Chemical Studies of Tephrosia vogelii and Commiphora schimperi Occurring in Ethiopia

Full text

Modern Chemistry
2019; 7(3): 45-53
http://www.sciencepublishinggroup.com/j/mc
doi: 10.11648/j.mc.20190703.11
ISSN: 2329-1818 (Print); ISSN: 2329-180X (Online)
Chemical Studies of Tephrosia vogelii and Commiphora
schimperi Occurring in Ethiopia
Tegene Tesfaye Tole
*
, Belay Akino Neme
Chemistry Department, College of Natural and Computational Sciences, Hawassa University, Hawassa, Ethiopia
Email address:
*
Corresponding author
To cite this article:
Tegene Tesfaye Tole, Belay Akino Neme. Chemical Studies of Tephrosia vogelii and Commiphora schimperi Occurring in Ethiopia. Modern
Chemistry. Vol. 7, No. 3, 2019, pp. 45-53. doi: 10.11648/j.mc.20190703.11
Received: February 12, 2019; Accepted: March 10, 2019; Published: September 19, 2019
Abstract:
The objective of this study is to extract, screen, isolate, and characterize the chemical constituents of Tephrosia
vogelii and Commiphora schimperi. In the course of this study the stem bark of T. vogelii and resin of C. schimperi were
collected from Areka Agricultural Research institute, Wolaita Zone and Konso, Gamo Gofa Zone, respectively. Phytochemical
screening of the crude extract of T. vogelii revealed the presence of tannins, saponins, flavonoids, and terpenoids.
Chromatographic separation of the methanolic crude extract of T. vogelii yielded the compound 8 (4, 5-dihydro-5, 5-dimethyl-
4-oxofuran-3yl)-5-hydoxy-7-methoxy-2-phenyl-4H-chrome-4-one. The essential oil from the resin of C. schimperi was
isolated by hydro-distillation and some of the components identified by means of GC and GC/MS analysis. The main
components of the essential oil from C. schimperi were: α-pinene (73%) and β-pinene (17%). The resin was also subjected to
extraction by petrol, ethyl acetate, and methanol. The components of C. schimperi were isolated using chromatographic
techniques and attempts were made to identify the isolated substances. Analysis of the petrol extract of the resin of C.
schimperi showed to be an excellent source of the industrially important fragrant compounds, α- and β-pinene. The structures
of the isolated compounds were characterized by comparing IR,
1
H NMR, and
13
C NMR chromatographic data of the
compounds with literature.
Keywords:
Chemical Studies, Commiphora schimperi, Ethiopia, Resin, Stem Bark, Tephrosia vogelii
1. Introduction
Medicinal plants have been used for thousands of years to
flavor and conserve food, to treat health disorders and to
prevent diseases. The knowledge of their healing properties
has been transmitted over the centuries within and among
human communities [1]. The use of Medicinal plants has a
long history throughout the world. They are known to
provide a rich source of raw materials and have been used in
traditional medicine for numerous human diseases for
thousands of years, in different parts of the world,
particularly those living in rural areas of the developing
countries [2]. Traditional medicine has been brought into
focus for meeting the goals of a wider coverage of primary
healthcare delivery, not only in Africa but also, in all
countries of the world. It is the first choice healthcare
treatment for at least 80% of Africans who suffer from high
fever and other common ailments [3]. Ethiopians have used
traditional medicines for many centuries, the use of which
has become an integral part of the different cultures in the
country. The indigenous peoples of different localities in the
country have developed their own specific knowledge of
plant resource uses, management and conservation [4].
Originally teas or decoctions (aqueous extracts) or
tinctures or elixirs action protocols and applied and coupled
with modern isolation techniques were used to prepare and
administer herbal remedies. These were usually the starting
points for isolation work. These days, various extraction
protocols are applied and coupled with modern isolation
techniques which include chromatography, often guided by
bioassay, to isolate the active compounds.
In Ethiopia, about 80% of the people use medicinal plant
remedies selected over centuries. Exploration of the chemical
constituents of the plants and pharmacological screening is of
great importance which leads for developments of novel
agents [5]. A systematic study of a crude drug from

46 Tegene Tesfaye Tole and Belay Akino Neme: Chemical Studies of Tephrosia vogelii and
Commiphora schimperi Occurring in Ethiopia
medicinal plants embraces through consideration of primary
and secondary metabolites derived as a result of plant
metabolism. The compounds that are responsible for
medicinal properties of the drug are usually secondary
metabolites such as flavonoids, alkaloids, saponins, tannins,
terpenoids, etc. and their derivatives [6].
