especially in ASAL regions in the process interfering
with biodiversity, colonizing water bodies while affecting livestock by the blocking of their rumens, causing
diarrhoea and in some cases leading to the loss of
teeth and eventually death (Vijayakumar et al., 2016).
In fact surveys done in regions where the trees dominate such as Baringo county in Kenya showed that
up to 90% of the natives wanted the Prosopis plant
to be totally eradicated from their habitat (Mwangi &
Swallow, 2008).
Despite all the negative reports on the plant,
attempts to valorise the plants have shown that the plant
is full of valuable phytochemicals and has potent ability to be used as a source of food, medicine and lead
compounds for drug development with flavonoids,
phenolics, alkaloids, and terpenes considered as the
most significant (Henciya, Seturaman, & Rathinam,
2016). Previous literature suggests that the plant has
been used as a form of traditional medicine with
the leaf used for treatment of cancerlike conditions,
sore throat, diarrhoea, measles, eye infections and flu.
The bark has been used for the treatment of wounds
with research showing it has both antifungal and
antibacterial properties (Prabha, Dahms, & Malliga,
2014).
Several studies have been done on the medicinal
properties of the Prosopis juliflora plant with ethanolic extracts of the leaves showing positive antibacterial
and anticancer activities (Sathiya & Muthuchelian,
2008, 2011). The extracts of the plant’s bark have been
shown to have antifungal and antioxidant activities.
Previous work on the heartwood of the P. juliflora
plant shows that it contains several compounds of
interest such as mesquitol, catechin, and epi-catechin
among other flavonoids. These have been shown to
have antitermite, antifungal, and antioxidant activities
as well as radical scavenging properties.
3 MATERIALS AND METHODS
3.1 Sample collection
The P. juliflora plant samples were collected from
Marigat, Baringo county in Kenya (latitude 0
◦ , 28
0.01
N, longitude 35
◦ , 57
0.01
E) and air dried.
3.2 Extraction
Extraction was done based on the method previously
used by Odero et al. (Odero, Munyendo, & Munyendo, 2017) with some modifications. Briefly, the
heartwood was separated first from other parts of
the plant and ground into fine powder using a hammer mill. It was then dried under a shade until the
achievement of a constant weight. Approximately 15
grams was then serially extracted in a Soxhlet machine
with solvents of increasing polarity starting from hexane, dichloromethane, and then acetone. The extracts
were then evaporated under vacuum conditions and the
extracts put in pre-weighed bottles for future use.
3.3 Chemicals and equipment
All the chemicals used were of analytical grade procured by Moi University and supplied by Pyrex and
Kobian Laboratories.
3.4 Phytochemical screening
Phytochemicals were screened using previously discussed methods (Anyalogbu, Anyalogbu, & Nwalozie,
2013; Lakshmibai & Amirtham, 2018; Sivanandham,
2016; Wangila, 2017). The following phytochemicals
were tested: terpenoids, alkaloids, flavonoids, tannins,
saponins and steroids.
3.4.1 Test for alkaloids
Two millilitres of the extracts were dissolved in 2 mL
of Wagners reagent. The colour changes were observed
with the appearance of a reddish-brown precipitate
suggesting the sample is positive for alkaloids.
3.4.2 Test for saponins
This was done using the foam test. 2 mL of the extracts
were diluted in 20 mL of distilled water in a test tube
and shaken for 20 minutes. Formation of foam on top
of the test tube showed the presence of saponins in the
plant extracts.
3.4.3 Test for flavonoids
Two millilitres of the plant extract was put in a test tube
and two drops of dilute NaOH added. The appearance
of a yellow colour which becomes colourless after two
drops of dilute sulfuric acid were added confirmed the
presence of flavonoids in the plant extract.
3.4.4 Test for steroids
Two millilitres of the plant extract were put in a test tube
with 2 mL of chloroform. One millilitre of sulfuric acid
was added to it with a dark reddish colour confirming
the presence of steroids.
3.4.5 Test for tannins
Two millilitres of the plant extract were dissolved in
45% of ethanol. The test tube was then boiled for five
minutes and 1 mL of 15% ferric chloride solution was
added. The appearance of a greenish to black colour
confirmed the presence of tannins.
3.4.6 Test for terpenoids
To 5 mL of the plant extract was added 2 mL of chloroform and 3 mL of concentrated sulfuric acid. A reddish
brown colour at the interface confirmed the presence
of terpenoids.
3.4.7 Total Phenolic Content (TPC)
The total phenolic content in both P. juliflora and
A. succotrina were determined using a previously
discussed method (Singleton, Orthofer, & LamuelaRavent, 1999). Acetonic extracts of P. juliflora and
fresh extraction of A. succotrina were prepared and
read in a Beckman Coulter Model DU
R 720 UV/VIS
spectrophotometer. This was done by mixing 0.5 mL
171
with biodiversity, colonizing water bodies while affecting livestock by the blocking of their rumens, causing
diarrhoea and in some cases leading to the loss of
teeth and eventually death (Vijayakumar et al., 2016).
