PHARMACEUTICAL BIOLOGY, 2017

VOL. 55, NO. 1, 1394–1400

http://dx.doi.org/10.1080/13880209.2017.1302966

RESEARCH ARTICLE

Antimalarial and antiplasmodial activity of husk extract and fractions of Zea mays
Jude E. Okokona,b, Bassey S. Antiac, Dinesh Mohanakrishnanb and Dinkar Sahalb
a
Department of Pharmacology and Toxicology, Faculty of Pharmacy, University of Uyo, Uyo, Nigeria; bMalaria Research Laboratory, International
Centre for Genetic Engineering and Biotechnology, New Delhi, India; cDepartment of Chemistry, University of Uyo, Uyo, Nigeria

ABSTRACT ARTICLE HISTORY

Context: Zea mays L. (Poacae) husk decoctions are traditionally used in the treatment of malaria by vari- Received 11 April 2016
ous tribes in Nigeria. Revised 1 February 2017
Objective: To assess the antimalarial and antiplasmodial potentials of the husk extract and fractions on Accepted 2 March 2017
malaria parasites using in vivo and in vitro models.
KEYWORDS
Materials and methods: The ethanol husk extract and fractions (187–748 mg/kg, p.o.) of Zea mays were Herbal medicine;
investigated for antimalarial activity against Plasmodium berghei using rodent (mice) malaria models and ethnopharmacology;
in vitro activity against chloroquine sensitive (Pf 3D7) and resistant (Pf INDO) strains of Plasmodium falcip- Plasmodium berghei;
arum using the SRBR green assay method. Median lethal dose and cytotoxic activities against HeLa and plasmodium falciparum;
HEKS cells were also carried out. The GCMS analysis of the most active fraction was carried out. tropical diseases
Results: The husk extract (187–748 mg/kg, p.o.) with LD50 of 1874.83 mg/kg was found to exert significant
(p < 0.05–0.001) antimalarial activity against P. berghei infection in suppressive, prophylactive and curative
tests. The crude extract and fractions also exerted prominent activity against both chloroquine sensitive
(Pf 3D7) and resistant (Pf INDO) strains of P. falciparum with the ethyl acetate fraction exerting the highest
activity with IC50 values of 9.31 ± 0.46 lg/mL (Pf 3D7) and 3.69 ± 0.66 lg/mL (Pf INDO). The crude extract
and fractions were not cytotoxic to the two cell lines tested with IC50 values of >100 lg/mL against both
HeLa and HEKS cell lines.
Discussion and conclusion: These results suggest that the husk extract/fractions of Zea mays possesses
antimalarial and antiplasmodial activities and these justify its use in ethnomedicine to treat malaria
infections.

Introduction husk extract of Zea mays (Dong et al. 2014). Information on the
biological activities of the husk extract is scarce. We report in
Zea mays L. (Poacae), known as maize or corn, is an annual
this study the antimalarial and antiplasmodial activity of the
grass plant cultivated for human consumption and as animal
husk extract and fractions to confirm its use as malarial remedy
feed. It was introduced to Nigeria in the sixteenth century
in Ibibio ethnomedicine.
(Osagie & Eka 1998). It is tall with strong erect stalks and a
fibrous root system. The plant has long narrow leaves that are
spaced alternately on opposite side of the stem and bears ears Materials and methods
that are enclosed in modified leaves known as husks (Simmonds
1979). Besides its nutritive values, maize grains, leaves, cornsilks, Collection of plant materials
stalk, and inflorescence are also used in ethnomedicine for the Fresh husks of Zea mays were collected in August, 2015 from
treatment of several ailments. The corn silk is used as an antidia-
Farmland in Uyo in Uyo LGA, Akwa Ibom State, Nigeria. The
betic or diuretic, and decoction of the silk is consumed for the
husks were identified and authenticated as Zea mays by Dr.
treatment of urinary troubles and gallstones (Foster & Duke
Margaret Bassey, a taxonomist in the Department of Botany and
1990; Gill 1992; Abo et al. 2008). The ash of the cob is used for
Ecological studies, University of Uyo, Uyo, Nigeria. Herbarium
the treatment of cough (Gill 1992) and inflammatory diseases.
specimen (FPH, 614) was deposited at the Faculty of Pharmacy
The husks are used for the treatment of pains and arthritis
(Owoyele et al. 2010). Warm tea of the husks is used for the Herbarium, University of Uyo, Uyo.
treatment of malaria and diabetes in Ibibio traditional medicine.
Biological activities reported on the husk extract include; anal- Extraction
gesic, anti-inflammatory (Owoyele et al. 2010), and antioxidant
(Dong et al. 2014) activities. Arabinoxylan, which has immuno- The plant parts (husks) were washed, cut into smaller pieces and
logical effects, has been isolated from the husk extract (Ogawa air-dried for 2 weeks. The dried husks were pulverized using a
et al. 2005), while eight phenolic compounds (gallic acid, proto- pestle and mortar. The powdered husk was macerated in 50%
catechuic acid, chlorogenic acid, cafeic acid, femlic acid, rutin, ethanol for 72 h. The liquid ethanol extract obtained by filtration
resveratrol, and kaempferol) have also been detected in ethanol was evaporated to dryness in a rotary evaporator 40
C. The crude