The genus Tephrosia belongs to the family Leguminosae
and subfamily Papilionacea. There are approximately 400
species included in this genus. The plants in this genus are
widely distributed in tropical, sub-tropical and arid regions of
the world [7]. Many plants from this genus have been used
traditionally for the treatment of rheumatic pains, syphilis,
dropsy, stomach ache, diarrhea, asthma, abortifacient,
respiratory disorders, laxative, diuretic and inflammation [8].
In Pakistan the roots are used to treat typhoid fever [9].
In China, Tephrosia vogelii is used as a botanical
insecticide and fly repellent [10]. It has been shown to have
toxic and repellent effects against certain insect pests of
stored grains [11]. Tephrosia has been used as a rat poison by
compounding with ground nut [12]. Powders of Tephrosia
vogelii are effectively used in the Congo against the stored
ground nut pest Carye donserratus [9]. In Nigeria, it is used
as a seed dresser for cereals and legumes [13]. It is also
applied directly to treat head lice, fleas, scabies and other
eco-parasites [14].
Recently, naturally occurring elata-dihydrochalcone,
isopongaflavone, obovatin methyl ether, and deguelin were
isolated from seed pods of Tephrosia elata [15]. Purpuritin
and semiglabrin are isolated from Tephrosia purpurea seed
extract [16]. Phytochemical analysis of Tephrosia candida,
dichloromethane: methanol (1:1) extract of the air-dried
aerial parts afforded new prenylated flavonoids, namely,
Tephro-candidins, candida chalcone, a sesquiterpene and a
flavonoid phytoalexin, pisatin [17]. Semiglabrin, apollinine,
pseudo semiglabrin, and lanceolatin A are isolated from the
roots of Tephrosia nubica [18]. Emoroidenone, tephrone,
ovalichacone, and hildecarpin are isolated from the root bark
of Tephrosia aquilata [19].
The sesquiterpene (1β, 6α, 10α)-guai-4 (15)-ene-6, 7, 10-
triol, the lignan (+)-lariciresinol 9’-stearate, the larvicidal
rotenoid deguelin and the flavonoid quercetin are secondary
metabolites isolated from Tephrosia vogelii [20, 21]. Chen et
al., reported the isolation of four flavonols, four flavans, a
flavanonol and a sesquiterpene from T. vogelii [22]. The
rotenoid and deguelin content of the leaf extracts of the white
and purple varieties of T. vogelii showed similar acaricidal
activity [22]. A 100% larval mortality of Rhipicephalus
sanguineus was observed at the concentration of 20% the
ethanolic extract of the leaf of T. vogelii [23]. Rotenoids from
T. vogelii Hook. F. (Leguminosae) including deguelin,
rotenone, sarcolobine, tephrosin and α-toxicarol are required
for pest control and soil enrichment agent for poorly
resourced small scale farmers in Southern and Eastern Africa
[24, 25].
The ethyl acetate extract of the fruit of Piper aduncum and
T. vogelii leaf extract showed a strong synergistic toxic effect
on larvae of C. pavonana at 5:1 mixture respectively [26].
Similarly the ethyl acetate-methanol (9:1) and ethyl acetate
extract of the leaf of T. vogelii showed insecticidal activity
against the cabbage head caterpillar, Crocidolomia pavonana
[27].
Although investigation of phytochemicals and biological
activities of the leaf of T. vogelii was done in different places,
there is no report on stem bark of T. vogelii, thus far. More
specifically to the best of our knowledge there is no
investigation that has been done in Ethiopia. Thus it is
worthwhile to investigate the phytochemicals found on the
stem bark of the medicinally important T. vogelii occurring
in Ethiopia.
The Burseraceae is a large family with 17-20 genera and
500-600 species, widespread in tropical and subtropical
regions [28, 29]. Engler [30] subdivided the Burseraceae into
three tribes on the basis of fruit structure: Protieae (4 genera),
Boswelliaea or Bursereae (eight genera which include
Boswellia and Commiphora) and Canarieae (9 genera).
The genus Commiphora with 150-200 species is wide
spread in the drier parts of tropical Africa and Madagascar,
also from Arabia to India and with few species occurring in
South America. The genus is a very conspicuous and
dominant element in the dry bush lands of North East Africa,
where a large number of species are endemic to this area
[31].
