South African Journal of Chemical Engineering 40 (2022) 126–133

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South African Journal of Chemical Engineering

journal homepage: www.elsevier.com/locate/sajce

Extraction of oil from Maesa lanceolata seeds and evaluation of its

antimicrobial activities
Yigezu Mekonnen Bayisa, Mohammed Seid Bultum *
School of Chemical Engineering, Jimma Institute of Technology, Jimma University, P.O.BOX 378, Jimma, Oromia, Ethiopia

A R T I C L E I N F O A B S T R A C T

Keywords: Now a day utilization of plants’ extract as alternative medicine becomes a common practice all over the world
Traditional medicine merit many scientists to examine the antimicrobial activity of several medicinal plants. Maesa lanceolata is one
Maesa lanceolata seed of such ethnomedicinal plant traditionally utilized as a cure for the treatment of pain against various diseases.
Solvent extraction
Besides, there is plenty of research done on the determination of which potential constituents of this plant are
Bioactive component
Antimicrobial activity
responsible for such treatment. In this study, an attempt has been made to extract oil from Maesa lanceolata seed
using n-hexane solvent, evaluate its physio-chemical properties, and antimicrobial activity on a selected strain of
bacterial and fungal. The maximum oil yield of 35.3% was obtained at the optimum parametric values;
extraction temperature of 70 ◦ C, extraction time of 6 h, solute to solvent ratio of 0.06 g/ml, and particle size of
0.25 mm. The chemical compositions and functional group of the extracted oils were determined using Gas
Chromatography mass spectroscopy and Fourier Transform Infrared Spectroscopy analysis. The result shows that
the abundant fatty acid was monounsaturated oleic acid C18:1 (53.49%) and poly-unsaturated linoleic acid
C18:2 (31.48%). The qualitative analysis test of the extracted m. lanceolata seed oil shows the presence of
different phytochemical constituents which are microbial resistant and having an antimicrobial activity like
flavonoids, alkaloids, tannins, saponin, Terpenoids, and phenols.

1. Introduction many African countries (Theunis et al., 2007), including Central Africa,
Ethiopia, and Kenya, where it has been used as an ethnomedicine for a
Traditional medicine often includes herbal medicines, which consist long time (Shin et al., 2014). Its medicinal value is evident from its use
of biologically active compounds from plant materials (Brunning et al., for the treatment of pain against various diseases including infectious
2010). The utilization of plants as medicine is an old practice common to hepatitis, bacillary dysentery, impetigo, bacterial infections in the small
all societies particularly the developing nations (van Wyk and Prinsloo, intestine, viral infections in the liver and throat, as well as for treatment
2020), and currently, the number of people using traditional or alter­ of some types of dermatoses and neuropathies (Okemo et al., 2003; Shin
native medicine is rapidly increasing all over the world (Gebrehiwot et al., 2014; Theunis et al., 2007; Timothy et al., 2018). The presence of
et al., 2018). The key reasons behind using plants as sources of thera­ bioactive components such as polyphenols, tannins, and alkaloids in this
peutic medicines arise from the fact that they considering these products plant makes it highly wanted due to their beneficial effects on the human
to be both safe and effective (Amarasiri et al., 2020), and this has body (Uoonlue and Muangrat, 2019).
prompted scientists to investigate the numerous bioactive compounds The recognition of traditional medicine as an alternative form of
available in whole plants or a certain portion of it (Gagnon et al., 2012; health care by the general public and the development of microbial
Fabricant and Farnsworth, 2001). On the other hand, many underutil­ resistance to the available antibiotics has led many scientists to examine
ized medicinal plants have potential sources of therapeutic agents and the antimicrobial activity of several medicinal plants currently.
most of which have remained unexploited (Haile et al., 2019). For Furthermore, emerging antimicrobial drug-resistant bacterial strains
instance, Maesa lanceolata is one such medicinal plant traditionally (Finch and Hunter, 2006), merit research into novel therapeutic alter­
utilized as a cure for different treatments (Shin et al., 2014) . natives and medicinal plants contemporaneous potential candidates
Maesa lanceolata is a shrub or small tree that belongs to the genus (Essawi and Srour, 2000). Studies have stated that seeds of M. lanceolata
family of Maesacea with 150 species. (Shin et al., 2014). It is growing in are sources of valuable oil (Okemo et al., 2003). However, there is plenty

