Enhancing The Hydrolysis of Africa Pear Seed Oil

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Volume 1 Issue 3 2022 International Journal of Scientific Research and Modern Technology

Enhancing the Hydrolysis of Africa Pear Seed Oil


M. A. Otache*, T. S. Arerefagha and S. C. Amagbor
Department of Industrial Chemistry, Michael and Cecilia Ibru University, Ughelli, Delta State, Nigeria.
(Corresponding author: M. A. Otache*)

Abstract:- Recent trend in terms of waste seed oil detergents, greases, cosmetics, and a variety of other things
application has shown vast industrial relevance. [11].
Therefore, the high fatty acid concentration of African
Pear (Dacryodes edulis) Seed-oil (APSO) is the reason for One of the most important steps in the manufacturing of
its utilization, thereby according it the possible potential seed oil is extraction as reported [12]. Therefore pretreatment
as feedstock for industrial applications. However, of the oil extract is key to its functionality as feedstock in
pretreatment of the oil extract is key to its functionality as industrial processes. Therefore, these pretreatment practices;
feedstock in industrial processes. Therefore the process of enzyme digestion, ultrasonication, microvaving, pulse
hydrolysis as a pretreatment route based on the types of electric fields etc., have been reported and grouped based on
catalysts, and a variety of reaction circumstances, chemical, physical, physicochemical and biological as
including concentration, temperature as well as reaction captured in figure 1.
time was adopted and characterization was affirmed with
FTIR and GC-MS analytical methods. The results showed
that the highest percentage Free Fatty Acid (% FFA)
hydrolysis was obtained at optimum concentration (2 M),
temperature (60 oC) and reaction time (2 hrs). Rapid
hydrolysis was observed at 1.5 M for both ethanolic KOH
and NaOH, thereby reporting a percentage yield of 70.58
% and 63.37 % of the % FFA for KOH and NaOH
respectively. Further characterization findings by GC-
MS confirmed a positive identification of % FFA
composition. Likewise the FTIR analysis also exhibited
high carboxylic acid peak absorption for NaOH and KOH
catalyst at 1781 cm-1 and 1179 cm-1 respectively.
Therefore, the result is a pointer that catalyst and the
choice of a suitable reaction condition has the potential to
influence the rate of hydrolysis in oil. Fig 1 Pretreatment techniques of oil [5, 13]

Keywords:- Hydrolysis, Fatty Acids, Pretreatment, Catalyst, In seed oil processing, physicochemical properties are
Physicochemical. essential quality criteria that has the potential to enhance
oxidative stability and functional components of seed oil
I. INTRODUCTION thereby improving its commercial value and shelf life [5, 14,
15]. Although pretreatment of seeds has been shown to have
The continuous search for an alternative industrial raw a considerable impact on factors such as oil output and colour
material for sustainable green practices, has unveiled several [5, 16, 17 18].
quest towards the use of waste sources [1]. The rapid global
growth in the area of industrialization and modernization has The alkaline hydrolysis of triacylglycerol is known as
triggered the high dependence on synthetic products that has saponification as described in Figure 2. Most oleo-chemical
created more cause of concern in the ecosystem. Evidence in productions begin with FFA, which are produced by these
this practices is a reflection in the dwindling petroleum processes. To maximize the economics of large-scale
reserves and numerous environmental pollution challenges production, reaction pathways and conditions with effective
occasioned by emission of greenhouse gases. Adopting glycerol recovery are necessary [8, 19]. Nowadays,
Africa Pear Seed-oil (APSO) as a potential feedstock for researchers employ a one-way rather than reversible alkaline
industrial application is based on the high oil yield as well as saponification as the catalytic route to the hydrolysis of esters
important FFA content [2]. It also contains bioactive based on the ease via which the product is separated. [11].
chemicals with a variety of functional qualities, including furthermore, it has been reported that FFA hydrolysis is
antioxidant and anti-cancer effects, antiinflammatory, etc. [3, affected by the type of catalysts, as well as the oil type [11,
4, 5]. These functional properties have revealed new potential 20, 21]. It is also affected by variables such as temperature
application in pharmaceuticals and foods [5, 6]. and time to accomplish 100 percent hydrolysis. Findings from
research using NaOH and KOH revealed that when methanol
The extraction of FAs and glycerol from oils is critical, and NaOH were used, the experiment was carried out with
particularly in the oleo-chemical industry [7]. This reaction the best molar ratio (6:1). However, when KOH was used, the
can be catalyzed via acid, base, or enzymatic route [8, 9]. experiment was carried out with the best molar ratio (8:1)
These techniques is employed in food, cosmetics, and while maintaining the catalyst concentration (1 percent
pharmaceutical industries [8, 10], as well as soap, synthetic

