Advance Journal of Food Science and Technology 5(3): 230-235, 2013

ISSN: 2042-4868; e-ISSN: 2042-4876

© Maxwell Scientific Organization, 2013
Submitted: July 27, 2012 Accepted: October 09, 2012 Published: March 15, 2013

Optimization of the Soxhlet Extraction of Oil from Safou Pulp (Dacryodes Deulis)
1
D. Mampouya, 1R. Kama Niamayoua, 1S. Goteni, 1A.N. Loumouamou,
1, 2
T. Kinkela and 1, 2T. Silou
1
Multidisciplinary Team for Research in Food and Nutrition (EPRAN), (French)
2
School of Technology Cataractes (EST Cataractes) Regional Centre of
Excellence in Food and Nutrition, B.P. 389, Brazzaville, Congo (French)

Abstract: The aim of this study was to investigate Soxhlet extraction of oil from safou using various organic
solvents. The safou, fruit of the safou tree (Dacryodes edulis) is very fragile. Post-hervest spoilage, essentially due
to the softening of the pulp, can affect 50% of production. Extracting oil from the pulp could offer a way to reduce
losses. The safou contains 30-70% of oil in its pulp and about 10% in its seeds. It is a major oilseed plant in the
countries of the Congo Bassin, where unfortunately it is still underexploited. One possibility is to extract fresh oil by
cold pressing. This oil would be characteristic of a geographical area, the Congo Bassin, much like olive oil is of the
Meditarranean. Soxhlet extraction of oil from safou using various organic solvents was carried out to obtain
optimization data for the assessment of cold pressing extraction rates. Using a 23 factorial design and a centred
composite design for the sample studied, we obtained an optimal yield of 52% after 2 h of extraction from a finely
ground safou powder containing 8% of residual moisture and with a ratio of pulp mass to solvent volume of 45
g/250 mL.

Keywords: Dacryodes edulis, FA, oil, organic solvents, safou, soxhlet extraction, TAG

INTRODUCTION included in the optimization process with a 23 factorial

and a centred composite design.
The reference method for the extraction of plant
oils is by organic solvents in the soxhlet apparatus MATERIALS AND METHODS
(AFNOR, 1981). We used this method to extract oil
from safou pulp (Dacryodes edulis) using a range of Plant material: This study was conducted 2010, the
organic solvents. plant material used was safou from Pointe Noire in
The safou tree, a major oil bearing plant that is still Congo-Brazzaville. Chemical and nutritional
underexploited in the Congo Basin is a very fragile characteristics of oils from this origin have already been
fruit. Post-harvest spoilage, due essentially to the studied (Kinkela, 2003). The safou pulp was dried and
softening of the pulp, can affect 50% of production in powdered.
less than one week of storage. The extraction of oil
from the pulp could offer a way to reduce these losses. Solvent: Trichloroethane (density 1.29-1.35, boiling
The optimization of soxhlet extraction to determine the point 74-76°C), chloroform (boiling point 61.2°C,
maximum oil content of samples is a preliminary step density 1.47-1.49), hexane (boiling point 78°C) and
in assessing cold pressing extraction rates. Knowledge petroleum ether (boiling point 35-60°C, density 0.63-
of these rates is needed to study the cost effectiveness 0.64), supplied by Prolabo (France) were used as
of cold pressing. extraction solvents.
First we defined the main parameters expected to
influence the extraction yield. These were identified Extraction: The oil was extracted from dried ground
as the extraction time, the temperature, the particle size safou pulp by standard methods using 500 mL Soxhlet
of the plant material and the nature of the solvent and extractors (AFNOR, 1981).
its recovery rate. We then assessed the influence of the
nature of the solvent and the extraction time on the Physical characteristics: Specific gravity and viscosity
yield, in relation to the quality of the oil obtained. were determined using AFNOR standard methods
Preliminary results enabled us to limit the factors (AFNOR, 1981).

