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Agriculture and Agricultural Science Procedia 7 (2015) 255 – 259

Farm Machinery and Processes Management in Sustainable Agriculture, 7th International

Scientific Symposium

Effects of thermal treatment of seeds on quality and oxidative

stability of oils
Magdalena Kachel-Jakubowska*, Artur Kraszkiewicz, Edmund Lorencowicz,
Milan Koszel, Artur Przywara

University of Life Sciences in Lublin, Department of Machinery Exploitation and Management of Production Processes
Głęboka 28, 20-612 Lublin, Poland

Abstract

Sustainable development, among other things, means seeking to improve the life quality associated with raising the requirements
both for farmers and manufacturers producing and processing the agricultural goods. One aspect of a healthy diet is getting the
good-quality agricultural products and their correct processing to maintain the highest quality and nutritional value. For this
purpose, an attempt to assess the oxidative stability of cold-pressed vegetable oils, was undertaken. We analyzed two types of oil
seeds and the oil produced therefrom. The study involved winter rape seeds and pumpkin seeds. Prior to the study, seeds were
subjected to chemical analysis in order to determine the water, fat and protein contents in seeds and then oils were extruded and
contents of carotenoids and chlorophylls, as well as acidic and peroxide numbers were determined. Value of the oxidative stability
was determined using Rancimat accelerated oxidation test applying Metrohm 670 device. Among the tested varieties of oilseeds,
the highest oxidative stability distinguished the oil made of two varieties of both rapeseed (Abakus and Bellevue) and pumpkin
seeds (Miranda and Olga).

©©2015
2015The
TheAuthors.
Authors.Published
Published by
by Elsevier
Elsevier B.V.
B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the Centre wallon de Recherches agronomiques (CRA-W).
Peer-review under responsibility of the Centre wallon de Recherches agronomiques (CRA-W)
Keywords: oilseeds; oxidative stability; vegetable oil; carotenoids; fat.

*
Corresponding author. Tel. +48 81-531-97-31; fax: +4881-531-97-29.
E-mail address: magdalena.kacheljakubowska@up.lublin.pl

2210-7843 © 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the Centre wallon de Recherches agronomiques (CRA-W)
doi:10.1016/j.aaspro.2015.12.041
256 Magdalena Kachel-Jakubowska et al. / Agriculture and Agricultural Science Procedia 7 (2015) 255 – 259

1. Introduction

Sustainable development, among other things, means seeking to improve the life quality associated with raising
the requirements both for farmers and manufacturers producing and processing the agricultural goods. One of the
requirements is to search and use the production techniques that can reduce the negative impact of an agriculture on
the environment and use the natural resources in a more efficient manner, taking into account the welfare of people
and animals. This idea comes not out from fashion, but also an objective economic necessity and the need to preserve
life on Earth (Cymerman and Hopfer, 1998; Woś, 1998).
Due to the ever-increasing environmental pollution and ubiquitous processed foods, and fast life pace, people tend
towards finding a way of healthy eating. This problem affects both the selection of an appropriate food and its
processing methods. Focusing on oil plants and products obtained from them, we can say that there are two ways of
their processing, particularly the oil extraction: cold and hot. The oil obtained by cold pressing generally affects better
keeping of its properties. This method allows for better performance and availability of desirable nutrients such as
phytosterols, carotenoids or tocopherols. Therefore, there is an increasing demand of the market dealing with cold-
pressed oil, but this method is less efficient in comparison with the method commonly used in the oleochemical plants
(Neᵭeral at al., 2012). Cold-pressed oils, due to the presence of antioxidants, are characterized by the highest oxidative
stability.
The oxidative stability is an important factor affecting the quality of oil and is particularly important in the case of
its use for frying due to the temperature and duration of the process. Oxidative changes occurring during frying
adversely affect the nutritional value, as well as sensory qualities of a fried product. Each vegetable oil has its
characteristic stability to oxidation depending on the content of fatty acids, antioxidants and other minor ingredients
(Nogala-Kalucka et al., 2005; Przybylski and Eskin, 2006). The scope of any oxidative conversion in oils also depends
on the storage conditions such as temperature, oxygen, light and type of packaging (Szukalska, 2003). Applying an
appropriate packaging in a form of metal can or a dark-glass bottle, the access of oxygen and light is limited, which
can impair the process of oxidation by singlet oxygen characterizing by extremely high reactivity – 1450 times greater
than the triplet oxygen (Bartosz, 2003; Tańska and Rotkiewicz, 2003). Plant oils, e.g. rapeseed oil or pumpkin seeds
oil, are the most important plant-origin fats on the global market of edible fats. Seeds of these plants and crude oil
contain high amounts of bioactive compounds: polyphenols, phytosterols, tocopherols, and other antioxidants, that
play an important role in the prevention and treatment of certain chronic diseases such as heart disease,
neurodegenerative diseases, aging, cancer, and arthritis (Caili et al, 2006; Kim et al, 2006; Stevenson et al, 2007;
Zuhair et al., 2000; Jian et al., 2005). Their presence gives a protection of the active compounds against destruction
of free radicals, that are responsible for the oxidative damage of lipids, proteins, and nucleic acids (Bouzid et al.,
2005; Younis et al., 2000; Ghani, 2003). Fats are the most concentrated source of energy delivered to an organism
with food, as well as being a carrier of essential unsaturated fatty acids (UFAs) (Kruszewski et al., 2013). Their
presence makes the plant oils are relatively unstable products that are subject to oxidation processes. They are readily
converted leading to the emergence of significant adverse changes in the quality known as “fat rancidity” (Krygier,
1997; Krygiet et al., 1998).
The purpose of this paper was to compare the impact of cold-pressing oil extraction on the oxidative stability of
two types of oils produced from oilseeds. The raw material for the study included three varieties of winter rapeseeds
and three varieties of pumpkin seeds. In the first stage, the analysis of physicochemical composition was carried out
taking into account the water, protein, and fat contents in seeds, and then the seeds were subjected to “cold pressing”
to achieve the edible oil and to determine chlorophyll and carotenoids contents, as well as peroxide and acidic
numbers, and to estimate the duration of oxidative stability of these oils due to oxidation.