* Corresponding author.

https://doi.org/10.1016/j.sajce.2022.02.007
Received 16 May 2021; Received in revised form 19 August 2021; Accepted 28 February 2022
Available online 1 March 2022
1026-9185/© 2022 The Author(s). Published by Elsevier B.V. on behalf of Institution of Chemical Engineers. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/).
Y. Mekonnen Bayisa and M. Seid Bultum South African Journal of Chemical Engineering 40 (2022) 126–133

of research done on the determination of the antimicrobial activity of according to the method reported in (Ayoola et al., 2008).
the oil produced from the portion of this plant. Test for alkaloids: To 0.25 g of the crude extract was stirred with 5
The current research focuses on the extraction of oil from the seed of ml of 1% HCl on a steam bath. 1 ml of the filtrate was treated with a few
M. lanceolata, and the evaluation of its antimicrobial properties against drops of Mayer‟s reagent and another 1 ml was similarly treated with
different bacterial and fungi strains. Moreover, the study investigates Dragendorff‟s reagent. Turbidity or precipitation with both reagents was
the extraction of oil from Maesa lanceolata seeds using solvent extraction taken as preliminary evidence for the presence of alkaloids.
method, and evaluates the effects of different process parameters such as Test for saponins: To 0.25 g of the crude extract, 5 ml of distilled
particle size, extraction time, solid to solvent ratio, and temperature on water was added to a test tube. Then, the solution was shaken vigorously
the oil yield, and also determines the optimum operating condition for and observed for a stable persistent froth. The formation of froth indi­
oil extraction. Furthermore, the physicochemical properties, fatty acid cated the presence of saponins.
compositions, and antimicrobial activities of the extracted oil were Test for polyphenols (phenolic compounds): About 0.25 g of
evaluated. ethanol extract was treated with few drops of 5% neutral ferric chloride
solution; the appearance of a greenish precipitate indicated the presence
2. Materials and methods of phenol (Shetty et al., 2016).
Test for flavonoid: Ten ml of ethyl acetate was added into a test tube
2.1. Materials and reagents having 0.25 g of ethanol extract and heated on a water bath for 3 min.
The mixture was cooled and filtered. Then, about 4 ml of the filtrate was
Maesa lanceolata oil was extracted from Maesa lanceolata seed ac­ taken and shaken with 1 ml of dilute ammonia 24 solutions. The layers
quired from Jimma Agricultural Research centre, Jimma area, Ethiopia. were allowed to separate and the yellow color in the ammonia layer
The chemical n-hexane with a purity of 99% was collected from Chem- indicated the presence of flavonoids.
Supply Kirkos Ltd. in Addis Ababa, Ethiopia. The chemical (n-hexane) Test for Terpenoids: Two ml of chloroform was added into a test
used was of pure analytical grade. tube having 0.25 g of ethanol extract. Then, 3 ml concentrated sulfuric
Maesa lanceolata seeds were first cleaned from dirt, dust, sand, small acid was carefully added to form a layer. A reddish-brown coloration of
stones and washed manually. Then Maesa lanceolata seeds were dried for the interface indicated the presence of Terpenoids.
two days by sunlight. The cleaned and dried seeds were weighted and 1 Test for tannins: About 0.25 g ethanol extract was boiled in 10 ml of
kg Maesa lanceolata seeds were further dried in an oven at 105 ◦ C for an water in a test tube and then filtered. Three drops of 0.1% ferric chloride
hour to remove the rest of their moisture content. Then the dried seed were added to the filtrate. The presence of tannins was confirmed by the
was crushed using a crusher to a particle size ranging from 0.25 to 1.25 formation of brown greenish or blue-black color.
mm and stored in a plastic bag for further solvent extraction analysis. Gas Chromatography mass spectroscopy analysis was performed
The prepared proximate analysis of Maesa lanceolata seed was then with Agilent GC – system – 7820A (Palo Alto, CA, USA). Sample analysis
