Article

Reused Plant Fried Oil: A Case Study with Home-Made Soaps

Bojan Antonic 1, Dani Dordevic 1, Simona Jancikova 1, Bohuslava Tremlova 1,*, Marcela Nejezchlebova 2,
Kristýna Goldová 2 and Jakub Treml 2

1 Department of Plant Origin Food Sciences, Faculty of Veterinary Hygiene and Ecology, University of
Veterinary and Pharmaceutical Sciences, Palackého tř. 1, 612 42 Brno, Czech Republic;
H19001@[Link] (B.A.); DORDEVICD@[Link] (D.D.); JANCIKOVAS@[Link] (S.J.)
2 Department of Molecular Pharmacy, Faculty of Pharmacy, Masaryk University, Palackeho tr. 1946/1,

612 00 Brno, Czech Republic; nejezchlebovam@[Link] (M.N.); 507373@[Link] (K.G.);

tremlj@[Link] (J.T.)
* Correspondence: TREMLOVAB@[Link]

Abstract: The study aimed to analyze the possibility of waste frying oil utilization in home-made
soap production. Soaps were made from unheated and fried rapeseed, sunflower and palm oils that
had total polar material (TPM) values up to 24%. Physicochemical and microbial analyses were per-
formed on produced samples to check their quality. The hardness increased with the degradation
level of rapeseed and palm oils, and opposite findings were obtained for sunflower-made soaps.
The highest malondialdehyde (MDA) contents were recorded for sunflower oil-made samples, with
the maximum of 6.61 µg/g, and the lowest for the palm oil-made samples, with the maximum of
0.94 µg/g. The antimicrobial assessment showed no significant (p > 0.05) differences between control
soap samples and soaps made of oils with the highest TPM value. Gram-positive bacteria (methicil-
lin-resistant Staphylococcus aureus: MRSA) were the most sensitive chosen microorganisms, com-
pared to Gram-negative bacteria and yeasts. The obtained results did not show exact differences
between experimentally produced soap samples from fried or not fried oils; these findings highlight
Citation: Antonic, B.; Dordevic, D.;
the potential of home-made soap production from this byproduct.
Jancikova, S.; Tremlova, B.;
Nejezchlebova, M.; Goldová, K.;
Keywords: waste frying oil; home-made soap; waste management and utilization; soap texture;
Treml, J. Reused Plant Fried Oil: A
soap microbiology
Case Study with Home-Made Soaps.
Processes 2021, 9, 529.
[Link]

Academic Editor: Carl Schaschke 1. Introduction

The amount of waste is increasing together with the world’s population growth,
Received: 13 February 2021 meaning that reverse logistics systems have been gaining in importance [1,2]. Waste fry-
Accepted: 11 March 2021 ing oil represents one of the most important byproducts due to its hazardous impact on
Published: 16 March 2021
the environment [3]. Food frying is the main cooking method around the world and usu-
ally highly accepted by consumers [4]. Approximately 200 million tons of frying vegetable
Publisher’s Note: MDPI stays neu-
oils are being produced annually in the world with an increasing trend [5,6]. According
tral with regard to jurisdictional
to some assumptions, 1 L of waste oil poured into water can pollute up to 500,000 L of
claims in published maps and insti-
tutional affiliations.
fresh water [3].
A larger portion of waste frying oil that comes from restaurants and industry is being
collected and utilized. According to Greenea, in the European Union, 51% of frying oil
waste comes from households and only a few percent is being collected [7]. Thus, a large
Copyright: © 2021 by the authors. Li- amount of this waste ends up in sewage, causing environmental and economic problems.
censee MDPI, Basel, Switzerland. In sewage cleaning systems, oil sticks to the apparatuses and causes clogging and corro-
This article is an open access article sion. Such cleaning processes are low in efficiency and require a lot of energy [8].
distributed under the terms and con- There is a great potential today for waste frying oil to be used as a low-cost raw ma-
ditions of the Creative Commons At- terial in the production of biodiesel, lubricants, resins, soaps and many other applications
tribution (CC BY) license ([Link] (bitumen rejuvenator or fermentation media component) [3,9]. It is of great importance to
[Link]/licenses/by/4.0/). find the solution for the utilization of household-originated waste frying oils. The possible

Processes 2021, 9, 529. [Link] [Link]/journal/processes

Processes 2021, 9, 529 2 of 12

solution could be the production of home-made soaps. Their production (saponification

process) requires a minimal amount of energy and practically no byproducts are created
[10]. Consequently, this production is also described as a technology with a green pro-
spective [11].
By some studies, soaps degrade up to 4 times faster than oils. This is mainly due to
their higher accessibility to microorganisms [12,13]. In addition, supportive data come
from the studies that include saponification as the one step in waste frying oil-rich
wastewater treatment [14].
Literature findings are limited to just a few articles describing home-made soap pro-
duction from waste frying oil, but with no clearly defined degradation level of fried oil
from which the soap samples were made [13,15,16]. The study aim was to evaluate the
quality of home-made soaps produced from waste frying oils, and to perceive the possible
more environmentally friendly utilization.

