial Wastes

Produced in Colombia—cont’d
Crop Agroindustrial
Agroindustrial Production Wastes (tons/
Waste (tons/year) year) References
Soursop peel and 13.029 2.840 Márquez (2009)
seed
Goldenberry calyx 8.211 821 Yepes et al. (2008), Cerón et al. (2010), Hernández
and López (2007), Ministerio de Agricultura y
Desarrollo Rural and Corporación Colombia
Internacional (2001), Cedeño and Montenegro (2004),
and Fischer et al. (2005)

In Colombia, cutting and transformation processes of the differ-

ent crops lead to the generation of several million tons of wastes per
year (Duque et al., 2015; Escalante et al., 2007), as illustrated in the
table. These agricultural crop residues can be classified as field and
processing residues (Escalante et al., 2007; Hadar and Faraco, 2013).
According to the Ministerio de Minas y Energía (Escalante et al.,
2007), 75% of the total biomass produced in Colombia corresponds
to crop residues, while processing residues represent the remain-
ing 25%. In the case of fruit residues, its pharmacological and nutri-
tional properties have attracted the attention of food, cosmetic, and
pharmaceutical industries for obtaining valuable products (Gupta
et al., 2015).

5.2.1 Valuable Compounds of Tropical Fruits and Its

Residues
Fruits contain substances such as vitamins and secondary me-
tabolites (polyphenols, carotenoides, sterols, flavors, dyes, essential
oils, flavonoids, alkaloids, tannins, coumarins, lactones, terpenes, and
saponins, among others) (Ramful et al., 2010; Genovese et al., 2008;
Contreras-Calderón et al., 2011), with a biological activity which can
protect the human body against cellular oxidation reactions. Besides,
fruit wastes, which usually represent an environmental problem due
to the current disposal methods (dumping, burning, or land filling),
are an attractive sources of several phytonutrients.
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 155

5.2.1.1 Antioxidant Compounds

The oxidative stress contributes to a wide variety of deceases such
as cancer, neurological disorders, atherosclerosis, hypertension, isch-
emia, diabetes, acute respiratory distress syndrome, idiopathic pul-
monary fibrosis, chronic obstructive pulmonary decease, heart failure,
chronic inflammation, hemorrhagic shock, kidney damage, rheuma-
toid arthritis, aging, and asthma, among others. In order to counteract
the harmful effects of the oxidative stress, antioxidants can be either
naturally produced in the body (endogenous antioxidants) or supplied
externally through the diet (exogenous antioxidants) (Dávila et al.,
2014). Under specific low concentrations, the antioxidants can inhibit
or delay the oxidative process or even inactivate the ROS at cellular
and molecular levels by interrupting the radical chain reaction (Flora,
2009). Plants contain different types of phytochemicals with antioxi-
dant properties. Some of the most important exogenous antioxidants
and its benefit effects over the human health are explained below.
Vitamin C is also called ascorbic acid and cannot be synthetized
by the human body. It is a water-soluble vitamin essential for the bio-
synthesis of neurotransmitters, collagen, and carnitine (Dávila et al.,
2014). In the cells the ascorbic acid reacts with glutathione for main-
taining its reduced form (Lobo et al., 2010). Health benefits associated
with vitamin C are its antioxidant, antiatherogenic, anticarcinogenic,
and immunomodulator properties. Several studies have demon-
strated the positive effects of vitamin C in reducing the incidence of
stomach cancer and the prevention of lung and colorectal cancers
(Dávila et al., 2014).
Vitamin E is a fat-soluble vitamin and a chiral metabolite with eight
stereoisomers: α-, β-, γ-, and δ-tocopherol and α-, β-, γ-, and δ-tocotrienol,
α-tocopherol being the most bioactive for humans. It is also consid-
ered to be the major membrane bound antioxidant used by the cell
(Dávila et al., 2014). The main function of vitamin E is to protect
against lipid peroxidation by reacting with lipid radicals produced in
the peroxidation chain reaction (Dávila et al., 2014). During this anti-
oxidant reaction, α-tocopherol radical is obtained and can be reduced
to its original form (α-tocopherol) by vitamin C.
Flavonoids are metabolites with variable phenolic structures that
can be found in different sources such as fruit, vegetables, grains,
bark, roots, stems, flowers, tea, and wine. The antioxidant effect of
flavonoids lays over its ability of scavenging the oxygen-derived free
radicals as well as its antiinflammatory, antiallergic, antiviral, and an-
ticarcinogenic properties (Dávila et al., 2014).
Carotenoids have several biological activities to help promote
health and can be synthetized by plants and microorganisms. Studies
have demonstrated that these pigments are involved in the scaveng-
ing of two of the ROS: molecular oxygen (O2) and peroxyl radicals
156 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

(Dávila et al., 2014). Beneficial effects of carotenoids on health have

been related to the decreasing the risk of certain cancers, cardiovas-
cular diseases, and disorders related to the eyes (Dávila et al., 2014).
Antioxidants are always present in fruits as very complex mixtures
and its activity is attributed to a group of compounds rather than one
single compound. Therefore, knowing the composition and charac-
terization (antioxidant capacity and total polyphenolic compounds)
is important to generate agroindustrial uses. The overall nutritional
value depends on the specific plant genotype and interactions of culti-
vation conditions; Table 5.2 summarizes the polyphenolic concentra-
tion and antioxidant activity contents of different tropical fruits.

Table 5.2 Polyphenolic Compounds and Antioxidant

Capacity Contents of Tropical Fruits
Polyphenolic Antioxidant capac-
compounds (mg ity (μmol Trolox g−1
Fruit GAE 100 g−1 fw) sample FW) Country References
Tamarillo pulp 78 ± 2 2.3 ± 0.1 Ecuador Vasco et al. (2009)
Tamarillo peel 387 ± 8 22 ± 4 Vasco et al. (2009))
Tamarillo seed 94 ± 1 3.8 ± 0.6 Vasco et al. (2009)
Naranjilla pulp 58.3 ± 2.39 12.2 ± 0.85 Colombia Acosta et al. (2009)
Naranjilla peel 83.6 ± 0.64 21.1 ± 0.23 Acosta et al. (2009)
Naranjilla pulp 48 ± 3 NR Costa Rica Acosta et al. (2009)
Naranjilla peel 91 ± 17 14.02 Ecuador Acosta et al. (2009)
Goldenberry fruit 40.45 ± 0.93 2.11 ± 0.09 Colombia Restrepo Duque (2008)
Goldenberry fruit 39.15 ± 5.43 1.93 ± 0.30 Colombia Botero Echeverri (2008)
Goldenberry peel 154 ± 3 NR Peru Repo-de-Carrasco and Encina
Zelada (2008)
Araza frozen pulp 64 ± 3 3.7 ± 0.1 Brazil Genovese et al. (2008)
Araza fruit 129 ± 9 4.1 ± 0.2 Brazil Genovese et al. (2008)
Araza fruit 111 ± 3.64 20.2 ± 2.44 Colombia Contreras-Calderón et al.
(2011)
Araza seed 1624 ± 44.9 440 ± 7.77 Colombia Contreras-Calderón et al.
(2011)
Cupuazu fruit 4.03 ± 0.57 9.59 ± 0.25 Colombia Contreras-Calderón et al.,
(2011)
Cupuazu peel 497 ± 17.8 145 ± 0.09 Colombia Contreras-Calderón et al.
(2011)
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 157

Table 5.2 Polyphenolic Compounds and Antioxidant

Capacity Contents of Tropical Fruits—cont’d
Polyphenolic Antioxidant capac-
compounds (mg ity (μmol Trolox g−1
Fruit GAE 100 g−1 fw) sample FW) Country References
Cupuazu peel 252 ± 28.7 65.3 ± 1.00 Colombia Con treras-Calderón et al.
(2011)
Algarrobo peel 1712 ± 42.5 428 ± 9.38 Colombia Contreras-Calderón et al.
(2011)
Coastal sapote 1488 ± 20.1 377 ± 8.06 Colombia Contreras-Calderón et al.
peel (2011)
Buriti 378.07 ± 3.12 NR Koolen et al. (2013)
Gabiroba fruit 851.03 ± 40.79 8027.52 ± 378.63 Brazil Malta et al. (2013)
Açaí 454 ± 44.6 15.1 ± 4.1 Brazil Rufino et al. (2010)
Acerola 1063 ± 53.1 96.6 ± 6.1 Brazil Rufino et al. (2010)
Camu camu 1320 ± 102 167 ± 11 Peru Chirinos et al. (2010)
Borojo fruit 41.8 ± 1.54 6.29 ± 0.86 Colombia Contreras-Calderón et al.
(2011)
GAE, Gallic acid equivalent.