characterized using the ASTM procedure to determine the parameters was carried out on a packed column- Agilent Technologies (30 m ×
like moisture content, seed density, crude fat, and ash content. 0.250 mm, 0.25 μm). Fatty acid (FA) profile was determined according
to the AOCS (1998) official methods through the preparation of fatty
acid methyl esters (FAME) and GC analysis. The data, obtained using MS
2.2. Extraction of oil
- Agilent Technologies EMS detector and processed using Chemstation
software, were used to obtain fatty acid composition of oils.
To extract the Maesa lanceolata seed oil the Soxhlet apparatus (model
Fourier transform infrared (FTIR) spectrometer (Bruker Tensor 270,
DW-SF-06C, shanghai, China) was used. A grinded sample was placed in
Ettlingen, Germany) was used to determine various functional groups
a thimble and loaded into the Soxhlet extraction unit. A solid: solvent
contained in the extracted oil, and the spectra were recorded from 4000
ratio from 0.02 to 0.1 g/ml, extraction time from 2 to 6 h., temperature
to 400 cm− 1. The analysis will be done using KBr for extracted m. lan­
from 65 to 75 ◦ C, and particle size from 0.25 to 1.25 mm were employed.
ceolata oil.
Finally, the solvent and oil were separated using a rotary evaporator and
the solvent was recovered using a condenser. Once the oil is extracted
using n-hexane, purification to remove free fatty acids was carried out 2.4. Determination of antimicrobial activity
using standard procedures and characterized with different techniques
(Jiao et al., 2014). The percentage oil yield was calculated using Antimicrobial activity of hexane extracted of m. lanceolata oil was
tested against the bacterial and fungal strain according to(Ayoola et al.,
percentage of oil yield =
MO
∗ 100 % 2008). Those activity tests of seed oil and its five fractions were per­
MS formed using the disk diffusion method and the zone of inhibition was
measured in millimeters (mm). The dimethyl Sulfoxide (DMSO) was
where Mo is the weight of extracted oil (g), and Ms is the weight of the used as negative control against bacteria and positive for fungi and
seed sample before extraction (g). Ciprofloxacin was used as a positive and negative control for bacteria
and fungi respectively. Reliable cultures of bacteria and fungi namely
2.3. Determination of the properties of oil Bacillus cereus, Staphylococcus aureus, Escherichia coli, Aspergillus
Niger, and Candida albicans were obtained from “Microbial Type Cul­
The quality and purity of the extracted m. lanceolata oil was con­ ture collection”, biotechnology research center, Jimma, Ethiopia. Bac­
ducted by different spectroscopic and physicochemical properties ana­ terial cultures were maintained on the Nutrient Agar medium whereas
lyses were. fungal cultures were maintained on the Sabouraud Dextrose Agar me­
The Physicochemical properties (kinematic viscosity, color, acid dium. All bacterial strains were incubated for 24 h at 37 ◦ C and fungal
value, iodine value, saponification value, and specific gravity) of the strains were incubated at 27 ◦ C for 48 h. The test organisms were grown
extracted oil were analyzed according to the American Society of Testing in respective broth media i.e.; Nutrient Broth for bacterial strains and
and Materials (ASTM) procedure: specific gravity (ASTM D4052, using Sabouraud Dextrose broth for fungal strains.
10 ml pycnometer), kinematic viscosity (ASTM D445), acid value Antibacterial activity following 24 h of incubation of the test plates
(titration method), iodine value (glass stopper conical flask, ASTM the clear zones was measured using a ruler. This was done by measuring
D5554) and saponification value (titration method). the entire diameter of the clear zone and the results were recorded.
The presence of phytochemicals like alkaloids, flavonoids, saponins, The antifungal activity of the oil and its fractions was assessed daily
tannins, terpenoids, and phenol in crude M. lanceolata oil was evaluated for three consecutive days through the measurement of the diameter of

