CSN 2016 Book of Proceedings

Table of Content
S/N Topics and Authors Page

In Vitro Study Of Isolated Compounds From The Leaf Of Flabellaria Paniculata Against Staphylococcus Aureus,
Pseudomonas Aeuruginosa And Candida Albicans; Joseph Adebisi O. Olugbuyiro1*, Olugbenga Samson Taiwo2,
1 Olaronke Temitope Olaniyi1 And Francis Adolphusobi1 1
A Comparative Evaluation/analysis Of Air Quality In Some Selected Oil Producing/bearing Communities In
2 Delta State Of Nigeria.; Ogodo Aemurerhimen Dickson (snr.) (chartered Chemist) 6
Batch Adsorption Of Cd2+, Pb2+and Ni2+ With Coconut (cocos Nucicera L.) Shell Activated Carbon Modified
3 With 1, 2-dihydro-1, 5-dimethyl-2, Phenyl- 4- (e) - (2, 3, 4- Trihydrophenyl) -3h-pyrazol-3-one (ddptp) 22
Ukoha, Pius. O, Anierobi, Peter. O And Nicholas, Eno-obong S.
4 Delonixregia Extracts As Non – Toxic Corrosion Inhibitor For Acid Corrosion Of Mild Steel In Hcl Solution 33
Olusegun K. Abiola1 *,abdulraman O.c. Aliyu2and Suleiman Muhammed3
5 Assessment Of Some Metals In Roadside Dust From Damaturu, Yobe State, Nigeria 38
Zaynab Muhammed Chellube, Joseph Clement Akan And Emmanuel Mshelia
6 Anti-oxidant And Elemental Analysis Of The Fruit Of Sarcocephalus Latifolius (smith Bruce)
43
Isah Yinusa, Woli Ajoke Balkees And Hassan Adam Adeiza
7 Proximate, Mineral And Anti-nutrient Composition Of Allium Sativum (garlic) Consumed In Uyo, Akwa Ibom 47
State, Nigeria; E. I. Uwah And E. A. Moses
8 Phytoconstituents, Proximate Composition And Mineral Contents Of Soybean (glycine Max) Flour; Useh, M. 52
U.,1* Adebiyi, A. B.,1 And Dauda, M. S.2
Equilibrium, Kinetics And Thermodynamic Properties Of Methylene Blue Adsorption By Termite Mound;
9 1
anebi, P. O., 2ugbe, F.a., 3ikudayisi, A.v. 57
10 Petrochemicals: Upstream To Plastic Industry Using Ideas, Methods, Developments And Procedures In
Developing Countries.; Ogodo Aemurerhimen Dickson (snr.) (chartered Chemist) 63
11 Kinetics Of The Photocatalytic Degradation Of Chlorendic Acid (flame Retardant) In Aqueous Tio2
Suspension; Boisa N., And Obunwo C.c. 78
12 Gas Chromatography-mass Spectrometric Analysis Of Methanolic Leaf Extract Of Blighia Sapida;
83
Theresa I Edewor And Nimotalai O Kazeem
13 Characterization And Distributions Of Aliphatic And Polyaromatic Hydrocarbons In Soils Of Oil Sand
Deposits Area Of Ondo State, Nigeria; T.a .adedosu1*, O.k. Adeniyi2, O.h. Adedosu3 89
14 Comparative Assessment of the Environmental Dynamics of Dissolved Organic Carbon (DOC), Nitrogen
(DON), Phosphorus (DOP) and Characterization of DOM in Subtropical Wetlands of Some Northern Nigerian 98
States.
Synthesis, Characterization And Kinetic Studies Of Fe (ii) And Cu (ii) Complexes Of Nicotinic Acid
15 Hydrazide.; *1o.w. Salawu, 2m.s. Iorungwa And 3m.u. Adaji. 109
16 Elemental Analysis And Cyanic Acid Contents Of Selected Cassava Based Foods In Nigeria; Orishadipe Abayomi,
115
Afolayan Michael, Odukomaiya Doyinsola, Aguzue Onyinye & Bwai Macham David
1 2
17 Production Of Methyl Ester (biodiesel) Using Melon Seed Oil As A Raw Material agbo, I.u. & onyezeka, E.G. 119
2+ 3+ 2+
Studies On Biosorption Of Co , Cr And Cd From Aqueous Solutions By Piliostigma Malabaricum Seed Pod.
18 Onwu, F. K And Amadi, O. K.
121
Corrosion Inhibition Properties Of Commiphora Africana (a. Rich.) Engl. Gum Exudates On Mild Steel In
19 Alkaline Medium.; 1odoemelam, S.a., 2onukwube, N.d. And 3eddy, N.o. 127
Ogodometrics:iso Approval/certifications/product Design, Innovation Using Globally Harmonized
20 International Reference Standards In Songhai Delta Amukpe Sapele As Targeted World Collaborating 134
Centre.; Ogodo Aemurerhimen Dickson (snr.) (chartered Chemist)
21 Chemical Constituents Of Sandbox Tree (hura Crepitans Linn.) And Anti- Patotoxic Activity Of The Leaves
And Stem Bark Extracts; Ganiyat K. Oloyede1*, Oluwatosin A. Adaramoye2 And Mutairu B. Olatinwo1 161
Studies On The Molar Ratios Of Optimal Interactions Of Nr-pvc Blend Using Ir Spectroscopy 175
22 1
okoye, O. N. N., 2eboatu, A.n
ANTIBACTERIAL AND ANTIOXIDANT ACTIVITY OF JUSTICIA SPICIGERA EXTRACTS: ACTIVITY
23 ENHANCEMENT BY ADDITION OF METAL SALTS. 180
Ohenhen O. N1., Njoku P. C2., Igara C. E3.
S/N Topics Page
Kinetics And Mechanism Of The Oxidation Of Potassium Trisoxalatoferrate(iii) By Permanganate Ion In 187
24 Aqueous Hydrochloric Acid Medium; 1b.o. Ogori, 2y.n Lohdip And 2j.n Egila
The Importance Of Chemistry Practice And Research In The Oil And Gas Sector Ogodo Aemurerhimen
Dickson (snr.) (chartered Chemist)(a2652) (hnd, Pgd, M.sc., Ph.d (in- View), Aislt, C.chem., Miccon, Mcsn, 193
25 Mnes, Mspe)
The Effect Of Extracts Of Crinum Jugas On The Acute Toxicity Of The Vernonia Amygdalina Root Poisoned Guinea 200
26 Pigs; Ogbuanu Cyril C, Amujiogu Steve N, Nwagu Lauretta N, Chime Charlse C, And Arinze-nwosu Uche L
27
Kinetics And Sorption Models Of Unmodified And Modified Okra Stem Derived Cellulose For The Removal 205
And Recovery Of Hg And As Ions From Aqueous Environment; Obidiozor, C.j. And Okoro I.a
Synthesis, Characterization And Corrosion Inhibitory Of Co (ii), Ni (ii), Cu (ii), And Zn (ii) Complexes
28 Derived From Nicotinic Acid Hydrazide.; *1o.w.salawu, 2r.a. Wuana, 3j.t.ashimom 215
Preparation And Characterization Of Activated Carbon From Hura Crepitans Linn Seed Shells
29 1
e. W. Nsi1 , A. E. Akpakpan1 And U. D. Akpabio 221
Phytochemical & Antimicrobial Studies Of Crossopteryx Febrifuga Leaf Extract Against Methicillin-resistant
30 Staphylococcus Aureus Skin Infections; Nma N. Y.*,1, Mann A.2, Ndamitso M. M.2 And Muhammad B. M.3 227
Antibacterial And Antioxidant Activity Of Justicia Spicigera Extracts: Activity Enhancement By Addition Of
31 Metal Salts.; Ohenhen O. N1., Njoku P. C2., Igara C. E3. 234
32 Comparison Of Heavy Metals Pb, Cu, Ni, Zn And Al In Human Scalp Hair: A Case Study Of Port-harcourt
And Okigwe; Ngwu, Comfort M And Onwu, Francis K. 240
Geochemical Assessment Of The Petroleum Generative Potential Of Ekenkpon And Nkporo Shale Formations
33 In The Calabar Flank South Eastern Nigeria; P. A Neji1; H A Neji2; Tnganje3;o.o Oli4;u J Ibok5 249
Synthesis, Characterization And And Biological Activity Of New Mannich Base 1-((4-chlorophenyl)(2,5-
34 dioxopyrrolidin-1-yl)methyl)urea; M.r. Abdullahi *1,s. Rajeswari2 262
35 Nanotechnology And Implementation In Natural Products Green Chemistry Processes; M.m. Adeyemi*1, P.e.
Omale1; O.a. Olawuyi1 A. Ngokat1 And A. Bhattacharya2 267
36 Sorption Kinetics Of Methylene Blue On Adsorbents Derived From Delonix Regia Seed Pods And Vigna 275
Subterranea Fruit Hulls; L.k. Akinola1*; Ali Ibrahim2 And Mohammed Mohammed2
37 Quantum Chemical Studies On The Efficiency Of 2-hydroxyethylethylendiamine And Diethylenetriamine As
Corrosion Inhibitors K. J. Uwakwe1, P. C. Okaforǂ,, 285
Comparative Studies Of The Inhibitive Potentials Of Red Peanut Skin Extract And Lasianthera Africana P.
38 Beauv. (nkanka Leaf) Mucilage On Mild Steel Corrosion In Sulphuric Acid Solution.
A
a.o. James, Bkalu Nnennaand Cakaranta O. 291
39 In Vitro Antioxidant Activity Of Heinsia Crinata Root Extracts.; *ita, Basil N And Willie, Akaninyene H. 298
Evaluation Of Medicinal Composition Of Cyathea Latebrosa Leave Extract
Ikpa, C.b.c1*., Ikezu, U.j. M2 And Ugochukwu, M.i3 304
40
Determination Of Heavy Metals In Telfeiria Ocidentalis (pumpkin Leaves) Grown At Orimekpang – Ochor
Village, Boki Local Government Area, Cross River State, Nigeria.; 1ikezu, U. J., 2udeozo, I.p. And 2odok, J.m. 309
41
42 Proximate Analysis And Production Of Bioethanolfrom Post-harvest Pineapple And Watermelon Waste Fruit 314
G.u. Igelige, P. Ekwumemgbo, G.i, Ndukwe And J.i Achika
43 Evaluation Of Chemical Composition Of Murraya Koenigii (linn) Spreng Leaf 320
1
c. E. Igara, 2d. A. Omoboyowa, 3a. A. Ahuchaogu, 1n. U. Orji And 1m. K. Ndukwe
Studies On The Energy Properties And Fuel Potentials Of Selected Indigenously Processed Plant-based Bio-
44 resources Used As Sources Of Sustainable Flame In North Central Nigerian Communities 324
J. S. Gushit*1, J. T. Shimuan1, M. O. Ajana1
45 Pollution Index Of A Crude Oil Producing Community In Niger Delta: A Case Study Of Ibeno Local
Government Area Of Akwa Ibom State, Nigeria; 330
*godwin Asukwo Ebong1 , Helen Solomon Etuk1 And Imeh Edet Essien2
Synthesis And Biological Activities Of Metal Complexes Of Some Organic Bases
46 Emwanta D. O. And Ngochindo R. I. 339
S/N Topics Page
Assessment Of Rainwater Quality Of Bichi Local Government Area Of Kano State Of Nigeria
47 Emmanuel Bernard
343
Effect Of Advance Organizers In The Teaching Of Chemistry In Secondary Schools: Anambra State As A Case
48 349
Study; Enekwechi E.e.
49 Removal Of Contaminants From Industrial Wastewater Using Photocatalyst; Ekwere, Ifiok Okon 358
Levels, Spatial Distribution And Ecological Risk Assessment Of Cadmium And Lead In Surface Soil Of
50 Nsukka Industrial Cluster Areas, South-east Nigeria N.r.ekere *, B.u. Ngang, J.n Ihdioha, P.o.ukoha 361
51 Low Cost/waste Catalyst For Fatty Acid Methyl Ester Production
366
M. O. Ekeomaa,*, P. A. C. Okoyeb, V.i.e., Ajiweb
52 Coordination Chemistry In Oil Industry; ehirim, Appolinus I. And Ogwuegbu, Martin O.c.; 378
53 Using Index Models For Heavy Metal Pollution Estimation Of Sediments From Bomu And Oginigba Rivers
Marcus, A. C. And Edori, O. S. 389
54 Effect Of Seasonal Water Fluctuation Of A Water Body On Antioxidant Activity Of Selected Plants Of Lower 395
Phylum(a Case Study Of Nchestream)
1
duru M.k.c; 2akubugwo E.i; 2chinyere G.c; 3amadi B.a; 4agomuo E.n; 5alisa, C. O; 6njoku V. O; And 4amadi
55 Characterization Of Violet Tree (securidaca Longepedunculata) Root Powder
1
403
donatus, R.b‫٭‬.,1 Barminas, J.t., 1maitera,o.n.,2riki, .e.y
56 Determination Of The Saccharin Content In Some Ice Creams Consumed In Port Harcourt 405
Dibofori-orji Amalo Ndu & Didia Lucky Ejikeme
57 Evaluation Of Chemical Composition Of Murraya Koenigii (linn) Spreng Leaf 410
1
c. E. Igara, 2d. A. Omoboyowa, 3a. A. Ahuchaogu, 1n. U. Orji And 1m. K. Ndukwe
58 Phytochemical Studies And Aphrodisiac Activity Of Byrsocarpus Coccineus Root Exract