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Volume 1 Issue 3 2022 International Journal of Scientific Research and Modern Technology

KOH), 70°C reaction temperature as well as a reaction  Hydrolysis


duration of 31/2 h. APSO hydrolysis for the production of FFA was
obtained as described by A.O.A.C. [26] with some
modification. In a typical experiment, APSO (25 g) was
mixed in the reactor with 150 mL of saponifying solution
comprising of ethanolic KOH and NaOH concentration (0.5-
2.00 M), and ethanol (150 mL: 90% v/v). The hydrolysis was
carried out in a 250 mL temperature-controlled reactor at
different reaction temperature 40-70°C and different reaction
time 1-2.5 h. Following hydrolysis, 20 mL of water was
added to the mixture. Thereafter, separation by extraction
with 50 mL of hexane was done to remove unsaponifiables
components. The soap-containing aqueous alcohol phase was
acidified to pH 1 with HCl 6N, and the FFA was extracted
Fig 2 Alkaline hydrolysis of triglycerides [8] with hexane. To achieve a neutral pH, the extract was rinsed
with distilled water. A separating funnel was used to remove
To improve the nutritional quality of seed oil, maximum the lowest layer, which was then discarded. Anhydrous
fatty acid recovery during oil extraction is required [22]. magnesium sulfate was used to dry the FFA-containing top
Pretreatment of seeds has a little influence on fatty acid layer, and the solvent was evaporated at 35°C in a vacuum
content, according to scientists, which could be favourable rotary evaporator. Following that, the FFA percentage was
from a nutritional standpoint. This assertion was based on calculated.
study conducted to examine the effect of microwave
pretreatment (2450 MHz, 100, 250, 600 W, 2, 6 min) on fatty  Determination of % Free Fatty Acid (FFA)
acids composition [5, 23, 24]. According to their report, Method according to Mahesar et al., [27] was adopted
microwave pretreatment did not affect the FFA content [17]. for the determination of the percent FFA in the hydrolyzed
However, in another finding by Mohseni et al. [25], reported seed oil. In a flask, 50 mL isopropanol was added, followed
that pretreatment procedures strongly affected the FFA by 0.5 mL phenolphthalein, which was then neutralized with
content. They discovered that the amount of oleic and linoleic sodium hydroxide (NaOH, 0.5 N) until a permanent pink
acids had reduced considerably. Thermal deterioration, colour was obtained. The neutralized isopropanol was added
according to the authors, was the cause of the drop in oleic to the 5 g of FFA, which will be placed into an Erlenmeyer
acid and linoleic acid. Therefore as a means of sorting an flask, and about 0.5 mL of phenolphthalein was added. After
effective catalytic route for the production of the required shaking the mixture gently, the mixture is neutralized by the
FFA for industrial application, this study will focus on a addition of NaOH, 0.5 N until the first permanent pink colour
comparative study involving the use of KOH and NaOH in is obtained. The % FFA was calculated by using the equation.
enhancing an effective hydrolysis of APSO.
% FFA as oleic = 28.2 × N × V
II. EXPERIMENTAL METHODS W