2. Materials and Methods

2.1. Preparation of Soap Samples
The rapeseed, sunflower and palm oils were used for preparation of soap samples.
Rapeseed and sunflower oils were produced in the Czech Republic and palm oil was bot-
tled in Austria. The frying of French fries served as the simulation of the oil frying process
in restaurants. The French fries (Hearty Food Co., Tesco, Praha, Czech Republic) in
batches of 100 g were fried in approximately 3.3 L of oil in the fryer FR 2035 (Concept,
Choceň, Czech Republic) for 5 min at the approximate average temperature of 175 °C.
Sampling of the fried oil for soap making purposes was conducted after three batches of
frying and, further, when the TPM (total polar matter) reached values of 10, 15, 20 and
24%. TPM values were measured by TPM meter testo 270 (Testo SE & Co. KGaA, Titisee-
Neustadt, Germany). The unheated oil was taken for making control soap samples. The
samples were labeled according to Table 1.
The soaps were made using the cold saponification method reported by Adigun et
al. [17] with slight modifications. An amount of 130 g of filtered oil was mixed with 66.92
g of 26% (w/w) NaOH water solution using the blender (3–5 min). The mixture was poured
into the molds and after 24 h was taken out of the filter paper to mature in the air. After 4
weeks, the soap samples were subjected to the analysis.

Table 1. Description of soap sample abbreviations and corresponding oil TPM% levels.

Oil TPM Level (%)

Sample Abbreviation
R S P
Controls R0/S0/P0 6 7.5 5
Sample 1 sets R1/S1/P1 6.5 9 6.5
Sample 2 sets R2/S2/P2 10 10 10
Sample 3 sets R3/S3/P3 15 15 15
Sample 4 sets R4/S4/P4 20 20 20
Sample 5 sets R5/5S/P5 24 24 24
R—rapeseed oil sample; S—sunflower oil sample; P—palm oil sample.

2.2. Assessment of the Physicochemical Properties of the Soap

The physicochemical parameters studied on soap samples included: pH, moisture,
total alkali, total fat matter, total fat content, MDA (malondialdehyde), foaming, hardness
and stickiness.
pH value was obtained by measurement in a 1% soap solution in distilled water [18],
using the pH meter Orion 4 star (Thermo Scientific, Waltham, MA, USA).
For determination of moisture content, 5 g of soap sample were dried in an oven at
105 °C until achievement of the constant mass. The mass of the sample before and after
Processes 2021, 9, 529 3 of 12

drying was weighed on the analytical balance ALS 250-4A (Kern, Frankfurt am Main, Ger-
many), with precision of 0.1 mg.
The total alkali was determined by acid base titration. An amount of 10 g of soap
sample was dissolved in ethanol and supplemented with 5 mL of 1N H2SO4 (aq). The ex-
cess of acid was titrated with 1N NaOH using the phenolphthalein indicator. The total
alkali was calculated using the formula [19]
% Total alkali = (V(Acid) − V(Base))/(m(sample)) × 3.1. (1)
For total fat matter determination, 10 g of soap sample was dissolved in hot neutral-
ized ethanol and filtered. The remaining residues on the filter represented matter insolu-
ble in alcohol (MIA). The MIA value was obtained by drying and weighing the filter, and
total fat matter was calculated using the formula [20]
% Total fat matter = (100 − (moisture content + MIA))/1.085 (2)
The MDA was determined with the TBA (thiobarbituric acid) method according to
Khalifa et al. [21] with slight modifications. An amount of 1.5 g of soap sample was ho-
mogenized with 1 mL of EDTA (ethylenediaminetetraacetic acid, 0.3% water solution), 5
mL of BHT (butylated hydroxytoluene, 0.8% solution in hexane) and 8 mL of TCA (tri-
chloroacetic acid, 10% water solution) and centrifuged for 5 min at 3000 RPM. The lower
layer was filtered into the 10 mL volumetric flask and supplemented with 10% TCA to the
mark. Mixture containing 4 mL of this solution and 1 mL of TBA was incubated for 90 min
at 70 °C in the dark. This was followed by rapid cooling to room temperature in the ice
bath and 45 min laying down at room temperature prior to the measurements. Measure-
ments were conducted at 532 nm on the spectrophotometer CE7210 (Cecil Instruments,
Milton, UK). The results were calculated using the standard calibration curve.
Total fat was obtained by the Soxhlet method on the instrument B-811 (Büchi, Flawil,
Switzerland). Extraction was conducted using the petrol ether as solvent from the 5 g sam-
ple. The program consisted of 90 min of extraction, followed by 30 min of washing and 20
min of solvent evaporation.
Foaming capacity and foam stability were assessed according to Kempka et al. [22]
An amount of 20 mL of 0.5% soap solution was homogenized for 30 s in a 400 mL glass
beaker (low form) using the homogenizer HG-15A (Witeg, Wertheim, Germany). Homog-
enization was performed at approximately 13,500 RPM with the dispersing tool HT1025.
The foam volume was measured in the 100 mL graduated cylinder right after mixing and
after 30 min. The foaming capacity was calculated using the formula
% Foaming capacity = (V(after homogenization) − V(initial solution))/V(initial solution) × 100 (3)
The foam stability was calculated using the formula:
% Foam stability = V(foam after 30 min)/V(foam after homogenization) × 100 (4)
The textural parameters, hardness and stickiness, of soap samples were measured on
the texturometer [Link] plus (Stable Microsystems, Godalming, UK). The results were ob-
tained using the stainless P/5 cylinder probe to penetrate into the soap bar dimensions 50
× 50 × 20 mm on 5 different places. The probe diameter was 5 mm, penetration depth was
set to 5 mm and test speeds were adjusted to 1 mm/s. A 50 kg load cell was fitted in the
moving arm of the instrument. The hardness was defined as the force (g) needed for a
probe to make a 5 mm deep hole in the soap sample; stickiness represented the force for
probe retraction from the sample.