1
balarabe M. M., 2mann A., 2 Ndamitso M. M. Adeyemi O. Y. H. And 3nma N. Y. 415
59 Gc-ms Analysis Of Leaf, Stem-back And Root Extracts Of Alstonia Boonei
421
Babatunde O*.1; Godwin R. E.1; Wahab N.o.2
60 Isolation And Characterization Of 2, 4 - Dihydroxycinnamic Acid From The Stem Bark Of Adina 425
Microcephala Delile; 1,2r.g. Ayo, 2j.d. Habila, 2j. I. Achika And 2o.o. Akinwande
61 Groundwater Quality In Okitipupa Township Of Ondo State, Nigeria; ‫٭‬1ayedun, H. And Ediagbonya T. F 428
Preparation And Characterization Of Activated Carbon From Hura Crepitans Linn Seed Shells
62 E. W. Nsi1 , And A. E. Akpakpan2* U. D. Akpabio3
434
Characterization Of Beach Water And Its Effect On Nearby Groundwater Quality In Eti-osa And Ibeju-lekki 440
63 Local Government Areas, Of Lagos State, Nigeria.; V. E. *agbazue And B. U. Ukazu
Comparative Studies Of The Inhibitive Potentials Of Red Peanut Skin Extract And Lasianthera Africana P.
64 Beauv. (nkanka Leaf) Mucilage On Mild Steel Corrosion In Sulphuric Acid Solution. 452
A
a.o. James, Bkalu Nnennaand Cakaranta O.
Assessment Of Physicochemical Properties Of Clay Samples Obtained From Ashaka, Potiskum And Tango 459
65 (nigeria) And A Commercial Bentonite Obtained From Kofar Ruwa Market.; Abdullahi S. L1 And A.a Audu2
Extraction And Characterization Of Magnesium Chloride From Different Brackish Water Sources In Rivers
66 State; 1konne, J.l.,2ujile, A.a.and1ogolo, J.j. 465
Neutron Activation Analysis (naa) And X-ray Fluorescence Analysis (edxrf) Of Corchorus Tridens Linn
67 1
umar S. Gwarzo, 2gimba, C.e., 3adeyemo, D.j. And 2paul, E.d. 470
IN VITRO STUDY OF ISOLATED COMPOUNDS FROM THE LEAF OF FLABELLARIA PANICULATA
AGAINST Staphylococcus aureus, Pseudomonas aeuruginosa and Candida albicans
1* 2 1
Joseph Adebisi.O. Olugbuyiro , Olugbenga Samson Taiwo , Olaronke Temitope Olaniyi and Francis
AdolphusObi1
1
Department of Chemistry, Natural Products Unit, Covenant University, P. M. B. 1023, Ota, Nigeria.
2
Department of Biological Sciences, Microbiology Unit, Covenant University, P.M.B. 1023, Ota, Nigeria.
*
Corresponding author. Tel.: +2348034112751
<>; <olugbuyiro@yahoo.com>
ABSTRACT
This study carried out activity-guided fractionation of Flabellaria paniculata against Staphylococcus aureus,
Pseudomonas aeuruginosa and Candida albicans and isolated the active principles. The methanol extract was
partitioned into hexane, ethyl acetate and aqueous layers successively. The active ethyl acetate fraction was
subjected to repeated column and thin layer chromatography for isolation process. The fractions and isolated
compounds were tested for antimicrobial activities against Staphylococcus aureus, Pseudomonas aeuruginosa,
and Candida albicans. Two pure compounds (P2a and P2b) demonstrated significant antibacterial activities. P2a had
MIC values of 0.8 mg/mL and 0.4 mg/mL against Staphylococcus aureus and Pseudomonas aeruginosa
respectively. P2b had MIC values of 0.1 mg/mL and 0.05 mg/mL against Staphylococcus aureus and Pseudomonas
aeruginosa respectively. Also, P2a had MBC values of 1.6 mg/mL and 0.8 mg/mL against Staphylococcus aureus
and Pseudomonas aeruginosa respectively while P2b showed MBC values at 0.2 mg/mL and 0.1 mg/mL against
Staphylococcus aureus and Pseudomonas aeruginosa respectively. The findings have proved the vegetable drug,
Flabellaria paniculata, as a potent anti-infective candidate useful in combating notorious S. aureus and P.
aeruginosa.
Keywords: Flabellaria paniculata, S. aureus, P. aeruginosa, Candida albicans.
INTRODUCTION
Infections are a major health challenge. Nature provides agents that accumulate new molecules capable of
combating health threatening infectious diseases. Staphylococcus aureus is a Gram-positive bacterium which
causes many infections such as impetigo, boils, carbuncles, abscesses, cellulitis, wound infections, toxic shock
syndrome, pneumonia, food poisoning etc. In contrast, Pseudomonas aeruginosa is a gram-negative.
Pseudomonas aeruginosa causes infections like urinary infections, respiratory infections, dermatitis, soft tissue
infections, bacteremia, bone and joint infections, gastrointestinal infections and variety of systemic infections
particularly patients with severe burns who are immunosuppressed (Kenneth, 2012). Candida is a yeast infection
which results from an overgrowth of yeast (a type of fungus) anywhere in the body. The most common type of
yeast infection is candidiasis. In women, yeast infections are the second most common reason for vaginal
burning, itching, and discharge. Yeasts are found in the vagina of 20% to 50% of healthy women and can
overgrow if the environment in the vaginal changes, pregnancy, menstruation, diabetes and birth controls also
can contribute to getting a yeast infection. Also, yeast infections are common after menopause (Jerry, 2013).
Medicinal plants represent a rich source of anti-infective agents as well as many potent drugs (Cowan, 1999).
Flabellaria paniculata is a climber that is indigenous to Tropical Western Africa. Traditionally, the plant found
uses in the treatment of wounds, dysentery, sore, snake bite and skin infections (Burkill, 1997). The preliminary
antibacterial activity of the extracts has been reported (Abo and Olugbuyiro, 2004). Also, the activities of the
extracts on wounds infected with Staphylococcus aureus and Pseudomonas aeruginosa has been previously
1
studied (Olugbuyiro et al., 2010). Study has also shown that the essential oil from leaf extract has anti-
inflammatory effect (Oladosu et al., 2012). As a follow up to earlier anti-infective investigation, the present
study carried out bioassay-guided isolation of the antimicrobial agents in the active fraction of the plant extract.
Materials and Methods

