KIMIKA 10: 47-52 (1994)
Printed in the Philippines
REVIEW PAPER
Bioactive metabolites from Moringa oleifera Lam.
IRENE M. VILLASENOR
Institute of Chemistry
University of the Philippines at Diliman
Diliman, Quezon City 1101, Philippines
(Accepted March 10, 1994)
Moringa oleifera Lam. is a well-known Philippine medicinal plant.
the plant as well as the known chemical composition of the seeds with emphasis on the bioactive metabolites. These metabolites
are: 4(a-L-rhamnosyloxy)benzylisothiocyanate, an antimicrobial
rhamnosyloxy)phenylacetonitrile, a mutagenic compound from the roasted seeds.
Key Words: Moringa oleifera, malunggay, 4(a-L-rhamnosyloxy)benzylisothiocyanate,
4(a- L-rhamnosyloxy)phenylacetonitrile,
MoRJNGA OLEIFERA LAM. IS CLASSIFIED UNDER THE DIVISION
Empryophyta Siphonogama, class Oppositifoliae
(Dicotyledoneae), subclass Strobiloideae, superorder
Apopetaeae-Polycarpellatae, order Rhoedales, family
Moringaceae (1). It is commonly. known as horse radish
tree, drumstick tree, radish tree, or West Indian Ben (2)
and is locally known as malunggay.
The plant is a small tree, growing up to 8 m high
with corky bark and soft, white wood. The roots have a
pungent taste. The leaves are alternate, usually thrice
pinnate, and 25-50 em long. There are 3 to 9 leaflets on
the ultimate pinnules, pale beneath, thin, ovate to ellip­
tic, and each is 1-2 em long. The pod is 15-30 em long,
pendulous, three-angled, and nine-ribbed. Its seeds are
about 1 em in diameter, three-angled, and winged on the
angles. It consists of an outer husk and an inner kernel
rich in oil (3).
Medicinal properties
Roots
The decoction of the root is used internally for fever,
epilepsy, hysteria, palsy, tetanus, paralysis, leprosy, and
high blood pressure (4); in relaxing the throat, curing
hoarse voice, and relieving sudden abnormal involuntary
contraction of the muscles; is an effective gargle; and is
used to clean sores and ulcers. The young roots are em­
ployed as a stimulant, diuretic, antidote for snake bites,
and for ear aches, tooth decay (5) and inflammatory swell­
ings; as an antiscorbutic, rubefacient, and counterirritant.
The juice of the root, when mixed with milk is also help­
ful as a decoction in hiccoughs, asthma, gout, lumbago,
rheumatism, enlarged spleen or liver, internal and deep­
seated inflammations, and calculous affections (6). The
root also has antibacterial activity (7-14).
47
This paper reviews the medicinal properties of all parts of
constituent from the non-roasted seeds; and 4(a-L­
antimicrobial, mutagen,
Leaves
The young leaves are rich in calcium, iron, phospho­
rus, and Vitamin C (15). When eaten as vegetables, the
leaves are laxative; galactogogue (16), and hypotensive
(4); promote digestion; and alleviate hysteria, flatulence, •
and gonorrhea. As a poultice, they are useful in reducing
glandular swellings (6) and expel intestinal worms (5).
Pods I Wood I Bark
The pods have anthelmintic property and are admin­
istered in affections of the liver and spleen, and in articu­
lar pains (6). The bark is used as a cardiac stimulant (5),
an emmenagogue and an abortifacient (17), a rubefacient
and a vesicant, and an antiscorbutic; as a cure for asthma
and cough; and as a relief for pain (5). A decoction of the
root-bark relieves spasms and is useful in calculous af­
fections. The gum is used for the relief of otalgia and
intestinal complaints (6).
Flowers I Seeds
The flowers are stimulant, tonic, diuretic, increase
the flow of bile (5) and banish cough that lingers after
influenza (17). When boiled in milk, the flowers are used
as aphrodisiacs (5). The seed oil has a curative effect on
skin diseases (18). For the treatment of rheumatism and
acute gout, the seeds are roasted, powdered, and applied
on the affected part (16). The seeds have also been used
in cases of ascitis (4). Antibacterial activity has also been
ascribed to the seeds (19). The paste made from th�
crushed seeds can be used to clear and purify water (20-
22).
48 IRENE M. VILLASENOR