2.3. Assessment of the Antimicrobial Properties of the Soap

To test antimicrobial efficacy, a control set of soaps (S0, R0, P0) and set 5 (S5, R5, P5)
were selected. The soaps were dissolved in distilled water and 20% solutions of the indi-
vidual soaps were prepared. To determine the antimicrobial efficacy of soaps, a modified
method according to EUCAST (European Committee on Antimicrobial Susceptibility
Processes 2021, 9, 529 4 of 12

Testing) was used to determine the minimum inhibitory concentration by the microdilu-
tion method. One plate was used to test one type of soap (control and set 5) in 6 concen-
trations (10%, 5%, 2.5%, 1.25%, 0.625%, 0.3125%), each with one microorganism. Lines A–
D contained soaps in various concentrations and with microorganisms, lines E–G con-
tained only soaps in various concentrations without microorganisms (blank), line H (1–6)
contained broth without soap and without microorganisms (control of possible broth con-
tamination) and line H (7–12) contained broth without soap with microorganisms (nega-
tive control). The reference strains of microorganisms used in the experiment were Staph-
ylococcus aureus subsp. aureus CCM 7110 (methicillin-resistant S. aureus; MRSA), Esche-
richia coli CCM 3954 and Candida albicans CCM 8261 from the Czech Collection of Micro-
organisms of the Department of Experimental Biology, Faculty of Science, Masaryk Uni-
versity. Experiments that included cultivation of S. aureus and E. coli were conducted on
MUELLER-HINTON broth (MiliporeSigma, formerly Sigma-Aldrich, Munchen, Ger-
many) and C. albicans was inoculated on Malt Extract Broth (Sigma-Aldrich, Germany).
Inoculum concentration was adjusted to approximately 1–2 × 108 CFU/mL corresponding
to suspension standard of 0.5 McFarland degrees. Final concentration in the wells was
adjusted by dilution to 104–105 CFU/mL. The inoculated plates were incubated for 18 h at
35–37 °C. After incubation, the absorbance (turbidity) in plates was measured spectropho-
tometrically using a microplate reader and the inhibition was calculated from the obtained
data. To visualize the results and to stain metabolically active cells, 1% TTC (triphenyl
tetrazolium chloride) solution was used. The reaction reduces tetrazolium chloride to red
formazan when bacterial and yeast dehydrogenases are present and active.

2.4. Statistics
The results are presented in the tables, together with the mean values and standard
deviations. Parameters moisture, total fatty matter, total alkali and total fat content were
assessed in duplicate; pH, foaming capacity, foam stability and MDA in triplicate; textural
parameters in quintuplicate. Statistical analysis was performed using one-way ANOVA
for finding the differences within the sample group. For better overview of differences
between the samples, principal component analysis (PCA) was used. IBM SPSS software
was used for conducting statistical analysis.

3. Results and Discussion

3.1. Physicochemical Analysis
The results for physicochemical analysis performed on soap samples are presented
in Tables 2 and 3. The obtained pH values were similar in all tested samples (Table 2). The
pH ranged from 9.48 in sample 4S to 10.16 in sample P1. From the obtained values, it can
be said that the pH value was not affected by the degradation level of fried oils. Significant
(p < 0.05) differences were obtained between the samples, but an unambiguous trend of
difference between control samples (R0, S0 and P0) and samples made from the oil with
the highest TPM% (R5, S5 and P5) was not observed. The obtained results are in line with
the study of Sanaguano-Salguero et al. [15], where pH values varied from 9.96 to 11.30.
The selected commercial soaps tested in the work of Tarun et al. [23] had pH values be-
tween 9 and 10. The study of Mendes et al. [17] confirmed that commercial soap bars in-
tended for kids had pH values up to 11.34. The normal human skin pH is between 5.4 and
5.9 and any introduction of soaps with a high pH can affect its protective role and micro-
biome [23].
Processes 2021, 9, 529 5 of 12

Table 2. Results for pH, moisture, total fatty matter, total alkali, total fat content and MDA for produced soap samples.