Commiphora myrrha is the chief source of myrrh today.
The plants grow wild in the North-Eastern province of Kenya
and adjoining areas of Somalia and Ethiopia. These plants
yield economically important gum exudates which have been
collected for centuries as medicinal and perfumery
substances [32].
Holmes [33, 34] apparently was the first to propose that
the myrrh of the Bible was the perfumed myrrh or "bissabol"
and not the medicinal myrrh or "heerabol" from C. myrrha.
Common myrrh (heerabol) is obtained from C. myrrha; this
is the species from which "oil of myrrh", or stacte, was
obtained. Other species sometimes passing as myrrh or
bdellium include C. africana, C. anglosomaliae, C.
gileadensis (C. opobalsamum), C. hildebrandtii, C. kataf, C.
mukul, and C. schimperi [35]. The odor of myrrh is described
as warm-balsamic, (sweet, and somewhat spicy aromatic,
sharp and pungent when fresh [36].
The gifts presented by the Maji to the infant Christ
symbolized His life: "gold for royalty, rank incense for
divinity, and myrrh for suffering" [36]. Myrrh was also in the
final drink offered to Christ on the cross: "And they gave
Him to drink wine mingled with myrrh; but He received it
not" (Mark 15:23). Myrrh was in addition, used to embalm
the body of Christ: "And there came Nicodemus, which at
first came to Jesus by night, and brought a mixture of myrrh
and aloes, about a hundred pound weight" (John 19:39).
Myrrh was also included in the "oil of holy ointment"
(Exodus 30: 23-24). Many herbalists recommend tincture of
myrrh as an astringent for the mucous membranes of the
mouth and throat [37]. Myrrh is found in salve used in
treating bed sores, hemorrhoids, and wounds. Internally,
myrrh is also used for indigestion, ulcers, and bronchial

Modern Chemistry 2019; 7(3): 45-53 47
congestion and as an emmenagogue [35].
Among local African traditional medicines, the resinous
gums of C. myrrha and C. guidottii; which are locally known
as "malmal" and "habak-hadi" in the Somali vernacular,
respectively, are used on livestock against ticks [32]. The
major use of myrrh is for burning as incense in religious
ceremonies. The resin is distilled to yield volatile oils and
these have their own characteristic balsamic odor which finds
use in perfumery [29]. The resin of C. guidottii is the second
major type of myrrh and it is commonly known in commerce
as gum "bissabol" (Hindi) or "opopanax". Opopanax occurs
also widely in Ethiopia from which the resin is collected for
export to India, China and Europe. Thulin et al. [38]
suggested that the myrrh of the Bible and the incense of the
ancient Egyptians and of Classical times [35] was most likely
the "perfumed myrrh" or "bissabol" from C. guidottii and not
the medicinal myrrh from C. myrrha. "Bissabol" according to
Holmes [39] meant buffalo myrrh, "as it is mixed with food
given to milch cows and buffaloes to improve quality and
quantity of their milk." Thulin and Claeson [38] confirmed
that the tree called "hadi" in Somali is C. guidottii and
produces the resin "habak-hadi" also known by several other
names including "opopanax", "bissabol", "scented myrrh",
"abeked" (Amharic). Tucholka [40], in a thesis on the
chemical composition of "bissabol", reported that "habak-
hadi" was used during female circumcision, by bathing in
water in which the resin was emulsified. A similar bath was
taken by Somali women after giving birth to a child. "Habak-
hadi" is used in Somalia for the treatment of stomach
complaints and diarrhea [41]. It is also used topically for the
treatment of wounds.
Extraction of the resins by organic solvents furnishes a
"resinoid" or an "absolute." The "resinoid" is prepared by
extraction of the crude resin with a hydrocarbon solvent such
as hexane or petrol. The "resinoid" contains almost all the
available essential oils of the resin. The "absolute" is
prepared by extraction of the resins with alcohol [29].
Essential oils on the other hand are separated by steam or
hydro-distillation at atmospheric pressure.
Gas chromatography is an excellent tool for the separation,
characterization, and quantitative analysis of essential oils.
The combined gas chromatogram-mass spectrometer
(GC/MS) method provides a facile, sensitive and convenient
system for the separation and identification of complex
mixtures [42]. Spectroscopic methods like UV, IR,
1
H and
13
C NMR are among the most powerful techniques for the
characterization of isolated compounds.
