For. Stud.

China, 2011 , 13(4): 263-269

DOI 1O.1007/s11632-013-0402-8
Selected physical and chemical properties of mechanically extracted
neem seed oil sourced as a preservative for ligno-cellulose in south-
western Nigeria
Andrew A. ERAKHRUMEN'
Department ofForest Resources Management, Faculty of Agriculture and Forestry, University ofIbadan, Ibadan, Nigeria
Forestry University and Springer-Verlag Heidelberg 2011
Abstract Most of the seeds produced by neem (Azadirachta indica A. Juss) trees in Nigeria are currently underutilized. Hence,
relevant literature provides only limited information conceming many of the seed oils from this country, especially where it con-
cems the potential applications of these oils as preservatives for ligno-cellulose against bio-deterioration. U sing standard procedures
therefore, this study was carried out to evaluate and document selected physical and chemical properties of neem seed oil (NSO),
mechanically extracted using a cold press at 31.03 N.mm-
2
pressure and a room temperature of

The results show that oil

yield was 38.42% with a specific gravity The amount ofacid was 18.24 :l: 1.31 mg KOH.g-
1
and that ofiodine
2.01 g.100 g-l , while saponification and peroxide values were and mg.g-
1
respectively. The implication of
the values obtained, particularly those for the chemical as they concem the potential application ofNSO as a preservative
for ligno-cellulose, is likely that it may be useful in this regard since the values may support some of the documented anti-microbial
properties of the oil, although other physical and properties that may affect this potential are recommended for investiga-
tions. Conclusions and other recommendations follow in line with the results ofthe
Key words neem oil, physical and chemical properties, mechanical extraction, cold press
1 Introduction
The neem (Azadirachta indica A. Juss) tree is known
to be a very important species in many parts of the
world. It is popularly known as the village pharmacy,
since all parts of this plant have, for centuries, been
used to prepare medications for several types of dis-
eases (Venmalar and Nagaveni , 2005). Extracts of
leaves and seeds exhibit, among others, anti-bacterial,
anti-fungal, anti-viral (Tewari, 1992) and anti-malarial
(Chavan and Nikam, 1988) properties. Its leaf extracts
is also known to inhibit the growth of plant pathogens
(Bhatnagar and McCormick, 1988; Kurucheve et
1997).
Other parts of the tree have many other beneficial
uses such as ingredient in cosmetics and in the pro-
duction of soap and toothpaste (OIA and PGA, 1992;
Puri, 1999; NF, 2006). Neem wood is known to be
durable against wood rot (Rao, 1990) and it is used
for many agricultural implements and in construc-
tion. Studies have also shown that extracts from dif-
ferent parts of this plant can be used to protect ligno-
for correspondence. E-mail: erakhrumen@yahoo.com
cel1ulose against certain insects and microbial attacks
(Subbaraman and Brucker, 2001; Dhyani et
Swathi et 2004; Islam et 2009).
Neem seeds are a part of the plant commonly used
for various Oils from the seeds have been
used in many different industries, especially the cos-
metics and pharmaceutical industries (as a supporting
ingredient in medicines). Neem seed oil (NSO), noted
to contain mainly Azadirachtin, among other organic
chemicals, is also used as insect repellent, feeding in-
hibitors, egg laying deterrents, growth retardants and
as sterilizer. It also has systemic action on plants for
controlling fungal diseases.
A series of studies have also shown that NSO has
certain anti-bacterial, anti-infl.amatory, antiseptic, anti-
viral, anti-fungal, anti-parasitic, gennicidal, pesticidal,
insecticidal, anti-microbial and other related properties
(Parveen and Alam, 1993; Locke, 1995; Mishra et al.,
1995; Puri, 1999; NF, 2006; Wikipedia, 2007). Studies
such as that by Venmalar and Nagaveni (2005) have
been used to demonstrate that NSO, containing toxic
metal, could be used to protect non-durable wood and
fibres against wood rot and termites. However, there
264
are increased concems regarding the limits of expo-
sure to many of these metals and other toxic substanc-
es to humans and the environment.
As a result of environmental and health implica-
tions of these inclusions coupled with documented
information conceming anti-microbial properties of
NSO without inclusions of toxic substances, the ne-
cessity for studies such as that by Erakhrumen (2010)
has been demonstrated. That study showed that NSO,
without such inclusions, could be used as a preserva-
tive for naturally non-durable ligno-cellulose such as
bamboo and presented efforts aimed toward the use
of plant-derived environmentally benign biocides. It
should be pointed out that presently the seeds pro-
duced by these trees in many parts of Nigeria have
little or no socio-economic value (Erakhrumen, 2010).
Regardless of the documents referred to ear1ier, in-
fOl mation is still scarce conceming many of the physi-
cal and chemical properties West Africa,
that relate these properties to the observed and docu-
mented anti-microbial properties of NSO. Owing to
the paucity of documented infolmation in this regard,
this study was therefore carried out to evaluate and
document some physical and chemical properties of
mechanically extracted NSO with a view to highlight
some salient information relating these properties to
the documented application of NSO as a preservative
for ligno-cellulose.
2 Materials and methods
2.1 Source of neem seeds
Ripe neem seeds from which oil was mechanically
extracted in this study were obtained from A. indica
trees on the campus of the University of Ibadan, 10-
cated at the northem edge of Ibadan City, Nigeria (1at-
itude 7
0
20'N and longitude