5.2.2 Potential of Fruits for Preparing Beverages

Currently the world trade of fresh fruits is earning approximately
20 billions dollars every year (Almeida et al., 2011). In addition, pro-
duction of functional products has been promoted by the demand
in the market by sustained product rich in natural compounds. The
United States is the largest importer of fresh tropical fruits followed
by the European Union, Japan, and China. In the case of functional
beverages, its market represents the largest and fastest growing seg-
ment of the functional foods sector, with an annual growing rate of al-
most 20% in the United States (Shahidi and Alasalvar, 2016). Besides,
production and consumption of fruit juices, beverages and hot drinks
have gained much importance since they are inversely associated with
morbidity and mortality from degenerative diseases.
Currently several researchers have dedicated their efforts to study
the nutritional value of different fruit beverages rich in antioxidant
compounds. For instance, Carlsen et al. (2010) studied 283 beverages,
including fruit-based products (see Table 5.3). According to this study,
red wine presented the highest antioxidant contents. Some fruits cur-
rently used in commercial beverages, due to the attractive antioxidant
158 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Table 5.3 Antioxidant Capacity of Different Fruit

Beverages
Fruit Beverage Antioxidant Content (nmol/100 g)
Apple juice 0.27
Cranberry juice 0.92
Grape juice 1.20
Orange juice 0.64
Pomegranate juice 2.10
Tomato juice 0.48
Red wine 2.50
Data from Carlsen MH, et al. The total antioxidant content of more than 3100 foods, beverages, spices, herbs and supplements used
worldwide. Nutr. J. 9:3–3, 2010.

content, include cranberry, pomegranate, black currant, acai, gua-

rana, mango, bilberries, grapes, cherries, kiwi, strawberries, feijoa,
peach, watermelon, sapodilla, orange, and plums (Corbo et al., 2014).
Different studies have established that the consumption of fruits
and vegetables are beneficial to human health such as reducing car-
diovascular and coronary heart diseases (Franzini et al., 2012; Wang
et al., 2011; Egan et al., 2001), diabetes (McCune and Johns, 2002),
inflammatory processes (Wu et al., 2006; Gene et al., 1992), neurode-
generative diseases (ArunaDevi et al., 2010; Kris-Etherton et al., 2002),
and cancer (Kris-Etherton et al., 2002; Collins, 2005; de Mesquita et al.,
2009; Forster et al., 2013; Habib et al., 2013; Tsai et al., 2013; Zhang
et al., 2013), among others. The constituents responsible for these pro-
tective effects are bioactive compounds known as antioxidants. An an-
tioxidant is a metabolite capable of preventing, reducing, or delaying
the effect of the oxidative stress of target biomolecules. The oxidative
stress is considered a consequence of an insufficiency of the antioxi-
dant defense system and/or the overproduction of free radicals (Dávila
et al., 2014). A free radical is defined as molecule with one or more un-
paired electron in its outer shell. The free radicals can be derived from
the essential metabolic processes in the human body as well as from
external exposure (Lobo et al., 2010). The most studied free radicals
are those derived from molecular oxygen and nitrogen (ROS and RNS,
respectively). The ROS includes oxygen radicals [e.g., superoxide an-
ion radicals (O 2 ) and hydroxyl radicals (OH−)] and some nonradicals
which are oxidizing agents and/or can be converted to free radicals
(e.g., H2O2 and O2) (Dávila et al., 2014). The ROS and RNS are gen-
erally the result of radiation by UV light, metal-catalyzed reactions,
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 159

Table 5.4 Physicochemical Composition of Andes

Berry (Dávila et al., 2014)
Feature Value Feature Value
Physicochemical characterization Total phenolic, vitamin C, and anthocyanin
content (mg/100 g FW)
Ash (%) 0.29 Total phenolics 499.0
Moisture (%) 90.88 Antioxidant capacity 71.0
pH 3.42 Vitamin C 21.0
TSS (°Brix) 7.60 Anthocyanins 125.6
TA (% citric acid) 1.26
Carbohydrates (%) 5.04
Total soluble sugars (g/100 g FW) Flavonoids content (mg/100 g FW)
Glucose 2.44 Quercetin 19.0
Fructose 1.99 Cyanidin 142.0
Total 4.35 Epicatechin 52.0
Total 213.0

atmospheric pollutants, neutrophils, and macrophages during in-

flammation and mitochondria-catalyzed electron transportation re-
actions, among other causes (Dávila et al., 2014).

5.2.2.1 Andes Berry (Rubus glaucus Benth)

Andes berry is a perennial climbing shrub from the Rosaceae fam-
ily. The fruit is a berry ellipsoid with a size between 15 and 25 mm
and with a weight of 3–5 g approximately. This fruit is rich in vitamin
C, calcium, and phosphorus and used commonly for juices, nectars,
jams, jellies, ice cream, pastries, and confectionery. Two annual pro-
ductions are there. Several studies have demonstrated the presence of
antioxidants in Andes berry (Cerón et al., 2012; Hassimotto et al., 2008;
Dávila et al., 2017). The chemical composition of the blackberry fruit is
summarized in Table 5.4.

5.3 Industrial Production of Beverages

5.3.1 State of the Art about Beverage Production
Scientific studies about fruit-based beverages have increased in
the last years, as illustrated in Table 5.5, due to the increasing interest
in healthy food consumption around the world. There is a wide range
160 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Table 5.5 Active Compounds Present in Fruit-Based

Beverages
Fruit-Based Beverage Active Compounds
Fortified-strawberry beverage Polyphenols
Grape-based fermented beverage Polyphenols
Fortified blackcurrant juice Polyphenols
Fortified fruit juice Antioxidants
Pomegranate fermented juice Phenolic compounds
Apple-based beverage Secoiridoid glycosides
Data from Corbo MR et al.: Functional beverages: the emerging side of functional foods. Compr. Rev. Food Sci. Food Saf.:13(6): 1192–1206, 2014.

of functional beverage in the market such as beverages based on dairy

as well as beverages with probiotics and minerals/ω-3, vegetable and
fruit beverages, and sports and energy drinks (Corbo et al., 2014). The
most typical beverages based on dairy drinks are fresh milk, fermented
milk, and yogurt. In the case of fruits, the most used are cranberry,
blueberry, black currant, acai, acerola, guarana, mango, bilberries,
grapes, cherries, kiwi, strawberries, feijoa, peach, watermelon, sapo-
dilla, orange, and plums. On the other hand, sport drinks are flavored
beverages to be consumed before or during exercise. This chapter is
focused on the production of fruit-based beverages with antioxidant
capacity.
There are different processes to obtain functional beverages from
fruits and can be divided into five groups: (1) exploration of function-
ality of microorganism, (2) production of new functional beverages,
(3) use of prebiotics and synbiotics, (4) use of natural ingredients for
beverages, and (5) use as functional ingredients, by-products based
on fruits (Corbo et al., 2014). This chapter is focused on the last two
methods, in which fruits and its by-products are used as functional in-
gredients for producing beverages.