The components of the essential oils obtained from only
few Commiphora species have been investigated and these
include: C. terebinthina and C. cyclophylla [43], C. myrrha
[44, 45] and C. rostrata [46]. The oils from C. rostrata are
distinguished by the presence of the homologous ketones
starting from 2-octanone, 2-nonanone, 2-decanone etc [46].
The other Commiphora species are rich both in the structures
of monoterpenes and sesquiterpenes. Oxygenated terpenoids
are the components of essential oils most often responsible
for their distinctive aroma and flavor, even though they are
often minor constituents of the oil [47].
It is interesting to note that as most of the previous reports
on resin of C. myrrha are based on the study of materials
obtained from commerce, it is highly likely that the resins are
derived from different Commiphora species. This shows the
significance of working on resins from properly identified
trees.
2. Materials and Methods
2.1. General
Hydro-distillation of resin was done at atmospheric
pressure using 4 L round bottom flask fitted with Clevenger
apparatus and glass condenser. Optical rotation of the hydro-
distillate was measured with Perkin-Elmer 241, Polarimeter,
at room temperature using sodium D line.
GC was run using Hewlett-Packard 6890 GC series
equipped with FID and HP-5 capillary column (cross
linked 5% diPh, 95% dimethylpolysiloxane, 30 m x 0.32
mm i.d. x 0.25 µm film thickness). The column
temperature was programmed at 50-210
ο
C at a rate of
3
ο
C/min. The injector and detector temperatures were
220
ο
C and 270
ο
C, respectively. Samples (0.5 µL of the oil
solutions in CHCl
3
, 2 mg/mL) were injected by the
splitless technique. Nitrogen was used as carrier gas (10
Psi or 2.3 mL/min).
GC/MS was performed on a Fisons GC model 8000 series
coupled to a mass spectrometer, MD 800 quadrupole
analyzer operating at 70 eV. The capillary column type was
DB-17 (50% Ph, 50% methylpolysiloxane, 30 m x 0.25 mm
i.d. x 0.25 µm film thickness) with helium as the carrier gas
(5 Psi or 1.15 mL/min). Samples (0.6 µL of the oil solutions
in CHCl
3
5 mg/mL) were injected by the split technique.
Identification of the constituents of the essential oils
was performed by matching MS data of each constituent
With Wiley, NIST and user generated mass spectral
libraries.
Refractive index was measured at room temperature using
Atago Abbe refractometer, No 99996, Japan.
Column chromatography (CC) was done with column
size 3 cm x 30 cm packed with silica gel 60, size 0.063-
0.200 mm (70-230 mesh ASTM) and thin layer
chromatography (TLC) on aluminum sheets, silica gel 60
F
254
, and layer thickness 0.2 mm (Merck). Preparative thin
layer chromatography (PTLC) plates were prepared on 20
cm x 20 cm glass, silica gel 60 PF
254+366
, 7748 (Merck)
layer thickness 0.5 mm. Spot detection on TLC was
performed by using UV (254 nm, 365 nm) and spray
reagent 1% vanillin in H
2
SO
4
.
NMR data was generated with 300 MHz for
l
H and 75
MHz for
l3
C, for compounds isolated from C. schimperi and
400 MHz for
1
H NMR and 100 MHz for
13
C NMR, for the
compound isolated from T. vogelii, TMS as internal standard
and CDCl
3
(for C. schimperi) and DMSO-d
6
(for T. vogelii)
solvent. IR spectrum was measured with Perkin-Elmer, 1600
series FTIR, using KBr pellets.

48 Tegene Tesfaye Tole and Belay Akino Neme: Chemical Studies of Tephrosia vogelii and
Commiphora schimperi Occurring in Ethiopia
2.2. Extraction and Compound Isolation from the Stem
Bark of Tephrosia vogelii
2.2.1. Plant Materials
The stem-bark of Tephrosia vogelii was collected in
November, 2017 from Areka Agricultural Research institute,
Wolaita Zone, Ethiopia. The plant was identified by botanist
Retta Regasa Department of Biology, Hawassa College of
Teachers Education. Plant specimen was deposited at the
herbarium of Hawassa College of Teacher Education, with
voucher number MB/0087.