Total Fatty Total Fat

Sample pH Moisture (%) Total Alkali (%) MDA (µg/g)
Matter (%) Content (%)
R0 9.67 ± 0.00 a 7.43 ± 0.17 82.34 ± 1.34 0.00 ± 0.00 1.63 ± 0.10 2.09 ± 0.05 a
R1 9.80 ± 0.01 b 7.06 ± 1.73 81.96 ± 1.56 0.00 ± 0.00 2.38 ± 0.11 2.58 ± 0.04 b
R2 10.08 ± 0.01 c 9.91 ± 0.52 80.00 ± 2.20 0.00 ± 0.00 1.74 ± 0.27 0.68 ± 0.00 c
R3 10.08 ± 0.00 acd 9.82 ± 1.48 81.61 ± 1.12 0.00 ± 0.00 1.43 ± 0.15 0.71 ± 0.00 d
R4 10.11 ± 0.01 d 9.21 ± 1.44 83.31 ± 1.10 0.00 ± 0.00 1.37 ± 0.06 0.83 ± 0.00 e
R5 10.08 ± 0.01 c 8.16 ± 0.87 84.33 ± 0.65 0.00 ± 0.00 0.95 ± 0.08 0.96 ± 0.00 f
S0 9.58 ± 0.01 x 6.44 ± 0.67 85.98 ± 0.51 x 0.00 ± 0.00 1.61 ± 0.15 4.66 ± 0.04 x
S1 9.85 ± 0.00 y 8.28 ± 1.11 84.35 ± 0.84 0.00 ± 0.00 1.29 ± 0.04 4.46 ± 0.04 y
S2 10.06 ± 0.01 z 9.32 ± 0.17 83.32 ± 0.14 y 0.00 ± 0.00 1.30 ± 0.13 4.86 ± 0.02 z
S3 10.07 ± 0.01 z 7.87 ± 0.82 84.67 ± 0.62 0.00 ± 0.00 1.37 ± 0.00 x 2.91 ± 0.00 u
S4 9.48 ± 0.00 y 6.26 ± 1.45 86.31 ± 1.09 0.00 ± 0.00 1.58 ± 0.08 5.73 ± 0.01 v
S5 9.59 ± 0.01 x 6.83 ± 1.32 85.77 ± 0.99 0.00 ± 0.00 0.62 ± 0.02 y 6.61 ± 0.02 w
P0 10.14 ± 0.01 op 3.73 ± 0.18 88.53 ± 0.14 0.00 ± 0.00 o 1.41 ± 0.01 o 0.58 ± 0.00 o
P1 10.16 ± 0.01 p 3.88 ± 0.13 88.35 ± 0.11 0.01 ± 0.01 1.01 ± 0.17 0.45 ± 0.00 p
P2 10.07 ± 0.00 opq 3.90 ± 0.06 88.25 ± 0.14 0.00 ± 0.00 o 2.20 ± 0.06 0.52 ± 0.01 o
P3 10.13 ± 0.01 op 3.88 ± 0.25 88.25 ± 0.23 0.00 ± 0.00 o 1.85 ± 0.14 0.78 ± 0.00 q
P4 10.00 ± 0.00 oq 3.74 ± 0.29 88.22 ± 0.25 0.03 ± 0.01 p 2.15 ± 0.02 p 0.81 ± 0.00 r
P5 9.97 ± 0.01 q 3.57 ± 0.17 88.71 ± 0.13 0.00 ± 0.00 o 1.38 ± 0.10 0.94 ± 0.00 s
Different lowercase letters indicate statistically significant differences (p < 0.05) between rows: (a, b, c, d, e, f) for rapeseed
oil-made samples, (x, y, z, u, v, w) for sunflower oil-made samples and (o, p, q, r, s) for palm oil-made samples. The results
are presented as the mean values ± standard deviation.

Table 3. Results for foaming capacity, foam stability, hardness and stickiness for produced soap samples.

Parameter Foaming Capacity (%) Foam Stability (%) Hardness (g) Stickiness (g)
R0 320 ± 35 53 ± 7 a 2320 ± 130 a −622 ± 116 a
R1 285 ± 18 65 ± 3 2424 ± 257 a −430 ± 34 bc
R2 340 ± 69 71 ± 3 1654 ± 235 b −318 ± 59 b
R3 350 ± 66 67 ± 3 1256 ± 183 c −326 ± 65 b
R4 262 ± 35 71 ± 8 1414 ± 128 bc −405 ± 38 bc
R5 283 ± 13 74 ± 1 b 1521 ± 185 bc −457 ± 31 c
S0 325 ± 75 54 ± 8 x 786 ± 51 x −411 ± 92 xz
S1 293 ± 32 10 ± 2 y 856 ± 133 x −299 ± 31 x
S2 408 ± 24 x 23 ± 2 yz 457 ± 23 y −148 ± 34 y
S3 318 ± 51 52 ± 10 x 804 ± 105 x −153 ± 69 y
S4 303 ± 25 33 ± 4 zw 1352 ± 155 z −442 ± 20 z
S5 250 ± 13 y 50 ± 6 xw 1272 ± 118 z −471 ± 45 z
P0 215 ± 41 72 ± 0 o 4990 ± 402 o −794 ± 101 o
P1 123 ± 8 61 ± 2 p 3866 ± 276 p −346 ± 85 pq
P2 175 ± 48 68 ± 6 3243 ± 363 pq −287 ± 96 q
P3 182 ± 51 71 ± 1 3023 ± 296 q −427 ± 127 qr
P4 235 ± 87 78 ± 8 3037 ± 370 q −568 ± 75 r
P5 205 ± 61 73 ± 2 3319 ± 348 pq −496 ± 146 pr
Different lowercase letters indicate statistically significant differences (p < 0.05) between rows: (a, b, c) for rapeseed oil-
made samples, (x, y, z, w) for sunflower oil-made samples and (o, p, q, r) for palm oil-made samples. The results are
presented as the mean values ± standard deviation.
Processes 2021, 9, 529 6 of 12