Plant Material
The plant leaf was collected at Olosan in Ibadan, Nigeria in September 2013. The plant was identified as
specimen species previously authenticated (FHI NO. 106122) at Forestry Research Institute of Nigeria, Ibadan.
Extraction and Isolation
The oven dried plant material was macerated with methanol. The extract was strained, filtered and dried in vacuo
at 450C. The crude extract was partitioned into hexane, ethyl acetate and aqueous layers successively. The layers
were dried and subjected to bioassay.The active portion was subjected to gravity column chromatography over a
silica gel (70-230 mesh) column (110.0 x 3.0 cm) and successively eluted with solvents of increasing polarity
starting from n-hexane to 100 % methanol. Further purification was done by repeated column chromatography to
afford two active pure compounds P2a (8.0 mg) and P2b (12.0 mg) respectively.
Antimicrobial Assay
Test Organisms: The following clinical organisms were used Staphylococcus aureus, Pseudomonas
aeruginosa, and Candida albicans.The test organisms were supplied by the Department of Microbiology,
Covenant University, Ota, Nigeria.
Susceptibility Test
All the microbes were checked for purity and maintained in slants of nutrient agar for the bacteria and the fungi
were maintained in slants of SDA. The antimicrobial activities of fractions were evaluated against the clinical
isolates of Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans at a concentration of 20
mg/mL. The well diffusion method was used for the study. 18 ml of nutrient agar plates were inoculated with 0.1
ml of an overnight broth culture of each bacterial isolate (~3x108cfu/mL) Mcfarland standard in sterile petri-dish.
A standard cork borer of 6 mm diameter was used to bore a well and equal volumes of the plant extracts were
transferred into the well with micropipette. Gentamicin and Tioconazole were used as reference drugs for
bacteria and fungi respectively. The extracts were allowed to diffuse into the agar media for 1 hour. The plates
were incubated at 37 0C for 24 h for the bacteria, the plate for Candida albicans was incubated at 25 0C for 3 days.
Three replicates were made for each condition. Zone of inhibitions were measured in millimetres and the average
values were calculated and recorded.
Determination of Minimum Inhibitory Concentration (MIC)

The minimum inhibitory concentration was carried out on Staphylococcus aureus and Pseudomonas aeruginosa
8
using broth dilution method. 1.5 x10 cfu/ml concentration of the bacteria was prepared using # 0.5 Mcfarland
turbidity standard. Two-fold serial dilutions of the test compounds were made to obtain the concentrations of
1600-, 800 -, 400 -, 200 -, 100 - and 50 ìg/ml. The cultures were incubated at 37° C for 24 h, with shaking. Having
obtained different concentration of the compounds (P2a and P2b) in the broth, 0.1ml of the standard inoculums of
the micro-organisms in the normal saline was then inoculated into different concentration in the test tubes and the
test tubes were incubated at 37O C for 24 hours. MIC values were determined from the lowest concentrations that
induced 100% inhibition.
Minimum Bactericidal Concentration (MBC)
The minimum bactericidal concentrations of the active compounds (P2a and P2b) were determined. Blood agar
2
O
was prepared according to manufacturer's instructions and sterilized at 121 C for 15 minutes and poured into
sterile petri dishes to cool and solidify. The content of the MIC in the serial dilution were sub-cultured onto the
O
agars, the blood agar plates were incubated at 37 C for 24 hours after which the plates were observed for growth.
The MBC was evaluated as the plate with the lowest concentration without colony growth.
RESULTS AND DISCUSION

The susceptibility test (Table1) proved the ethyl acetate layer active against the bacteria compared with hexane
and aqueous layers. The three layers were inactive against Candida albicans. The findings demonstrated the mid
polar layer to possess the highest antimicrobial activity. The previous work (Abo and Olugbuyiro, 2004) reported
that the chloroform layer demonstrated the highest antibacterial potential against S. aureus, P. aeruginosa, K.
pneumonia, and E. coli while pet-ether layer was completely inactive. In this study, hexane was used in place of
pet ether while chloroform batch was substituted with ethyl acetate to capture more mid polar solutes. The results
still supported the previous report with respect to choice of solvent- polarity.
The antimicrobial screening results (Table 2) of the further purified ethyl acetate portion revealed fraction R1 to
be most potent against Staphylococcus aureus and Pseudomonas aeruginosa with zones of inhibition of 50 mm
and 30 mm respectively. With the exception of fraction R3 that had weak inhibition (8 mm) none of the column
fraction exhibited activity against Candida albicans. Further purification of R1 afforded two active pure
compounds- P2a and P2b. The two compounds (P2a and P2b) demonstrated significant antibacterial activities. P2a
had MIC values (Table 3) of 0.8 mg/mL and 0.4 mg/mL against Staphylococcus aureus and Pseudomonas
aeruginosa respectively. P2b had MIC values (Table 3) of 0.1 mg/mL and 0.05 mg/mL against Staphylococcus
aureus and Pseudomonas aeruginosa respectively. Table 4 demonstrated the MBCs of the isolated compounds.
Compound P2a had MBC value of 1.6 mg/mL against Staphylococcus aureus and 0.8 mg/mL against
Pseudomonas aeruginosa while P2b showed MBC values at 0.2 mg/mL and 0.1 mg/mL against Staphylococcus
aureus and Pseudomonas aeruginosa respectively. Compound P2a is a colourless needle crystal while P2b is a
yellow amorphous solid.
Table 1: Susceptibility test of the extracts of Flabellaria paniculata
Test organism Zone of inhibition (mm)
HL EL AL GENT TIOC
Staphylococcus 20 49 44 37 -
aureus
P. aeuruginosa 0 10 0 0 -
Candida albicans 0 0 0 - 40
HL – Hexane layer; EL– EtOAc layer; AL – Aqueous layer; GENT– Gentamycin; TIOC– Tioconazole;
Assays run @ 20 mg/ml
3
Table 2: Susceptibility test of the column fractions of Flabellaria paniculata
Test organism Zone of inhibition (mm)

R1 R2 R3 R4
S. aureus 50 0 0 34
P. aeuruginosa 30 0 6 0
C. albicans 0 0 8 0
R1 – Hexane/EtOAc 80: 20; R2 - Hexane/EtOAc 70: 30;R3 – EtOAc;
R4 – MeOH; Assays run @ 20 mg/ml
Table 3: MIC of P2a and P2b isolates of Flabellariapaniculata

Compound Concentration (µg/ml) Test organism
S. aureus P. aeuruginosa
1600 - -
+
P2a 800 O -
400 + O+
200 ++ +
100 +++ ++
50 +++ +++
800 - -
400 - -
P2b 200 - -
100 O+ -
50 + O+
+
O , MIC value; -, No growth; + , Light growth
++, Moderate growth; +++, High growth
Table 4: MBC of P2a and P2b isolates of Flabellaria paniculata
Compound Concentration (µg/ml) Test organism
S. aureus P. aeuruginosa
1600 O+ -
P2a 800 + O+
200 O+ -
P2b 100 + O+
MBC value;-, No growth;+, Light growth
4
CONCLUSION
The findings revealed leaf extracts of Flabellaria paniculata to be active against Staphylococcus
aureus and Pseudomonas aeruginosa but inactive against Candida albicans.The two active principles
isolated from the leaf of F. paniculata demonstrated significant activities against Staphylococcus
aureus and Pseudomonas aeruginosa. The compound P2b proved to be more potent than P2a against the
test bacteria. The anti-infective property supports the traditional use of the plant for treatment of
infections. This study supports the projection of the previous work reported by Abo and Olugbuyiro
(2004) that further purification will enhance the antimicrobial potency of the plant.
REFERENCES
Abo, K.A and Olugbuyiro, J.A.O. (2004). Phytochemical and antibacterial studies of extracts of
Flabellaria paniculata. African Journal of Biomedical Research 7: 35-36.
Burkil, H.M. (1997). The useful plants of West Tropical Africa. Vol. 3. 2nd ed. Royal Botanic Gardens,
Kew.
Cowan, M.M. (1999). Plant products as antimicrobial agents. Clin microb Rev.12: 564 – 582.
Jerry, R. (2013). Candidiasis (Yeast Infection). Retrieved June 10, 2013, from
http://www.nlm.nih.gov/medlineplus/ency/article/000880.html.
Kenneth, T. (2012).Pseudomonas aeruginosa. Retrieved May 24, 2013, from
http://textbookofbacteriology.net/pseudomonas.html.
Oladosu, I. A., Moronkola, D. O., Osho, A. and Mohammed S. A. (2012).Volatile oil constituents and
anti-inflammatory potential of the essential oil of the leaves and roots of Flabellaria paniculata
Cav (Malpighiaceae). Elixir Pharmacy 50: 10272-10274.
Olugbuyiro, J. A. O., Abo, K. A. and Leigh, O. O. (2010). Wound healing effect of Flabellaria
paniculata. Journal of Ethnopharmacology 127: 786-788.
5
A COMPARATIVE EVALUATION/ANALYSIS OF AIR QUALITY IN SOME SELECTED OIL
PRODUCING/BEARING COMMUNITIES IN DELTA STATE
OF NIGERIA.
OGODO AEMURERHIMEN DICKSON (Snr.) (Chartered Chemist)