2. Materials and methods

Experimental material consisted of 2 kg samples of seeds from two oilseed plants such as Winter rape Abakus,
Bellevue and Abakus, Seed-less pumpkin Olga, Miranda and Junona. All the plants were grown in South-East Poland
in Lublin Province. All the crops were collected in 2014 year. Prior the experiments seeds were dried under natural
conditions and stored under laboratory conditions at 20 0C and a relative humidity of 60-70%. Determinations of fat
(CLA/GC/3b/2011) and protein (CLA/PSO/13/2013) were carried out at the Central Agro-Ecological Laboratory of
University of Life Sciences in Lublin. Seed moisture content was determined using a moisture analyzer Radwag max
50/1/WH (oven-dry test). Analysis of moisture was carried out during drying at the sample load of 4g at 120 0C.
 Magdalena Kachel-Jakubowska et al. / Agriculture and Agricultural Science Procedia 7 (2015) 255 – 259 257

The oil was pressed using a screw press with variable nozzle with a diameter of 8 mm and a set of microscopic
strainers from Farmet DUO (Czech Republic) in the continuous operation mode. Before starting the press was heated
to a temperature of 600C. The press was started and after the stabilization of the working temperature the oil pressing
process was commenced. The stabilization was achieved after the pressing of oil from a mass of about 1 kg of seeds
when the temperature was about 700C. The temperature during pressing was measured with aim of ama-digit
thermometer.
The oil after extrusion was stored in dark glass bottles at a temperature of 50C, in order to obtain natural decantation
for 6 days and then subjected to analyses.
The contents of chlorophyll and carotenoids were determined by spectrophotometry with fresh oil using a double-
beam UV - VIS Jasco V -630 spectrophotometer. Oil samples were diluted 20 times with acetone and the spectrum
measured between wavelengths 350 and 700 nm. Calculations of concentrations of chlorophyll a and b and total
carotenoids content (in mg/ml) were made in accordance with the procedure of Lichtenthaler and Buschmann (2003).
All the measurements were done with three independent replicates.
Evaluation of the quality of oils included determination of the acid number (AN) by titration in accordance with
DIN EN ISO 660: 2005 and the number of peroxide (LOO) by titration in accordance with DIN EN ISO 3960: 2005.
The oxidative stability was measured using a Rancimat 670 apparatus (Metrohm AG). Oil samples (2.5 g) were
weighed into reaction vessels and heated at 120 0C under a dry air flow of 20 l/h. The volatile compounds released
during oxidation were collected into a cell containing distilled water, and the increasing water conductivity was
continually measured. The time taken to reach the conductivity inflection point was recorded as the induction period
(IP), and it is expressed in hours. All determinations were carried out in triplicate.
Results are presented as means ± standard deviation from three replicates of each experiment. A significance level p
< 0.05 was used to denote significant differences between mean values determined by the analysis of variance, Pearson
correlation, post–hock Tuckey tests (ANOVA) with the assistance of Statistica 10 software.