an area of about
10.4 km
2
Ibadan lies at 200 m above sea level with
a humid tropical climate (27
0
C average), a March
October rainy season (1 250 mm) followed by a mild
dry season. Collection of the seeds was done by plac-
ing nylon sheets around the stems in such a way that it
covered a substantial cross sectional area of the crown
in order to collect the seeds as they fall. The seeds
were sourced in the months of June to ear1y August of
2008.
2.2 Drying and storage of neem seeds
The neem seeds obtained from the field were first
thoroughly washed using disti11ed water to remove
dirt and other impurities and then air dried in an open
Forestry Studies in China, VoI.l 3, No.4, 2011
space with regular movement for aeration for proper
drying as suggested by Mitra (1 963), a method also
applied by Soetaredjo et al. (2008) to reduce the mois-
ture content for proper crushing and to facilitate high
oil volume recovery during mechanical extraction.
The seeds were stored in nylon lined jute bags at room
temperatures of

and kept away from rodents

and other animals that could consume them and also
to prevent contamination. They were daily air dried
with proper monitoring to prevent damage as a resu1t
of possible moisture fluctuation.
2.3 Extraction of neem seed oil
Oil extraction was carried out in this research using
a cold press to force out the seeds at a room
of

This allows for easy adop-

tion by most of the target end-users, particular1y those
in rural areas. The mechanical extraction has several
advantages compared to other methods, i.e. , simple
equipment, low investment and low operating costs,
and the oil does not undergo a solvent separation pro-
cess (Fasina and Ajibola, 1989).
The seeds were first decorticated, cleaned from
dirt and then air dried. Dried kemels were carefully
ground into smaller particles using a seed grinder en-
suring no significant loss of oil. Mechanical extraction
of oil was carried out by cold pressing using an oil
expeller at 31.03 N.mm-
2
pressure or 31.03 MPa until
the oil stopped flowing, in line with the recommenda-
tions by Liauw et al. (2008).
2.4 Selected physical and chemical properties of
extracted NSO
2.4.1 Percentage yield for extracted NSO
Extracted oil yield is defined as the percentage of oil
expelled at the pressing stage on a total oil extractable
basis. Oil expelled is defined as the difference be-
tween total oil in the original seeds and the oil in the
residual cake (Uquiche et al., 2008). The method used
in estimating the amount of oil obtained was based on
the ratio of percentage of weight of oil obtained to the
total weight of crushed seeds in line with Mwithiga
and Moriasi (2007) and Uquiche et al. (2008). In this
method, the dried seed kemels were weighed before
grinding and cold pressing and after the oil had been
removed to satisfaction; the neem seed cake obtained
was also weighed. The percentage yield of NSO was
obtained using the following equation, based on the
definition and modification by Uquiche et al. (2008).
Andrew A. ERAKHRUMEN: Selected physical and chemical properties of mechanically neem seed oil... 265
the total weight of dried crushed neem seeds). The
amount of oil obtained fel1 within the range obtained
in some published studies such as that of
et al. (2008) on NSO. The literature on oil yields from
neem seed kernels varies widely, i.e. , from 25% to
45% (Wikipedia, 2007). It should also be noted that
this range may be increased or decreased depending
on the pressure applied during mechanical extraction
as in other studies (Fasina and Aji-
bola, 1989; Adeeko and Ajibola, 1990; Abu-Arabi et
al., 2000).
Furthermore, studies have shown that there are
threshold pressures at which oil differ-
ent types of seed kernels during mechanical extrac-
tion. These still have to be detennined. The maximum
pressure at which mechanical extraction was carried
out in this study was 31.03 N'mm-
2
or 31.03 MPa, for
increasing the pressure beyond 27.58-34.47 N'mm-
2
may not necessarily result in an increase in oil yield
(Liauw et al., 2008). It has also been noted that higher
temperatures of extraction may yield higher volumes
of oil but the quality may be lowered at these higher
(Liauw et al., 2008). This is the reason
why the temperature of extraction in this study was
kept constant at a room temperature of

Nev-
ertheless, it is to be noted that quality is
the point of view of the end user.
It should also be noted that variation in oil yield
may be due to the differences in the methods used to
produce the seed, such as cultivation climate, ripen-
ing stage, the time of harvesting the seeds, kernel
prperties and the extraction method used (Dhingra
and Kapoor, 1985). It is also worth mentioning that
the method of processing is likely to affect the com-
position of the oil, since the methods used, such as
pressing (expel1ing) or solvent extraction are unlikely
to remove exactly the same mix of components in the
same proportlOn.
The mean value obtained for the specific gravity of
oil was similar to the result of similar studies on this
physical property of NSO (Majumder et al., 1997;
Akpan, 1999; Puri, 1999). Knowledge ofthis property
might be important for potential users of this oil for
treating ligno-cel1ulose, since it has been noted that
absorption of liquid by ligno-cellulose such as bam-
boo varies with density of the liquid (Sobrinho et al.,
2009). Density/specific gravity of vegetable oil has
been observed to decrease as temperature of exposure
increases, thereby leading to inc
AU AU