Moisture content analysis revealed low values in all soap samples. No significant (p
> 0.05) differences were observed among the samples made from the same oil (rapeseed,
sunflower or palm). The moisture ranged from 3.57% (P5) to 9.91% (R2). The range of the
soap moisture content in the work of Sanaguano-Salguero et al. [15] was from 24.90% to
43.24%. The tested commercial soaps had moisture values around 30–35% [15]. On the
other hand, laundry soaps made from waste frying oils in the work of Adane [16] had
moisture content values from 6.67 to 14.47%. This can be explained by the different recipe
and method for the preparation of soap samples. Lower moisture content in soaps inhibits
the hydrolysis and alteration inside soaps [16]. Some soap producers declare on their
products’ maximal 14% of moisture [24].
One of the most important factors of the soap quality is the total fatty matter. A higher
value indicates a better quality of the soap since fatty acids positively affect skin rehydra-
tion and cleansing [22]. Values ranged from 80.00% (R2) to 86.31% (S4) and changes in
values among the samples did not correlate to the degradation level of frying oils from
which the soaps were made. The total fatty matter of soaps made from waste frying oils
in the work of Adane [16] ranged from 75.42 to 88.53%. Moisture contents measured in
commercial soaps ranged from 82.10 to 88.42% and are in accordance with the present
study [16]. Total alkali was 0.00% in all samples except P1 (0.01%) and P4 (0.03%). Accord-
ing to the literature, the lower total alkali values mean a better quality of the soap [24].
The total fat content revealed low values that ranged from 0.62% (sample S5) to 2.38%
(sample R2). In general, no significant (p > 0.05) differences were obtained within the same
sample groups (the same oil type).
Foaming is one of the main properties that take part in soap cleaning properties [25].
The following parameters are used for the estimation of soap foaming properties: foaming
capacity and foam stability. From the obtained results, one can see that the lowest foaming
capacities were found in the palm oil soaps (Table 3), with the range from 123% (P1) to
235% (P4). Samples made from sunflower and rapeseed oils showed similar values, with
the highest value obtained for sample S2 (408%), though no statistically significant (p >
0.05) differences were observed between the samples belonging to the same oil group. The
foam stability revealed the highest and statistically significant (p < 0.05) differences be-
tween the sunflower oil-made samples. The lowest value was found in sample S1 (10%),
while the highest was found in sample S0 (54%), but there was no clear trend that would
correlate with the degree of fried oil. The first (S0) and the last (S5) sample did not differ
significantly (p > 0.05). The foam stability of rapeseed oil-made soaps ranged from 53%
(sample R0) to 74% for sample R5 and a significant (p < 0.05) difference was obtained be-
tween those two samples too. The palm oil-made soaps revealed similar values between
all the samples.
MDA determination cannot be found in the literature as the soap quality parameter.
Peroxidation of oil occurs during the oil frying, and MDA represents a secondary product
of oxidation [26]. The soaps made of palm oil showed the lowest values, ranging from 0.45
(sample P1) to 0.94 µg/g (sample P5). This can be explained by the fact that palm oil is
more stable than the other two oils because of the structure rich in saturated fatty acids
and without the presence of trans fats [27]. Soap samples from rapeseed oil had higher
MDA values for the samples R0 and R1 (2.09 and 2.58 µg/g, respectively) than for the
samples made from fried oil with a higher TPM. The reason might be found in some reac-
tions during and after the saponification process that have still not been studied [13]. Soap
samples made from sunflower oil had values ranging from 4.46 (S1) to 6.61 µg/g (S5).
Consequently, it can be concluded that the sunflower oil had the highest level of oxidation
during the process of frying.
Concerning other measured parameters of soaps (such as textural properties, hard-
ness and stickiness), there was a lack of information in the literature on methods applied,
so they are not comparable to the other studies. In the samples made from rapeseed and
palm oils, hardness decreased with the increased oil degradation level. In the rapeseed
oil, the value decreased from 2424 (sample R1) to 1256 g (sample R3). Hardness of the
Processes 2021, 9, 529 7 of 12

palm oil decreased from 4990 (sample P0) to 3023 g (sample P3). Oppositely, in the sam-
ples made from sunflower oils, the hardness of the soap samples increased with the deg-
radation level (unheated oil: 786 g; the highest level of frying: 1272 g).
The recorded stickiness for soaps prepared with rapeseed and palm oils revealed the
highest values in samples made from unheated rapeseed (R0) and palm oils (P0), −622 and
−764 g, respectively. Regarding the sunflower oil-made soaps, no statistical (p > 0.05) dif-
ference was obtained between the S0 and S5 samples.