5.3.2 Supercritical CO2 Extraction of Valuable

Compounds
Different extraction technologies have been evaluated for perform-
ing the extraction of antioxidants from plants. Although, in general
four main steps are involved in the antioxidant production process:
pretreatment, extraction, concentration, and stabilization. In the
pretreatment stage, the feedstock is dried allowing a rapid cell wall
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 161

breakdown during the subsequent milling and homogenization pro-

cesses and minimizing the enzymatic degradation of the fruits and the
antioxidant compounds (Dávila et al., 2014). The most used technol-
ogies in the pretreatment stage for the extraction of antioxidants are
air drying, freeze drying or lyophilization, and vacuum drying (Dávila
et al., 2014).
In the second stage, the extraction of antioxidant-rich extracts is
carried out. This process have been performed through different ex-
traction technologies such as conventional solvent extraction (CSE),
supercritical fluids (SFE), and ultrasonic- and microwave-assisted ex-
traction (MAE) (Dávila et al., 2014).
In the concentration stage the most used technologies are the vac-
uum distillation (Chumsri et al., 2008), ultrafiltration, and nanofil-
tration (Dávila et al., 2014). In the concentration process, low oxygen
presence, low temperatures and low residence time are required in
order to preserve the high quality of the extract.
Since the antioxidants extracted from fruits are sensitive to many
environmental conditions such as pH, light exposure, temperature,
and water activity (Dávila et al., 2014), the fourth stage involves the
stabilization of the concentrated extracts. The microencapsulation is
the most used technology for performing the stabilization of the an-
tioxidants leading to preservation their effectiveness (Dávila et al.,
2014).

5.3.2.1 Supercritical Extraction of Antioxidants

In the supercritical extraction process, a solvent at supercritical
conditions is used to separate the extractant from the matrix, which
is usually solid but that can be liquid or even viscous. There is a pos-
sibility of modifying the selectivity of the process when the density of
the fluid is changed, which is the main advantage of the supercritical
extraction process (Dávila et al., 2014). A general flow diagram of a su-
percritical extraction process is illustrated in Fig. 5.1.
In the supercritical extraction process, the carbon dioxide is first
subcooled in order to guarantee the liquid phase before entering into
the pump to avoid cavitation. Then the pressurized CO2 is heated
above its critical temperature, while the extractor is maintained at the
operating temperature through electricity or hot water. The CO2, at su-
percritical conditions, flows through the feedstock into the extractor
extracting its soluble compounds. Then the solvent together with the
extract leaves the extractor, which is at the operating pressure, and
goes to the separator (at atmospheric pressure). In the separation ves-
sel due to the depressurization the extract is precipitated.
The supercritical fluid extraction using CO2 as solvent has been
widely used for extracting antioxidants from tropical fruits such as
blackberry (Cerón et al., 2012), citrus (Moncada et al., 2013), zapote
162 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Fig. 5.1 Simplified flow diagram of the supercritical fluid extraction with CO2 (Dávila
et al., 2014).

(Cerón et al., 2014), tamarillo (Cerón, 2013), goldenberry (Cerón,

2013), and Andes berry (Dávila et al., 2014; Dávila et al., 2017).

5.4 Supercritical CO2 Extraction for

Phenolic Compound-Rich Extracts
5.4.1 Supercritical CO2 Extraction of Valuable
Compounds From Andes Berry Residues
The extraction of supercritical fluid has been considered a green
method due to a substitute with advantageous results with respect to
a traditional extraction. The low viscosity and the higher diffusion co-
efficient allow reaching a great thermal compressibility. This process
involves a simple separation, changing the temperature or pressure,
which generates a dissolution capacity similar to a liquid, which has
an effect of a solvent with high power (Sanchez et al., 2017). This kind
of process combines advantages of distillation and extraction with liq-
uids since it works at low temperatures, in this case a process at low
temperatures and pressures carried out with CO2 will be analyzed
(Sanchez et al., 2017; Brunner, 2005).
The CO2 improves the condition of the extraction since the criti-
cal point is reached at low temperatures and pressures (Sahena et al.,
2009; Machado et al., 2015). The CO2 is an ideal solvent due to the low
viscosity and high density that allows a better mass transfer in su-
percritical conditions. This solvent is nontoxic and nonflammable;
besides, the obtained extracts have a higher purity in comparison
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 163

with other extraction methods such as solvent extraction tech-

niques. Because CO2 is a nonpolar molecule, it is necessary to use
a cosolvent to increase the solubility of polar components to obtain
high yields of anthocyanins. In this case the water-ethanol mixture
helps in the production of upper extracts with a great ranking of
purity, improving the polarity of the supercritical CO2 which allows
to dissolve polar compounds (Domínguez and Parzanese, 2005;
Zulkafli et al., 2014a).
This technique uses supercritical conditions to modify the solu-
bility between target compounds and the solvent by the fluid density.
CO2 is accentuated as the first component subcooled in all the process
to make it sure that it stay in liquid phase and avoid a cavitation. Thus,
CO2 must be pressurized to a temperature near or above the critical
temperature, thus, supercritical conditions should be maintained in
order to allow a flow through the soluble components contained in the
vegetable matrix (Brunner, 2005; Sahena et al., 2009). Then, the sol-
vent comes out with the extract at a low pressure through the separator
at an atmospheric pressure. Thus, the precipitated extract is obtained
in a separator vessel and at the same time, CO2 is cooled to −25°C to
have a liquid phase. Theoretically, the pumps maintain an approxi-
mate pressure of 200 bar looking for the supercritical conditions to
enter the extraction; the cosolvent is added to increase the yield in the
extraction, thus improving the anthocyanins content in the final ex-
tracts (Cerón et al., 2012).
The company Frugy S.A gave samples of Andes berry residues
(Manizales, Colombia). These samples were dried by a conventional
method (Chemical Industries FIQ LTDA, Colombia y Memmert, 854
Schwabach, Germany) to get a regular weight between every sample
at a regular temperature of 35°C that avoid the degradation of anti-
oxidants and phenolic compounds. Then, the sample was processed
by a hammer mill (Raymond No. 82, China) and sifted by a series of
Tyler-type sifts (ASTM E-11) obtaining an average of 756 μm diameter
particle. The reagents used were CO2 at 99.9% (Linde Colombia S.A.),
ethanol at 99.8% (Merck, Germany), which was acidified with distilled
water and citric acid to have a pH of 2.0 obtaining a mixture of water-
ethanol at 57%v/v, chloride acid at 37% (Alquera), sodium acetate, and
potassium chloride (Bioquigen) (Garzón, 2008).
The supercritical fluid extraction equipment is located at the high
pressures in the laboratory of the university Jorge Tadeo Lozano,
Bogota Colombia. This system has a pressurization pump (piston
bomb-Milroyal B Pump), a tank extractor (1.3 L), and a separator ves-
sel (1 L) with a capacity to operate at a pressure of 4500 psi to obtain all
the extracts. Besides, this system has an automatic control (SuperView
2.9.9) where it is possible to verify the temperatures and pressures of
the extractor and separator vessel (Acosta, 2017; Santacruz, 2016).
164 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

An amount of 30 g of samples were used and placed in the 550 mL

extractor while the cosolvent (acidified ethanol) were added in the
extraction chamber and both the pressure and the temperature were
fixed at the desired supercritical conditions. In parallel, the refriger-
ant (propilenglicol) was recirculated using a chiller at a temperature
of approximately −20°C to allow CO2 (99.9% of purity) to remain in a
liquid state (Acosta, 2017). There is necessary for the system, 1 hour
to stabilize thermally the system and starting the pressurizing to the
desired pressure. The extractions have been carried out in static mode,
which means the batches of approximately 15 min was followed for
depressurization and then pressurized again to the desired pressure.
Once the operating conditions have been reached, the extraction time
is calculated, waiting 15 min to start another discharge, which is nec-
essary to renovate the CO2, thus, improving the sample-mass transfer
(Santacruz, 2016).
After, the equipment is depressurized conducting the CO2 and
the sample toward the tank separation. As the CO2 passes through the
valve, it expands and losses all its proprieties as solvent, therefore,
the extract (rich in anthocyanins) tends to precipitate, facilitating the
recovery process (Acosta, 2017; Santacruz, 2016).
After the depressurization of CO2, the solute compounds (an-
thocyanins) are obtained free of solvent and because of the photo-
sensitive character of the extracts, the samples were collected and
protected from light and stored at −10°C to further analyze the phys-
icochemical properties (del P Garcia-Mendoza et al., 2017; De Cássia
Rodrigues Batista et al., 2015) and to avoid any type of degradation.
The process flow diagram of the supercritical extraction is depicted
in Fig. 5.2.