Corresponding Author: D. Mampouya, Multidisciplinary Team for Research in Food and Nutrition (EPRAN), Regional Centre
of Excellence in Food and Nutrition, B.P. 389, Brazzaville, Congo
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 Adv. J. Food Sci. Technol., 5(3): 230-235, 2013

Analysis of Fatty Acids (FAs): An oil sample (100 (Goupy, 2004). To reduce the number of experiments,
mg), weighed to within 5 mg in a test tube was four variables, namely the solvent (hexane), the volume
dissolved in 5 mL of hexane. To this was added 0.2 mL of the solvent (250 mL), the sample mass (50 g) and the
of transesterification agent (11.2 g of potassium temperature (boiling point of hexane) were kept
hydroxide dissolved in 100 mL of methanol). The tube constant, reducing the experiment to the following three
was stoppered and the contents vigorously mixed with a
variables:
vortex-type blender for 1 min. The mixture was left for
5 min and 0.5 g of solid NaHSO 4 was added.
The sample was homogenized and centrifuged for X 1 = Particle size of safou pulp
3 min. at ambient temperature and an aliquot of X 2 = Residual water content of safou pulp
supernatant was sampled for analysis. X 3 = Extraction time
The methyl esters obtained were analysed by Gas
Phase Chromatography (GPC) using a Perichrom The safou pulp oil extraction yield Y depends on
(France) type PERI 2000 apparatus, equipped with a the variables X 1 , X 2 and X 3 , i.e., mathematically:
glass capillary column 30 m long and 0.4 m in internal
diameter, impregnated with Carbowax 20 M (Applied Y = f (X 1 , X 2 , X 3 )
Science Labs, State College, PA, USA).
The analysis was carried out at a constant
temperature of 195°C with a nitrogen flow rate of where,
3 mL/min and a pressure of 0.5 bar. The injector Y : The yield or response
temperature was 220°C and the flame ionization f : The response function
detector temperature was 215°C (Kinkela, 2003). X 1 , X 2 & X 3 : The variables, or factors
Data acquisition (peak areas and mass fraction of
FAs), was carried out with Azur v. 2.0 software The purpose of the experiments was thus to determine
(Datalys, Saint-Martin-d’Hères, France). the effects of certain variables or factors on the
The response of the detector to the different methyl response. In effect, the following two-part question was
ester was studied with quantitative mixtures (Nu Chek- addressed: does a particular factor have a specific effect
Prep. Elysian, MN, USA). on the response and if so what is the relation between
that factor and the response?
Determination of the Triacylglycerol (TAG)
composition: The TAGs purified by column The two-level factorial design as developed by
chromatography were analysed by reversed-phase Davies (1954) appeared well suited to this type of
High-Performance Liquid Chromatography (HPLC) problem and it offered the advantage of requiring only
using a stainless steel column 250 mm long and 4.6 mm very basic mathematical knowledge (Ortigosa, 1993)
in internal diameter packed with silica (4 µm diameter) and no cumbersome machine calculation.
grafted with octadecyl radicals (HibarLichrospher 100 The general formula for the number (N) of
CH-18, Merck). experiments for a complete factorial design is:
The column was set up on a Waters 717 Plus N = 2k, where k is the number of variables in the
Autosampler equipped with a differential refractometer factorial design.
(Waters 996 Photodiode Array Detector, Waters,
If k = 3, then N = 2K = 23 = 8 experiments
Milford, MA, USA). The analysis conditions were as
follows: isocratic analysis at ambient temperature To construct the DOE matrix, we defined:
(21°C), using an acetone-acetonitrile (47:33, v/v)
mobile phase with a flow rate of 1 mL/min). • Reduced variable x i such that:
Data acquisition (peak area, mass fraction of TAG) o x i = (X i - X i0 ) /∆X
was carried out with Azurv. 2.0 software o X i0 = The base value, at the centre of the
experimental domain (level 0)
Optimization by the Design of Experiment (DOE) o ∆X = The variation increment, i.e., the unit of
method: variation of the variables
23 factorial design: The DOE method uses a factorial
• Two levels of the variables: High (+1) and low (-1)
design, i.e., simultaneous variation of all the factors.
The influence of each factor is then determined by This operation enables us to replace the
calculating its principal and interactive effects. experimental domain by the domain (-1, +1), with
The listed variables that influence the oil extraction eight responses described by the matrix (Table 1)
yield were: time, temperature, particle size of plant after randomization.
material, residual moisture content of pulp, mass of the
sample and nature and volume of solvent. For a first-degree model with interactions, the
A model with seven variables, even for a first points representative of a three-variable experimental
degree model, would need 27 = 128 experiments design are located in three-dimensional space (cube).
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Table 1: Design of experiment matrix (23 factorial design) Table 4: Variation in extraction yield (%) according to the extraction time
x1 x2 x3 Time (min) Yield (%) (1/t) 103
43 11.36 23.3
-1 -1 -1 114 27.60 8.8
+1 -1 -1 147 54.40 6.8
-1 +1 -1 221 55.40 4.5
+1 +1 -1 343 56.96 2.9
-1 -1 +1 427 57.72 2.3
+1 -1 +1
Table 5: Effect of the solvent on the extraction yield
-1 +1 +1
Solvent Trichloro ethane chloroform Petroleum ether Hexane
+1 +1 +1 Yeild (%) 54.1 50.0 56.5 51.6