 Materials and Apparatus Where; V = Volume in ml of 0.5 N NaOH required for


African Pear Seeds (Dacryodes Edulis) oil, Wij’s titration in ml.
solution, n-hexane as well as other chemicals are products of W = Weight in g of sample taken.
Sigma-Aldrich, USA and Merck Co Ltd. Laboratory oven
(DHG 9030) magnetic stirrer with hotplate (UNICON). A  FTIR spectroscopic analysis
three-necked round-bottom flask, a measuring cylinder, a For this investigation, we used a SHIMADZU FTIR-
beaker, a separating funnel, a burette, a density bottle, 8400S equipped with deuterated triglycine sulphate (DTGS)
funnels, thermometers, and a measuring flask were all as the detector and potassium bromide (KBr) as the beam
utilized. splitter, as described by Liang et al., [28]. The measurements
were at room temperature in the IR range of 4000–450 cm-1
 Sample Collection and Treatment by aggregating 40 scans with a resolution of 4 cm-1.
Samples were purchased from Otovwodo market in
Ughelli North Local Government Area of Delta State,  GC-MS Analysis
Nigeria. To separate the seed from the pulp, the seeds were On a GCMS-3800 equipment, gas chromatography-
dehulled using a sharp stainless knife. The produced pulp mass spectrometry analysis was done. This technique was
sample was then dried for 48 hours in a Gallenkamp hot air adopted from Adams [29]. Hexane (99 percent, Sigma–
oven at 70 0C. The powdered dried materials were pulverized Aldrich, Germany) was used to dilute 20 microliters of the
into a fine powder. extracted oil sample to 1 mL. The flow rate of helium carrier
gas (99.999 percent, AGA Lithuania) was set at 1.23 mL/min.
After injection, the oven temperature was maintained at 40 °C
for 2 minutes before being programmed at 3 °C/min to 210
°C, where the column was kept for 10 minutes. The split ratio
was set at 1:10 with a mass detector electron ionization of 70

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Volume 1 Issue 3 2022 International Journal of Scientific Research and Modern Technology

eV. A search of a mass spectra database was used to identify 77 66.5


volatile chemicals (NIST 14).
76 66
III. RESULTS AND DISCUSSION 75
65.5
74
 Percentage of free fatty acid (% FFA)
73 65
Experiments on percent FFA for KOH and NaOH under
reaction conditions of concentration, temperature and 72 64.5
reaction time as captured in Figure 3, 4 and 5 respectively 71
yielded the following findings. The findings demonstrate the 64
70
hydrolysis performance of these catalysts as well as their
69 63.5
impact under various experimental settings. The results
showed that increasing the KOH concentration as shown in 68 63
figure 3 indicates a corresponding increase in the % FFA. 40 50 60 70
Results from this study is in agreement with findings from
observation involving C. rugosa lipase hydrolysis of various KOH NaOH
vegetable oil [30-31]. However, increasing the concentration
of NaOH did not result in any significant changes in the Fig. 4 Effect of Temperature (oC) on % FFA
percentage of FFA. Furthermore, the impact of temperature
as captured in figure 4, showed that when the temperature was
raised from 50 to 70°C, the percent FFA increased, with a 77 68.5
maximum yield obtained at 60°C. This is also an important
indicator that the hydrolysis of APSO is influenced by
68
temperature. The result also in figure 5, describe the effect of 76.5
reaction time (1-2.5 hrs) on the % FFA, which showed an
increased in % FFA with an increase in the reaction time for 67.5
both catalyst. However, the highest yield of % FFA was 76
recorded after 2 hrs for both catalyst. 67
75.5
74 64.2 66.5
73 64 75
66
72
63.8
71 74.5 65.5
63.6
1 1.5 2 2.5
70
63.4
69 KOH NaOH
63.2
68 Fig. 5 Effect of Reaction time (h) on % FFA
67 63
 FTIR Analysis of Fatty Acids
66 62.8 The hydrolysis of APSO sample revealed by FTIR
0.5 1 1.5 2 spectroscopy as summarized in Table 1, displayed the main
peaks as well as their functional groups. For the FTIR study,
KOH NaOH the reaction conditions that yielded the highest yield (2 M, 60
o
C, and 2 hours) were used.
Fig. 3 Effect of Concentration (M) on the % FFA