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Y. Mekonnen Bayisa and M. Seid Bultum South African Journal of Chemical Engineering 40 (2022) 126–133

Table 1 3.2. Statistical analysis

Analysis of variance for response surface quadratic model of m. lanceolata seed
oil yield. Table 1 shows the models and their significant coefficients of the
Sources Sum of Degree of Mean of F- P-value responses, where all the models are significant at a level of less than 0.
squares freedom squares value 0001. The statistical analyses show that quadratic models fit very well
Model 371.37 14 26.53 143.64 <0.0001 into the data for the response and the response was highly significant (p
A-temperature 19.51 1 . 19.51 105.63 < < 0.0001). The Model F value of 143.64 implies the model is significant
0.0001 for extracted m. lanceolata seed oil yield. Form. lanceolata oil yield A, B,
B-solute 6.75 1 6.75 36.55 <
C, D, AB, AC, AD, CD, A2 B2, C2, and D2 were found to significantly affect
solvent ratio 0.0001
C-extraction 87.48 1 . 87.48 473.69 < the yield of extracted. lanceolata oil, while, BC, and BD were not signif­
time 0.0001 icant as is shown in (Table 1). The analysis of variance for the lack of fit
D-particle size 150.52 1 . 150.52 815.04 < test did not show the inadequacy of the model concerning them. lan­
0.0001 ceolata oil yield (P > 0.05), indicating that the model could adequately
AB 1.21 1 . 1.21 6.55 0.0227
AC 1.21 1 1.21 6.55 0.0227
fit the experimental data.
AD 6.50 1 6.50 35.21 <
0.0001 3.3. Influence of process parameter on extracted oil
BC 0.1225 1 0.1225 0.6633 0.4290
BD 0.5625 1 0.5625 3.05 0.1028
3.3.1. Effect of extraction temperature
CD 1.32 1 1.32 7.16 0.0181
A2 74.73 1 74.73 404.63 < During the extraction process, temperature has a high impact on oil
0.0001 yield as shown in Fig. 1a. From this figure, it is evident that an increment
B2 14.24 1 14.24 77.11 < in the temperature favored the extraction process, and a maximum of
0.0001
2
35.5% oil yield value was obtained at 70 ◦ C after a reaction time of 6 h
C 35.26 1 35.26 190.95 <
0.0001
and 0.25 mm particle size. Nevertheless, as the temperature increased
D2 6.74 1 6.74 36.48 < from 70 to 75 ◦ C the oil yield was slightly decreased which shows that
0.0001 the temperature around 70 ◦ C is an optimum temperature. This shows
Residual 2.59 14 0.1847 that increasing the temperature is always related to counter effects on
Lack of Fit 2.35 10 0.2353 4.04 0.954
the oil yield, which is due to increasing the rate of diffusivity through the
cell walls of the seed, decreasing solvent density, and increasing mass
inhabitation zones in millimeters using a transparent ruler. transfer and vapor pressure of the solutes (Jadhav et al., 2016).

2.5. Statistical analysis 3.3.2. Effect of solute to solvent ratio

The other parameter of the extraction process which has a significant
Response Surface Methodology was used to determine the effects of effect on the extracted oil was the solute (g) to solvent (ml) ratio and the
extraction factors and to optimize the percentage oil yield response. A obtained result was presented in Fig. 1b. From this figure, as the solute
Box-Behnken Design (BBD) with four numerical factors on three levels to solvent ratio increased from 0.02 to 0.06 g/ml the extracted oil was
was used (Yolmeh and Jafari, 2017) according to extraction was done in increased, High solvent rate leads to a higher diffusion in mass transfer
Section 2.2 that considering the four factors such as solid: solvent ratio, and is attributed to the rapid solution of the oil on the solid’s surface
extraction time, temperature, and particle size were investigated as in­ which resulted in high initial extraction rate, whereas it was decreased
dependent variables. from 0.06 to 0.1 g/ml. The reason behind this decrement in oil yield is
related to the mass transfer process. On the other hand, the lower solvent
3. Result and discussion concentration has a lower motive force resulted in a lower extraction
rate and formulation of agglomeration which produces a high amount of
3.1. Results of proximate analysis of m. lanceolata seed the cake (Reshad et al., 2015).