3. Results and discussion

The content of fat and protein in tested seeds along with their moisture content is presented in Table 1. A
preliminary analysis of the chemical composition of seeds confirmed the significant difference in fat and protein
levels. Among considered seeds, pumpkin seeds of Olga cv. were characterized by higher average fat content (47.5%),
while the highest average percentage for rapeseed was 44.0% for Adriana cv. However, these are minor differences
that may result from the varietal difference and habitat conditions. A similar situation occurred also for the protein
content. The average protein concentration in pumpkin seeds ranged from 30.8% to 35.3%, which was remarkably
higher than for rapeseed ranging from 20.40% to 22.30%. The statistical analysis referring to the correlation
coefficient at the level of p<0.05 showed that there was a weak correlation between water content in seeds and fat
percentage both in the rape and pumpkin seeds. A strong positive correlation at the level of 0.931 was recorded in the
case of the fat content in both seed groups and duration of oxidative stability induction, which confirmed the
relationship between the fat content and oxidative stability level for analyzed products.

Table 1. Profile of seeds.

Specification Winter rape Pumpkin
Seed variety Abakus Bellevue Adriana Junona Miranda Olga
Water content in
6.89±0.26 7.35±0.29 6.19±0.42 9.57±0.12 9.33±0.29 10.10 ±0.35
seeds (%)
Fat (% d.m.) 43.87±0.25 42.63±0.15 44.00±0.10 46.9±3.3 41.3±2.9 47.5±3.3
Protein (% d.m.) 20.40±0.30 21.90±0.26 22.30±0.44 34.2±2.4 30.8±2.2 35.3±2.5

During the extraction of fat from the seeds, chlorophylls and carotenoids are transferred to the oil in quantities
depending on the species of raw materials, environmental conditions, degree of ripeness, storing method, and
processing techniques. These factors affect the color of oil, which is one of the basic qualitative characteristics of the
resulting oil (Rotkiewicz et al., 2002; Gandul-Rojas et al., 2000). Chlorophyll content has negative impact on the
quality, especially the odor and stability of the oil – it has pro-oxidative properties and makes the oil darken (Fornal
et al. 1994). Presence of carotenoids in the oil is equally important due to their antioxidant and pro-vitamin activities
(Sujak et al., 2000; Sujak and Kachel-Jakubowska, 2012).
Table 2 shows detailed analyzes of the content of carotenoids and chlorophyll “a” and “b” in oils. The highest
258 Magdalena Kachel-Jakubowska et al. / Agriculture and Agricultural Science Procedia 7 (2015) 255 – 259

chlorophyll “a” content characterized the rapeseed oil, which comprised within the range from 1.76 to 6.68 μg/ml.
Pumpkin seed oil was characterized by a low content of the chlorophyll.
The highest content of pigments in the form of carotenoids was recorded in the oil produced from pumpkin seeds. Its
largest quantities were observed for Olga cv. (22.46 μg/ml). In the case of rapeseed oil, the largest amounts of
carotenoids were observed in Bellevue cv. (18.10 μg/ml) and Adriana cv. (18.67 μg/ml). Statistical analysis of the
correlation coefficient at significance level of p<0.05 demonstrated slight relationship between content of carotenoids
in oils and the oxidative stability induction time. The post-hock tests also showed statistical differences in the content
of chlorophyll “a” in the case of rapeseed Bellevue cv. in relation to other rapeseed and pumpkin seed oil types.
Statistically significant differences were found for chlorophyll “b” content between pumpkin seed and rapeseed oil.

Table 2. Chlorophyll and carotenoid content of the oils.