5.4.2 Anthocyanins Analysis (Measurement

and Analysis of the Anthocyanins Content in the
Obtained Extracts)
For the determination of anthocyanins, the colorimetric method
was used. This method is defined by Giusti and Wrolstad (2001). Two
types of buffer were required for the extraction solutions: sodium ac-
etate with a pH of 4.5 and potassium chloride with a pH of 1.0 (Giusti
and Wrolstad, 2001). The anthocyanins concentration is expressed in
mg/L of extract determined as cyanidin 3-glucoside because this type
of anthocyanin is predominant in this fruit. The calculation of antho-
cyanins concentrations have been carried out by Acosta (2017) where
the absorbance method was used with a wavelength between 300 and
700 nm for each of the buffers. The concentration is expressed accord-
ing to Eq. (5.1) where Abs is the absorbance calculated in Eq. (5.2),
 CO2 recovered

CO2
CO2

Pump Water
CO2 solvent
Mix

Product (beverage)

Mixer

Blackberry waste
Mill Collector
Moisture
Extract
AIR Extract Antoxidants rich
Dryer extract
Extractor Valve
Solids

Evaporator
SOLIDS

Ethanol Ethanol
Ethanol
Ethanol recovered
Pump
co-Solvent

Fig. 5.2 Process flow diagram for the supercritical extraction.

Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES
165
166 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

MW is the molecular weight (449.2 g/mol), and ε is the molar extinc-

tion coefficient (26,900 L × mol−1 × cm−1) (Giusti and Wrolstad, 2001).
The anthocyanins concentration for all the samples is presented in
Table 5.6.
Abs × MW × DF × 1000
[ Antocianinas ] = 5.1
µ× 1
Abs = ( A 515 nm − A 700 nm ) − ( A 515 nm − A 700 nm ) 5.2
pH1 pH 4.5

The maximum concentration of anthocyanins (20.74 mg/L) cor-

responds to a temperature of 40°C and a pressure of 200 bar. This
value is in agreement with other authors, who have found the value
of 68.42 mg/L (Acosta, 2017). Other authors have found that low pres-
sures can enhance the extraction yield of anthocyanins for it generates
low CO2 flows, which in turn can increase the mass transfer from veg-
etable matrix to the solvent (Porto and Natolino, 2017).
The relation between the temperature and the cosolvent/sample
established the postulation that at high temperature properties such
as pigmentation is lost due to loss of the glycosylated sugar, which is in
the third position in the molecule and produces colorless chaconnes,
therefore, the yield of the extraction should be carried out at low tem-
peratures (Gorriti Gutierrez et al., 2009; Porto and Natolino, 2017;
Colchao et al., 2011).

Table 5.6 Experimental Results of Anthocyanins

Concentration
Factor Results
Pressure Temperature Cosolvent/Sample Anthocyanins (mg/L)
1 −1 −1 12.847
1 1 −1 7.586
1 −1 1 12.919
1 1 1 6.278
−1 −1 −1 20.740
−1 1 −1 15.329
−1 −1 1 16.810
−1 1 1 15.580
0 0 0 12.874
0 0 0 15.607
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 167

5.4.3 Total Phenolic Compound Analysis

(Measurement and Analysis of the Total Phenolic
Compounds in the Obtained Extracts)
The TPCs turn out to be of a great importance in the secondary
metabolites of vegetables where they have multiple functions. These
compounds are considered antinutritive substances with the effect
of digestibility of proteins (Cueva et al., 2010; García et al., 2015). The
amount of polyphenols varies according to the species, sun exposure,
culture conditions, degree of maturity, storage that makes them pres-
ent with different rings of benzenes or hydroxyl groups in the structure
(García et al., 2015).
These molecules can be combined with sugars such as galactose,
xylose, or galacturonic acids. These type of compounds can be respon-
sible for sensory properties.
The method used in this process was Folin-Ciocalteu that stand-
out for the capacity to react with any type of phenol. The phenolic
compounds react with the Folin reagent which is formed by sodium
tungstate and sodium molybdate in acid medium. Thus, this reaction
produces a yellow complex that can be reduced with the phenolic
compounds and identified with a spectrophotometric method at a
wavelength of 765 nm (Cueva et al., 2010).
Thus, the method proposed by Singleton and Rossi Jr. (1965) was
used, with some modifications suggested by Cicco et al. (2009), where
it was necessary multiple dilutions were made (1:100, 1:50, 1:20, 1:10)
due to the concentration of phenols in the samples and was not al-
lowed to the ranges of the spectrophotometer as neither perform ac-
cording to the Lambert and Beer laws that has to be carried out when
using a spectrophotometric method.
For the analysis of TPCs, the samples obtained from supercritical
fluid extraction method were protected from light and stored at −20°C.
Then, each sample was taken to a centrifuge for a minimal time of
15 min and 4000 rpm for the particles to settle in suspension. Every re-
action was carried out in Eppendorf tubes of 2 mL each one perform-
ing the following steps:
The standard solution to this method was gallic acid, which has to
be diluted to values of 0–500 mg GA/L which is shown in calibration
curve (Fig. 5.3). It is used to realize the total concentration of phenols
in the samples analyzed. Then, 10 μL of the diluted sample was taken
and added to 1600 μL of distilled water finally completing with 100 μL
of the Folin reagent (1 N). These samples were shaken in a vortex and
kept at room temperature for approximately 8 min. After that, 200 μL
of sodium carbonate (20% W/V) was shaken and placed in the dark
for 60 min. At the end of this time, the absorbance reading was per-
formed on a UV/VIS spectrophotometer (Thermo Fisher Scientific,
168 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Fig. 5.3 Calibration curve for total phenolic compound measurements.

United States) at a wavelength of 765 nm. Phenolic compound content

is expressed as milligrams of gallic acid equivalent (GAE) per dry mass
unit (mg GAE/g). To know the phenolic content, it was determined in
units of milligrams of equivalent gallic acid for each unit of dry mass
(mg AGE/g).
The contents of TPCs are illustrated in Table 5.7. The higher con-
centration of TPC was obtained at 4200 psi and cosolvent to solid ratio
of 180 mL. According to the results, at higher temperatures the TPC
have to be solubilized because high temperatures increase the volatil-
ity of the solutes (TPC) present in the vegetable matrix, which in turn,
increases the solubility of the solvent. The high pressures (2400 and
4200 psi) used allow to increase the diffusivity in the solvent medium
and thus, improving the mass transfer since the cell walls and the

Table 5.7 Phenolic Compounds Results

Cosolvent: Solid Phenolic Content (mg A.G.E/g de
Relation (mL) Pressure (Psia) Temperature (°C) pulp)
60 4200 40 11.19
60 4200 60 19.75
180 4200 40 21.93
180 4200 60 24.16
60 2900 40 4.15
60 2900 60 4.93
180 2900 40 4.49
120 3550 50 12.44
180 2900 60 7.33
120 3550 50 13.45
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 169

molecules bond become weak due to the desorption process of the

solvent next to the solute tend to magnify.
The yield of the extraction depends strongly on the combination
of variables such as temperature, pressure, and cosolvent charge, to
obtain the best results for solubility and mass transfer (Espinosa-
Pardo et al., 2017). Thus, from this research, it is recommended that
intermediate pressures as well as the use of a cosolvent for is neces-
sary to obtain higher yields. In spite of the fact that high pressures can
improve the mass transfer due to best penetration in the plant matrix
and their pores, for generating a better drag of the solutes from the
inside of the matrix, it is necessary to have optimal conditions of all
variables that generate maximum solubilization of solutes, and thus,
attain higher yields (del P Garcia-Mendoza et al., 2017; Mustafa and
Turner, 2011).