The moisture content of the sample, which is an important factor that 3.3.3. Effect of extraction time
affects the oil yield and quality, was obtained after drying the seed in the Extraction time is another parameter that affects both the extraction
oven at 105 ◦ C for 24 h. The average moisture content was found to be efficiency of the solvent and oil yield. As shown in Fig. 1c, as the reaction
5.02 ± 0.13%% which was comparable with the value reported by time increases the oil yield was extremely increased until it reaches its
(Abdelaziz et al., 2014), for castor seed and lower than the value re­ optimum value and slightly increased beyond that which shows that the
ported by (Mohamed et al., 2014). Moisture content in the seeds de­ equilibrium point is reached above the limit center. Initially, the oil
pends upon the maturity and quality of the seed. The moisture contents diffuses quickly from the grinded sample to the liquid phase due to the
of seed determine the ability of all seeds to be stored well and the critical high concentration gradient (Sayyar et al., 2009). This enhances the oil
moisture level for the beginning of rapid spoilage is relatively higher in extraction rate due to the rapid penetration of solvent into the seed
seeds of low oil contents and relatively low for high oil content seeds sample inducing high oil yield (Reshad et al., 2015; S Zhao and Zhang,
(Zerihun and Berhe, 2020) The result shows that the sample is suitable 2013.). However, after the optimum extraction time the extraction rate
for oil extraction without further drying of the seeds. In addition to this, was diminished which indicates that the sample was left with a residual
the obtained value of volatile, ash content, and fixed carbon of m. lan­ amount of oil, and also may be due to the saturation of the solvent with
ceolata seed were 86.46± 0.24, 2.85± 0.11, and 5.73± 0.09% respec­ oil.
tively, Whereas the density of m. lanceolata seed was 1.02 g/ml. This
refers m. lanceolata seed was rich in volatiles (86.46%) but low in ash 3.3.4. Effect of particle size
content (2.25%). The value obtained for fixed carbon content is very low Particle size has a significant effect on the extraction process Fig. 1.
this due to high contained of volatile mass dry based. d shows the effects of particle size on the oil yield. The result shows that
fine particle produces a high yield of oil for the seed sample, (35%). This
may be due to the reason related to the surface area of contact and oil
diffusion (Jalili et al., 2018; Reshad et al., 2015; Yusuff, 2021; S Zhao
and Zhang, 2013.). This means that it is more difficult for the solvent to

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Fig. 1. Influence of process parameter on extracted oil.

diffuse freely through the larger particles due to the larger distance to enhanced the solubility of the oil and its mass transfer was more rapid,
the solvent entrance and exposure to lower oil yield due to small surface thus resulting in high extraction yield up to 70 ◦ C. But, further increment
contact. Similarly, for smaller particle sizes the solvent is freely or highly of temperature leads to increasing in the rate of diffusivity through the
dispersed or diffuse through the seed due to larger surface contact which cell walls of the seed, decreasing of solvent density, and increasing mass
resulted to give a high oil yield using the solvent. transfer and vapor pressure of the solutes which resulted in a slight
decrement in oil yield (Jadhav et al., 2016). On the other hand, a high oil
3.3.5. Interaction effects yield favors a small solute-solvent ratio. The same observation was re­
The 3-D Response Surface diagram Plotted in Fig. 2 illustrates the ported by Reshad et al. (2015), where the maximum oil yield of 49% was
interaction effects of all experimental variables on oil extraction yield obtained with a 0.08 solute to solvent ratio.
Fig. 2.a shows that the effect of the Solute-solvent ratio and Temperature Fig. 2b displays the interaction effect of extraction time and extrac­
on oil yield for 0.75 mm particle size and 4 h. extraction time. As it is tion temperature on the oil yield for a solute-solvent ratio of 0.06 g/ml
observed from the plot the oil yield increased until some optimum and particle size of 0.75 mm. The plot shows that as the reaction time
temperature is reached, i.e., 70 ◦ C, and slightly decreased beyond that increases the oil yield was extremely increased until it reaches its opti­
point. The reason for this observation was that increasing temperature mum value and slightly increased beyond that which shows that. This

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Y. Mekonnen Bayisa and M. Seid Bultum South African Journal of Chemical Engineering 40 (2022) 126–133

Fig. 2. Interaction effects of process variable on M. lanceolata of oil yield.