3.2. Antimicrobial Assessment

The results of the antimicrobial assessment are presented in Table 4. Inhibition (anti-
microbial activity) is given in percentages. According to the approximate determination
of the antimicrobial efficacy of soaps made from various oils, the soaps made from sun-
flower oil samples had the highest antimicrobial effect, while the lowest effectiveness can
be observed for the palm oil soap. Sunflower oil soaps completely inhibit the growth of
MRSA in all concentrations, regardless of the decreasing pH values. The growth inhibi-
tory effect on E. coli was also the highest among the soap samples produced from the
sunflower oil. The high alkalinity of the highest concentrations in all types of soaps un-
doubtedly potentiates the antimicrobial activity. However, it seems likely that inhibitory
substances from the original oils pass into the soaps. It can also be stated that the results
of control set 0 and set 5 do not show a significant difference.
The sunflower oil contains mainly unsaturated fatty acids, such as linoleic and oleic
acids [28]. Jang et al. [29] proved the antibacterial effect of potassium salts of oleic and
linoleic acids at a concentration of 5 mmol/L. After 1 h of incubation, the concentration of
S. aureus, expressed as colony-forming units (CFU)/mL, was significantly reduced [29].
Moreover, Kawahara et al. [30] showed that potassium oleate was able to significantly
reduce numbers of CFU/mL of S. aureus and E. coli after 10 min in incubation. The effect
was comparable to the effect of alcohol-based disinfectant. Furthermore, the fatty acids
themselves possess an antibacterial effect, as it was demonstrated by Huang et al. [31].
Unfortunately, no results were found for soaps containing sodium salts of linoleic
and oleic acids in the literature. However, it can be assumed that the change in the salt
cation would have little effect. The antibacterial effect of soaps is generally attributed to
their detergent properties in water solutions. From this point of view, our results demon-
strate that Gram-positive bacteria (MRSA) are more susceptible to the antimicrobial effects
of soap solutions compared to Gram-negative bacteria and yeasts.

Table 4. Results for antimicrobial activity of soap samples.

Soap Inhibition of Inhibition of Inhibition of pH of Soap

Sample Name
Concentration E. coli (%) MRSA (%) C. albicans (%) Solution
R0 100 100 100 10.0
0%
R5 100 100 100 10.2
R0 100 100 100 10.1
5%
R5 100 100 100 10.2
R0 80 100 100 10.1
2.50%
R5 65 100 100 10.2
R0 79 100 90 10.2
1.25%
R5 35 88 89 10.1
R0 75 100 76 8.1
0.63%
R5 29 85 84 8.3
R0 58 90 69 7.2
0.31%
R5 24 79 67 6.6
S0 100 100 100 9.9
10%
S5 100 100 100 9.8
Processes 2021, 9, 529 8 of 12

S0 100 100 100 9.9

5%
S5 100 100 100 9.8
S0 100 100 100 10.0
2.50%
S5 63 100 100 9.8
S0 49 100 95 10.0
1.25%
S5 50 100 90 9.8
S0 41 100 39 8.5
0.63%
S5 38 100 53 8.5
S0 14 100 37 7.7
0.31%
S5 21 100 29 8.0
P0 100 100 100 10.5
10%
P5 100 100 100 10.3
P0 100 100 100 10.5
5%
P5 80 100 100 10.4
P0 78 100 100 10.5
2.50%
P5 71 100 100 10.3
P0 77 92 55 10.0
1.25%
P5 63 100 53 9.9
P0 54 83 47 8.3
0.63%
P5 49 80 29 8.2
P0 46 36 35 7.8
0.31%
P5 49 76 26 7.7

3.3. PCA Analysis

The principal component analysis (PCA) of all parameters in all samples is presented
in Figure 1, and for each sample group separately in Figures 2–4.

Figure 1. Principal component analysis for all analyses on the obtained samples. * Samples’ de-
scription is given in Table 1.

The analysis revealed the highest overall differences between sunflower samples and
the lowest among the palm oil samples (Figure 1.). Among all tested parameters, MDA
and textural parameters (hardness and stickiness) can be identified as the main parame-
ters distinguishing soap samples according to the TPM oil content. Palm oil-made samples
revealed the smallest changes between the extreme values (MDA and textural parameters)
Processes 2021, 9, 529 9 of 12

in those parameters in comparison to the other two sample groups (prepared with sun-
flower and rapeseed oils). The reason might be, in fact, that the sunflower oil contains the
highest content of polyunsaturated fatty acids (up to 70%), following the rapeseed oil
(around 20%) and palm oil with 15%. The highest content of saturated fatty acids is in
palm oil (approximately 50%) [27,32–35]. Rapeseed oil samples, according to PCA, created
three clusters (R0 and R1, R2 and R3, R4 and R5) and differences between them partially
correlate with the degradation level of fried oil, which can be observed in Figure 2, though
the samples made from sunflower and palm oils do not have such relation (Figures 3 and
4). The sunflower-made control sample (S0) is the closest to sample S4. For palm oil sam-
ples, P0 (as a control) is the closest to samples P3 and P5, among this sample group. PCA
figures show unambiguous differences between samples prepared with unheated oils and
soap samples prepared with fried oils; these differences are better emphasized between
control samples (soaps prepared with unheated oils) and soaps with the highest TPM con-
tents.

Figure 2. Principal component analysis of soap samples made of rapeseed oil. * Samples’ descrip-
tion is given in Table 1.

Figure 3. Principal component analysis of soap samples made of sunflower oil. * Samples’ descrip-
tion is given in Table 1.
Processes 2021, 9, 529 10 of 12

Figure 4. Principal component analysis of soap samples made of palm oil. * Samples’ description is
given in Table 1.