(HND, PGD, M.Sc., Ph.D (In- View), AISLT, MCSN, MICCON, C.CHEM., MNES, MSPE)
School of Applied Science, Department of Science Laboratory Technology, Chemistry Laboratory, Delta
State Polytechnic, P.M.B 1030, Ogwashi – Uku, Delta State, Nigeria: Tel: 08030738401, 08094921724, E-
mail:dicksonogodo@yahoo.com
Abstract
The Concentration levels of some Air Quality Parameters of interest namely SOx, NOx, CO, O3, TSPM, VOC, and
heat radiation were determined in samples collected around selected flare sites and Control/Reference Sites in
Six study areas which includes Amukpe – Sapele, Ugbekoko – Sapele, Ekpan (Uvwie), Ozoro, Ogwashi-Uku and
Issele-Uku in other to monitor such gaseous emissions and Comparatively Analyse/Evaluate the Air Quality in
the selected Oil Producing/Bearing Communities in Delta State of Nigeria. Thirteen Air Parameters were
monitored. Appropriate Approved Methodologies for the aforementioned Parameters were determined. The
Ambient Air Quality was determined by the aid of Lamotte Air Pollution Sampling and Test Equipment. Each
Pollutant to be tested for has its own test equipment. The Monthly Average Results generated were also
compared with selected parameters which had DPR /FMEnv limits. The various Air Quality Parameters varied
from site to site. There is marked variations between Air Quality Parameters Indices determined for both the
Reference Sites and immediate industrial areas of the flare sites. Results available indicate that the average
concentration values of SOx ranged from 0.009+0.010 to 0.043+ 0.048ppm while NOx ranged from 0.011+0.010
to 0.029+ 0.031ppm. CO valuesranged from 0.007+ 0.016 to 0.064+ 0.070ppm while TSPMranged from
15.800+10.047 to 45.003+31.973m g / m3 . The site at Ekpan (Uvwie) recorded the highest mean concentration
of VOC of 31.901+ 30.085 m g / m.3 . Sox, NOx, CO, O3, TSPM, VOC, and heat radiations were generally below
the DPR/FMEnv Standards in all the study areas. The outstanding parameter that exceeded DPR/FMEnv
threshold are the VOCs. The air quality parameters in all the study areas have abnormally high values of VOC
which ranged from 2.944+3.224 to 31.910+30.085 m g / m3 . Highly significant correlations were obtained from
many of the Thirteen Parameters determine in the study area. The possible ways to reduce air pollution in the
study areas were critically investigated.
Keywords: Air Quality Parameters, Study Areas, DPR/FMEnvStandards,Sources of Air Pollution, Pollution
Reductions Strategies. 1
Introduction
Air pollution has been defined as any atmospheric condition in which substances are present at concentrations
high enough and above their normal ambient levels to produce a measurable effect on man, animals, vegetation,
or materials 1. The health and environmental negative impacts of air pollutants is now an issue of global concern.
Gaseous Pollutants for example can impair health, reduce visibility, bring about vast economic losses, as well
as, cause cardiac spasm, odour, heart and brain damage, lower birth weight, deterioration of aesthetic structure
like building, monuments and cars2,3,4.5,6,7,8,9,10. Air pollution arises from sources of very great variety. Many
industrial activities contribute as well as domestic and agricultural practices. Air is polluted when the presence of
a foreign substance or a variation in its components is liable to harmful effects or cause nuisance. Natural gas
6
yield combustion products such as Carbon dioxide and Carbon monoxide (CO2 and CO), Nitrogen oxide (NOx),
Water vapour and Soot or Carbon particles.
Also such polluting emissions could directly damage green foliages on which they settled, deteriorate air quality
by causing breathing problems and lung illness, spoil laundry and other valuable property susceptible to dirty
particles. Air pollution through gas flaring also causes heat radiation and thermal conduction into the environment
and high noise level. Polluted air brings fumes of Sulphur and Nitrogen oxides, Sulphuric acid and photochemical
oxidants. These leads to watery eyes, irritate the respiratory system, causing coughing, chest discomfort and
impair breathing. Carbon monoxides interfere with the ability of the body cells (Red blood Cells, RBC) to carry
Oxygen to the heart. Nerves tissues are particularly susceptible to deficiency and this can even impair the
nervous system seriously 11. The best way to control air pollution is to prevent contaminations from entering the
atmosphere in the first place.
12.
Several industrial and automobile accidents have been traced to the problems of poor air quality
In general, there are various air quality parameters which can be determined in the cause of effective and detail
o
research as far as this research work is concerned. This include, Air Temperature ( C), Noise Level [dB(A)],
Smoke Density, (Ringleman Unit), Pressure(mm Hg), Wind Speed m/s Wind Direction, Oxides of Sulphur,
(ppm), Oxides of Nitrogen (ppm), Oxides of Carbon (ppm), Ozone (ppm), Total Suspended Particulate Matter
(µg/m3), Volatile Organic Compounds (µg/m3), and Heat Radiation. (KW/m2)
In several studies, which have been carried out, it is found that a very close association exists between
quality of air and such diseases as bronchitis, asthma, lung cancer and other diseases. Other studies have also
identified a strong causal link between poor air qualities and reduced agricultural productivity. Poor air quality
(say with a large amount of Ozone) poses a very serious threat to agricultural productivity 13.
However, little or no work has been reported on the Comparative Evaluation of the Statistical Analysis of the
Air Quality in the Environment of Some Selected Oil Producing /Bearing Communities in the Niger-Delta (Delta
State) Areas of Nigeria. This is what my project Area is addressing. The knowledge of the Comparative Evaluation
and Statistical Analysis of the Air Qualities in the Environment of Some Selected Oil Producing /Bearing
Communities is necessary because it helps in understanding the details of the Health and Environmental
1
Negative Impacts of Air Pollutants on Man, Animals, Vegetations or Materials .
From the Comparative Analysis, the details such as the measurement of the various Air Quality parameters which
determines the desire to have an articulated engineering control method of gas flares would be established
14
through a proposed model of hydrocarbon pyrolysis during flaring .
Location of Study
The area for this study is Delta State which have Twenty-four (24) Local Government Areas. The Study shall
cover Delta Central, Delta South and Delta North Senatorial Districts of Delta State, Nigeria of which Four Local
Government Areas namely Uvwie, Sapele, Isoko North and Aniocha South Local Government Areas of Delta
State, are selected for the study. Issele-Uku and Ogwashi-Uku in Aniocha South Local Government Areas of Delta
State will serve as the Control/Reference Stations.
In Delta Central Senatorial Districts, the proposed selected sites in (1) Uvwie Local Government Areas of Delta
State for the study is Warri Refining and Petrochemical Company (WRPC)/ Nigerian Gas Company (NGC) and
(2) Sapele (Amukpe Sapele NNPC – Shell flow Station/Ugbekoko (Sapele West) NNPC – Shell Flow Station/
Sapele Gas Plant.
7
In Delta South Senatorial Districts, the proposed selected sites in Isoko North Local Government Areas of Delta
State for this study are: Ogini NNPC – Shell Flow Station in Okpaile in Ozoro Kingdom.
In Delta North Senatorial Districts. The proposed selected site in Aniocha North Local Government Area of
Delta State for this study which will serve us Control/Reference stations are (1) Issele – Uku and (2) Ogwashi-
Uku (for comparison of the results obtained in this study. Detailed statistical Analysis of the results of the Air
Quality in the Environment of some selected Oil Bearing/Producing Communities in the flared and non-flared
Areas of Delta State were computed and compared with approved World Health Organization (WHO) Standards
/ Department of Petroleum resources ((DPR) Standards/Federal Ministry of Environment (FMenv) Standards.
Methodology (Experimental) (Materials and Method)