Table 2: Experimental domain (centred composite design)

Variable levels Variable X 1 Variable X 2 where,
Centre (0) 45 g/250 mL 80 min N F : The number of experiments in relation to the
Low level (-1) 20 g/250 mL 50 min number of variables
High level (+1) 70 g/250 mL 110 min N A : The number of experiments in relation to the
Very low level (-α) 9.65 g/250 mL 37.58 min
Very high level (+α) 80.35 g/250 mL 122.42 min
variation increment
N 0 : The number of experiments at the centre of the
Table 3: Experimental design (centred composite design) matrix
Experiments x1 x2
1 -1 -1 With base points of 45 g/250 mL for X 1 and 80 min
2 +1 -1 for X 2 and increments of 25 g/250 mL and 30 min, we
3 -1 +1
4 +1 +1 arrive at the levels of variation and the experimental
5 -α 0 design presented respectively in Tables 2 and 3.
6 +α 0
7 0 -α RESULTS AND DISCUSSION
8 0 +α
9 0 0
10 0 0 Influence of extraction time on yield: Finely ground
11 0 0 powdered safou pulp (50 g) was extracted in a 500 mL
Soxhlet apparatus with 250 mL of solvent. Extraction
The corresponding response function is a first times extended to 2 h 30 min. The results obtained are
degree polynomial for each factor taken independently. given in Table 4.
It is noted: Figure 1, which gives the plot of the variation in
yield according to the extraction time, presents the
y = b 0 + b 1 x 1 + b 2 x 2 + b 3 x 3 + b 12 x 1 x 2 + b 13 x 1 x 3 + usual profile of a substance being gradually extracted to
b 23 x 2 x 3 + b 123 x 1 x 2 x 3 exhaustion from a plant matrix.
The function Y = f (1/t) takes the form Y (%) =
As the mathematical model associated with the 62.8-2.3 (1/t), with coefficient R2 = 0.85. This equation,
factorial design is built with centred, reduced variables, by extrapolation to 1/t = 0, gives the maximum yield of
the coefficients of the polynomial have very extractable oil contained in the sample, i.e., 62.8%.
This relation is also important for technological
straightforward meanings: mean: b 0 , principal effects:
applications, because it provides the extraction rate,
b i , interactions: b ij and b ijk (Goupy, 2004). The
which is the ratio of the real yield to the maximum
experimental data were processed using NemrodW theoretical yield obtained by extrapolation of the Y = f
software ([email protected]). (1/t) plot. Also, this result shows that almost 90% of the
oil contained in the pulp is extracted after an extraction
Centred composite design: The model used here was a time of 2 h 30 min longer extraction times are therefore
second degree one with two variables: not worthwhile.
• X 1 = Ratio of mass of plant material to solvent Effect of the solvent on the extraction yield: Four
volume organic solvents were used in this work. Trichloro
• X 2 = Reaction time or duration of extraction ethane and petroleum ether gave yields of respectively
54.1 and 56.5% (Table 5). These were the highest
The response is Y, the percentage of oil extracted. extraction yields. Chloroform had the weakest
The mathematical expression of the response using extractive power, with a yield of 50.0%. Hexane gave
the reduced variables takes the form: yields between these two extremes (51.6%). All the
yields were greater than 50%.
y = a 0 + a 1 x 1 + a 2 x 2 + a 12 x 1 x 2 + a 11 x 1 2 + a 22 x 2 2 Given that chlorinated solvents are toxic and
petroleum ether is a mixture that is difficult to eliminate
The number of experiments is given by the expression: from extracts and is also extremely volatile, hexane was
selected as a solvent to model the soxhlet extraction of
N = N F + N A + N 0 = 22 + (2 × 2) + 3 = 11 safou pulp oil.
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70