Table 1 FTIR wavelengths of functional groups of APSO before and after hydrolysis
Wavelengths Functional group
S/N Fresh oil Hydrolyzed oil sample Hydrolyzed oil sample
sample with KOH with NaOH
1 3468 cm-1 - 3468 cm-1 The vibration of O-H stretching
2 2917 cm-1 2912 cm-1 2917 cm-1 Unsaturated C-H stretching of carbon chain
3 2851 cm-1 2849 cm-1 2851 cm-1 Saturated carbon chain C-H stretching vibration
3 1736 cm-1 1738 cm-1 1734 cm-1 C=O stretching
4 1472 cm-1 1472 cm-1 1472 cm-1 Bending symmetric vibration of CH2 (methylene)
groups' C–H bonds
5 1179 cm-1 1179 cm-1 1181 cm-1 The stretching vibration peak of C-O in triglyceride
6 723 cm-1 - 723 cm-1 Carbon skeleton vibration peak

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Volume 1 Issue 3 2022 International Journal of Scientific Research and Modern Technology

7 718 cm-1 718 cm-1 716 cm-1 The out-of-plane vibration of the cis –HC=CH
group of disubstituted olefins overlaps with the
rocking vibration of CH2

The comparison between APSO hydrolyzed by KOH stretching asymmetric was observed for NaOH with no
and NaOH as captured in table 1 reveal the spectra and key change observed for KOH. The result also indicated the
peaks, as well as the functional groupings to which they absence of ester functional groups at 1746 cm-1 for both
belong. Peaks for carboxylic acid carbonyl functional groups catalyst. Other peaks at 2912 cm-1 and 2917 cm-1 denote the
(C = O) were found at 1733 cm-1 for KOH while there was presence of CH2 and CH3 scissoring for both catalyst. The
increased intensity with NaOH at 1734 cm-1. Absorption absorption bands for (C-H) group vibration in the NaOH
bands of hydrolysis at 1472 cm-1 for stretching vibration was catalyst were also seen in the FTIR spectrum at 7233 cm -1.
same for both catalyst. A higher intensity at 1181 cm-1 for

Table 2 GC-MS characterization of APSO after hydrolysis


S/N Fatty acids Systematic names Composition (%)
KOH NaOH
1 Palmitic acid; C16:0 Hexadecanoic (C16H32O2) 9.37 2.87
2 Palmitoleic acid; C16:1 cis-9-hexadecenoic C16H30O2 3.62 2.02
3 Stearic acid; C18:0 Octadecanoic (C18H36O2) 14.35 12.21
4 Oleic acid; C18:1 cis-9-octadecenoic (C18H34O2) 51.32 43.19
5 Linoleic acid; C18:2 cis-9-cis-12-octadecedianoic (C18H30O2) 9.65 16.09
6 Linolenic acid; C18:3 cis,cis,cis-9,12,15-octadactrienoic 2.76 3.15
C18H30O2
SAFA 23.72 15.08
UFA 67.35 64.45
others 8.93 35.55

Further characterization study by GC-MS as captured in hampered as seen in the case of increase in reaction time and
table 2, confirmed a positive identification of FFA temperature.
composition. Fatty acids identified from this study for KOH
catalyst include the following: palmitic acid (9.37 %) Conflict of Interest. The author declare that he has no
Palmitoleic acid (3.62 %), Stearic acid (14.35 %), Oleic acid conflict of interest.
(51.32 %), Linoleic acid (9.65 %) and Linolenic acid (2.76
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