2.2.2. Extraction
The collected stem bark of Tephrosia vogelii was chopped
into small pieces and air dried under shade for 30 days and
milled to suitable size by using mortar and pestle. About 500
g of the powdered stem bark of Tephrosia vogelii was
soaked in 3 L petrol in a round bottom flask, at room
temperature. The round bottom flask was put on an orbital
shaker and left for 24 hours at a speed of 120 revolutions
per minute. After 24 hrs the solution was filtered using
Whatman filter paper. The filtered solution was
concentrated using rotary evaporator at reduced pressure and
a temperature of about 40
ο
C. The marc left was further
extracted with chloroform and methanol consecutively, likewise.
2.2.3. Phytochemical Screening
Phytochemical screening tests were carried out on the
crude extracts (petroleum ether, chloroform and methanol)
following the standard procedures of Ganesh and Vennila
[48] and Prashant et al., [49] in order to investigate the types
of secondary metabolites present in the plant species under
investigation.
2.2.4. Compound Isolation
TLC profile analyses were carried out in chloroform and
methanol extract in different solvent systems to identify the
appropriate solvent for column chromatography. TLC profile
of methanolic extract was conducted in n-hexane-ethyl
acetate, ethyl acetate-chloroform, and methanol-ethyl acetate,
solvent combinations by varying solvent ratios. Ethyl acetate-
n-hexane combination showed good TLC profile. Twelve
gram of methanolic extract was adsorbed on 15 g of silica gel
and was subjected to column chromatography. The column
was packed by n-hexane to achieve least polarity to the
mobile phase during the beginning of elution. Elution was
conducted with increasing gradient of n-hexane-ethyl acetate.
A total of 65 fractions (25 ml each) were collected and
analyzed by TLC for identification of the pure components.
Fractions 16-27 showed two spots and were combined and
further fractionation was conducted in small column in n-
hexane-ethyl acetate (6:4, 4:6, 7:3, 3:7, ethyl acetate) solvent
system and a total of 34 fractions were collected. TLC
analysis of fractions 10-17 showed a single spot with Rf
value (0.63) in (7:3) n-hexane-ethyl acetate solvent system
having minor impurity. These fractions were combined and
further purified by washing in n-hexane for several times. A
white solid compound (40 mg) was obtained and coded BA-
5. Spectroscopic data of BA-5 was generated using UV, IR,
1
H and
13
C NMR techniques, for structure elucidation.
2.3. Extraction and Compound Isolation from the Resin of
C. schimperi
2.3.1. Plant Material
Plant materials of C. schimperi were collected on two
occasions in January, and April, 2016 from Gamo Gofa
(Konso). Konso is located 587 km South of Addis Ababa.
The local name of all the trees at Konso is "Qahatita". The
trees are approximately 4-6 m, with bark that peels into
flakes. When bark is incised milky exudate flows out and
solidifies within 30 min and becomes dark-brown after one
day. Naturally exuded resins were collected. Leaves and bark
were collected to aid in the botanical identification of the
species. The plant was identified by Kaj Vollesen (Kew
Royal Botanical Garden, U.K.) and voucher specimens have
been deposited at the National Herbarium, Addis Ababa
University with collectors’ number Tegene 3 and herbarium
numbers 072773 and 072774.
2.3.2. Ethnobotany
Ethnobotanical information was gathered by interviewing
the local people and through observation.
2.3.3. Hydro-distillation
The dry resin (15 g) was first crushed as much as
possible into smaller pieces and placed in a round bottom
flask fitted with Clevenger apparatus and was hydro-
distilled for 3 h at atmospheric pressure. The strongly
aromatic oil was separated from the water layer by
separatory funnel and dried by adding anhydrous Na
2
SO
4
and then weighed.
2.3.4. Extraction
Resin of C. schimperi (62 g) was first crushed and
extracted with petrol (200 mL) to give crude extract (30 g,
48%). The marc was soaked with MeOH (150 mL), filtered,
and the solvent removed to give 3.5 g (6%). The total amount
of extract obtained was 33.5 g (54%).
2.3.5. Compound Isolation
The petrol extract (28 g) of the resin of C. schimperi was
applied on column packed with silica gel (petrol) and eluted
with petrol-CHCl
3
gradients. Out of the 31 fractions
collected, fractions number 4 and 5 (petrol: CHCl
3
, 49: 1)
gave nearly pure colorless liquid compound coded 82-1A (1
g). Subsequent fractions gave colorless liquids with two spots
having the same Rf values and was coded 82-1B.