nu
w
=
% W PA
where PW is approximate weight of oil obtained (%),
W
c
total weight of crushed neem seeds (kg) and W
sc
total weight of neem seed cake obtained (kg).
2.4.2 Determination of specific gravity ofNSO
The specific gravity bottle used for this experiment
was first washed and dried in an oven. It was allowed
to cool to room weighed and the weight
recorded. It was then filled with distilled water and
corked with a stopper ensuring no entrapment of air
bubbles. The bottle containing the distilled water was
weighed and the weight recorded. The distilled water
was then removed and the bottle was again oven dried
and al10wed to cool to room The bottle
was then filled with an equal volume ofNSO and also
corked with a stopper ensuring no entrapment of air
bubbles. As for disti l1ed water, the bottle containing
the NSO was weighed and the weight recorded. The
specific gravity ofthe oil was calculated using the fol-
lowing equation in line with Kurian (2010).
Specific
"2 2)
where W] is the weight of NSO in the total volume
and W
z
is the weight of deionised water in the total
volume.
2.4.3 Detennination of acid, saponification, iodine
and peroxide values for extracted NSO
The method for determining the acid value for NSO
in this study conformed to the standard of ASTM D
974-08 while the test method for determining saponi-
fication value for NSO in this study conformed to the
standard of D 5558-95. Furthermore, the stan-
dard method for determining the iodine value for NSO
in this study conformed to the standard of ASTM D
5768-02 while the test method for determining per-
oxide value for NSO in this study conformed to the
AOAC Official Method 965.33.
3 Results and discussion
Table 1 shows the selected physical properties of the
mechanically extracted NSO. The amount of oil ob-
tained using this method of extraction was 38.42%
(w/w, percentage ratio of weight of oil obtained to
266
Forestry Studies in China, Vo1.l 3, No.4, 2011
Table 1 Selected physical of mechanically extracted neem seed oil
Properties
Volume (%)
gravity
Color
Physical state at room
Mean values
38.42
0.910
Darkbrown
Liquid
Standard deviation
0.01
Values are means for 10 test samples and obtained at room temperature of 25 :l: 2
0
C. The same comrnent applies to Table 2.
in but unlike specific gravity it can also
be altered by agitation. The color of the extracted oil
for this study was initial1y yel10wish but later turned
dark brown as a result of oxidation when exposed to
oxygen from the air, as also observed in earlier similar
studies such as by Porter et al. (1 981) and Porter and
(1984).
The amount of acid indicates the amount of free fat-
ty acids present due to enzymatic activities in a samp1e
of oil and is a good indicator of oil degradation caused
by hydro1ysis. It can be used to quantify the amount
of acid present in a sample of oil. It is a basic quan-
tity, expressed in mi11igrams of potassium hydroxide,
which is required to neutralize the acidic constituents
in 1 g of an oil sample. The probable reason for the
high mean value of 18.24 (mg KOH.g-
1
) obtained for
the acid in this study might be because the extracted
NSO was not first neutralized before this test, simi-
lar to the conditin noted by Abdul1ah and Salimon
(2009) in their study. Similar1y, SathyaSelvabala et al.