Analytical Procedures
The summary of the results obtained are shown in tables. Data from the relevant literature were analyzed.
The following parameters are analyzed using the following analytical techniques. As part of Air Quality Studies,
the following parameters were monitored.
i. Determination of Air (Flare) Temperature, 0C
ii. Determination of Noise Level, dB(A)
iii. Determination of Relative Smoke Density, Ringlemann unit
iv. Determination of Absolute Pressure, mm Hg
v. Determination of Wind Speed, m/s
vi. Determination of Wind Direction
vii. Determination of SOx, (Sulphur Oxides), ppm
viii. Determination of NOx,(Nitrogen Oxides), ppm
ix. Determination of CO, (Carbon Monoxide), ppm
x. Determination of O3,(Ozone), ppm
3
xi. Determination of TSPM, (Total Suspended Particulate Matter), ìg/m
xii. Determination of VOC, (Volatile Organic Compounds), ìg/m3
xiii. Determination of Heat Radiation, KW/m2
Methodology
1.1: Sampling Collection and Handling
1.1.1 Stationary Source Monitoring
Samples were collected, handled and preserved according to the relevant test methods. Samples were
collected at the locations determined in the contract arrangement. When determining compliance
with the emission limits of pollutants, sampling was carried out during normal daily activities
1.2: Ambient Air Monitoring

15,
When setting ambient air sampling units, method followed was adopted. Monitoring sites are
classified into three types: peaks, neighborhood and background. When selecting a monitoring site,
many parameters including possible chemicals or physical interference, locality, terrain, services and
local activities were considered. Wind speed and wind direction were located with ambient air
monitoring instruments.
1.3: Methods for Analysis of Ambient Air Pollutants

Methods approved by ASTM for specific parameters are listed in the tables below:
8
Table 1: General Methods for Ambient Air Monitoring
Parameter Measured Method
15
Guide for sitting of sampling units
16
Guide for Laboratories engaged in sampling and analysis of
atmospheres and emissions
Table 2:Specific Methods for Ambient Air Monitoring
Parameter Measured Method
Carbon Monoxide 18
Nitrogen Oxides 18
Ozone 19
Particulate Matters – TSP High Volume 16
Sampler
Sulfur Dioxide 18
Volatile Organic Compounds 18
1.4: Fieldwork
1.4.1: Distance Mapping of Sampling Point

Sample points (60m, 200m and 500m) from the flare point source were measured with a GPS Garmin
® GPS 12.
1.4.2: Heat Radiation

Heat radiation was measured with a Pyranometer equipped with an automatic logging system. (Skye
Data Hog 2). The sensor was focused on the flare for ten minutes to record radiation every ten seconds
interval. The mean reading taken over the ten minutes exposure time was then recorded as the heat
radiation value at each station. The values were recorded in W/m2.
1.4.3: Noise Levels

Noise levels at each point were measured with a pre-calibrated digital readout noise meter. The sensor
of the noise meter was directed towards the source of noise and the average readings over a period of
two minutes were taken to be the Noise-level at each point. The noise levels were measured in decibels
(dB).
1.4.4: Relative Smoke Density

Relative smoke density at each point was measured using the Ringlemann smoke chart approved by
20
the British Standard Institute . This was done by visual comparison of the density of the smoke at the
tip of the flare with the chart's standard shades of grey to black on a scale of 0-4.
1.4.5: Wind Speed and Direction

A combined wind vane and digital Anenometer were used in determining wind direction and speed.
The wind speeds were measured in m/s.
9
1.4.6: Flare Temperature
Flare Temperature was determined with a Testo 860 Infra Red Thermometer.
1.4.7: Air Quality

(a) Ozone: Sampling was carried out at each sample point using an air sampler mounted on a platform
at a height of 1.5 – 2.0 meters from the ground. Air was drawn at a flow rate regulated by a calibrated
critical orifice into a series of glass scrubbers filled with absorbent solutions to absorb specific gases.
Sampling was carried out for a period of thirty minutes at each point. The absorbent solutions were
then subjected to analysis in the laboratory to determine the concentration of O3 in air.
(b) Hydrocarbons/Volatile Organic Compounds (CxHy/VOC), Nitrogen Oxides (NOx), Sulphur

Oxides (SOx) & Carbon Monoxide (CO)
A TESTO 350- flue gas analyzer was used. It performs automatic analysis of ambient air through the
use of physical properties, giving cyclic or continuous output signal. Air is continuously extracted
from the atmosphere and the sample is sent to the analyzer for the determination of the pollutant gas of
interest.
1.4.8: Suspended Particulate Matter (SPM)

A mini-volume portable air sampler (Airmetric ®) with a pre-weighed membrane filter (47µm) was
o
used to collect particulate matter. After sampling, the membrane filter was dried at 105 C, cooled in a
dissicator and weighed to the nearest milligram. The mass concentration was calculated by measuring
the mass of particulate matter and dividing by the volume of air.
The volume of Air was determined as follows:
Volume of Air (m3) = flow rate x time of sampling
V = q x t
Where q= flow rate in liters per minute
t = time of sampling in minutes
0.001(the conversion from liters to cubic meters)
Vm3 = 0.0001m3/l x q l/min x t min
3
Conc. of SPM (g/m ) = 1000g/mg x Wn (mg)
Vm3
Where: Wn (mass change of the filter)
3
Vm (volume of air sample)
1000 (conversion from milligrams to micrograms).
1.4.9: Absolute Pressure
A logger (Testo 450) was used for Atmospheric Pressure. The logger is equipped with an
atmospheric pressure probe (barometer). The logger measures and stores the values.
3.4.10 Data Analysis

The analysis are based on the data collected from Air Monitoring which will now be used to assess
environmental Air Pollution, health hazards and socio-economic problems as a result of gas flaring on the
Oil Bearing /Producing Communities in the selected Areas of Delta State of Nigeria for:
A. Agricultural/fishery activities
10
B. Implications of the pollution in the ecosystems
Thirteen parameters will be determined in this work whereas the relatively few investigations (mainly from
Ekpan (Uvwie) near Warri and Sapele, Delta State, Nigeria) done to date contains not more than 7
Table 6: Air Quality Statistical Analysis for the Year 2013 (Mean and Standard
Deviation)
Table 7: Air Quality Statistical Analysis for the Year 2014 (Mean and Standard
Deviation)
11
Results and Discussion
In order to protect the environment and ensure sound and good industrial practice, the Department of Petroleum
Resources (DPR) and Federal Ministry Environment (FMENV) put in place a number of regulations to control
levels of gaseous emissions into the Nigerian environment.
Air pollution may be defined as the presence of any air-contaminant (solid, liquid, gaseous or colloidal
mixtures), radioactive materials or noise in the atmosphere, which is in excess of the permissible level.
Gaseous emissions from oil producing industries are very significant sources of air pollution. Such emissions
emanate from gas flaring as well as electric power generating facilities used for the operational activities.
The present research work is designed to monitor such gaseous emissions that may be produced in the following
areas:
ü SPDC Amukpe
ü WRPC
ü NGC
ü Ogini Isoko
ü Ugbekoko
ü Ogwashiuku
ü Issele uku
Thirteen air quality parameters were monitored namely air temperature, noise level, smoke density, pressure,
wind speed, wind direction, oxides of nitrogen, carbon and sulphur, ozone, suspended particulates, volatile
organic carbons and heat radiation. Appropriate approved methodologies for the aforementioned parameters
have also been stated (Table 1).
Parameters Units Methods DPR Limit
Flare/Heat Radiation KW/m2 Infrared thermometer 6310
S/Density Ringlemann chart 2
Ambient Tempt. Portable analyser 30
Atm. Pressure Mbar Digital barometer N/A
Wind Velocity m/s Anemometer N/A
Wind Direction N/A Wind vane N/A
Air Turbulence N/A Portable analyser N/A
Sun Radiation KW/m2 Infrared thermometer 6310
Precipitation mm Rain guage 90
Rel. Humidity % Hydrometer N/A
Noise Level dB(A) Noise meter 80 - 100
SPM ug/m3 Portable analyser 250
NOx ppm Lamotte AQ kits 0.04 – 0.06
SOx ppm Lamotte AQ kits 0.1
Carbon monoxide ppm Lamotte AQ kits 10.00-20.00
Hydrogen Sulphide ppm Lamotte AQ kits N/A
O3 ppm Lamotte AQ kits 0.06
VOC ppm ASTM D2972-97 0.16
12
TEMPERATURE
A portable thermometer was used.
NOISE LEVEL
The noise level was measured using a Sound Level Meter calibrated to 94.0dB (A).
The table below gives the various duration of exposure per day for different permissible noise level.
Table 2: National Air Quality Guidelines for Maximum Exposure.
Hours of Exposure Per Day FMENV Permissible Exposure Limit
dB(A)
8 90
6 92
4 95
3 97
2 100
1.5 102
1 105
0.5 110
0.25 or less 115
SMOKE DENSITY
A graduated Ringelmann chart was used to determine the smoke density of the flares.
The chart is numbered from 0-4 with 0 being the lowest density, while 4 is the highest. The chart is held high, at
about 20 meters from the flare and the smoke coming from it compared with the chart.
The relative smoke density of emitted from the flare should not exceed 2 Ringlemann Number as stipulated in the
DPR guidelines. The table below shows the relationship between Ringlemann Number/Percent Light
Transmission and Smoke Density.
Table 3: Ringlemann Number/Percent Light Transmission and Smoke Density.

Ringlemann % Light Transmission Smoke Density
Number/Units Through Smoke (Percent)
0 100 0
1 80 20
2 60 40
3 40 60
4 20 80
WIND SPEED AND DIRECTION

The wind speed was measured in m/s using an anemometer (Kestrel Pocket Wind Meter 273847A). The direction
of the wind was noted using a wind vane.
AMBIENT AIR QUALITY

The ambient Air Quality was determined by with the aid of LaMotte Air Pollution Sampling and Test Equipment.
Each pollutant to be tested for (SOx, NOx, CO, H2S, O3 and SPM) has its own test equipment.
Volatile Organic Carbon (VOC) was measured by using a VOC meter manufactured by ELE, United Kingdom.
The table below gives the Nigerian Ambient Air Quality Standard.
Table 4: Nigerian Ambient Air Quality Standard.