5.4.4 Total Antioxidant Activity Analysis

(Measurement and Analysis of the Total Antioxidant
Activity in the Obtained Extracts)
The DPPH technique (2,2-diphenyl-1-picrylhydracil-Sigma
Aldrich) proposed by Brand-Williams et al. (1995) was used in this re-
search. Diluted samples in ethanol (1:10, 1:100, 1:1000) were shaken
on a vortex using a 60-μM DPPH and taken to an ultrasound bath for
5 min at 40 kHz (Branson 3800, United States). It is important to keep the
samples in the dark to protect the samples from sunlight. Afterwards,
75 μL of diluted samples were taken and mixed with 1425 μL of the
solution of DPPH previously prepared (saved and kept in the dark for
60 min), which allowed a reading at 515 nm in Evolution 300 UV/VIS
spectrophotometer (Thermo Fisher Scientific, United States). To know
the inhibition percent Eq. (5.3) was used where the Abs control is the
absorbance of the DPPH solution (60 μM) and Abs sample is the ab-
sorbance of the sample plus the DPPH solution (60 μM).
Abs Control − AbsSample
%Inhibition = ×100 5.3
Abs Control
The berry has been identified by its antioxidant capacity, the ranges
of antioxidant capacity of the extracts varied from 23.83 to 112.89 μmol
of Trolox/g pulp that is derived using high pressure (4200 psia), high
temperature (60°C), and 180 mL of cosolvent (ethanol). These con-
ditions improve significantly the extraction yield of phenolic com-
pounds using supercritical fluid extraction. Other authors have
evaluated the antioxidant capacity of extracts of Andes berry using
the DPPH method. Pasquel Reátegui et al. (2014) obtained 99.11 μmol
of Trolox/g pulp. Because the final characteristics of the extracts ob-
tained from the supercritical fluid extraction depend on the nature of
170 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

the raw material as well as the operational conditions (temperature,

pressure, flows, cosolvent use, etc.), it is necessary to explore a wide
range of the variables. Therefore, an optimal combination of these
variables results in an economic process as well as in an improvement
of the final characteristic of the extracts for human consumption.
Table 5.8 illustrates the antioxidant activities where the maximum
value registered was 112.89 μmol of Trolox/g pulp that corresponds to
the higher conditions of temperature and pressure. Therefore an in-
crease in pressure is directly related to the solvation power of CO2 and
thus, high pressures are suggested to obtain extracts rich in phenolic
compounds with high antioxidant capacities (Zulkafli et al., 2014b).
Fig. 5.4 depicts the relation between TPCs and antioxidant capacity
of the extracts of Andes berry. According to these results, 81.66% of the
antioxidant activity can be attributed to the phenolic compounds in
the extracts. Similar results have been found for Andes berry extracts
where 91% of the antioxidant activity is due to the phenolic com-
pounds content (Cerón et al., 2012).
According to the results (Table 5.8), the antioxidant activity of the
recovered extract increased as the temperature, pressure, and cosol-
vent ratio levels increase, similar to the behavior of the phenolic com-
pounds content. The extracts with the highest phenolic compounds
content also exhibited the maximum antioxidant activity, thus sug-
gesting again that high pressures and temperatures should be used to
obtain extracts with valuable characteristics. Besides, it became ev-
ident from the relation between TPCs and antioxidant capacity that
the phenolic compounds including anthocyanins should be extracted

Table 5.8 Antioxidant Capacity Results

Cosolvent: Solid Antioxidant Activity (μmol de
Relation (mL) Pressure (Psia) Temperature (°C) Trolox/g Pulp)
60 4200 40 39.75
60 4200 60 56.17
180 4200 40 97.56
180 4200 60 112.89
60 2900 40 23.79
60 2900 60 28.62
180 2900 40 23.83
120 3550 50 31.79
180 2900 60 30.93
120 3550 50 36.88
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 171

Fig. 5.4 Correlation between the phenolic compounds and the antioxidant activity.

because they are associated with the total antioxidant activity of the
extracts as other authors have also confirmed a correlation between
antioxidant activity and phenolic contents of 96.50% (Da Fonseca
Machado et al., 2015).

5.5 Conclusions
Beverage preparation can be carried out by several techniques,
however, supercritical fluids extraction is a promising technology
to take advantages of agroindustrial wastes since it allows to obtain
phenolic-rich extracts that also contains anthocyanins and important
antioxidant activity that suggest applications in different fields such as
pharmaceutical, food, and cosmetic industries. As it was shown here,
extracts obtained from supercritical fluid extraction technique have
valuable characteristics that can be used for beverage preparation and
should be refined according to the level of concentration and final pre-
sentation. Thus, processes such as evaporation, microfiltration, and
nanofiltration can be utilized for adding value to these extracts and
thus obtaining beverages with valuable applications.

References
Acosta, Ó., Pérez, A.M., Vaillant, F., 2009. Chemical characterization, antioxidant prop-
erties, and volatile constituents of naranjilla (Solanum quitoense Lam.) cultivated
in Costa Rica. Archivos Latinoamericanos de Nutrición. Arch. Latinoam. Nutr. 59
(1), 88–94.
Acosta, S.R.A., 2017. Obtencion de extractos ricos en antiocianinas a partir de los
residuo de mora (Rubus glaucus benth) usando Co2 supercritico. Facultad de cien-
cias naturales e ingenieria. Universidad Jorge Tadeo Lozano, Bogotá.
Alarcón García, M.Á., López Vargas, J.H., Restrepo Molina, D.A., 2015. Agro-industrial
fruit co-products in Colombia, their sources and potential uses in processed food
industries: a review. Rev. Facul. Nac. Agronomía 68, 7729–7742.
172 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Almeida, M.M.B., et al., 2011. Bioactive compounds and antioxidant activity of fresh ex-
otic fruits from northeastern Brazil. Food Res. Int. 44 (7), 2155–2159.
ArunaDevi, R., et al., 2010. Neuroprotective effect of 5,7,3′,4′,5′-pentahydroxy
dihdroflavanol-3-O-(2″-O-galloyl)-β-d-glucopyranoside, a polyphenolic com-
pound in focal cerebral ischemia in rat. Eur. J. Pharmacol. 626 (2–3), 205–212.
Bonilla-Lavado, H.A., Vasquez-Acosta, N.B., Rubiano-Rodríguez, J.A., 2006. Evaluación
de Residuos Orgánicos (Coco y Aserrín) como Sustratos para la Producción de
Pleurotus ostreatus (Jacq: Fr.) en Buenaventura. Revista Institucional. Universidad
Tecnológica del Chocó D. L. C. 24, 54–59.
Botero Echeverri, A.I., 2008. Aplicación de la Ingeniería de Matrices en el desarrollo
da la uchuva mínimamente procesada fortificada con calcio y vitaminas C y E.
Facultad de química farmacéutica, Magíster en ciencias farmacéuticas énfasis en
alimentos, Medellín185.
Brand-Williams, W., Cuvelier, M.E., Berset, C., 1995. Use of a free radical method to
evaluate antioxidant activity. Food Sci. Technol. 28, 25–30. https://doi.org/10.1016/
S0023-6438(95)80008-5.
Brunner, G., 2005. Supercritical fluids: technology and application to food processing.
J. Food Eng. 67, 21–33.
Carlsen, M.H., et al., 2010. The total antioxidant content of more than 3100 foods, bever-
ages, spices, herbs and supplements used worldwide. Nutr. J. 9, 3.
Castellanos, F.J., Lucas, J.C., 2011. Caracterización Física del Fruto en Variedades
de Plátano Cultivadas en la Zona Cafetera en Colombia. Acta Agronóm. 60 (2),
176–182.
Cedeño, M.M., Montenegro, D.M., 2004. Plan Exportador, Logistico y de Comercialización
de Uchuva al Mercado de los Estados Unidos para Frutexpo S.C.I. LTDA. Pontificia
Universidad Javeriana.
Cerón, I.X., 2013. Design and Evaluation of Processes to Obtain Antioxidant-Rich
Extracts from tropical fruits cultivated in Amazon, Caldas and Northern Tolima
Regions. Faculty of Engineer and Architecture. Department of Electronic and
Electric Engineers. Universidad Nacional de Colombia at Manizales, Manizales.
Cerón, I.X., Higuita, J.C., Cardona, C.A., 2010. Capacidad antioxidante y contenido
fenólico total de tres frutas cultivadas en la región andina. Vector 5, 7–16.
Cerón, I.X., Higuita, J.C., Cardona, C.A., 2012. Design and analysis of antioxidant com-
pounds from Andes Berry fruits (Rubus glaucus Benth) using an enhanced-fluidity
liquid extraction process with CO2 and ethanol. J. Supercrit. Fluids 62, 96–101.
Cerón, I.X., et al., 2014. Process synthesis for antioxidant polyphenolic compounds pro-
duction from Matisia cordata Bonpl. (zapote) pulp. J. Food Eng. 134, 5–15.
Chirinos, R., et al., 2010. Antioxidant compounds and antioxidant capacity of Peruvian
camu camu (Myrciaria dubia (H.B.K.) McVaugh) fruit at different maturity stages.
Food Chem. 120 (4), 1019–1024.
Chumsri, P., Sirichot, A., Itharat, A., 2008. Studies on the optimum conditions for
the extraction and concentration of roselle (Hibiscus sabdariffa Linn.) extract.
Songklanakarin J. Sci. Technol. 30 (1), 133–139.
Cicco, N., Lanorte, M.T., Paraggio, M., Viggiano, M., Lattanzio, V., 2009. A reproducible,
rapid and inexpensive Folin-Ciocalteu micro-method in determining phenolics of
plant methanol extracts. Microchem. J. 91 (1), 107–110. https://doi.org/10.1016/j.
microc.2008.08.011.
Colchao, V., Miranda, A., Castañeda, D., 2011. Evaluación de la estabilidad de antocian-
inas durante el almacenamiento de zanahoria púrpura (Daucus carota). Chimbote.
Collins, A.R., 2005. Antioxidant intervention as a route to cancer prevention. Eur. J.
Cancer 41 (13), 1923–1930.
Contreras-Calderón, J., et al., 2011. Antioxidant capacity, phenolic content and vitamin
C in pulp, peel and seed from 24 exotic fruits from Colombia. Food Res. Int. 44 (7),
2047–2053.
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 173