was due to the reason that initially the oil diffuses quickly from the seed particle size increased. This shows that higher extraction time with
grinded sample to the liquid phase due to the high concentration lower particle sizes give a higher yield and higher particle sizes with
gradient which enhances the oil extraction rate due to the rapid pene­ lower extraction time can give a lower yield. This observation agreed
tration of solvent into the seed sample inducing high oil yield. However, with the result reported by Reshad et al. (2015).
after the optimum extraction time of around 4 h. there is a slight change
in the extraction rate which may be due to the saturation of the solvent 3.4. Physiochemical properties of extracted M. lanceolata seed oil
with an oil (S Zhao and Zhang, 2013.).
Fig. 2c also illustrates the effect of M. lanceolata powder particle size M. lanceolata seed oil obtained from M. lanceolata seed was charac­
and extraction temperature on oil extraction yield for a solute-solvent terized for its physicochemical properties according to the standard
ratio of 0.06 g/ml and 4 h. extraction time. The figure showed that method. The color of oil obtained during the extraction process was
the oil yield decreased as the seed particle size increased. The reason is golden yellowish whereas the specific gravity of the extracted oil was
that seed particle with smaller particle size has a high specific surface 0.92±0.14 which was lighter than the pure water. To determine the
area, which facilitates the solvent diffusivity within the seed powder and rancidity and stability of the oil for long time storage the acid value and
enhances mass transfer rate. This observation agreed with most of the peroxide value of the oil were investigated respectively. The acid value
studies on oil extraction such as extraction of oil from Moringa oleifera of obtained oil was 1.25 ± 0.18 mg of KOH/ g of an oil which cannot
seeds (S Zhao and Zhang, 2013.), from rubber seeds (Reshad et al., undergo rancidity. The peroxide value extracted oil was 1.45±0.11
2015), and Canola Seeds (Jalili et al., 2018). Meanwhile, the oil meq/kg resulted in a low level of oxidative and lipolytic activities
extraction yield increased with increasing extraction temperature. (Uoonlue and Muangrat, 2019). To understand alkali reactive groups in
The influence of the interaction of extraction time and particle size fats and oils and predicting the type of glycerides the saponification
on the M. lanceolata seed yield was investigated, and the result is value of the oil was determined and the value was 126±0.15 KOHg− 1
depicted in Fig. 2d. The figure revealed that oil yield increased rapidly which was fall in range and cannot leads into soap formulation due to
with increasing extraction time, whereas the oil yield decreased as the lower than 200 ± 0.12 KOHg− 1. In addition to these, the iodine value of

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Y. Mekonnen Bayisa and M. Seid Bultum South African Journal of Chemical Engineering 40 (2022) 126–133

Table 2
Results for the antimicrobial activity m. lanceolata oil.
Zone of inhibition (mm)
Sample tested Serial dilution Bacterial strain Fungal strain
% v/v Bacillus cereus Staphylococcus aureus E. coli Aspergillus Niger Candida albicans
NA
M. lanceolata oil 1/32 12 7 11 11
NA
1/16 13 9 13 12
NA
1/8 16 11.5 15 14
NA
1/4 18 13.4 16 15
NA
½ 20 16 18 17
NA NA NA
Ciprofloxacin 20 25
NA NA NA
Dimethyl Sulfoxide (DMSO) 20 25

NA = not active.

obtained oil was 110.5 ± 0.02 g I2/ 100 g of an oil which falls in the
Table 3
range of semi-drying oil and indicates that the oil has high unsaturated
A fatty acid profile of m. lanceolata oil.
fatty acid (Haile et al., 2019). The obtained values of physicochemical
properties extracted oil were important for making medicinal raw since Fatty acid Carbon structure Saturation level %total abundance
it falls in ranges and indicating a minimal degree of triglyceride hy­ Palmitic 18:0 Saturated 5.14
drolysis to free fatty acid and glycerol, and minimal triglyceride was Heptadecanoic acid 17:0 Saturated 1.1
Pentadecanoic acid 15:0 saturated 1.03
oxidized.
Oleic 18:1n-9 Monounsaturated 53.49
Linoleic 18:2n-6 Polyunsaturated 31.48
3.5. Phytochemical properties of oil extracted from M. lanceolata seed Linolenic 18:3n-3 Polyunsaturated 3.87
Stearic 16:0 Monosaturated 4.93