Composition of the seeds �-0- CH2·N:C:S

ACID
HYDROLYSIS
L, RHAMNOSE

Fatty Acid Composition HO OH

(D]
The seed consists of 35.8% tegmentum and 64.2% nut.
The nut has a high content of oil and protein while the
MUSTARD OIL

j CoH1COSH
� CH COSH
1
OACETIC ACID!

tegmentum is extremely rich in cellulose (2,23) (Table 1). I THIOBENZOIC ACID I

The seed contains 25-50% of an oil which when refined is ""1c:;cooJ20

o-0-cH2NHCOCoHs
� RIDINE
slightly yellowish and quite odorless and has a sweet

V
pleasant odor (24). The usual physico-chemical constants HO

(18,25-27) (Table 2) of this Ben or Behen oil were already '\:.-10

AcO
!o:;: o .....F;;\_
\J -v---CH2NHCOCH1

determined. The fatty acids (15,18,25,28-30) are palm­ H O OH

[Ill] AcO OAc
itic, stearic, behenic, lignoceric, oleic, linoleic (28,30) lau­
1
BENZAMIOE
�
ric, palmito-oleic (15), and myristic (29) (Table 3). H , H20 [mJ

0
I ACID HYDROLYSIS)
PERACETYL DERIVATIVE

The Parent Glucosinolate H CH2NHCOC0H0

Badgett obtained 4(a-L-rhamnosyloxy)benzylglucosino­ PH[7JoL

late [1] as the heptaacetate after acetylation of M. oleifera

seed extracts (31). He established the structure and ste­ Figure 1. Derivatization of 4(a-L-rhamnosyloxy)benzyl­
reochemistry of the alkyl group of the parent glucosinolate isothiocyanate [Badgett B.L. (1964) DissertationAb­
,
by derivatization and by synthesis (Figure 1). stracts:XXV(3), 1556-7].

Glucosinolates are thioglucosides mainly found in the

Table 1. Composition of the seeds of Moringa
Cruciferae. The side chain R can represent a number of
pterygosperma
structural possibilities, but all are derived from or elabo­
rated from the side chains of naturally occurring a-amino
Seed Tegmentum Nut
acids (32,33).
Moisture 9.60 12.16 8.16
Ethyl ether extract 22..42 0.96 34.39 R �
N 2.87 0.70 4.08
C=N-0- S-O·
?
Crude protein 17.94 4.38 25.50
Crude cellulose
Ash
10.86
3.12
27.45
2.87
1.61
3.26
13-glucosyl-S / 6
N-free extract 36.06 52.18 27.08
The glucosides are split by the action of an enzyme
Ref.: Garcia de Orta, 9 (Part 2), 384 (1961). known as myrosinase or thioglucosidase. Simple hydroly­
sis will yield a hydroxamic acid, glucose, and inorganic
sulfate. The hydroxamic acid usually undergoes an im-

Table 2. Physico-Chemical Characteristics of Ben Oil

ref.18 ref.25 ref.26 ref.27

Specific gravity 0.8984 0.8435

Refractive index 1.46524 1.4650 1.4490 1.4598
Acid value 3.48 160.9 1.777
Eq. to FFA,% 6.5 80.8
Saponification value 182.20 188.1 191.6 182.1
Saponification eq. 298.3
Ester value 30.7
Iodine value, % 64.20 67.1 68.2
Thiocyanogen value, % 64 4 .
Unsaponifiable Matter, % 3.05 1.1 0.7 1.769
Ash 0.01
Sat'd Fatty Acids 22.0
Reichert-Meisel value 0.44 0.58
Hehner value 91.59
Polenske value 0.28