Corbo, M.R., et al., 2014. Functional beverages: the emerging side of functional foods.
Compr. Rev. Food Sci. Food Saf. 13 (6), 1192–1206.
Cuéllar, O.A., 2010. Obtención del Extracto Polar Etanol: Agua (1:1) de la Cáscara de Cacao
y Evaluación de su Actividad Antibacteriana. Universidad Tecnológica De Pereira.
Cueva, A., Medina, R.A., Lorach, R., Urpi-Sarda, M., Khan, N., Chiva-Blanch, G., Zamora-
Ros, R., Rotches-Ribalta, M., Larnuela-Raventós, R.M., 2010. Phenolic compounds:
chemistry and occurrence in fruits and vegelables. In: De la Rosa, L.A., Alvarez-
Parrilla, E., González-Aguilar, G.A. (Eds.), Fruit and Vegetable Phytochemicals:
Chemistry, Nutritional Value and Stability. first ed.. Blackwell Publishing, lowa.
Da Fonseca Machado, A.P., Pasquel Reátegui, J.L., Fernández Barbero, G., Martínez, J.,
2015. Pressurized liquid extraction of bioactive compounds from blackberry (Rubus
fruticosus L.) residues: a comparison with conventional methods. Food Res. Int. 77,
675–683. https://doi.org/10.1016/j.foodres.2014.12.042.
Dávila, J.A., Rosenberg, M., Cardona, C.A., 2017. A biorefinery for efficient processing
and utilization of spent pulp of Colombian Andes Berry (Rubus glaucus Benth.): ex-
perimental, techno-economic and environmental assessment. Bioresour. Technol.
223, 227–236.
Dávila, J.A., et al., 2014. Design of processes for supercritical extraction and microencap-
sulation of antioxidants from fruits. In: Osborne, J. (Ed.), Handbook of Supercritical
Fluids. Fundamentals, Properties and Applications. Nova Publishers, New Delhi.
De Cássia Rodrigues Batista, C., De Oliveira, M.S., Araújo, M.E., Rodrigues, A.M.C., Botelho,
J.R.S., Da Silva Souza Filho, A.P.…Carvalho, R.N., 2015. Supercritical CO2 extraction of
açaí (Euterpe oleracea) berry oil: global yield, fatty acids, allelopathic activities, and
determination of phenolic and anthocyanins total compounds in the residual pulp. J.
Supercrit. Fluids 107, 364–369. https://doi.org/10.1016/j.supflu.2015.10.006.
de Fonseca, E., 2009. In: Metano En Residuos Domésticos. II Simposio Iberoamericano
de Ingeniería de Residuos : Barranquilla, Colombia.
de Mesquita, M.L., et al., 2009. Cytotoxic activity of Brazilian Cerrado plants used in
traditional medicine against cancer cell lines. J. Ethnopharmacol. 123 (3), 439–445.
del P Garcia-Mendoza, M., Espinosa-Pardo, F.A., Baseggio, A.M., Barbero, G.F.,
Maróstica Junior, M.R., Rostagno, M.A., Martínez, J., 2017. Extraction of phenolic
compounds and anthocyanins from juçara (Euterpe edulis Mart.) residues using
pressurized liquids and supercritical fluids. J. Supercrit. Fluids 119, 9–16. https://
doi.org/10.1016/j.supflu.2016.08.014.
Díaz, C.A., 2011. Categorización de la latencia en semillas de mora (Rubus glaucus
Benth.), para el apoyo a programas de mejoramiento y conservación de la especie.
Facultad de Ciencias Agrarias. Universidad Nacional de Colombia, Manizales.
Domínguez, L., Parzanese, M., 2005. Fluidos supercríticos. Aliment. Argentino. 1, 1–12.
Duque, S.H., Cardona, C.A., Moncada, J., 2015. Techno-economic and environmental
analysis of ethanol production from 10 agroindustrial residues in Colombia. Energy
Fuel 29 (2), 775–783.
Durán, J.D., Méndez, G.A., 2008. Plan de Negocios para Exportar Maracuyá y Cholupa
como Fruta Fresca y/o en Pulpa hacia Canadá. Pontificia Universidad Javeriana.
Egan, B.M., Greene, E.L., Goodfriend, T.L., 2001. Insulin resistance and cardiovascular
disease. Am. J. Hypertens. 14 (6), S116–S125.
Escalante, H., et al., 2007. Atlas del Potencial Energético de la Biomasa Residual en
Colombia. Ministerio de Minas y Energía.
Espinal, C.F., Martínez, H.J., Peña, Y., 2005. La Cadena de Cítricos en Colombia,
Documento de Trabajo No. 107. Ministerio de Agricultura y Desarrollo Rural
Observatorio Agrocadenas Colombia.
Espinosa-Pardo, F.A., Nakajima, V.M., Macedo, G.A., Macedo, J.A., Martinez, J., 2017.
Extraction of phenolic compounds from dry and fermented orange pomace using
supercritical CO2 and cosolvents. Food Bioprod. Process. 101, 1–10. https://doi.
org/10.1016/j.fbp.2016.10.002.
174 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Fischer, G., et al., 2005. Avances en cultivo, Poscosecha y Exportación de la Uchuva