The qualitative analysis test of the extracted m. lanceolata seed oil

shows the presence of different phytochemical constituents which are concluded that the presence of active components in m. lanceolata seed
microbial resistant and having an antimicrobial activity like flavonoids, oil would help develop the extract as medicinal raw material.
alkaloids, tannins, Terpenoids, and phenols. The phytochemical
screening of the plants studied showed the presence of these all com­ 3.7. Fatty acid profile of M. lanceolata seed oil
pounds. The presence of the flavonoids indicates the contribution of
their share for the observed antimicrobial activities especially against A fatty acid profile is the main determinant of oil quality. As shown
gram-positive bacteria for damaging or disruption of the cell membranes in Table 3, the obtained m. lanceolata oil compose seven main compo­
and inhibition of the synthesis of nucleic acids which can lead to the nents from these three saturated fatty acids, two monounsaturated fatty
death of the susceptible bacterium (Uoonlue and Muangrat, 2019). acids, and two polyunsaturated fatty acids were identified. Among them,
The presence of phenolic compounds have been found to have a the most abundant fatty acid was monounsaturated oleic acid C18:1
growth inhibition effect against different bacteria as well as Terpenoids (53.49%), poly-unsaturated linoleic acid C18:2 (31.48%), linolenic acid
have the ability to have antibacterial activities against various patho­ (3.87%), saturated palmitic acid C16:0 (5.14%), stearic acid C18:0
genic microbes and to the disruption of the cytoplasmic cell membrane (3.9%), heptadecanoic acid (1.1%) and pentadecanoic acid (1.03%). It
(Jiao et al., 2014). Both saponins and alkaloids also have an antimi­ is, however, important to note that the high percentage of mono-
crobial activity to inhibit the growth of both gram-positive and unsaturated oleic acid provides a high degree of oiliness (L Zhao et al.,
gram-negative test bacterial strains (Gharsallah et al., 2021), meanwhile 2016.). Acid’s degree of unsaturated fatty acid leads to solidification at
tannic acid also plays a role in inhibiting acyl-homoserine lactones and low temperature or cloud formation. Oils rich in unsaturated free fatty
chelate metal ions like Fe (II) and interfere with one of the reaction steps (e.g. oleic acid and linoleic acid) are generally more stable to oxidative
in the Fenton reaction thus retarding oxidating. The presence of those rancidity and stable as deep-frying oils (Wu et al., 2018).
phytochemical constituents makes the extract suitable for pharmaco­ The total percentage of unsaturated fatty acid in this study of
logical action on the human body. extracted oil of m. lanceolata oil was 88.83% which was lower than
sunflower oil, but higher than the total percent of unsaturated fatty acid
3.6. Antimicrobial activity value of Soybean oil, Mustard oil, Palm oil, and Coconut oil for values
81.14, 86.18, 53.30, and 7.12% respectively (S. Feleke, F. Haile, 2012).
The results of the initial antimicrobial screening assay of the crude This indicates that the extracted m. lanceolata oil has the potential to
extracted m. lanceota seed oil on selected microbial strains in different produce an alternative healthy oil since it contains high polyunsaturated
concentrations was shown in Table 2. The result suggests that m. lan­ fatty acid percentages.
ceolata seed oil was active on both phytogenic strains. A similar result
was also reported by (Bourgou et al., 2020). As the concentration of m. 3.8. Functional groups found in M. lanceolata seed oil
lanceolata seed oil has increased the zone of inhibition was increased on
both phytogenic strains. The bacillus cereus was more sensitive than The presence of various organic functional groups in the m. lanceo­
Staphylococcus aureus which shows more zone of inhibition with lata oil is identified through the FT-IR spectrum shown in Fig. 5. The
respective concentration and for E. coli is not active. FTIR analysis of the extracted oil showed the presence of different
On other hand, as the concentration of the serial dilution or Mini­ functional groups that belongs to different compounds. The weak ab­
mum inhibition concentration increased, the obtained oil activities sorption band that appeared in the range of 3250–3550 cm− 1 indicates
against the Aspergillus Niger and Candida albicans also increased. In the presence of –OH stretching vibrations (Yusuff, 2021), which in turn
general, the extracted oil from M. lanceolata seed was active against confirm the presence of phenols and alcohols in the m. lanceolata oil.
tested microorganisms, showing the lowest MIC values (1/32 and 1/ Fatty acids groups were identified by the presence of C–H vibrations
32% (v/v) against all strains except for E. coli (Jiao et al., 2014). The with a very sharp peak between 2950 and 2850 (Yusuff, 2021) and
antimicrobial activity of the m. lanceolata oil may be attributed to carbonyl bands at 1750 and 2800 cm− 1 (Yusuff and Ewere, 2020). Ar­
various phytochemical constituents present in the oil extracts. It can be omatic C = C skeletal vibrations at 3120 and 1460 cm− 1 are typical of

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Y. Mekonnen Bayisa and M. Seid Bultum South African Journal of Chemical Engineering 40 (2022) 126–133

Fig. 3. Effect of m. lanceolata oil on selected microbial.