(2010) also obtained NSO with an acid value of 24.4
(mg KOH.g-l) that was reduced to 1.8 (mg KOH-g-
1
)
during esterification.
High acidic content of preservatives has been noted
to be effective against wood destroying organisms.
For instance, since the 19th tannin and tan-
nic acid have been used to improve wood properties
and durability (Yang, 2009). A series of patents have
been filed in this area (Mitchel1 and Sleeter, 1980;
Laks et al., 1988; Lotz, 1993; Lotz and Hol1away,
1993). Likewise, the natural resistance of western red
cedar (Thuja and eastern white cedar (Thuja
occidentalis) to biodegradation has been attributed to
the presence of thuja plicins and acid in their
heartwood. It is estimated that 0.1 %-0.3% of such
compounds in heartwood can inhibit fungal growth
(Gripenberg, 1949; Stir1ing et al., 2007). Thus, a high
acid content in bio-oil to be used as for
ligno-cel1ulose wil1 most likely aid the anti-microbial
properties of oil.
The saponification value indicates the average mo-
lecular weight of triglycerides in the oil. The mean
value obtained in this study was similar to those ob-
tained by O'Brien (2004) for sunflower, corn and
flower oils and by Adebowale and Adedire (2006) for
Jatropha curcas seed oil. A high saponification value
is an indication that the oil consists of a high propor-
tion of low molecular weight fatty acids. An increase
in may increase the saponification value
because higher temperatures cause lipids to break
down. A reduction therefore in the average molecular
weight of oil results in a reduction of oil viscosity.
Viscosity is a measure of resistance of a fluid to de-
form under shear stress. It is commonly perceived as
thickness, or resistance to pouring. Viscosity describes
the internal resistance of a fluid to flow and may be
thought ofas a measure offluid friction (Nzikou et
2009). Viscosity increases with molecular weight but
decreases with increasing unsaturated levels of fat and
oils and with temperature (Nourredini et al., 1992).
Knowledge of this property when affected by saponi-
fication is important for vegetable oil to be used as a
preservative for ligno-cel1ulose, particular1y at a low
regime since it is likely to affect oil ab-
sorption which, in turn, affects some other properties
such as penetration and distribution of preservatives in
treated material. Penetration and distribution are some
of the treatment factors upon which the effectiveness
of preservatives depends.
The iodine value expresses the non-saturation leve1
of oil. The mean value obtained in this study com-
pares with some other oils in the literature such as that
ofperah seed oil (1 06 g.100 g-l) (Yong and Jumat,
2006), cotton seed oil (99-119 g.100 g-l) and corn
seed oil (103-128 g.lOO g-l) (Gunstone et al., 1994),
rubber seed oil (l35.79 g.100 g-l) (Abdul1ah and Sali-
mon, 2009) and Jatropha curcas seed oil (1 11.6 g.100
g-l) (Adebowale and Adedire, 2006), al1 considerably
higher than that for palm oil (52 g.100 g-l) (Onyeike
and Acheru, 2002).
High iodine values for vegetable oil may be due
to the high content of unsaturated fatty acids in these
oils, as observed by Abdullah and Salimon (2009).
Higher may reduce the iodine value (Li-
auw et al., 2008), since higher temperature initiates
the breakdown of carbon chin bonding, leading to
the fonnation of saturated carbon chains, thus result-