KIMIKA • Volume 10 • 1994

 BIOACTIVE METABOLITES FROM MORINGA OLEIFERA LAM. 49

Table 3. Percent Fatty Acids from Seeds of Moringa oleifera

ref.15 ref.18 ref.25 ref.28 ref.�9 ref.30

oleic 73. 3 65. 7 75 0

. 71. 0 68. 9 67.48
linoleic 0. 8 0. 08 38
. 3.4 0
myristic 1.5
palmitic 9.6 9.3 5.5 3.1 3.6 3.40
stearic 3.0 7.4 7.7 8. 0 10.8 10.5
behenic 8.6 1.2 3. 5 6.3
lignoceric 5.3 5.8 0.13
lauric 12.5
palmitoleic 1.6
arachidic 7.8

mediate Lossen rearrangement to form an isothiocyanate

(34). Bioactive metabolites

R R 4(a-L-Rhamnosyloxy)benzylisothiocyanate
\ thioglucoside \
C=N-0-SO2O· ------i> C=N-OH 4(a-L-Rhamnosyloxy)benzylisothiocyanate [2] was
I glucohydratase / identified as an active antimicrobial agent from the seeds
13-glucosyl-S HS of M. oleifera (19,39,40). It is formed from the hydrolysis
·of the parent glucosinolate (Figure 2). This reaction is
Lossen catalyzed by the endogenous seed enzyme myrosinase.
R-N=C=S + �0 The compound acts on several bacteria and fungi. It was
reanangement found to be active against B. subtilis but inactive against
E. coli (40). When left to stand in aqueous methanol solu­
The other degradative routes comprise the fragmen­ tion, it decomposes and loses its antibacterial activity.
tation to nitriles, and the production of thiocyanates (35). Compound £21 was toxic to mice even at low dosages of
1.26 mg and 0:55 mg I 25 g mouse . At the non-toxic dose
R-NC S + S0;2 of0.22 mg /25 g mouse, compound [2] was not mutagenic
R-CN + S + S04oo2 (41).
(R+) + SCN + S04•2
·

4(a-L-Rhamnosyloxy)phenylacetonitrile
The nitrile is supposedly formed by a protonation
mechanism and is therefore favored at lower pH values. The seeds of M. oleifera Lam., when roasted, are used
Between pH 3 and 5, a mixture of nitrile and externally for the treatment of gout and rheumatism (16).
isothiocyanate is expected. But above pH 5 only the However, studies documenting the mutagenicity of the
isothiocyanate should be obtained (36). Studies on the roasted seeds after metabolic activation exist in the lit­
thermal degradation of glucosinolates (37 ,38) showed that eratlp'e (42,43). Mutations have been attributed to DNA
the effect of heat is to promote nitrile formation. damage or misrepair (44-48).

fSO,.K0H HO
OH

HO � -0-cH2-c-s N
n
0
CHzOH
GLUCOSINOLAS J H*-0\IJ:\._
v�-vCHaN=C=S
v HO OH
H20

CH OH

[I] [II]
MUSTARD OIL GLUCOSIDE MUSTARD OIL + GLUCOSIDE

Figure 2. Formation of 4(a-L-rhamnosyloxy)benzylisothiocyanate [Badgett, B.L. (1964) Dissertation Abstracts XXV (3), 1556-7].