3. Results and Discussion
3.1. Chemical Analysis of the Stem Bark of Tephrosia
vogelii
3.1.1. Extraction
The yields of the crude extracts of Tephrosia vogelii are
presented in Table 1 below.

Modern Chemistry 2019; 7(3): 45-53 49
Table 1. The percent yield of crude extracts.
Solvent Extract (g) Yield (%)
Petroleum ether 2 0.4
Chloroform 5 1
Methanol 12 2.4
As can be seen in Table 1 above the crude extract yield
increased up on increasing the solvent polarity. This indicates
that most of the chemical constituents in the plant stem bark
are polar. The small yield of petroleum ether and chloroform
extracts only allowed phytochemical screening.
3.1.2. Phytochemical Screening
A variety of herbs and herbal extracts contain different
secondary metabolites with biological activity that can be of
valuable therapeutic index. These secondary metabolites are
natural products that are responsible for medicinal properties
of plants to which they belong. Plants of the genus Tephrosia
are known as rich source of secondary metabolites such as
alkaloids, terpenoids, saponins, tannins and other aromatic
compounds [50]. Phytochemical screening tests of different
solvent extract of stem bark of Tephrosia vogelii revealed the
presence of medicinally active secondary metabolites
including tannins, saponins, terpenoids, and flavonoids.
Anthraquinones, steroids and alkaloids are absent, however.
The phytochemical screening results of the methanolic and
aqueous extracts of the leaf of T. vogelii by Inalegwu and
Sodipo [51] showed the presence of cardiac glycosides,
alkaloids, saponins, and phlobatannins and absence of
anthraquinones and tannins. Similarly the ethanolic leaf
extract of T. vogelii showed the presence of catechol tannins,
saponins, sugars, leuco-anthocyanins, polyterpenes, and
sterols [32].
3.1.3. Structure Elucidation of Compound BA-5
Compound BA-5 is deep yellow solid, melting point
128°C and Rf
0.63(n- hexane-ethyl acetate; 70:30). The UV-
VIS spectrum showed λ
max
343 nm (in CHCl
3
) attributed to
the presence of n-π* transition of carbonyl.
In the IR spectrum, a broad absorption band at 3397 cm
-1
attributed to hydroxyl group; 1743 cm
-1
presence of carbonyl
group. Absorption bands at 2965 cm
-1
and 2918 cm
-1
shows
the presence of sp
3
and sp
2
C-H stretching vibrations. The
medium bands at1618 and 1516 cm
-1
attributed to aromatic
ring C=C stretching. The absorption bands at 1138 cm
-1
and
1092 cm
-1
indicated C-O stretching.
1
H-NMR (400 MHz, DMSO-d
6
) δ = 3.96 (3H, s, H-7’) and
1.55 (3H, d, H-6’’) correspond to methyl and methoxy
groups, respectively. The signals at δ = 7.70 - 6.20 are
aromatic; 7.66 (1H, d, J = 8.8 Hz, H-4’’), 7.33 (1H, d, J = 8
Hz, H-2’and 6’), 6.92 (1H, d, J = 7.6 Hz, H-3’ and 5’), 6.80
(1H, d, J = 8.4 Hz, H-5), 6.51 (1H, d, J = 6.4 Hz, H-6), 6.34
(1H, m, H-5’’).
The
13
C NMR and DEPT-135 spectra of compound BA-5
shows a total of nineteen carbon signals δ: 24.8 (C-6′′) and
56.2 (C-7′) for the methyl and methoxy carbons, respectively;
δ: 147.99 (C-4′′), 130.8 (C-4′), 128.7 (C-5), 127.5 (C-3′ &
5′), 126.3 (C-2′ &C-6′), 110.9 (C-6), 103.0 (C-3), 79.7(C-5′′)
are CH carbons; δ: 190.6 (C-4), 174.9 (C-2′′), 165.1 (C-7),
163.6 (C-2), 158.1 (C-8a), 130.3 (C-1′), 128.7 (C-3′′), 115.6
(C-4a), 111.6 (C-8) are quaternary carbons.
Based on UV, IR,
1
H and
13
C NMR spectroscopic data and
comparison with previous reports (Table 2), compound BA-5
shares most of the spectroscopic data of 8 (4, 5-dihydro-5, 5-
dimethyl-4-oxofuran-3-yl)-5-hydoxy-7-methoxy-2-phenyl-
4H-chrome-4-one (Figure 1 below). This compound was
isolated for the first time from Tephrosia vogelii.