CONTACT Jude E. Okokon [email protected] Department of Pharmacology and Toxicology, Faculty of Pharmacy, University of Uyo, Uyo, Nigeria
ß 2017 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
 PHARMACEUTICAL BIOLOGY 1395

ethanol husk extract (20 g) was dissolved in 200 mL of distilled Evaluation of in vivo anti-malarial activity of ethanol crude
water and further partitioned successively into each of petroleum extract
ether, chloroform, ethyl acetate and n-butanol to give the corre-
sponding fractions of these solvents, while the residue was taken Evaluation of suppressive activity of the extract (4-day test)
as aqueous fraction. The extract (yield 2.83%) and fractions were This test was used to evaluate the schizontocidal activity of the
stored in a refrigerator at 4
C until they were used for the extract and artesunate against early P. berghei berghei infection in
experiments reported in this study. mice. This was done as described by Knight and Peters (1980).
Thirty mice were randomly divided into five groups of six mice
Phytochemical screening each. On the first day (D0), the 30 mice were infected with the
parasite and randomly divided into various groups. These were
Phytochemical screening of the crude husk extract was carried orally administered with the extract and artesunate. The mice in
out employing standard procedures and tests (Trease & Evans group 1 were administered 187 mg/kg, group 2, 374 mg/kg
1989; Sofowora 1993), to reveal the presence of chemical constit- and group 3, 748 mg/kg of crude husk extract, while group 4
uents such as alkaloids, flavonoids, tannins, terpenes, saponins, was administered 5 mg/kg of artesunate (positive control), and
anthraquinones, reducing sugars and cardiac glycosides. 10 mL/kg of distilled water was administered to group 5 (negative
control) for four consecutive days (D0–D3) between 8 am and
Animals 9 am. On the fifth day (D4), thin blood film was made from tail
blood of each mouse. The film was then stained with Giemsa
Thirty Swiss albino mice (18–25 g) of either sex divided into five stain to reveal parasitized erythocytes out of 500 in a random
groups of 6 mice each per model were used for these experi- field of the microscope. The average percentage suppression of
ments. The animals were housed in standard cages and were parasitaemia was calculated in comparison with the controls as
maintained on a standard pelleted feed (Guinea feed) and water follows:
ad libitum. Permission and approval for animal studies were
Average % parasitaemia in negative control
obtained from the College of Health Sciences Animal Ethics
Committee, University of Uyo.  Average % parasitaemiain Positive groups
 100
Average % parasitaemia in negative control
Parasites
A chloroquine sensitive strain of Plasmodium berghei (ANKA) Evaluation of prophylactic or repository activities of extract
was obtained from the National Institute of Medical Research
(NIMER), Yaba Lagos, Nigeria and was maintained by sub-pas- The repository activity of the extract and artesunate was assessed
sage in mice. While Plasmodium falciparum strains Pf 3D7 and by using the method described by Peters (1965). The mice were
Pf INDO were obtained from the International Center for randomly divided into five groups of six mice each. Groups 1–3
Genetic Engineering and Biotechnology, New Delhi, India. were orally administered 187, 374 and 748 mg/kg/day of the husk
extract respectively, Groups 4 and 5 were respectively adminis-
tered 5 mg/kg/day of artesunate (positive control) and 10 mL/kg
Determination of acute toxicity of crude husk extract of distilled water (negative control). Administration of the
The median lethal dose (LD50) was determined for estimating extract/drug continued for three consecutive days (D0–D2). On
acute toxicity of the crude husk extract in Swiss albino mice the fourth day (D3), the mice were inoculated with P. berghei
model using the method of Lorke (1983). This involved intraperi- berghei. The parasitaemia level of each mouse was assessed by
toneal administration of different doses of the extract blood smear 72 h later.
(1000–5000 mg/kg) to groups of three mice each. The animals
were observed for manifestation of physical signs of toxicity such Evaluation of curative activities of extract (Rane’s test)
as writhing, decreased motor activity, decreased body/limb tone,
decreased respiration and death. The number of deaths in each This was used to evaluate the schizontocidal activity of the
group within 24 h was recorded. extract, and artesunate in established infection. This was done as
described by Ryley and Peters (1970). Plasmodium berghei berghei
was injected intraperitoneally into another 30 mice on the first
Parasite inoculation day (D0). Seventy-two hours later (D3), the mice were divided
Each mouse used in the experiment was inoculated intraperito- randomly into five groups of six mice each. Different doses of
neally with 0.2 mL of infected blood containing about 1  107 P. the extract, 187, 374 and 748 mg/kg were orally administered,
berghei berghei parasitized erythrocytes. The inoculum consisted respectively, to mice in groups 1–3, 5 mg/kg/day of artesunate
of 5  107 P. berghei berghei erythrocytes per mL. This was pre- was administered to the group 4 (positive control) and group 5
pared by determining both the percentage parasitaemia and the was given 10 mL/kg of distilled water (negative control). The
erythrocytes count of the donor mouse and diluting the blood extract and drugs were administered once daily for 5 days.
with isotonic saline in proportions indicated by both determina- Giemsa stained thin smears were prepared from tail blood sam-
tions (Odetola & Basir 1980). ples collected on each day of treatment to monitor parasitaemia
level. The mean survival time (MST) of the mice in each treat-
ment group was determined over a period of 29 days (D0–D28).
Drug administration
No of days survived
The drug (artesunate) and extract used in the in vivo antiplasmo-  100 ¼ MST
Total No: of days ð29Þ
dial study were orally administered with the aid of a stainless
metallic feeding cannula.
1396 J. E. OKOKON ET AL.