KIMIKA • Volume 10 • 1994

50 IRENE M. VILLASENOR

Spectral analyses showed that the structure of the
isolated mutagen is 4(a.-L-rhamnosyloxy)phenylaceto­
nitrile [3] (49). The Micronucleus test (50-53), an in vivo
method, using albino mice as the test system, showed
that the number of micronucleated polychromatic eryth­
rocytes (MN-PCE) per 1000 PCE for compound [3) (11.38
± 2.12) is higher than of the solvent control, dimethyl­
sulfoxide (3.93 ± 0.39), and approximates that of the posi­
tive control, tetracycline (12.20 ± 1.09). Compound [3]
did not exhibit antimicrobial activity (40).
The parent glucosinolate was not isolated in the
previous study (49). There was no experimental proof
that the precursor of the mutagen is the glucosinolate al�
though it is reasonable to assume the thermal degrada­
tion of the glucosinolate to the nitrile based on existing
literature (37,38). Instead an experiment was done to
investigate whether the precursor of the mutagen might
be the isothiocyanate itself (54).
Comparison of the HPLC chromatograms of the
crude ethyl acetate extracts of non-roasted and roasted
seeds showed that [2] is present in both non-roasted and
roasted seeds while [3] is present only in the extracts of
the roasted seeds (54). The results were confirmed by
co-injection experiments.
In order to determine whether [2) is the precursor
of [3], compound [2] was heated to 180•C and sampled
at different time intervals and analyzed by HPLC. No
trace of compound [3] was detected in any of the chro­
matograms. Thus, it can be concluded that [3] was
·not formed from [2] but may have arisen from the
thermal degradation of the parent glucosinolate [1]
(Figure 3).
Hydrolysis Products of [8]
A number of biosynthetically and chemically related
compounds were isolated from the roasted seeds of M.
oleifera. The structures of 4-hydroxybenzylcyanide [4], 4-
hydroxyphenylacetamide [5], and 4-hydroxyphenylacetic
acid [6], were established by comparison of their spectral
data with literature values (41).
Structure-activity correlation studies using the Mi­
cronucleus test showed that compounds [4] (9.93 ± 0.68)
and [5] (8.07 ± 0.93) exhibited mutagenic activity while
compound [6] (3.53 ± 0.81) di,d not (55) (Figure 4). This
suggests that [4] is the proximate mutagenic agent since
the deoxy sugar moiety of [3] is readily cleaved by a
glycosylase while [5] is converted to [4] by protonation
and subsequent dehydration (Figure 5). Compound [4] is
then converted to 4-hydroxybenzaldehyde by a.-hydroxy­
lation and subsequent release of cyanide (56,57). The mu­
tagenicity of several aldehydes is well-documented.
Conclusion
This review gives us insights into the effect of heat­
ing on the chemical composition of the seeds of M. oleifera.
The non-roasted seeds contain an antimicrobial constitu­
ent, 4(a.-L-rhamnosyloxy)benzylisothiocyanate [2]. Heat­
ing produces 4(a.-L-rhamnosyloxy)phenylacetonitrile [3]
which is mutagenic. Compound [3] is not present in the
non-roasted seeds nor is it produced from the pyrolysis of
[2]. Both compounds arise from the parent glucosinolate
[1], the former by enzyme-catalyzed hydrolysis and the
latter by thermal degradation.

� �-oso1K .
�- CH2-C MYROSINASE •
H20
CH1 J.GLUCOSE
HO
(N-OSO,K
r-0- cH2-�
CH�
3 �:1 + GLUCOSE + H +

HO
H OH

/LOS SEN
REARRANGEMENT

D<·I:Rham- 0
{'�
-0- cH2 .(J
n
5

.c-L - Rham-o -0- cH2 N=c-s

+ S + HS04'

Figure 3. Mechanisms of formation of the isothiocyanate and nitrile from the parent glucosinolate.

KIMIKA • Volume I0 • 1994

 BIOACTIVE METABOLITES FROM MORINGA OLEIFERA LAM . 51

Compounda from Roaated Seeda This review also highlights the possibility that even
if some plant constituents may pro\jde the desired me­
Ave. No. of Mlcronucleated PCE/1000 PCE
dicinal effects; some negative actions on the genetic ma­
c
terial can take place. Hence. it is imperative that a pre­
liminary bioassay ofthe mutagenic potential of bioactive
natural products be done.

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HO

HO-o-�-H

[HO� OH�-H <-+HO . r"-- C-H

�I
�I 1 OH

[tn-G=r-H + OH

Figure 5. Proposed metabolic activation pathway for 4(u-L-rhamnosyloxy)phenylacetonitrile, its aglycone, and 4-
hydroxyphenylacetamide [Villasenor, I.M. et.al. (1989) Mutation Research 224, 209-212].

KIMIKA • Volume 10 • 1994

52 IRENE M. VILLASENOR

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KIMIKA • Volume 10 • 1994