Contemporary Engineering Sciences, Vol. 11, 2018, no.

37, 1837 - 1844

HIKARI Ltd, www.m-hikari.com
https://doi.org/10.12988/ces.2018.84158

Statistical Analysis of the Supercritical Fluid Oil

Extraction from Grape Seed

Diofanor Acevedo Correa1, Piedad Montero Castillo1

and Marlene Duran-Lengua2
1
Faculty of Engineering, Research Group Innovation, Agricultural and
Agroindustrial Development, University of Cartagena
Av. El Consulado, St. 30 No. 48-152, Colombia
2
School of Medicine, Zaragocilla Campus, Universidad de Cartagena
130015 Cartagena de Indias, D. T. y C., Colombia

Copyright © 2018 Diofanor Acevedo Correa, Piedad Montero Castillo and Marlene Duran-
Lengua. This article is distributed under the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work
is properly cited.

Abstract

Traditional methods for the extraction of edible oils from natural matrices
regularly require more than one extraction step with organic solvents.
Furthermore, additional process steps are necessary during the refining.
Consequently, faster processing methods that are compatible with food production
are extremely important. Supercritical carbon dioxide extraction of edible oils is
an alternative to solvent extraction because it provides a high-speed extraction
process with a simple purification stage. In the study, the effect of the main
operating variables during the supercritical fluid extraction (SFE) of oil from
grape seeds was statically analysed. For this, the oil extraction yield oil was
determined as a function of pressure (200 bar and 300 bar), temperature (40 °C
and 60 °C), solvent flow (1 g/min and 5 g/min) and particle size (0.4 mm and 0.80
mm). The statistical analysis showed that the extraction yield increased with an
increase in pressure and mass flow, and on the contrary, with a reduction in
temperature and size; pressure and temperature being the most influential factors,
in that order. The results demonstrate that it is necessary to work at a pressure of
300 bar and a temperature of 40 °C to obtain a significant yield in the extraction
of seed oil.

Keywords: p-value, analysis of variance, factorial design, Vitis labrusca,

statistical effects
1838 Diofanor Acevedo Correa et al.

1. Introduction
Grapes are one of the fruit crops grown widely in many areas of the world and 46
% of the fresh grapes produced are accounted for in wine production [1]. Since
about 80 % of the total amount is used in winemaking, some 10 million tons of
grape pomace arise within a few weeks of the harvest campaign. The seeds
constitute a considerable proportion of the pomace, amounting to 38 % –52 % on
a dry matter basis [2].

Grape seed oil is a product resulting from the processing of grape marc. It is ever
more appealing for the cosmetic and pharmaceutical industries: the
pharmaceutical activities concern its antioxidant properties, due to the large
content of unsaturated fatty acids [3], which represents positive benefits on the
human health, such as antimutagenic and antiviral activity or inhibition of the
oxidation of low-density lipoproteins [4]. Apart from being a rich source of a
high-value fatty oil, grape seeds have also been appreciated because of their
content of phenolic compounds such as gallic acid, catechin and epicatechin, and
a wide variety of procyanidins. The latter is also referred to as condensed tannins.
Grape seed extracts and procyanidins have been a matter of intense investigations
with respect to their potentially beneficial effects on human health [2].

Usually, the production of edible oils from vegetable seeds comprehend several
stages during the industrial processes, where the extraction with n-hexane is an
important one [5]. However, this organic solvent is non-selective and
simultaneously removes non-volatile pigments and waxes, giving rise to dark
coloured and viscous extracts contaminated with solvent residues [6].

The application of supercritical carbon dioxide (sc-CO2) has received much

attention in the food industry in the last few years, taking into account that this
procedure offers extraction yields comparable with those obtained by
conventional extraction methods using organic solvents [7]. Additionally,
differently than with organic solvents such n-hexane, CO2 is non-toxic, non-
flammable, non-corrosive, cheap and readily available in large quantities with
high purity. Furthermore, CO2 also has a relatively low critical pressure (73.77
bar) and critical temperature (30.98 °C), it can be considered an ideal solvent for
the treatment of natural products [8].