Table 2. Comparison of
1
H-NMR and
13
C-NMR spectra of compound BA-5 with previous report [50].
Position
13
C-NMR (δ in ppm)
1
H-NMR (δ in ppm) DEPT Reported [50]
δ
1
H-NMR δ
13
C-NMR
1 - - - - -
2 163.63 - Quaternary 161.69
3 103.04 7.01 (1H, s) CH 6.74(s) 107.35
4 190.64 - Quaternary - 177.72
4a 115.61 - Quaternary - 118.04
5 128.7 6.5(1H,d,H-5) CH 6.4(d) 128.13
6 110.98 7.3(1H, d) CH 7.08(d) 109.36
7 165.08 - Quaternary - 163.18
8 111.58 - Quaternary - 109.7
8a 158.05 - Quaternary - 154.87
1′ 130.33 - Quaternary - 131.92
2′, 6′ 126.3 7.63(1H, d) CH 7.43(m) 126.2
3′, 5′ 127.52 7.3(1H, d) CH 7.74(m) 128.99
4′ 130.80 6.9(1H, d) CH 7.43 (m) 131.5
6′′ 24.8 1.55(3H, s) CH
3
1.65(6H,s) 25.83
7′ 56.1 3.96 (3H,s) OCH
3
3.94(3H,s) 56.6
2′′ 174.99 - Quaternary 170.62
3′′ 128.69 - Quaternary 124.24
4′′ 147.99 7.1(1H, s) CH 7.52(s) 159.89
5′′ 79.71 6.34 (1H,s) CH 6.29 (s) 84.92 (q)

50 Tegene Tesfaye Tole and Belay Akino Neme: Chemical Studies of Tephrosia vogelii and
Commiphora schimperi Occurring in Ethiopia
Figure 1. A proposed structure of compound BA-5.
3.2. Ethnobotany and Chemical Analysis of Commiphora
Schimperi
3.2.1. Ethnobotany
The Commiphora plant is widely grown in Arbaminch and
Konso because it is suitable for hedge and fence. In
Arbaminch the tree is known as "Tsedaki" (Amharic), the
Konso people call it "Qahatita". The name "Qahatita" means
that the plant drives away wild animals that destroy
vegetables in the garden. The ease with which the plant is
propagated from cutting accounts for its wide use for fences
and hedges in Konso and Arbaminch. Furthermore the leaves
are used for cattle feed and the wood for building purposes.
However, interview made with several residents in
Arbaminch and Konso revealed that the residents have very
little knowledge of the resins produced by the trees. The
resins are not collected for use as incense. The resin and plant
specimens used in this study were collected from Konso.
3.2.2. Isolation and Analysis of the Essential Oil
The resin is collected in such a way that the milky liquid
exudate coming out from the tree hardens on exposure to air
into droplets or "tears" which are then easily detached by a
collector. Occasionally, some "tears" are produced by
accidental injury or from splits which occur in the stems or
branches of the tree.
The essential oil of the resin of C. schimperi was obtained
by hydro-distillation. GC and GC/MS analysis of the oil was
undertaken and the result is presented below (Table 3).
Table 3. Physical data on the isolated essential oil of C. schimperi resin.
Species Resin (g)
Oil (mg)
% yield
[α]
D
Ref. Ind.
C. schimperi 14 662 5.0 -41.6
1.472
The total number of components is 15 (98.6%) with α-
pinene and β-pinene accounting for 89% of the oil, making
this resin an excellent source of these industrially important
chemicals. Other constituents found in the essential oil
include: camphene (0.6%), d, l-limonene (0.9%), β-
phelandrene (0.5%), p-cymene (0.9%), pinocarveole (0.5%),
t-verbenol (0.3%), p-menth-l-en-4-ol (0.5%), myrtenol
(0.2%), Z-citral (1.4%), 2-pinene-4-one (0.6%), E-citral
(1.5%), t-caryophyllene (0.8%), and caryophyllene oxide
(0.8%). α-Pinene (6.2%), β-pinene (6.7%), and t-
caryophyllene (33.4%) exist in the essential oil extracted
from Commiphora kataf resin [52].
1: α-pinene (72.5%) 2: β-pinene (16.8%)
Figure 2. Gas chromatogram of hydro-distillate of C. schimperi.