Evaluation of in vitro antiplasmodial activity (Mosmann 1983) using HeLa cells cultured in RPMI containing
10% foetal bovine serum, 0.21% sodium bicarbonate (Sigma) and
In vitro cultivation of Plasmodium falciparum 50 lg/mL gentamicin (complete medium) and human embroynic
CQ-sensitive strain 3D7 and CQ-resistant strain INDO of P. kidney 293 cells cultured in DMEM and supplemented with 10%
falciparum were used in this study to assess the antiplasmodial foetal bovine albumin. Briefly, cells (104 cells/200 lL/well) were
activity of the crude husk extract and fractions on erythrocytic seeded into 96-well flat-bottom tissue culture plates in complete
stages in vitro. The culture was maintained at the Malaria medium. Drug solutions were added after 24 h of seeding and
Research Laboratory, International Centre for Genetic incubated for 48 h in a humidified atmosphere at 37
C and 5%
Engineering and Biotechnology, New Delhi, India. Plasmodium CO2. DMSO (as positive inhibitor) was added at 10%. A stock
falciparum culture was maintained according to the method solution (20 lL) of MTT (5 mg/mL in 1  phosphate buffered
described by Trager and Jensen (1976) with minor modifications. saline) was added to each well, gently mixed and incubated for
Plasmodium falciparum (3D7) cultures were maintained in fresh another 4 h. After spinning the plate at 1500 rpm for 5 min,
O þve human erythrocytes suspended at 4% hematocrit in RPMI supernatant was removed and 100 lL of DMSO (stop agent) was
1640 (Sigma) containing 0.2% sodium bicarbonate, 0.5% albu- added. Formation of formazon was read on a microtiter plate
max, 45 lg/L hypoxanthine, and 50 lg/L gentamicin and incu- reader (Versa max tunable multi-well plate reader) at 570 nm.
bated at 37
C under a gas mixture of 5% O2, 5% CO2, and 90% The 50% cytotoxic concentration (TC50) of drug was determined
N2. Every day, infected erythrocytes were transferred into fresh by analysis of dose–response curves and IC50 estimator.
complete medium to propagate the culture. For Plasmodium fal-
ciparum (INDO strain) in culture medium, albumax was replaced Gas chromatography-mass spectrometry analysis
by 10% pooled human serum.
Quantitative and qualitative data were determined by GC and
GC-MS, respectively. The fraction was injected onto a Shimadzu
Drug dilutions GC-17 A system, equipped with an AOC-20i autosampler and a
split/splitless injector. The column used was an DB-5 (Optima-
Dimethyl sulfoxide (DMSO) was used to prepare the stock solu- 5), 30 m, 0.25 mm i.d., 0.25 lm df, coated with 5% diphenyl-95%
tions of husk extract, fractions and artemisinin, while water polydimethylsiloxane, operated with the following oven tempera-
(Milli-Q grade) was used in the case of CQ stock solution. ture program: 50
C, held for 1 min, rising at 3
C/min to 250
C,
Culture medium was used to dilute the stock solutions to their held for 5 min, rising at 2
C/min to 280
C, held for 3 min; injec-
required concentrations exception of CQ. The final solution of tion temperature and volume, 250
C and 1.0 lL, respectively;
each stock was constituted to contained nontoxic concentration injection mode, split; split ratio, 30:1; carrier gas, nitrogen at