POLLUTANTS TIME AVERAGE FMENV LIMIT
SOx 1- hour 0.1ppm
NOx Daily average of 0.04-0.06ppm
hourly value range
VOC 3 – hourly average 160ppm
13
HEAT RADIATION
A pyranometer (Data HOG Skye Instruments, USA) was used for this purpose. The meter was set to take
readings at thirty seconds (30s) interval for about five minutes (5min) and an average of the readings was then
calculated.
The gaseous emission monitoring results for the entire sampling duration is discussed as follows:
The Monthly average results generated were also compared with selected parameters, which had
DPR/FMEH&WHO limits using radial graph representations. These graphs were presented alongside the table
for each sampling location.
Table 1:
Parameters Units Methods DPR Limit
Flare/Heat Radiation KW/m2 Infrared thermometer 6310
S/Density Ringlemann chart 2
Ambient Tempt. Portable analyser 30
Atm. Pressure Mbar Digital barometer N/A
Wind Velocity m/s Anemometer N/A
Wind Direction N/A Wind vane N/A
Air Turbulence N/A Portable analyser N/A
Sun Radiation KW/m2 Infrared thermometer 6310
Precipitation mm Rain guage 90
Rel. Humidity % Hydrometer N/A
Noise Level dB(A) Noise meter 80 - 100
SPM ug/m3 Portable analyser 250
NOx ppm Lamotte AQ kits 0.04 – 0.06
SOx ppm Lamotte AQ kits 0.1
Carbon monoxide ppm Lamotte AQ kits 10.00-20.00
Hydrogen Sulphide ppm Lamotte AQ kits N/A
O3 ppm Lamotte AQ kits 0.06
VOC ppm ASTM D2972-97 0.16
Table 5A: Comparison of SPDC Amukpe With DPR Limits
SPDC SPDC
AMUKPE DPR AMUKPE DPR
PARAMETERS F/S LIMTS PARAMETERS F/S Limits
Noise Level 79.9 90 Noise Level 89% 100%
Smoke Density 0.63 2 Smoke Density 31.5% 100%
Temp. 31.3 30 Temp. 104% 100%
VOC 6.18 0.16 VOC 3863% 100%
O3 0.00 0.05 O3 0% 100%
SPM 25.01 260 SPM 9.62% 100%
Heat Radiation 0.405 6310 Heat Radiation 0.006% 100%
SOx 0.0288 0.1 SOx 29% 100%
14
Figure 5a:
Fig. 5A: Radial Graph Representation of SPDC Amukpe – Sapele
Table 5B: Comparison of WRPC With DPR Limits

DPR DPR
PARAMETERS WRPC LIMTS PARAMETERS WRPC Limits
Noise Level 80.7 90 Noise Level 89.7% 100%
Temp. 34.0 30 Temp. 113.3% 100%
VOC 31.91 0.16 VOC 100%
O3 0.00 0.05 O3 100%
SPM 45.00 260 SPM 17.3% 100%
SOx 0.043 0.1 SOx 43% 100%
Fig. 5B: Radial Graph Representation of WRPC in Ekpan (Uvwie)
15
Table 5c: Comparison of NGC with DPR Limits
DPR DPR
PARAMETERS NGC LIMTS PARAMETERS NGC Limits
Smoke Density 0.00 2 Smoke Density 100%
Temp. 31.6 30 Temp. 105.3% 100%
VOC 2.94 0.16 VOC 100%
O3 0.00 0.05 O3 100%
SPM 15.80 260 SPM 6.1% 100%
SOx 0.009 0.1 SOx 9% 100%
Fig. 5C: Radial Graph Representation of NGC in Ekpan (Uvwie)
Table 5D: Comparison Of Ogini Isoko North with DPR Limits

OGINI OGINI
ISOKO DPR ISOKO DPR
PARAMETERS NORTH LIMTS PARAMETERS NORTH Limits
Smoke Density 0.50 2 Smoke Density 25% 100%
Temp. 31.3 30 Temp. 104.3% 100%
VOC 7.31 0.16 VOC 100%
O3 0.00 0.05 O3 100%
SPM 35.16 260 SPM 13.5% 100%
SOx 0.0292 0.1 SOx 29.2% 100%
16
Fig. 5D: Radial Graph Representation of Ogini in Isoko North
Table 5E: Comparison o Ugbekoko with DPR Limits

DPR DPR
PARAMETERS UGBEKOKO LIMTS PARAMETERS UGBEKOKO Limits
Temp. 30.4 30 Temp. 101.3% 100%
VOC 3.09 0.16 VOC 100%
O3 0.00 0.05 O3 100%
SPM 41.92 260 SPM 16.1% 100%
SOx 0.033 0.1 SOx 33% 100%
Fig. 5E: Radial Graph Representation of SPDC in Ugbekoko (Sapele West)
17
Table 5F: Comparison of Ogwashiuku with DPR Limits
DPR DPR
PARAMETERS OGWASHI LIMTS PARAMETERS OGWASHI Limits
Temp. 29.5 30 Temp. 98.3% 100%
VOC 0.00 0.16 VOC 100%
O3 0.00 0.05 O3 100%
SPM 14.66 260 SPM 5.6% 100%
SOx 0.00 0.1 SOx 100%
Fig. 5F: Radial Graph Representation of Ogwashi -Uku (Reference/Control Station)
Table 5G: Comparison of Issele-Uku with DPR Limits

ISSELE DPR ISSELE DPR
PARAMETERS UKU LIMTS PARAMETERS UKU Limits
Noise Level 45.9 90 Noise Level 51% 100%
Temp. 30.1 30 Temp. 100.3% 100%
VOC 0.00 0.16 VOC 100%
O3 0.00 0.05 O3 100%
SPM 15.87 260 SPM 6.1% 100%
Heat Radiation 144 6310 Heat Radiation 0.002% 100%
SOx 0.00 0.1 SOx 100%
18
Fig. 5G: Radial Graph Representation of Issle -Uku (Reference/Control Station)
Discussion
Ambient Temperature
The average temperature at all locations ranged between 29.537+0.135 – 33.957+1.1500C. The high temperature
measured at 60m from the flare in WRPC is an indication of the effect of flare heat on the environment. The
o
average ambient temperature is above the set limit of 30 C by DPR. Please cite other international guidelines and
compare the results to related analysis done within the country and beyond.
Noise Level
The recorded average noise levels at all locations, irrespective of distances from the flare, pump and generator
areas were within the DPR range of 82.408+1.592 - 80.662+2.377 (A) for a period of 8hrs noise exposure per
day. The highest noise levels of 78.00dB (A) and 76.00dB (A) were recorded at NGC F/S Ekpan – (Uvwie) and
WRPC F/S Ekpan (Uvwie) respectively. No noise level exceeded 90.00dB(A) level of the FMenv. Permissible
exposure limit for a period of 8hours noise exposure per day.
Smoke Density
The smoke density of the flares monitored in all the locations complied with the DPR limits of 2 Ringelmann
Number. The highest average smoke density of 8.853+21.685Ringelmann Number were recorded at Ogwashi-
Uku follow by 2.792+0.534 for WRPC in Ekpan (Uvwie) this may be due to the smoke emitted from the flare
stack daily into the atmosphere. So various gaseous emissions are poured into the atmosphere near and around
dwelling places. In this way, most industrial time and cities in Nigeria are always polluted.
Air Turbulence and Wind Direction

The entire locations monitored fell under the stable atmospheric classified as F using the dispersion equations
quantification in a range of A to F. The air turbulence was generally stable and near calm.
Ambient Air Quality

When compared with the Federal Ministry of Environment (FMENV) standard for ambient air quality, all the
locations complied with the standards for all the gases monitored.
19
Heat Radiation Level
The heat radiation level decreases as the distance from the flare increased at all the locations monitored. The high
values obtained in the afternoon can be attributed to increase in the ambient temperature. The values were well
2
below the DPR allowable heat radiation of 6.31Kw/m at ground level during maximum flaring at a distance of
60m from the base of the flare.
VOLATILE ORGANIC CARBON

Emissions from fossil fuel combustion have long been known to have negative effects on air quality inducing
episodes of smog and contributing to increased levels of ozone and volatile organic compounds.
Conclusion
From the results above, the outstanding parameter that exceeded DPR and FMEnv threshold are the VOCs. They
include benzene, toluene, ethylbenzene, xylene and other monoaromatics. Quite disturbing is the serious
repercussions on human health. Prolonged exposure to benzene, for example which is emitted directly from
petrol – driven automobiles, is known to cause haematotoxicity, genotoxicity, and carcinogenicity, and can result
in leukaemia, anaemia and thrombocytopenia (WHO, 2000).
Air Pollution Abatement