Physalis peruvirana L. en Colombia. Universidad Nacional de Colombia.
Flora, S.J.S., 2009. Structural, chemical and biological aspects of antioxidants for strat-
egies against metal and metalloid exposure. Oxidative Med. Cell. Longev. 2 (4),
191–206.
Forster, G.M., et al., 2013. Rice varietal differences in bioactive bran components for in-
hibition of colorectal cancer cell growth. Food Chem. 141 (2), 1545–1552.
Franco, G., Giraldo, M.J., 1998. El Cultivo de la Mora. Corporación Colombiana de inves-
tigación Agropecuaria y Regional.
Franzini, L., et al., 2012. Food selection based on high total antioxidant capacity im-
proves endothelial function in a low cardiovascular risk population. Nutr. Metab.
Cardiovasc. Dis. 22 (1), 50–57.
García, A.M., Torres, R.G., 2003. Producción de Enzimas Lignolíticas por Basidiomycetes
Mediante la Técnica de Fermentación en Sustrato Sólido. Rev. Colomb. Biotecnol.
5, 56–64.
García M.E. , Fernández S. I., Fuentes L. A., 2015. Determinación de poli fenoles totales
por el método de Folin- Ciocalteu , ETSIAMN. Universitat Politècnica de València,
Departamento de Tecnología de Alimentos.
Garzón, G.A., 2008. Las antocianinas como colorantes naturales y compuestos bioacti-
vos: Revisión. Acta Biol. Colomb. 13 (3), 27–36.
Gene, R.M., Marlin, E., Adzet, T., 1992. Anti-inflammatory effect of aqueous extracts of
three spices of the genus baccharis. Planta Med. 58, 565–566.
Genovese, M.I., Da-Silva-Pinto, M., De-Souza-Schmid Gonçalves, A.E., 2008. Bioactive
compounds and antioxidant capacity of exotic fruits and commercial frozen pulps
from Brazil. Food Sci. Technol. Int. 14, 207–214.
Giusti, M.M., Wrolstad, R.E., 2001. Characterization and measurement of anthocyanins
by UV-Visible spectroscopy. Curr. Protocol. Food Anal. Chem. 1–13. https://doi.
org/10.1002/0471142913.faf0102s00.
González, I.A., 2010. Caracterización Química del Color de Diferentes Variedades de
Guayaba (Psidium Guajava L.) Colombiana. Universidad Nacional de Colombia.
Gorriti Gutierrez, A., Arroyo Acevedo, J., Cordova, A., Jurado Teixeira, B., 2009.
Extracción de antocianinas de las corontas de Zea mays L. Maiz Morado. Ciencia
E Invest. 12 (2), 64–74.
Gupta, P., et al., 2015. Food processing residue analysis and its functional components as
related to human health: recent developments. Austin J. Nutr. Food Sci. 3 (3).
Habib, H.M., et al., 2013. Camel milk lactoferrin reduces the proliferation of colorec-
tal cancer cells and exerts antioxidant and DNA damage inhibitory activities. Food
Chem. 141 (1), 148–152.
Sources for lignocellulosic raw materials for the production of ethanol. In: Hadar, Y.,
Faraco, V. (Eds.), Lignocellulose Conversion. 2013. Springer, Berlin Heidelberg, pp.
21–38.
Hassimotto, N.M.A., et al., 2008. Physico-chemical characterization and bioactive
compounds of blackberry fruits (Rubus sp.) grown in Brazil. Food Sci. Technol.
(Campinas) 28, 702–708.
Hernández, R.A., López, C.L., 2007. Evaluación del Crecimiento y Producción de
Pleurotus Ostreatus Sobre Diferentes Residuos Agroindustriales del Departamento
de Cundinamarca. Pontificia Universidad Javeriana.
Koolen, H.H.F., et al., 2013. Antioxidant, antimicrobial activities and characterization
of phenolic compounds from buriti (Mauritia flexuosa L. f.) by UPLC–ESI-MS/MS.
Food Res. Int. 51 (2), 467–473.
Kris-Etherton, P.M., et al., 2002. Bioactive compounds in foods: their role in the pre-
vention of cardiovascular disease and cancer. Am. J. Med. 113 (9, Supplement 2),
71–88.
Lobo, V., et al., 2010. Free radicals, antioxidants and functional foods: impact on human
health. Pharmacogn. Rev. 4 (8), 118–126.
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 175

López, A., et al., Comparación de diferentes procesos para el aprovechamiento de residuos

agroindustriales de naranja (citrus sinensis) variedad valencia mediante análisis mul-
tivariada, in II Simposio Internacional y III Nacional Agroalimentario 2011, Colombia.
Machado, A.P.D.F., Pasquel-Reátegui, J.L., Barbero, G.F., Martínez, J., 2015. Pressurized
liquid extraction of bioactive compounds from blackberry.(Rubus fruticosus L.) res-
idues: A comparison with conventional methods. Food Res. Int. 77, 675–683.
Malta, L.G., et al., 2013. Assessment of antioxidant and antiproliferative activities and
the identification of phenolic compounds of Exotic Brazilian fruits. Food Res. Int.
53 (1), 417–425.
Márquez, C.J., 2009. Caracterización Fisiológica, Físico-Química, Reológica,
Nutraceútica, Estructural y Sensorial De La Guanábana (Annona Muricata L. Cv.
Elita). Universidad Nacional de Colombia.
McCune, L.M., Johns, T., 2002. Antioxidant activity in medicinal plants associated with
the symptoms of diabetes mellitus used by the Indigenous Peoples of the North
American boreal forest. J. Ethnopharmacol. 82 (2-3), 197–205.
Mejía, L.F., et al., 2007. Aprovechamiento del residuo Agroindustrial del Mango común
(Mangifera indica L.) en la Obtención de Azucares Fermentables. Ing. Ciencia 3 (6), 41–62.
Ministerio de Agricultura y Desarrollo Rural and Corporación Colombia Internacional,
2001. Inteligencia de Mercados: Uchuva.
Ministerio de Agricultura y Desarrollo Rural and Corporación Colombia Internacional,
2002. Inteligencia de Mercados: Piña.
Ministerio de Agricultura y Desarrollo Rural and Corporación Colombia Internacional,
2003. Inteligencia de Mercados: Maracuyá.
Moncada, J., Cardona, C.A., Pisarenko, Y.A., 2013. Solubility of some phenolic acids con-
tained in citrus seeds in supercritical carbon dioxide: comparison of mixing rules,
influence of multicomponent mixture and model validation. Theor. Found. Chem.
Eng. 47 (4), 381–387.
Mongabay, 2010. Colombia: Tropical Rainforest. Avaliable from: http://rainforests.
mongabay.com/20colombia.htm.
Motato, K.E., Mejía, A.I., León, A., 2006. Evaluación de los Residuos Agroindustriales
de Plátano (Musa Paradisíaca) y Aserrín de Abarco (Cariniana Piriformes) como
Sustratos para el Cultivo del Hongo Pleurotus djamor. Rev. Vitae 13 (1).
Mustafa, A., Turner, C., 2011. Pressurized liquid extraction as a green approach in food
and herbal plants extraction: a review. Anal. Chim. Acta 703 (1), 8–18. https://doi.
org/10.1016/j.aca.2011.07.018.
Navarrete, C., et al., 2010. Extracción y Caracterización del Aceite Esencial de Mandarina
Obtenido de Residuos Agroindustriales. Dyna 77 (162), 85–92.
Ospina, S.M., Hernández, E., Lozano, C.A., 2012. Estudio Experimental del Proceso de
Fermentación de Residuos Agroindustriales del Mango (Mangifera Indica L) dsando
Saccharomyces cerevisiae. In: Bacteriología. Universidad Católica de Manizales.
Pandey, A., et al., 2000. Biotechnological potential of coffee pulp and coffee husk for
bioprocesses. Biochem. Eng. J. 6 (2), 153–162.
Pasquel Reátegui, J.L., Machado, A.P.D.F., Barbero, G.F., Rezende, C.A., Martínez, J.,
2014. Extraction of antioxidant compounds from blackberry (Rubus sp.) bagasse us-
ing supercritical CO2 assisted by ultrasound. J. Supercrit. Fluids 94, 223–233. https://
doi.org/10.1016/j.supflu.2014.07.019.
Porto, D., Natolino, A., 2017. Supercritical fluid extraction of polyphenols from grape
seed (Vitis vinifera): study on process variables and kinetics. J. Supercrit. Fluids
130, 239–245. Available from: http://www.sciencedirect.com/science/article/pii/
S0896844616304259.
Procolombia, 2016. The World Invests in Colombia. Investment in the Fruit and
Vegetable Sector. Procolombia.
Ramful, D., et al., 2010. Bioactive phenolics and antioxidant propensity of flavedo ex-
tracts of Mauritian citrus fruits: Potential prophylactic ingredients for functional
foods application. Toxicology 278 (1), 75–87.
176 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES

Repo-de-Carrasco, R., Encina Zelada, C.R., 2008. Determinación de la capacidad anti-

oxidante y compuestos bioactivos de frutas nativas Peruanas. Rev. Soc. Quím. Perú
74 (2), 108–124.
Restrepo Duque, A.M., 2008. Nuevas perspectivas de consumo de frutas: Uchuva
(Physalis peruviana L.) y Fresa (Fragaria vesca L.) mínimamente procesadas forti-
ficadas con vitamina E. Facultad de Ciencias Agropecuarias, Universidad Nacional
de Colombia, Medellin107.
Revelo, V.H., 2011. Evaluación de la calidad poscosecha en genotipos mejorados e injer-
tos de tomate de árbol (Solanum betaceum Cav.). Facultad de Ingeniería Química y
Agroindustria, Escuela Politécnica Nacional.
Rodríguez, N., 2009. Manejo de residuos en la industria cafetera. Seminario Internacional
Gestión integral de residuos sólidos y peligrosos.
Rodríguez, O.V., Hanssen, H., 2007. Obtención de Dextrano y Fructosa Utilizando
Residuos Agroindustriales con la Cepa Leuconostoc mesenteroides NRRL B512-F.
Rev. EIA 7, 159–172.
Rufino, M.D.S.M., et al., 2010. Bioactive compounds and antioxidant capacities of 18
non-traditional tropical fruits from Brazil. Food Chem. 121 (4), 996–1002.
Sahena, F., Zaidul, I.S.M., Jinap, S., Karim, A.A., Abbas, K.A., Norulaini, N.A.N., Omar,
A.K.M., 2009. Application of supercritical CO2 in lipid extraction: a review. J. Food
Eng. 95 (2), 240–253.
Sanchez, P.A., Ibañez, E., Cifuentes, A., Herrero, M., 2017. Chapter two—bioactives ob-
tained from plans, seaweed, microalgae and Food by-products using pressurized
Liquid extraction and supercritical fluid extraction. In: Comprehensive Analutical
Chemistry , Institure of Food sience Research (CIAL-CSIC) Madrid. vol. 76. pp.
27–51.
Sánchez, W., Murillo, E., 2010. Potencial Antioxidante de Residuos Agroindustriales de
Tres Frutas de Alto Consumo en el Tolima. Sci. Technica 17 (46).
Santacruz, C.J., 2016. Ing. Alimentos Evaluación De La Calidad Funcional Del Aceite
Obtenido A Partir De Granos De Café Verde Defectuoso (Coffea Arábica) Mediante
Co2 Supercrítico. Facultad de ciencias naturales e ingenieria. Universidad Jorge
Tadeo Lozano.
Serra Bonvehı́, J., Ventura Coll, F., 1999. Protein quality assessment in cocoa husk. Food
Res. Int. 32 (3), 201–208.
Shahidi, F., Alasalvar, C., 2016. Handbook of Functional Beverages and Human Health.
CRC Press.
Singleton, V.L., Rossi Jr., J.A., 1965. Colorimetry of total phenolics with phosphomolybdic-
phosphotungstic acid reagents. Am. J. Enol. Vitic. 16 (3), 144–158. https://doi.
org/10.12691/ijebb-2-1-5.
Torres, V.I., 2010. Determinación del Potencial Nutritivo y Funcional de Guayaba
(Psidium guajava L), Cocona (Solanum Sessiliflorum Dunal) y Camu Camu
(Myrciaria dubia Vaugh). Escuela Politécnica Nacional.
Tsai, C.-F., et al., 2013. Growth inhibition and antioxidative status induced by selenium-
enriched broccoli extract and selenocompounds in DNA mismatch repair-deficient
human colon cancer cells. Food Chem. 139 (1–4), 267–273.
Vasco, C., et al., 2009. Physical and chemical characteristics of golden-yellow and
purple-red varieties of tamarillo fruit (Solanum betaceum Cav.). Int. J. Food Sci.
Nutr. 60 (S7), 278–288.
Villegas, V., Pérez, A.M., Arredondo, C., 2007. Evaluación de la producción del Hongo
Lentinula edodes Pegler en Bloques sintéticos a base de residuos agroindustriales.
Ing. Ciencia 3 (6), 23–39.
Wang, S., et al., 2011. How natural dietary antioxidants in fruits, vegetables and legumes
promote vascular health. Food Res. Int. 44 (1), 14–22.
Wu, S.J., et al., 2006. Supercritical carbon dioxide extract exhibits enhanced antioxidant and
anti-inflammatory activities of Physalis peruviana. J. Ethnopharmacol. 108 (3), 407–413.
 Chapter 5 FRUIT AGROINDUSTRIAL WASTES FOR PREPARING BEVERAGES 177

Yepes, S.M., Montoya, L.J., Orozco, F., 2008. Valorización de Residuos Agroindustriales–
Frutas–en Medellín y el Sur del Valle del Aburrá, Colombia. Rev. Facult. Nacional
Agronomía Medellín 61 (1), 4422–4431.
Zhang, C.-Y., et al., 2013. Curcumin inhibits invasion and metastasis in K1 papillary thy-
roid cancer cells. Food Chem. 139 (1–4), 1021–1028.
Zulkafli, Z.D., Wang, H., Miyashita, F., Utsumi, N., Tamura, K., 2014a. Cosolvent-modified
supercritical carbon dioxide extraction of phenolic compounds from bamboo
leaves (Sasa palmata). J. Supercrit. Fluids 94, 123–129. https://doi.org/10.1016/j.
supflu.2014.07.008.
Zulkafli, Z.D., Wang, H., Miyashita, F., Utsumi, N., Tamura, K., 2014b. Cosolvent-modified
supercritical carbon dioxide extraction of phenolic compounds from bamboo
leaves (Sasa palmata). J. Supercrit. Fluids 94, 123–129. https://doi.org/10.1016/j.
supflu.2014.07.008.

Further Reading
Paes, J., Dotta, R., Barbero, G.F., Martínez, J., 2014. Extraction of phenolic compounds
and anthocyanins from blueberry (Vaccinium myrtillus L.) residues using su-
percritical CO2 and pressurized liquids. J. Supercrit. Fluids 95, 8–16. https://doi.
org/10.1016/j.supflu.2014.07.025.
Vatai, T., Škerget, M., Knez, Ž., 2009. Extraction of phenolic compounds from elder
berry and different grape marc varieties using organic solvents and/or supercrit-
ical carbon dioxide. Int. J. Food Eng. 90 (2), 246–254. ISSN 0260-8774 https://doi.
org/10.1016/j.jfoodeng.2008.06.028.
 LA FUNDACIÓN UNIVERSIDAD DE BOGOTÁ JORGE TADEO LOZANO

CERTIFICA

Que el libro titulado: “Production and Management of Beverages” con número de

ISBN: 978-0-12-815260-7, publicado en Nueva York 2021, en el que Javier Andrés
Dávila Rincón identificado con cédula de ciudadanía No. 9.975.578 y otro son
autores del capítulo de libro: “Fruit Agroindustrial Wastes for Preparing Beverages
for Medicinal Purposes by Supercritical Fluid Extraction Technology: Andes Berry
(Rubus glaucus benth) Case”, fue financiado en su publicación por la editorial
Elsevier.

Este certificado se expide a solicitud del investigador, en Bogotá D.C., a los

veintisiete (27) días del mes de agosto de 2021.

LINA MARÍA CEPEDA MELO

Secretaria General y Representante Legal Suplente