of DMSO (0.4%), which was found to be harmless to the para- 30 cm/s linear velocity and inlet pressure 99.8 KPa; detector tem-
site. Drugs, husk extract and fractions were then placed in 96- perature, 280
C; hydrogen, flow rate, 50 mL/min; air flow rate,
well flat bottom tissue culture grade plates. 400 mL/min; make-up (H2/air), flow rate, 50 mL/min; sampling
rate, 40 ms. Data were acquired by means of GC solution soft-
ware (Shimadzu). Agilent 6890 N GC was interfaced with a VG
In vitro antiplasmodial assays Analytical 70–250 s double-focusing mass spectrometer. Helium
The crude husk extract and fractions of this plant were evaluated was used as the carrier gas. The MS operating conditions were:
for their antiplasmodial activity against 3D7 and INDO strains of ionization voltage 70 eV, ion source 250
C. The GC was fitted
Plasmodium falciparum. For drug screening, SYBR green I-based with a 30 m  0.32 mm fused capillary silica column coated with
fluorescence assay was set up as described previously (Smilkstein DB-5. The GC operating parameters were identical with those of
et al. 2004). Sorbitol synchronized parasites were incubated under GC analysis described above.
normal culture conditions at 2% hematocrit and 1% parasitemia in
the absence or presence of increasing concentrations of husk Identification of the compounds
extract and fractions. CQ and artemisinin were used as positive
controls, while 0.4% DMSO was used as the negative control. After The identification of components present in the active fraction of
48 h of incubation, 100 lL of SYBR Green I solution (0.2 lL of the plants’ extract was based on direct comparison of the reten-
10,000  SYBR Green I (Invitrogen)/mL) in lysis buffer {Tris tion times and mass spectral data with those for standard com-
(20 mM; pH 7.5), EDTA (5 mM), saponin (0.008%, w/v), and pounds, and by computer matching with the Wiley and Nist
Triton X-100 (0.08%, v/v)} was added to each well and mixed twice Libraries (Adams 2001; Setzer et al. 2007).
gently with multi-channel pipette and incubated in dark at 37
C
for 1 h. Fluorescence was measured with a Victor fluorescence Statistical analysis and data evaluation
multi-well plate reader (Perkin Elmer) with excitation and emis-
sion wavelength bands cantered at 485 and 530 nm, respectively. Data obtained from this work were analyzed statistically using
The fluorescence counts were plotted against the drug concentra- Student’s t-test and ANOVA (One-way) followed by a post test
tion and the 50% inhibitory concentration (IC50) was determined (Turkey–Kramer multiple comparison test). Differences between
by analysis of dose–response curves and IC50 estimator. Results means was considered significant at 1% and 5% level of signifi-
were validated microscopically by examination of Giemsa-stained cance, that is p  0.01 and 0.05.
smears of extract/fraction-treated parasite cultures.
Results
Cytotoxic activity on HeLa and HEKS cells using MTT assay Phytochemical screening
The cytotoxic effects of extract and fractions on host cells Results of phytochemical screening of the crude ethanol husk
were assessed by functional assay as described previously extract revealed the presence of chemical constituents such as
 PHARMACEUTICAL BIOLOGY 1397