4. Conclusions
The present study emphasized the possibility of reusing waste frying oil from house-
holds and restaurant facilities, since this waste material is mostly discharged into the sew-
ers. Physicochemical analysis of soaps produced from unheated and fried rapeseed, sun-
flower and palm oils indicated no significant quality changes for most of the tested pa-
rameters. The antimicrobial assessment showed similar results, since no differences were
obtained between soaps produced from unheated and fried oils (oils with different levels
of degradation, expressed in TPM values). Overall significant differences between soaps
made from fried and unheated oils were not found and they cannot be distinguished un-
ambiguously. Certainly, any possible inequality between control soaps and soaps pro-
duced from fried oils could be overwhelmed by some soap additives that can be investi-
gated by future studies. It should be emphasized that among many applications for the
utilization of frying oils, soap production stands out as a promising solution since this
production can be adopted by small businesses and entrepreneurs.

Author Contributions: Conceptualization B.A., B.T., D.D.; methodology S.J., M.N., J.T.; software,
B.A., D.D.; validation B.A., D.D., J.T.; formal analysis B.A., M.N., K.G.; investigation, B.A., D.D.;
resources, B.T.; data curation, B.A., S.J.; writing—original draft preparation, B.A., B.T., D.D., M.N.,
J.T.; writing—review and editing, B.A., D.D., J.T.; visualization, B.A.; supervision, D.D., B.T.; project
administration, B.T.; funding acquisition, B.T. All authors have read and agreed to the published
version of the manuscript.
Funding: This research was funded by the Veterinary and Pharmaceutical University: Internal
Grant Agency IGA 228/2020/FVHE.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Data sharing not applicable.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Nikolaou, I.E.; Evangelinos, K.I.; Allan, S. A reverse logistics social responsibility evaluation framework based on the triple
bottom line approach. J. Clean. Prod. 2013, 56, 173–184.
2. Herva, M.; Neto, B.; Roca, E. Environmental assessment of the integrated municipal solid waste management system in Porto
(Portugal). J. Clean. Prod. 2014, 70, 183–193.
3. Panadare, D.C. Applications of waste cooking oil other than biodiesel: A review. Iran. J. Chem. Eng. (IJChE) 2015, 12, 55–76.
Processes 2021, 9, 529 11 of 12