Finally, the statistical analysis is a scientific approach to analysing numerical data

in order to enable us to maximise our interpretation, understanding and use [9].
This means that statistics help us turn data into information; that is, data that have
been interpreted, understood and are useful [10]. Therefore, considering that
statistics is the systematic collection and analysis of numerical data, to investigate
relationships among phenomena so as to explain, predict and control their
occurrence [11], the aim of this study was to analyse statistically the effect of the
main operating variables during the supercritical fluid extraction (SFE) of oil from
grape seeds.
Statistical analysis of the supercritical fluid oil extraction 1839

2. Methodology
2.1 Raw material

Grape (Vitis labrusca) from southwest Colombia were used for this work. At the
begging, stalks were separated from the seeds and skins at the winery. The
mixture of seeds and skins was stored at – 20 °C before drying. The samples were
dried at 60 °C for 48 h, and then the skins and seeds were separated by means of
sieves and further cleaned manually. The seeds were stored in dark under vacuum
at ambient temperature and finally, dried grape seeds were milled for two cycles
of 10 s and then sieved [5,12].

2.2 Installation and operation procedure

All the extractions resulted from the experimental design were done in a
homemade semi-continuous flow apparatus that consisted of a pure CO2 feed line
and an SFE equipment with 50 mL. Complete details of the equipment used were
described in a previous report [12]. A heating jacket was placed around it to
control the temperature inside. The CO2 flow rate was fixed with a pump, while
pressure was controlled by a backpressure regulator. The quantity of CO2 was
measured using a mass-flow meter that was placed at the end of the line. An
approximate amount of 5 g of sample was introduced in the vessel forming a
fixed. A wire mesh was placed on top of the bed to prevent particles entrainment.
The extractor was then closed and pre-heated. After that, the CO2 was pumped in,
and, once the desired pressure was reached, the back-pressure regulator (BPR)
was opened to provide a continuous flow of the CO2 through the bed. The CO2
was distributed through the bed and after the BPR, this was depressurized; the
solvent power of the supercritical fluid dropped, and the oil was precipitated in a
previously weighed glass flask. The amount of CO2 circulated was read by the
mass flow meter. When the operation was over, the apparatus was depressurized
and the exhausted sample unloaded. After each treatment, lines were cleaned
before to start the next experiment. The experimental error was deduced from
selected tests that were repeated six times. The maximum standard deviation in
extraction yield was 1.5 %.

For a better comparison of the oil yield extractions, the operation was over at the
same consumption of CO2 (3000 g) for all the treatments. This, to recollect the
sample at the end of the line stage of extraction. For this, a previous extraction
curve was done at the highest pressure (300 bar), the lowest temperature (40 °C),
a mass flow of 5 g/min and a particle diameter of 0.40 mm.

2.3 Experimental design

A general factorial design 24 was carried out. Pressure (200 bar and 300 bar),
temperature (40 °C and 60 °C), solvent flow (1 g/min and 5 g/min) and particle
diameter (0.4 mm and 0.80 mm) were considered as the design factors. A total of
1840 Diofanor Acevedo Correa et al.

16 experiments and its duplicates resulted. The only response variable was the oil
extraction yield, expressed as milligram of dry oil extract per gram of dry sample.
Table 1 shows the factorial experimental design, levels, density values at the
conditions tested and the results obtained for all the experiments.

2.4 Statistical analysis

Statistical analysis was made by using the Statgraphics Centurion XVII® software
with analysis of variance (ANOVA). All mean values were analysed by a
multifactorial ANOVA. When a factor had a p-value smaller than 0.05, its
influenced the process in a significant way for a confidence level of 95 %.

4. Results

The oil extraction yields are shown in Table 1 and are expressed as percent dry
weight/initial weight. These values were obtained for the same consumption of
CO2 (3000 g) for the different extraction conditions studied. For information, a
column including the CO2 density has been included.