60

Rendement (%)
50

40

30

20

10

0
0 50 100 150 200 250 300 350 400 450
Durée de l'extraction

Fig. 1: Variation in extraction yield according to the extraction time (Yeild = f (extraction time))

Table 6: Influence of the extraction solvent on: physical and chemical characteristics, FA and TAG composition
Trichloro ethane Petroleum ether Chloroform Hexane
Specific gravity (T °C) 0.923 (23.4) 0.924 (25.8) 0.924 (25.8) 0.914 (24.4)
Viscosity (T °C) 72.90 (40) 61.03 (40) 70.97 (40) 34.87 (40)
Fatty Acids (FA)
C16: 0 50.11 48.91 49.90 50.25
C18: 0 4.31 4.27 4.21 2.61
C18: 1 n-9 30.24 30.15 29.95 32.13
C18: 2 n-6 13.14 14.59 13.58 13.74
Triacylglycerols (TAG)
PLL 3.29 4.24 4.08 4.50
POL 9.01 10.59 10.43 -
PPL 19.37 21.22 20.36 15.00
POO 20.60 19.92 20.86 14.80
PPO 47.73 44.28 44.28 58.30

Influence of the extraction solvent on: physical and 1

chemical characteristics, FA and TAG composition 0.9
0.8
of the oils obtained: The oils obtained were semi-solid
0.7
at ambient temperature and presented very similar
0.6
physical and chemical characteristics. Their density 0.5
D.O.

varied only slightly according to the extraction solvent 0.4

used. Hexane gave oil that was half as viscous as that 0.3
obtained with the other solvents. It probably extracted 0.2
less wax and other heavy fractions than the other 0.1
solvents (Table 6). 0
All the oils were shades of green in colour. The 400 500 600 700 800
chlorophyll pigments, which predominated in the green Longueurs d'onde (nm)
oils and reddish-yellow coloured carotenoids absorb in
the visible range between 400 and 700 nm. The Fig. 2: Absorption spectrum in the visible range of safou pulp
absorption in this range is a measure of the intensity of oil (Absorbance = f (wave number (nm)))
the colour of the oil or fat studied. This intensity is in
absorption of carotenoids, which thus occur in small
principle proportional to the quantity of pigments
quantities and a double peak between 630 and 670 nm
present in the oil when the Beer-Lambert law is obeyed.
corresponding to the absorption of chlorophyll
In studies of oils extracted from citrus and gourd
pigments, responsible for its greenish colour (Fig. 2).
seeds, Helmy (1990) identified absorption maxima at The oils extracted with the different solvents also
400, 425, 455 and 480 nm for carotenoids, 610 and presented similar FA and TAG compositions (Table 6).
670 nm for chlorophylls and 525, 570 and 595 nm for These oils contained three major fatty acids
an unknown pigment, respectively. representing more than 90% of the total FAs. These
Whichever the extraction solvent used, the visible were: palmitic (P or C16:0), oleic (O or C18:1 n-9) and
spectrum of the safou pulp oil obtained presented a linoleic acids (L or C18:2 n-6). Then came stearic acid
small peak at about 500 nm corresponding to the (S or C18:0) at about 2-3% and then minor
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Fig. 3: Graphical illustration of the effects and interactions of

variables during soxhlet extraction of safou pulp oil
b 1 : Particle size effect; b 2 : Moisture effect; b 3 : Fig. 5: Optimum yield (%) of safou pulp oil extraction
Extraction time effect
%PPO>%POO>%PPL>%POL

This work thus enabled us to collect important data

for subsequent modelling of the process of safou oil
extraction with the Soxhlet apparatus, justifying the use
of hexane as solvent and the extraction time, generally
no longer than 3 h in the laboratory. We also
established that the physical and chemical quality of the
oil did not vary significantly with the nature of the
extraction solvent. Overall, this quality was compliant
with standards applicable to edible oils and was
consistent with the results available in the literature.