Specification Winter rape Pumpkin
Seed variety Abakus Bellevue Adriana Junona Miranda Olga
Chlorophyll a
1.76±0.47 6.68±0.98 3.52±0.19 1.13±0.92 1.21±0.10 1.26±0.11
(μg/ ml)
Chlorophyll b
2.02±0.79 3.14±0.41 2.37±0.92 5.89±3.22 7.17±1.18 7.83±0.84
(μg /ml)
Carotenoids
8.41±0.76 18.10±2.90 18.67±3.80 17.30±5.89 13.91±2.24 22.46±2.51
(μg /ml)

The oxidative stability time obtained during the analysis allows for estimating the oil resistance to oxidation under
accelerated conditions. The induction time for all the oils stored for 6-8 days was relatively low (Table 3). Analysis
of the pumpkin seed oil showed exceptionally lowest oxidative stability amounting 0.26 hours on average and for
Adriana cv. rapeseed oil to an average of 0.28 hours. For the same conditions, the highest oxidative stability was
shown by oil produced from Bellevue cv. rapeseed amounting to an average of 4.26 hours and Olga cv. pumpkin seed
– 4.75 hours. These results for rapeseed oil are comparable with those reported in the literature (Krygier et al., 1998).
The factor determining the durability can be a high content of chlorophyll pigments in oils or free fatty acid content
determined by the acidic and peroxide numbers. These are indicators that describe the hydrolytic and oxidative
changes in triglycerides both in the seeds and in the oil. Normal results were shown by cold-pressed oils produced
from particular rapeseed and pumpkin seed oils, for which the acidic number value did not exceed 4 mg KOH/g, and
the peroxide number amounted for rapeseed oil to 1.13 (Abakus cv.) and to 1.03 mg KOH/g (Adriana cv.), while for
pumpkin seed oil to 1.53mg KOH/g (Junona cv.). This means that all oils were characterized by a low content of AN
and they meet the quality requirements set by the Codex Alimentarius. The statistical analysis on the correlation
coefficient showed that in our case there is no association between the induction time of various oils and their acidic
numbers. The Pearson correlation at the level of 0.574 was observed between induction time vs. the peroxide number
for particular oils. There was no correlation between the carotenoid content and oxidative stability. The post-hock
tests showed no statistically significant differences between the acidic and peroxide numbers vs. oxidative stability of
various oils.

Table 3. Acid number, peroxide number and the induction time of oils.
Specification Winter rape Pumpkin
Seed variety Abakus Bellevue Adriana Junona Miranda Olga
AN [mgKOH/ g] 1.13±0.01 3.24±0.04 1.03±0.03 1.53±0.01 2.44±0.04 2.03±0.06
LOO [mmolO/ kg] 1.43±0.06 3.57±0.06 1.17±0.12 1.37±0.06 1.17±0.12 1.23±0.12
Oxidative Induction time 4.05±0.07 4.26±0.01 0.28±0.01 0.26±0.06 4.16±0.07 4.75±0.18
Stability [h] Normal time 21.56±0.45 22.74±0.08 1.52±0.07 1.39±0.32 22.18±0.38 25.33±0.95

4. Summary and conclusion

This study demonstrates that both the rape and pumpkin seeds are a rich source of many important nutrients, which
include fat and protein. Based on the results achieved, we can conclude that pumpkin seeds have a higher average
both of fat (47.5% for Olga cv.) and protein (35.3% also for Olga cv.) than rapeseed, for which the highest parameters
of these components were shown by Adriana cv. (44% fat, and 22.30% protein).
Oils produced due to cold pressing were characterized by good quality parameters both in terms of the peroxide
and acidic numbers. In the case of carotenoids, that are generally considered the compounds with antioxidant and pro-
 Magdalena Kachel-Jakubowska et al. / Agriculture and Agricultural Science Procedia 7 (2015) 255 – 259 259

vitamin properties, their largest amounts were recorded in the oil made from pumpkin seeds of Olga cv. (22.46 μg/ml).
In the case of winter rapeseed, the highest quantity of carotenoids was observed in Bellevue cv. (18.10 μg/ml) and
Adriana cv. (18.67 μg/ml). The statistical analysis showed no relationship between the content of this component and
oxidative stability of the oils studied.
Studies upon the oxidative stability of both groups of oils were comparable. Each group contained one variety with
very low oxidative stability of 0.28 h for oil made from rapeseed Adriana cv. and 0.26 h for Junona cv. pumpkin seed
oil.

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