GC-MS,
1
H and
13
C NMR spectra of the essential oil
confirmed that the major components are α-pinene and β-
pinene. Also fractionation of the petrol extract of the resin
resulted in isolation of α-pinene and a mixture of α-pinene
(69%) and β-pinene (20%).
α-Pinene is the main constituent of turpentine, widely
distributed in conifers and other plants. It is important
intermediate in the manufacture of synthetic fragrant
compounds and also used as a flavoring ingredient. It
undergoes cationic polymerization to give terpene resins. In

Modern Chemistry 2019; 7(3): 45-53 51
Kenya, C. schimperi, traditionally known as “Osilalei” is
used for treatment of malaria. The methanolic extract of the
resin of C. schimperi showed anti malarial activity [53]. The
stem extract of the same species exhibited antioxidant effect
in the DPPH assay [54].
3.2.3. Characterization of Compounds Isolated from the
Resin of Commiphora Schimperi
The resin of C. schimperi was first extracted with petrol
(48%) and then with MeOH (6%). Fractionation of the petrol
extract resulted in the isolation of the two major constituents
of the essential oil which make up 89%. The isolated
compounds are coded 82-1A and 82-1B.
IR,
1
H and
l3
C NMR values are compared with literature
[55] for characterization of the isolated compounds in
addition to GC.
i. Compound 82-1A
IR: v
max
(cm
-l
) 3024 (w) H-C str. of unsaturated
compounds, 2918 (s) H-C- str. of saturated compounds, 1380
and 1355 (both m) H-C- def. of branched saturated
compounds.
1
H NMR: δ 5.15 (1H, t) imply vinyl proton, 1.63 (3H, s),
1.23 (3H, s), and 0.79 (3H, s) are methyl protons at carbon
number 8, 9 and 10, respectively.
l3
CNMR: The total number of carbons in the spectra is 10.
Therefore the compound is a monoterpene. Comparing NMR
data with literature [55], all chemical shifts match with that of
α-pinene (1). Therefore the compound is α-pinene as a result of
its NMR data and by its retention time indicated in the GC.
i. Compound 82-1B:
1
H and
l3
CNMR data comparison with literature [55], GC,
and GC/MS data are used for characterization.
GC and GC/MS: The gas chromatogram and GC/MS
clearly showed that, fraction 82-1B is a mixture of α-pinene
(1, 69%) and β-pinene (2, 20%).
1
H NMR: 84.58 (1H, td) and 4.52 (1H, td) are vinyl
protons of β-pinene. In addition to these all the chemical
shifts of α-pinene are found in the spectrum.
Figure 3. Compounds isolated from the resin of C. schimperi.
13
C NMR: The chemical shift values of the
13
C NMR
spectra match with α-pinene (1) and β-pinene (2), when
compared to literature [55]. Therefore, 82-1B is a mixture of
α-pinene and β-pinene.
4. Conclusion
To the best of our knowledge there is no prior report on the
chemical constituents of stem bark of Tephrosia vogelii
contrary to its high traditional use. Preliminary
phytochemical screening of the extracts of the stem bark
revealed the presence of terpenoids, flavonoids, tannins, and
saponins and the absence of anthraquinones, steroids, and
alkaloids. Fractionation of the methanolic extract resulted in
the isolation of the terpenoids 8 (4, 5-dihydro-5, 5-dimethyl-
4-oxofuran-3yl)-5-hydoxy-7-methoxy-2-phenyl-4H-chrome-
4-one which is isolated for the first time, from the species. It,
however, is important to investigate its biological activity.
Analysis of the essential oil isolated from the resin of
Commiphora schimperi revealed the presence of α-pinene
(69%) which is an important intermediate in the synthetic
fragrant compounds and also used as a flavoring ingredient
and β-pinene (20%) as the major components. here is no
previous report on the analysis of the essential oil from the
resin of C. schimperi, in Ethiopia. Fractionation of the petrol
extract of the resin of C. schimperi further confirmed the
presence of α-pinene and β-pinene, which together make up
89% of the essential oil. In both species isolation and
characterization of other compounds needs to be done.
Conflict of Interest
The authors have not declared conflict of interest.
Acknowledgements
We acknowledge the Swedish Agency for Research Co-
operation with Developing Countries (SAREC) for the
financial aid obtained through the Ethiopian Science and
Technology Commission (ESTC) and Addis Ababa
University, Research and Publication Office (RPO). Hawassa
University, Research and Publication Office, is also
acknowledged for providing additional funding and material
support.
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