Optimization of safou pulp oil extraction by the

DOE method:
23 factorial design: The mathematical model
representing the yield of oil extraction (y) from safou
pulp as a function of reduced centred variables (x 1 , x 2 ,
x 3 ) in a 23 factorial design is:

y = b 0 + b 1 x 1 + b 2 x 2 + b 3 x 3 + b 12 x 1 x 2 + b 13 x 1 x 3 +
Fig. 4: Response diagram of safou pulp oil extraction b 23 x 2 x 3 + b 123 x 1 x 2 x 3
Extraction time = f (ration of pulp mass to solvent
volume) The effects (x i ) and interactions (x ij , x ijk)
determined using the NemrodW software, shown in
components, linolenic acid (Ln or C18:3 n-3), arachidic Fig. 3, gave the following coefficients:
acid (or C20:0) andeicosaenoic acid (or C20:1 n-9) at
levels less than 1%. y = 28.9 + 13.7x 1 + 4.67x 2 + 2.68x 3 + 0.97x 1 x 2 -
For the major FAs we find here the characteristic 1.42 x 1 x 3 + 1.38x 2 x 3 - 0.33x 1 x 2 x 3 .
profile of safou pulp oil as reported in the literature
(Silou, 1996; Mbofung et al., 2002; Ondo, 2004; Ondo- The fractioning of the pulp was by far the factor
Azi et al., 2009), i.e.: most strongly influencing the extraction. It was
followed, with a division factor of about 3, by the
%P>%O>%L moisture content of the pulp feedstock. The extraction
time came third.
We note the presence of four major TAGs in the
oils extracted from pulps with a cumulated content of Centred composite design: We repeated the
more than 95% of the total TAGs and the following optimization, keeping constant the most influential
profile: variables (fine grinding and 8% pulp moisture) using a
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 Adv. J. Food Sci. Technol., 5(3): 230-235, 2013

centred composite design, with the extraction time (X 1 ) Helmy, E.H., 1990. Studies on the pigments of some
and ratio of pulp mass to solvent mass (X 2 ) as variables. citrus, prune and cucurbit seed oils when processed
The results of automatic data processing with the with or without cottonseed oil. J. Amer. Oil Chem.
NemrodW software are given in Fig. 4 and 5. Soc., 67(6): 376-380.
These two figures show that the maximum yield, Kinkela, T., 2003. Study of the triglyceride fraction of
about 52% for this sample, corresponds to a pulp mass oïl safou (Dacryodes dulies) and in vitro evaluation
/solvent volume ratio of 45 g/ 250 mL and an extraction of its nutritional value for populations of central
time of 122.4 min. Africa State. Ph.D. Thesis, University Marien
Ngouabi, Brazzaville (French), pp: 130.
CONCLUSION Mbofung, C.M.F., T. Silou and I. Mouaragadia, 2002.
Chemical characterisation of safou (D. edulis) and
These two treatments show that the optimal evaluation of its potential as an ingredient in
conditions for oil extraction from this sample by hexane nutritious biscuits. Forests Trees Livelihoods, 12:
105-117.
in a soxhlet apparatus (Y max = 52%) were: fractionation,
Ondo, A.S.A., 2004. Physicochemical and
fine grinding, 8% residual moisture in the pulp, a ratio
morphological diversity and potential of the oil mill
of pulp mass to solvent volume of 45 g/ 250 mL and an
safous (Dacryodes edulis) of the Franceville region
extraction time of 122.4 min. (Southeastern Gabon). Ph.D. Thesis, University
Marien Ngouabi, Brazzaville (French), pp: 147.
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Goupy, J., 2004. Introduction to Experimental Design. Silou, T., 1996. The safoutier (Dacryodes edulis), a
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