The abatement of air pollution can be achieved as follows:
(i) Installation of an emission control device (e.g. the Catalytic Converter) at the Exhaust Pipe of a
Motor Vehicle or Smokestack or Chimney of an Industrial Plant.
(ii) Use of giant vacuum cleaners, or electrostatic precipitators or wet scrubbers for the removal of
particulates from Industrial exhaust; use of mist eliminators for the collection of liquid droplets.
(iii) Fewer Petroleum-powered vehicles are desired; in fact, electric cars could replace the present motor
vehicles. Electric cars are pollution-free almost noiseless, fumeless and more efficient in energy
conversion.
(iv) Use of harmless fuel as an alternative to gasoline.
(v) Acquisition of the necessary technology by industry to attain the zero emission level
(vi) Legislation. 24
References
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21
2+ 2+ 2+
BATCH ADSORPTION OF Cd , Pb and Ni WITH COCONUT (Cocos nucicera L.) SHELL
ACTIVATED CARBON MODIFIED WITH 1, 2-DIHYDRO-1, 5-DIMETHYL-2, PHENYL- 4- (E) -
(2, 3, 4- TRIHYDROPHENYL) -3H-PYRAZOL-3-ONE (DDPTP)
Ukoha, Pius. O, Anierobi, Peter. O and Nicholas, Eno-obong S.
Department of Pure and Industrial Chemistry, Faculty of Physical Sciences,
University of Nigeria, Nsukka, Enugu State, Nigeria.
*Corresponding author's e-mail: pius.ukoha@unn.edu.ng, eno-
obong.nicholas.pg78610@unn.edu.ng; +2348037171322,+2348062316565
Abstract
Batch adsorption of Cd2+, Pb2+ and Ni2+with coconut (Cocos nucicera L.) shell activated carbon modified with
1,2- dihydro-1,5- dimethyl-2-phenyl-4-(E)–(2,3,4-trihydrophenyl) -3H-pyrazol-3-one (DDPTP) were studied. It
was activated chemically using CaCO3 as the activating agent. Proximate analysis on the coconut shell showed
8.7% moisture content, 10.4% volatile matter, 3.2% ash content and 77.7% fixed carbon. The developed
adsorbent has bulk density of 0.46g/cm3, pore volume of 8.0 x 10-3cm3 and the conductivity was 37.9µS/cm.
Fourier Transform Infrared (FTIR) analysis showed that hydroxyl, carbonyl, amino and azo groups are present
on the surface of the adsorbent. Scanning Electron Microscope (SEM) showed the micro-pores in the modified
coconut shell activated carbon (MCSAC) while Energy Disperse X-ray Spectrum exposed carbon as the major
quantitative element with 57%. Batch adsorption was carried out and the results obtained showed that, MCSAC
2+ 2+ 2+
adsorbed Pb (98%), Cd (80%) and Ni (92.2%) ions more than unmodified coconut shell activated carbon
2+ 2+ 2+
which adsorbed Pb (79%), Cd (60.2%) and Ni (73.6%) ions from aqueous solutions. The analysis of
adsorption isotherm showed that adsorption of Ni2+ followed Langmuir Isotherm than Cd2+ and Pb2+; Ni2+ and
Cd2+ followed Freunlich isotherm than Pb2+;Ni2+ and Pb2+ followed Temkim isotherm than Cd2+. Kinetic studies
2+ 2+
showed that the adsorption of the metal ions can also be described by pseudo-first-order (Pb and Ni ), pseudo-
2+ 2+
second-order (Cd and Ni ) and intra-particle diffusion models for the three metals.
KEYWORDS: Heavy metal, Adsorption, pH, Carbonization, Coconut shell, Activation.
Introduction
The presence of trace heavy metals in natural water has aroused the interest of many Nigerian scientists as a result
of their environmental effects on the health of both plants and animals. More so, concern about environmental
protection has increased due to the technology1 development which keeps on changing, producing industrial
product, as well as waste. Manufacturing industries have played an important role for economic growth in major
countries and this sector provides services and product for better way and quality of life.
Heavy metals such as cadmium, chromium, lead, copper, manganese, zinc as well as mercury and nickel are
widely discharged in the wastewater from industries and are very toxic and harmful to living organisms by
lowering the reproductive success, preventing proper growth and even causing death2. Some of the heavy metals
are important for our body requirement; however exceeding the tolerance limit may create harm to body
functions.
Presence of phenols in water bodies caused carbolic odour to receiving water bodies, thus causing toxic effects
on aquatic flora and fauna 3.
2.0 Materials and Methods
2.1 Preparation of reagents: All reagents used for this study were analytical reagent grade. Acids and bases
were of the highest purity and were used as received. The analytical reagents grade, nitrate salts of lead and
cadmium and chloride of nickel were used without any further investigation. About 100mg/l aqueous solution of
22
the metal ions were prepared as stock solution from their salt [Pb(NO3), Cd(NO3)2.4H2O and NiCl2. 6H2O]. From
the stock solution, working solutions of 0.5, 1.0, 2.0, 3.0 and 4.0mg/l were prepared from appropriate aliquots
(portion) diluted to the appropriate concentration with de-ionized water.
pH adjustment was done by the addition of dilute nitric acid or sodium hydroxide into the solution of metal salts.
Activated Carbon (AC) was prepared from coconut shell and was modified with 1, 2-dihydro-1, 5-dimethyl-2-
phenyl-4(E)-(2,3,4-trihydroxyphenyl)-3H-pyrazol-3-one, DDPTP.
2.1.1 Preparation of 0.5 moldm-3 of CH3COOH (Acetic Acid): To prepare 0.5 moldm-3 of CH3COOH (99.5%)
was accurately measured into 1000ml (1L) standard volumetric flask containing some quantity of de-ionized
-3
water and was later made up to the mark with de-ionized water to give 0.5moldm solution of CH3COOH.
-3 -3
2.1.2 Preparation of 0.1 moldm of HNO3: To prepare 0.1moldm of HNO3, 3.17 ml concentrated HNO3 (70%)
was accurately measured into 1000ml (1L) standard volumetric flask containing some quantity of de-ionized
water and was later made up to the mark with de-ionized water to give 0.1 moldm-3of HNO3.
2.1.3 Preparation of 1000ppm Pb(NO3)2 solution: To prepare 1000ppm of Pb(NO3)2 solution, 1.599g of
3
Pb(NO3)2 was accurately weighed, dissolved and made up to the mark in 1000cm standard volumetric flash to
give 1000ppm Pb(NO3)2 solution from which 100ppmsolution was made. It was diluted to 10ppm from which
working solutions of 0.5,1,2,3 and 4ppm were made.
2.1.4 Preparation of 1000ppm Cd(NO3)2.4H2O solution: To prepare 1000ppm Cd(NO3)2.4H2O solution,
3
2.744g of Cd(NO3)2.4H2O was accurately weighed, dissolved and made up to the mark in 1000cm standard
volumetric flask to give 1000 ppm Cd(NO3)2.4H2O solution from which 100ppm solution was made. It was
diluted to 10ppm from which working solutions of 0.5, 1, 2, 3 and 4ppm were made.
2.1.5 Preparation of 1000pm NiCl2.6H2O solution: To prepare 1000ppm NiCl2.6H2O solution, 5.481g of
3
NiCl2.6H2O was accurately weighed, dissolved and made up to the mark in 1000 cm standard volumetric flask to
give 1000ppm NiCl2.6H2O solution from which 100 ppm solution was made. It was diluted to 10ppm from which
working solution of 0.5, 1, 2, 3 and 4 ppm were made.
2.2 Synthesis of the ligand: Reagent grade chemicals were used without further purification. 1,2-dihydro-1,5-
dimethyl-2-phenyl-4(E)-(2,3,4-trihydroxyphenyl)-3H-pyrazol-3-one, DDPTP was prepared following the
method of Heinosuke Yasuda4.
2.2.1 Preparatory stage (preparing it for carbonization): This stage includes washing, drying, grinding and
sieving. The coconut shells (coir) were soaked in water for 1hr before they were clean using sponge in water to
5 0 5
remove dirt and some stinky grease .They were dried in an oven (OV-160) at 105 C for 1h . The dried shells were
broken into pieces with hammer on a clean surface. The broken pieces were ground into tiny particles by a
grinding machine then different sieve sizes were used to obtain different particles sizes such as 0.25, 0.5, 1.4, and
2.8 mm particles.
2.2.2 Carbonization: The ground samples with 0.25, 0.5, 1.4, and 2.8mm particle sizes were all carbonized in a
vaster chesterfield furnace, Model L F 3 at 4000C for 1h. The carbonization was repeated with 0.25mm adsorbent
0 0 0
at varied temperatures; 300 C, 350 C and 450 C.
2.2.3 Activation (Chemical activation): The carbonized samples (coir) were activated with calcium carbonate
(CaCO3). About 10g of CaCO3 was dissolved in 50cm3of distilled water in a 10cm3 beaker 5. The resultant
solution was mixed with 50g of carbonized coir and allowed to stay for 24 h which helped to open the pores 7, 8, 5.
The mixture was filtered after 24 h and washed first with 0.5M acetic acid and later rinsed with de-ionized water
0 7
till the pH=7,dried at110 C for 3 h and was stored in an air-tight container.
2.3 Modification with 1,2-dihydro-1,5-dimethyl-2-phenyl-4(E)-(2,3,4-trihydroxyphenyl)-3H-pyrazol-3-
one, DDPTP.
1,2-dihydro-1,5-dimethyl-2-phenyl-4(E)-(2,3,4-trihydroxyphenyl)-3H-pyrazol-3-one, DDPTP (30 mg) of
23
3 3
ligand was dissolved in 2 cm of methanol and was diluted with 50cm of de-ionized water. A 5g of activated
3
carbon (CSAC) stirred with 50cm 0.06% of DDPTP (30 mg) solution with the aid of a mechanical shaker, Multi-
fix (M80) for 30 min 9. The resultant solution was allowed to stay for 24 h. It was thereafter dried at 600C for 1hr in
an oven 10. Four other samples, 5g each of CSAC were modified with varying quantity of ligand such as 10mg, 20
mg, 40 mg, and 50 mg for the determination of the effect of the amount of ligand used on the adsorption of the
metal ions.
2.4 Characterization of the modified Coconut Shell Activated Carbon, MCSAC: Some physical parameters
were determined so as to know the properties of the MCSAC;
11
2.4.1 Determination of the moisture content: This was done according to ASTM standard E711-87 by
weighing 2g of the adsorbent into a known weight crucible as w2.
2.4.2 pH measurement: It was determined according to ASTM standard E711-8711.
2.4.3 The Determination of bulk density: The bulk density, B.D of the sample of MCSAC was determined
11
according to ASTM standard E711-87 .
2.4.4 Determination of volatile matter: volatile matter was determined according to ASTM standard E711-
8711. The sample (2g) was pulverized in a crucible and placed in an oven until a constant weight, W1 was obtained.
The coconut shell samples were then kept in a furnace at a temperature of 5500C for 10 min. and weighed after
cooling in desiccators to obtain a weight W2. The loss in weight accounts for the volatile content of the coconut
12
shell sample .
2.4.5 Pore Volume Determination (PV): PV was determined according to ASTM standard E711-8711, the
sample, W0 was weighed into a beaker and the weight of sample plus beaker was noted, W1. A measured 50cm3 of
distilled water was added into the beaker containing the dry sample and the mixture was boiled for 15min. after
the air in the pores has been displaced, the sample was drained, dried superficially and weighed. The procedure
was repeated twice 13.
2.4.6 Determination of fixed Carbon: This is gotten by deducting the sum of the percentages of ash, volatile,
11
and moisture content from 100 as it was stated by ASTM standard E711-87 . It is expressed as: Fixed carbon% =
100- moisture% - ash% - volatile matter % 7.
2.5 Adsorption Procedure
2.5.1 Variation of initial metal ion concentration: The concentration of residual metal ions in the filtrates was
5
determined using AAS. The amount of metal ions adsorbed from the solution was determined by difference .
2.5.2 Variation of contact time, pH value, ligand amount, particle size, temperature of carbonization and
Competitive adsorption: Same as above procedure.
2.5.3 Variation of level of treatment of adsorbent on adsorption: Same as above
3.0 Results and Discussions
1, 2-dihydro-1, 5-dimethyl-2-phenyl-4-[E]-(2, 3, 4-trihydrophenyl)-3H-pyrazol-3-one (DDPTP) was
characterized and the following result were obtained.
Molar
Compound Colour Texture Melting Yield % yield conductivity
point(0C) (µS/cm)
Dark
Ligand brown Powdery 190 0.35 35.40 26.10
24
It was observed from Table 3.1 that the molar conductivity of the ligand was high compared to instrumental range
-3
at which a compound can be non conductor which is 0-0.3 µScm . The ligand showed slight conducting
properties probably due to dissociation occurring in the solution of the ligand.
3.1 Electronic Spectra of the ligand: The UV spectra of 1, 2-dihydro-1, 5-dimethyl-2-phenyl-4-[E]-(2, 3,4-
trihydroxylphenyl)-3H-pyrazol-3-one (DDPTP) is shown in figure 3.1. From the figure, it was observed that the
ligand showed maximum absorption at 376.50nm. The peak corresponds to what had been reported of this ligand
14
earlier .
3.2 FTIR Spectra of the ligand: The IR spectra of the ligand gives characteristics absorption band around
3249.20cm-1 due to intermolecular hydrogen bonding resulting from V (OH) group. The band at 1620.74cm-1 in
-1
the ligand is assigned to V(C=O) carbonyl group. Absorption band at 1498.74cm was due to azo group V
-1 1 13 14
(N=N). The band at 1234.48cm is assigned to V(C=O). The H and CNMR of the ligand had been reported .
3.3 Physico-Chemical properties of the Adsorbed: The data table is as follows;