alkaloids, flavonoids, tannins, terpenes, saponins, cardiac glyco- Effect on repository activity of ethanol husk extract of
sides and sugars. Zea mays
The ethanol husk extract of Zea mays showed a dose-dependent
Determination of acute toxicity of crude husk extract chemosuppressive effect (65.89–81.85%) on the parasitaemia dur-
ing prophylactic studies. These effects were statistically significant
The median lethal dose (LD50) of the crude husk extract was cal- relative to the control (p < 0.001), but weak compared to that of
culated to be 1874.83 mg/kg. The physical signs of toxicity the standard drug, artesunate, with chemosuppression of 90.92%
observed included excitation, paw licking, increased respiratory (Table 2).
rate, decreased motor activity, gasping and coma, followed by
death.
Antiplasmodial effect of ethanol husk extract of Zea mays
on established infection
Effect on suppressive activity of ethanol husk extract of
Zea mays The extract showed a dose-dependent schizonticidal effect on the
parasitaemia. There were reductions in the percentage parasit-
The extract showed a dose-dependent chemosuppressive effect aemia of the extract/artesunate-treated groups compared to that
on the parasitaemia. These effects were statistically significant of the control in which prominent increases were recorded. These
relative to the control (p < 0.05–0.001). The chemoinhibitory per- reductions were statistically significant relative to the control
centages ranged from 34.58 to 69.18 (Table 1). However, the (p < 0.05–0.001) (Figure 1). Though the extract showed a signifi-
effect of the extract was weak compared to that of the standard cant (p < 0.05–0.001) dose-dependent mean survival time on
drug, artesunate, with a chemosuppression of 98.82% (Table 1). established infection, the effect of the extract (187–748 mg/kg)
was weak compared to that of the standard drug, artesunate
Table 1. In vivo suppressive activity of crude ethanol husk extract of Zea mays (Table 3).
(4-day test) against P. berghei infection in mice.
Drug/extract Dose (mg/kg) Parasitaemia % Chemosuppression
Distilled water extract 10 mL/kg 44.33 ± 2.18 – In vitro antiplasmodial and cytotoxic activities
187 29.00 ± 1.02a 34.58
374 20.66 ± 0.33a 53.39 The results of the in vitro studies showed that the plant extract
748 13.66 ± 1.45a 69.18 and fractions displayed antiplasmodial activity against chloro-
Artesunate 5 0.52 ± 0.01a 98.82 quine sensitive Pf 3D7 and resistant Pf INDO strains of
Values are expressed as mean ± S.E.M. Significance relative to control.
a
p < 0.001: n ¼ 6.
Table 3. Mean Survival Time (MST) of mice receiving different doses of
ethanol husk extract of Zea mays during established infection.
Drug/extract Dose (mg/kg) MST (days)
Table 2. Repository/Prophylactic activity of ethanol husk extract of Zea mays
against Plasmodium berghei infection in mice. Distilled water extract 10 mL/kg 14.33 ± 0.33
187 15.33 ± 0.66
Treatments Dose(mg/kg) Parasitaemia % Chemosuppression 374 19.0 ± 1.00a
Normal saline 10 mL/kg 14.66 ± 0.66 – 748 24.00 ± 1.73b
Crude extract 187 5.00 ± 3.24a 65.89 Artesunate 5 30.00 ± 0.00b
374 4.66 ± 0.90a 68.21 Values are expressed as Mean ± S.E.M. Significance relative to control.
748 2.66 ± 1.33a 81.85 a
p < 0.05.
Artesunate 5.0 1.33 ± 0.66a 90.92 b
p < 0.001, n ¼ 6.
Values are expressed as Mean ± SEM. Significance relative to control.
a
p < 0.001, n ¼ 6.