In fact surveys done in regions where the trees dominate such as Baringo county in Kenya showed that
up to 90% of the natives wanted the Prosopis plant
to be totally eradicated from their habitat (Mwangi &
Swallow, 2008).
Despite all the negative reports on the plant,
attempts to valorise the plants have shown that the plant
is full of valuable phytochemicals and has potent ability to be used as a source of food, medicine and lead
compounds for drug development with flavonoids,
phenolics, alkaloids, and terpenes considered as the
most significant (Henciya, Seturaman, & Rathinam,
2016). Previous literature suggests that the plant has
been used as a form of traditional medicine with
the leaf used for treatment of cancerlike conditions,
sore throat, diarrhoea, measles, eye infections and flu.
The bark has been used for the treatment of wounds
with research showing it has both antifungal and
antibacterial properties (Prabha, Dahms, & Malliga,
2014).
Several studies have been done on the medicinal
properties of the Prosopis juliflora plant with ethanolic extracts of the leaves showing positive antibacterial
and anticancer activities (Sathiya & Muthuchelian,
2008, 2011). The extracts of the plant’s bark have been
shown to have antifungal and antioxidant activities.
Previous work on the heartwood of the P. juliflora
plant shows that it contains several compounds of
interest such as mesquitol, catechin, and epi-catechin
among other flavonoids. These have been shown to
have antitermite, antifungal, and antioxidant activities
as well as radical scavenging properties.
3 MATERIALS AND METHODS
3.1 Sample collection
The P. juliflora plant samples were collected from
Marigat, Baringo county in Kenya (latitude 0
◦ , 28
0.01
N, longitude 35
◦ , 57
0.01
E) and air dried.
3.2 Extraction
Extraction was done based on the method previously
used by Odero et al. (Odero, Munyendo, & Munyendo, 2017) with some modifications. Briefly, the
heartwood was separated first from other parts of
the plant and ground into fine powder using a hammer mill. It was then dried under a shade until the
achievement of a constant weight. Approximately 15
grams was then serially extracted in a Soxhlet machine
with solvents of increasing polarity starting from hexane, dichloromethane, and then acetone. The extracts
were then evaporated under vacuum conditions and the
extracts put in pre-weighed bottles for future use.
3.3 Chemicals and equipment
All the chemicals used were of analytical grade procured by Moi University and supplied by Pyrex and
Kobian Laboratories.
3.4 Phytochemical screening
Phytochemicals were screened using previously discussed methods (Anyalogbu, Anyalogbu, & Nwalozie,
2013; Lakshmibai & Amirtham, 2018; Sivanandham,
2016; Wangila, 2017). The following phytochemicals
were tested: terpenoids, alkaloids, flavonoids, tannins,
saponins and steroids.
3.4.1 Test for alkaloids
Two millilitres of the extracts were dissolved in 2 mL
of Wagners reagent. The colour changes were observed
with the appearance of a reddish-brown precipitate
suggesting the sample is positive for alkaloids.
3.4.2 Test for saponins
This was done using the foam test. 2 mL of the extracts
were diluted in 20 mL of distilled water in a test tube
and shaken for 20 minutes. Formation of foam on top
of the test tube showed the presence of saponins in the
plant extracts.
3.4.3 Test for flavonoids
Two millilitres of the plant extract was put in a test tube
and two drops of dilute NaOH added. The appearance
of a yellow colour which becomes colourless after two
drops of dilute sulfuric acid were added confirmed the
presence of flavonoids in the plant extract.
3.4.4 Test for steroids
Two millilitres of the plant extract were put in a test tube
with 2 mL of chloroform. One millilitre of sulfuric acid
was added to it with a dark reddish colour confirming
the presence of steroids.
3.4.5 Test for tannins
Two millilitres of the plant extract were dissolved in
45% of ethanol. The test tube was then boiled for five
minutes and 1 mL of 15% ferric chloride solution was
added. The appearance of a greenish to black colour
confirmed the presence of tannins.
3.4.6 Test for terpenoids
To 5 mL of the plant extract was added 2 mL of chloroform and 3 mL of concentrated sulfuric acid. A reddish
brown colour at the interface confirmed the presence
of terpenoids.
3.4.7 Total Phenolic Content (TPC)
The total phenolic content in both P. juliflora and
A. succotrina were determined using a previously
discussed method (Singleton, Orthofer, & LamuelaRavent, 1999). Acetonic extracts of P. juliflora and
fresh extraction of A. succotrina were prepared and
read in a Beckman Coulter Model DU
R 720 UV/VIS
spectrophotometer. This was done by mixing 0.5 mL
171