ing in a decrease in the iodine of oil. The iodine value
indicates the drying nature of oil. Natural oils can be
divided into drying, semi-drying and non-drying oils.
High iodine values > 140 g.l 00 g-l are typical of dry-
ing oils and low values < 125 g.100 g-l of non-drying
Andrew A. ERAKHRUMEN: Selected physical and chemical properties ofmechanically extracted neem seed oil... 267
Table 2 Selected chemical properties of mechanically extracted neem seed oil
Properties
Acid value (mg KOH.g-l)
Saponification value (mg.g-I)
lodine value (g.100 g-I)
Peroxide value (mg.g-
1
)
oils, with the iodine values of semi-drying oils lying
between the two (Wicks et al., 1992).
Moreover, studies such as by Lyon et al. (2007) re-
vealed that vegetable oils high in polyunsaturated and
monounsaturated fatty acids (with high iodine values)
are likely to be drying oi1s, a property that aids in pre-
venting the leaching of chemicals from
ligno-cellulose. The NSO used for this experiment
falls within the range of those characterized as slow or
non-drying oils. Similar results concerning vegetable
oils have been reported by Das (1 982) and Majid et al.
(2004). All the same, it is important to note that poly-
unsaturated fatty acids are considerably more reactive
than monounsaturated fatty acids, mainly because
their alpha hydrogens are much more acidic (Koski,
2008)
The peroxide value is used as an indicator of oil
rancidity. Rancidity is described simply as having the
strong disagreeable smell or taste of decomposing fats
or oils. High peroxide values of oil indicate poor resis-
tance of oils to peroxidation, particularly during stor-
age. Increasing the temperature during extraction may
also cause the oil to be rancid, thereby leading to a
reduction in its oxidative stability. Rancidity is caused
by aldehydes, ketones and oxidation (Mongkholkha-
jornsilp et al., 2004).
Improper or long storage may also lead to rancid-
ity. For instance, storage of crude tall oil has been ob-
served to lead to esterification of the sterols and other
alcohols with acids (Holmbom and Avela, 1971a,
1971 b; Holmbom and Stenius, 2000), which reduces
the number of acids contained and increases its hydro-
phobicity (Koski, 2008). This condition may lead to a
reduction in toxicity to agents of biodegradation and
reduction in waterproofing layer that prevents fungal
penetration and growth within the ligno-cellulose to
be treated with such oil.
4 Conclusions
The results obtained, particularly for the selected
chemical properties of neem seed oil investigated in
this study show that this oil might possess some anti-
microbial properties that can be exploited in prevent-
ing or controlling biodegradation, most especially
in ligno-cellulose. Nevertheless, it is important that
Mean values
18.24
172. 88
93.12
1.42
Standard deviation
1.31
2.06
2.01
0.04
other physical and chemical properties that may af-
fect these possible anti-microbial properties should
be investigated. Furthenllore, studies applying this oil
in treating ligno-cellulose of interest against associ-
ated biodegrading micro- and macro-organisms will
be necessary to ascertain the efficacies of the physical
and chemical properties of this oil.
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(Received April 20, 2011 Accepted August 26, 2011)