Figure 1. In vivo curative antiplasmodial activity of crude ethanol husk extract of Zea mays (Rane’s test) against P. berghei infection in mice.
1398 J. E. OKOKON ET AL.

P. falciparum (Table 4). The ethyl acetate fraction was found to Discussion
exhibit promising activity against both strains of P. falciparum
with IC50 values of 9.31 ± 0.46 lg/mL (Pf 3D7) and 3.69 ± 0.66 lg/ The husk extract of Zea mays used as malarial remedy by the
mL (Pf INDO). The potency order was ethyl acetate > chloro- Ibibio tribe of Nigeria was investigated for in vivo and in vitro
form > butanol > crude extract > petroleum ether. The crude antiplasmodial activities using standard models. Acute toxicity
extract and fractions were not cytotoxic to the two cell lines and cytotoxicity studies as well as phytochemical studies of the
tested with TC50 of >100 lg/mL against both HeLa and HEKS husk extract and fractions were carried out.
cell lines. The median lethal dose (LD50) which was determined to be
1874.83 mg/kg was found to be relatively safe with insignificant
toxicity (Homburger 1989). The results of the in vivo study
GCMS analysis
revealed that the crude extract significantly reduced parasitaemia
The GCMS analysis of the ethyl acetate fraction of Zea mays in prophylactic, suppressive and curative models in a dose-
revealed the presence of bioactive compounds with major and dependent fashion confirming the antimalarial potential of this
minor ones as represented in Table 5. extract. These findings were further supported by the results of

Table 4. In vitro antiplasmodial activities of crude husk extract and fractions of Z. mays.
Cytotoxicity
Crude extract/fraction IC50(lg/mL) Pf 3D7 IC50(lg/mL) Pf INDO Hela cells TC50(lg/mL) HEKS cells TC50(lg/mL)
Crude extract 45.52 ± 0.86 38.51 ± 0.18 >100 >100
Pet. ether 47.88 ± 0.36 49.11 ± 0.15 >100 >100
Chloroform 8.46 ± 0.37 3.84 ± 0.32 >100 >100
Ethyl acetate 9.31 ± 0.46 3.69 ± 0.66 >100 >100
Butanol 35.35 ± 0.16 44.81 ± 0.12 >100 25.0
Aqueous >100 >100 >100 >100
Values are expressed as mean ± S.E.M.

Table 5. GCMS analysis of ethyl acetate fraction of Zea mays husk.

Peak RT Compound name Formula Mol. mass
1. 4.474 1,2,3-Propanetriol C3H8O3 92
2. 8.875 2,3-Dihydro-3,5-dihydroxy-6-methyl-4h-pyran-4-one C6H8O4 144
3. 10.088 Butanedioic acid, hydroxy-, diethyl ester, (.þ/–.)- C8H14O5 190
4. 11.213 2,3-Dihydro-benzofuran C8H8O 120
5. 12.397 2H-Pyran-2-one, tetrahydro-4-hydroxy-4-methyl- C6H10O3 130
6. 13.261 2-Methoxy-4-vinylphenol C9H10O2 150
7. 14.920 1-Tridecene C13H26 182
8. 16.315 Ethyl .beta.-d-riboside C7H14O 178
9. 17.526 1R-Ethoxy-3-trans-methoxy-2-cis-methylcyclohexane C10H20O2 172
10. 18.092 Benzeneacetic acid, 4-hydroxy-, methyl ester C9H10O3 166
11. 18.258 Dodecanoic acid, methyl ester C13H26O2 214
12. 19.684 Dodecanoic acid C12H24O2 200
13. 19.833 Trifluoroacetic acid,n-tridecyl ester C15H27F3O2 296
14. 19.912 Dodecanoic acid, ethyl ester C14H28O2 228
15. 22.757 Delta.1,.alpha.-Cyclohexaneacetic acid C8H12O2 140
16. 23.820 4-((1E)-3-Hydroxy-1-propenyl)-2-methoxyphenol C10H12O3 180
17. 24.027 Tetradecanoic acid C14H28O2 228
18. 24.297 n-Tetracosanol-1 C24H50O 354
19. 24.351 Tetradecanoic acid, ethyl ester C16H32O2 256
20. 25.912 p-Hydroxycinnamic acid, ethyl ester C11H12O3 192
21. 27.149 Hexadecanoic acid, methyl ester C17H34O2 270
22. 27.451 Ethyl (2E)-3-(4-hydroxy-3- methoxyphenyl)-2-propenoate C12H14O4 222
23. 28.537 Pentadecanoic acid C15H30O2 242
24. 28.879 Hexadecanoic acid, ethyl ester C18H36O2 284
25. 31.944 Ethyl (9Z,12Z)-9,12- octadecadienoate C20H36O2 308
26. 32.111 9-Octadecenoic acid, methyl ester C19H36O2 296
27. 32.756 Ethyl (9Z,12Z)-9,12-octadecadienoate C20H36O2 308
28. 33.553 9-octadecenoic acid (Z)-, ethyl ester C19H36O2 296
29. 33.664 Octadecanoic acid, methyl ester C19H38O2 298
30. 34.161 Nonadecanoic acid, ethyl ester C21H42O2 326
31. 36.833 2-Hydroxy-3-[(9E)-9-octadecenoyloxy]propyl (9E)-9-octadecenoate C39H72O5 620
32. 37.072 1,2-Benzenedicarboxylic acid C24H38O4 390
33. 38.028 Docosanoic acid, ethyl ester C24H48O2 368
34. 38.784 1,1'-Biphenyl-3,4,4'-trimethoxy-6'-formyl- C16H16O4 272
35. 39.157 (R-(R,R))-4-(1,5-Dimethylhexyl)-1-cyclohexenecarboxylic acid C15H26O2 238
36. 41.091 Docosanoic acid, ethyl ester C24H48O2 368
37. 42.365 Pentacosane, 1-bromo- C25H51Br 430
38. 46.599 Stigmast-5-en-3-ol, (3.Beta.)- C29H50O 414
39. 49.936 Ergost-5-en-3-ol, (3.Beta.,24R)- C28H48O 400
40. 50.813 Stigmasterol C29H48O 412
41. 52.610 Stigmast-5-en-3-ol, (3.Beta.)- C29H50O 414
 PHARMACEUTICAL BIOLOGY 1399