4. Mannu, A.; Garroni, S.; Ibanez Porras, J.; Mele, A. Available Technologies and Materials for Waste Cooking Oil Recycling.
Processes 2020, 8, 366.
5. [Link]. Available online: [Link] (accessed on 5
May 2020).
6. Lopes, M.; Miranda, S.M.; Belo, I. Microbial valorization of waste cooking oils for valuable compounds production—A review.
Crit. Rev. Environ. Sci. Technol. 2019, 50, 2583–2616.
7. Hillairet, F.; Allemandou, V.; Golab, K. Analysis of the Current Development of Household UCO Collection Systems in the EU;
GREENEA: Coivert, France, 2016.
8. Maniak, B.; Szmigielski, M.; Piekarski, W.; Markowska, A. Physicochemical changes of post-frying sunflower oil. Int. Agrophys.
2009, 23, 243–248.
9. Gökalp, İ.; Uz, V.E. Utilizing of Waste Vegetable Cooking Oil in bitumen: Zero tolerance aging approach. Constr. Build. Mater.
2019, 227, 116695.
10. Maotsela, T.; Danha, G.; Muzenda, E. Utilization of Waste Cooking Oil and Tallow for Production of Toilet “Bath” Soap. Procedia
Manuf. 2019, 35, 541–545.
11. Félix, S.; Araújo, J.; Pires, A.M.; Sousa, A.C. Soap production: A green prospective. Waste Manag. 2017, 66, 190–195.
12. Lefebvre, X.; Paul, E.; Mauret, M.; Baptiste, P.; Capdeville, B. Kinetic characterization of saponified domestic lipid residues
aerobic biodegradation. Water Res. 1998, 32, 3031–3038.
13. Antonić, B.; Dordević, D.; Jančíková, S.; Tremlova, B.; Kushkevych, I. Physicochemical Characterization of Home-Made Soap
from Waste-Used Frying Oils. Processes 2019, 8, 1219.
14. Zhu, X.N.; Lyu, X.J.; Wang, Q.; Qiu, J.; Wang, S.S.; Liu, X.Y.; Li, L. Clean utilization of waste oil: Soap collectors prepared by
alkaline hydrolysis for fluorite flotation. J. Clean. Prod. 2019, 240, 118179.
15. Sanaguano-Salguero, H.; Tigre-Leon, A.; Bayas-Morejon, I.F. Use of waste cooking oil in the manufacture of soaps. Int. J. Ecol.
Dev. 2018, 33, 19–27.
16. Adane, L. Preparation of Laundry Soap from Used Cooking Oils: Getting value out of waste. Sci. Res. Essays 2020, 15, 1–10.
17. Adigun, O.; Manful, C.; Prieto Vidal, N.; Mumtaz, A.; Pham, T.H.; Stewart, P.; Nadeem, M.; Keough, D.; Thomas, R. Use of
Natural Antioxidants from Newfoundland Wild Berries to Improve the Shelf Life of Natural Herbal Soaps. Antioxidants 2019,
8, 536.
18. Mendes, B.R.; Shimabukuro, D.M.; Uber, M.; Abagge, K.T. Critical assessment of the pH of children’s soap. Jornal de Pediatria
(Versão em Português) 2016, 92, 290–295.
19. ČSN 68 1148 Methods of Test for Surfactants and Detergents-Surfactants-Analysis of Soaps-Determination of Free Corrosive Alkalis;
(Translated by Google Translate); Czech Standards Institute: Prague, Czech Republic, 1994.
20. Vivian, O.P.; Nathan, O.; Osano, A.; Mesopirr, L.; Omwoyo, W.N. Assessment of the physicochemical properties of selected
commercial soaps manufactured and sold in Kenya. Open J. Appl. Sci. 2014, 4, 433–440.
21. Khalifa, I.; Barakat, H.; El-Mansy, H.A.; Soliman, S.A. Improving the shelf-life stability of apple and strawberry fruits applying
chitosan-incorporated olive oil processing residues coating. Food Packag. Shelf Life 2016, 9, 10–19.
22. Kempka, A.P.; Horvath, F.J.; Fagundes, P.; Prestes, R.C. Foaming and emulsifying capacity, foam and emulsion stability of
proteins of porcine blood: Determination at different values of pH and concentrations. Revista Brasileira de Tecnologia 2015, 9,
1797–1809.
23. Tarun, J.; Susan, J.; Jacob Suria, V.J.S.; Criton, S. Evaluation of pH of bathing soaps and shampoos for skin and hair care. Indian
J. Dermatol. 2014, 59, 442.
24. Betsy, K.J.; Jilu, M.; Fathima, R.; Varkey, J.T. Determination of Alkali Content & Total Fatty Matter in Cleansing Agents. Asian
J. Sci. Appl. Technol. 2012, 2, 8–12.
25. Awang, R.; Ahmad, S.; Ghazali, R.A.Z.M.A.H. Properties of sodium soap derived from palm-based dihydroxystearic acid. J. Oil
Palm Res. 2001, 13, 33–38.
26. Gaweł, S.; Wardas, M.; Niedworok, E.; Wardas, P. Malondialdehyde (MDA) as a lipid peroxidation marker. Wiadomosci Lekarskie
2004, 57, 453–455.
27. Oboh, G.; Falade, A.O.; Ademiluyi, A.O. Effect of thermal oxidation on the physico-chemical properties, malondialdehyde and
carotenoid contents of palm oil. Rivista Italiana Delle Sostanze Grasse 2014, 91, 59–65.
28. Akkaya, M.R. Prediction of fatty acid composition of sunflower seeds by near-infrared reflectance spectroscopy. J. Food Sci.
Technol. 2018, 55, 2318–2325.
29. Jang, H.; Makita, Y.; Jung, K.; Ishizaka, S.; Karasawa, K.; Oida, K.; Takai, M.; Matsuda, H.; Tanaka, A. Linoleic acid salt with
ultrapure soft water as an antibacterial combination against dermato-pathogenic Staphylococcus spp. J. Appl. Microbiol. 2016, 120,
280–288.
30. Kawahara, T.; Takita, M.; Masunaga, A.; Morita, H.; Tsukatani, T.; Nakazawa, K.; Go, D.; Akita, S. Fatty Acid Potassium Had
Beneficial Bactericidal Effects and Removed Staphylococcus aureus Biofilms while Exhibiting Reduced Cytotoxicity towards
Mouse Fibroblasts and Human Keratinocytes. Int. J. Mol. Sci. 2019, 20, 312.
31. Huang, C.B.; George, B.; Ebersole, J.L. Antimicrobial activity of n-6, n-7 and n-9 fatty acids and their esters for oral microorgan-
isms. Arch. Oral Biol. 2010, 55, 555–560.
32. Matthaus, B.; Özcan, M.M.; Al Juhaimi, F. Some rape/canola seed oils: Fatty acid composition and tocopherols. Z. Naturforschung
C 2016, 71, 73–77.
Processes 2021, 9, 529 12 of 12

33. Aydınkaptan, E.; Mazı, I.B. Monitoring the physicochemical features of sunflower oil and French fries during repeated micro-
wave frying and deep-fat frying. Grasas Aceites 2017, 68, 202.
34. Aung, W.P.; Bjertness, E.; Htet, A.S.; Stigum, H.; Chongsuvivatwong, V.; Soe, P.P.; Kjøllesdal, M.K.R. Fatty acid profiles of
various vegetable oils and the association between the use of palm oil vs. peanut oil and risk factors for non-communicable
diseases in Yangon region, Myanmar. Nutrients 2018, 10, 1193.
35. Montoya, C.; Cochard, B.; Flori, A.; Cros, D.; Lopes, R.; Cuellar, T.; Espeout, S.; Syaputra, I.; Villeneuve, P.; Pina, M.; et al. Genetic
architecture of palm oil fatty acid composition in cultivated oil palm (Elaeis guineensis Jacq.) compared to its wild relative E.
oleifera (HBK) Cortés. PLoS ONE 2014, 9, e95412.