Fig. 4. FTIR spectroscopy.

aromatic structures in flavanols. Strong absorption at 1180 cm− 1 is and alcohols group in m. lanceolata seed oil is confirmed by FT-IR
characteristic of C–O vibrations in sugar units. The peak observed at analysis.
1575–1675 cm− 1 corresponds to C–H stretching vibrations of alkenes
and aromatics (Dutta et al., 2014; Yusuff, 2021; Yusuff and Ewere, 4. Conclusion
2020). The presence of mono, polycyclic, and substituted aromatic
groups is confirmed by the appearance of peaks at 1420–1610 cm− 1. The results of the present study provide substantial evidence that
Also, bands at 650 cm− 1 are ascribable to aromatic structures (Dutta Maesa lanceolata seed is a good source of oil enriched with bioactive
et al., 2014). Thus, the presence of various alkanes, aromatics, phenolic compounds. The study revealed that all four variables significantly

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Y. Mekonnen Bayisa and M. Seid Bultum South African Journal of Chemical Engineering 40 (2022) 126–133

affect the oil yield. Based on our finding extraction temperature of 70 ◦ C, Jalili, F., Jafari, S.M., Emam-Djomeh, Z., Malekjani, N., Farzaneh, V., 2018. Optimization
of ultrasound-assisted extraction of oil from canola seeds with the use of response
extraction time of 6 h, solute to solvent ratio of 0.06 g/ml and Particle
surface methodology. Food Anal. Methods 11 (2), 598–612, 10.1007/s12161-017-
size of 0.25 mm is the best combination of process parameters that result 1030-z.
in the highest oil yield of 35.5%. The phytochemical characteristic of the Jiao, J., Li, Z.G., Gai, Q.Y., Li, X.J., Wei, F.Y., Fu, Y.J., Ma, W., 2014. Microwave-assisted
extracted oil suggests the presence of bioactive compounds such as fla­ aqueous enzymatic extraction of oil from pumpkin seeds and evaluation of its
physicochemical properties, fatty acid compositions, and antioxidant activities. Food
vonoids, alkaloids, tannins, and phenols which are suitable for subse­ Chem. 147, 17–24, 10.1016/j.foodchem.2013.09.079.
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bacterial and fungal diseases. The total percentage of unsaturated fatty Applied Science and Agriculture Formation of Dihydroxystearic Acid from
Hydrolysis of Palm Kernel Oil-Based Epoxidized Oleic Acid. 9(11), 86–92.
acid of extracted oil was 88.83% of which 53.49% is monounsaturated Okemo, P.O., Bais, H.P., Vivanco, J.M., 2003. In vitro activities of Maesa lanceolata
oleic acid. The antimicrobial activity results of M. lanceolata seed oil was extracts against fungal plant pathogens. Fitoterapia 74 (3), 312–316. https://doi.
active against tested microorganisms, showing the lowest MIC values org/10.1016/S0367-326X(03)00039-X.
Reshad, A.S., Tiwari, P., Goud, V.V., 2015. Extraction of oil from rubber seeds for
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Declaration of Competing Interest Sayyar, S., Abidin, Z.Z., Yunus, R., Muhammad, A., 2009. Extraction of oil from Jatropha
seeds-optimization and kinetics. Am. J. Appl. Sci. 6 (7), 1390–1395, 10.3844/
ajassp.2009.1390.1395.
The authors declare that they have no known competing financial Shetty, S.B., Mahin-Syed-Ismail, P., Varghese, S., Thomas-George, B., Kandathil-, P.
interests or personal relationships that could have appeared to influence (2016). Antimicrobial effects of Citrus sinensis peel extract against dental caries
bacteria : an in vitro study. 8(1). 10.4317/jced.52493.
the work reported in this paper.
Shin, S.Y., Dekebo, A., Dinku, W., Terfa, A., Lee, Y.H., Lim, Y., Yong, Y., 2014.
Identification of an anticancer compound contained in seeds of Maesa lanceolata, a
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