the mean survival time of the extract-treated mice which was sig- Flavonoids are known to exert antiplasmodial activity by che-
nificantly prolonged compared to those of the control group lating with nucleic acid base pairing of the parasite (Lui et al.
demonstrating a significant protection potential of the extract. 1992), while the plasmodicidal activities of terpenes and their
The results of in vitro study revealed that the husk extract and derivatives such as monoterpenes and sesquiterpenes have been
fractions had antiplasmodial activity against both chloroquine linked to endoperoxidation (Hatzakis et al. 2007).
sensitive (3D7) and resistant (INDO) strains of Plasmodium fal- These compounds (flavonoids and terpenes) present in this
ciparum with ethyl acetate fraction as the most active fraction. plant extract may have contributed to the plasmodicidal activity
This suggests the localization of the active molecules in the ethyl of this extract and therefore explained the mechanism of anti-
acetate fraction. The results of the in vitro activity corroborated plasmodial effect of the extract.
that of the in vivo study and confirm the antimalarial and anti-
plasmodial potentials of the husk extract. These findings further Conclusion
lay credence to the use of the husk of Zea mays in the treatment
of malaria traditionally. Besides, the fact that the extract and The results of this study indicate that the husk of Zea mays plant
fractions were active against the chloroquine-resistant strain possesses significant in vivo antimalarial activity against P. ber-
(INDO) of P. falciparum implies that the husk extract can be an ghei infection in mice and in vitro antiplasmodial activity against
effective agent against chloroquine resistant malaria. The activ- chloroquine sensitive and resistant strains of P. falciparum. These
ities of the crude husk extract and fractions are predicated on findings justify and confirm the ethno botanical usage of this
the chemical constituents of the extract and fractions as revealed plant in the treatment of malaria. Therefore, further research on
by the results of phytochemical screening of the crude husk ethanol husk extract and fractions of Zea mays could be carried
extract and GCMS analysis of the active fraction. out in order to isolate, identify and characterize the active prin-
The husk extract of Zea mays was found in this study to ciple from this plant.
contain alkaloids, saponins, tannins, phlabatannins, flavonoids
and cardiac glycosides. Some secondary metabolites of plants
Acknowledgements
such as alkaloids, flavonoids and triterpenoids have been
reported to possess antiplasmodial activity (Kirby et al. 1989; Dr Jude Okokon is grateful to International Centre for Genetic
Philipson & Wright 1991; Christensen & Kharazmi 2001). These Engineering and Biotechnology (ICGEB) for financial support for
chemical compounds which are present in this extract and frac- Postdoctoral fellowship and ICGEB, Delhi, India for providing
tions may in part be responsible for the observed antimalarial research facilities.
and antiplasmodial activities. Also, the GCMS analysis of the
active ethyl acetate fraction revealed the presence of some
pharmacologically active compounds such as phenolics and Disclosure statement
polyunsaturated fatty acids (PUFA) especially C-18 fatty acids There is no conflict of interest to declare.
among others. Polyunsaturated fatty acids such as p-hydroxycin-
namic acid ethyl ester, stigmasterol, docosanoic acid ethyl ester,
octadecanoic acid methyl ester, 9-octadecenoic acid (Z)-ethyl References
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