Table 1. Factorial experimental design and oil extraction yield obtained

Run P T CO2 density Mass flow Particle diameter Yield
(bar) (°C) (kg/m3) (g/min) (mm) (%)
1 1 0.40 59.01
2 40 839.81 0.80 53.96
3 5 0.40 65.99
4 200 0.80 56.09
5 1 0.40 51.02
6 60 723.68 0.80 46.59
7 5 0.40 52.94
8 0.80 47.98
9 1 0.40 98.01
10 40 909.89 0.80 91.04
11 5 0.40 105.92
12 300 0.80 95.18
13 1 0.40 86.08
14 60 829.71 0.80 79.05
15 5 0.40 87.04
16 0.80 80.99

On the other hand, the estimated effects between the range of variables studied
and the analysis of variance of the extraction process are given in Table 2. The
sign associated with each of the effects indicates a positive or negative influence
on the yield of the dependent variable. The degree of significance of each factor is
represented in the table by its p-value. When a factor had a p-value smaller than
0.05 it influenced the process in a significant way for a confidence level of 95 %.
Statistical analysis of the supercritical fluid oil extraction 1841

Table 2. Estimated effects and the analysis of variance of the process for the oil
extraction yields with supercritical carbon dioxide

Variable Effects p-Value

Pressure 0.98 0.01
Temperature -0.56 0.02
Mass flow 0.12 0.04
Particle diameter -0.23 0.03

The results obtained in Table 2 show that pressure, temperature, mass flow and
particle diameter had a statistically significant influence on the oil extraction
process (p < 0.05). Pressure and mass flow had a positive influence on the yield.
On the other hand, temperature and particle size had a negative influence on the
extraction yield. This influence was increasing by the order: pressure >
temperature > particle diameter > mass flow.

It could be observed in Table 1 that, for a given temperature, the oil extraction
yield increased with pressure. The effect of sc-CO2 pressure on the yield
extraction is well known and there is an agreement in the research community that
increasing operating pressure has a positive effect on the oil yield extraction
[4,13]. The reason is that an increase in pressure at constant temperature makes
the density of sc-CO2 increase, which enhances its solvent power and, ultimately,
the oil yield extraction if all the other parameters are kept constant. However, it
should be taken into account that the economic viability of working at high
pressure has to be evaluated because any increase in pressure is related to a rise in
energy consumption [5]. In the case of seed oil extraction, it seems convenient to
use the sc-CO2 according to some works [5–7,12,14–16].

It should also take into account that the cellular walls of the sample used had not a
great resistance to the mass transfer. This phenomenon explains the lack of
variation observed in the yields obtained in the oil extraction within the pressure
range selected for each of the temperatures studied.

On the other hand, it could be seen that, for each pressure, an increase in the
temperature generally produced a reduction in the yield of the oil extraction. This
yield depended on the decrease caused in the density of the sc-CO2, which was
expected [17].

The higher was the flow rate, the higher the extraction yield, in line with an
increase in both the external and internal mass transfer parameters. This means
that the higher is the flow rate, the lower could be the extraction time.
Nevertheless, the specific consumption of the solvent increased at increasing sc-
CO2 flow rate, which would imply that, for commercial purposes, the solvent flow
rate should be optimized in terms of extraction time and solvent volume used per
operation [5].
1842 Diofanor Acevedo Correa et al.

Finally, in particles with lower diameter was higher the yield extraction, because
they had a large surface area per unit volume, contain a high percentage of “free
oil” and require less distance for the “tied oil” to reach the surface, which reduces
the internal mass transfer resistance [5].

5. Conclusions
The effect of the main process variables affecting the supercritical carbon dioxide
extraction of oil from grape seeds was investigated experimentally and analysed
statically. The statistical analysis showed that the extraction yield increased with
an increase in pressure and mass flow, and on the contrary, with a reduction in
temperature and size; pressure and temperature being the most influential factors,
in that order. The most appropriate operating conditions to obtain the best yield in
the extraction were 300 bar, 40 °C, 5 g/min and a particle size of 0.4 mm.

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Received: April 19, 2019; Published: May 28, 2018