Table 3.2: Physico-chemical properties of the adsorbent
Parameters Result
Colour Black
pH 8.6
Conductivity (1%) 37.9µS/cm
Pore volume 8.0x10-3cm3
Fixed Carbon 77.7%
Moisture content 8.7%
Volatie matter 10.4%
Ash content 12%
Bulk density 0.46g/cm3
3.4 Adsorption:
3.4.1 Effect of concentration on the removal of Pb2+, Cd2+ and Ni2+ from solution
The effect of initial concentration of metal ions on metal ion adsorbed, Qe (mg/l) by MCSAC was studied at room
temperature. It was found that the concentration of metal ions adsorbed Qe(mg/l) increased with increase in
initial metal ion concentration as shown in the Table 3.3. This is due to the fact that as the initial concentration is
increased, more metal ions are available in the solution for the adsorption process 5. Adsorption for the metal ions
increases with increase in metal ion concentration because at low concentrations, the active site on the surface of
the adsorbent are not completely covered, so more ions would probably get adsorbed on increment of the initial
concentration.
Table 3.3: Concentration of the metal ions adsorbed (Qe) at various concentration at room temperature and
pH 7.5 by MCSAC
Qe(mg/l)
Initial % % %
2+ 2+ 2+
concentration Pb Removal Cd Removal Ni Removal
Co(mg/l) of Pb2+ of Cd2+ of Ni2+
0.5 0.452 90.40 0.301 60.20 0.446 89.20
1 0.939 93.90 0.431 43.10 0.796 79.60
2 1.923 96.15 0.989 49.45 1.612 80.60
3 2.891 96.37 1.599 53.30 2.49 83.00
4 3.702 92.55 2.153 53.83 3.41 85.25
25
Fig.1. shows the variation of the concentration of metal ions adsorbed by MCSAC with initial concentration. It
was observed that the quantity of metals absorbed increased with increase in the initial metal ion concentration,
and that the rate of adsorption with respect to concentration was uniform and progressive in all the ions except for
cadmium ion which had a little delay at initial concentration probably due to a low activation energy need to kick
start the reaction.
Fig. 3.1: Variation of quantity of ions adsorbed by MCSAC with initial concentration of metal ions
2+ 2+ 2+
3.4.2 Effect of Contact time on the Removal of Pb , Cd and Ni from their solutions
The adsorption of Pb2+, Cd2+ and Ni2+ on MCSAC was studied as a function of time in order to find out the
equilibrium time required for maximum adsorption of metallic ions.
Table 3.4: % Removal of the metal ions adsorbed (Qe) at various time interval at room temperature and pH
7.5 by the adsorbent (MCSAC)
% Removal of metal ions, Qe

2+
Time(min) Pb Cd2+ Ni2+
10 30.4 19 23
30 46.2 28.4 35.54
50 70.8 38.4 58.8
70 80.2 44.6 77.8
90 88.4 59.8 82.4
Fig.3.2: Variation of % removal of ions adsorbed by MCSAC with time at room temperature
Table 3.4 and Fig.3.2 show the adsorption of the metal ions using MCSAC. It is observed that the concentration
of metals ions adsorbed on the MCSAC increased with increase in agitation time. This is also due to the migration
of higher fraction of the metal ions from the bulk solution through the adsorbent boundary layer onto the active
sites of the adsorbent as time progresses 5.The removal of metal ions by MCSAC could also be due to difference
in sorption processes: ion-exchange, adsorption (Physisorption/chemisorptions) co-ordination, complexion as a
result of the various types of ion binding groups such as hydroxyl, carboxylic, phenolic functional groups in
MCSAC.
26
3.4.3 Effect of temperature of carbonization on the sorption capacity of the adsorbent:
Temperature can affect the adsorption behaviour of metallic ions in solution. The effect temperature of
2+ 2+ 2+
carbonization on the adsorption of Pb , Cd and Ni from solutions containing MCSAC was studied at initial
2+
concentration of 0.5mg/l for 30 min contact time at room temperature. Table 3.5 showed that the removal of Pb ,
Cd2+ and Ni2+ from aqueous solution by MCSAC was dependent on the temperature of carbonization. Increase in
temperature of carbonization from 3000C to 4500C was found to result in a steady increase in the removal of metal
ions by the adsorbent. This implies that increase in temperature of carbonization creates a wider surface area of
the adsorbent for adsorption. This is a result of the decrease in sizes of the adsorbent particles as the temperature
of carbonization increases.
Table 3.5: % Removal of the metal ions adsorbed (Qe) at various temperature, pH 7.5 by coconut (MCSAC).
% Removal of metal ions

0 2+
Temperature ( C) Pb Cd2+ Ni2+
300 68 40.4 59.2
350 78.2 59.8 68
400 84 77.8 80.2
450 90 80.2 84.4
3.4.4 Effect of pH on the removal of Pb2+, Cd2+ and Ni2+ from solution:
2+ 2+ 2+
The effect of pH on the adsorption of Pb , Cd and Ni from solutions containing MCSAC was studied at an
initial concentration of 0.5mg/l for 30min and at room temperature. Table 3.6 shows that the concentration of the
metal ions increased as the pH moves from the acidic pH 2 to alkaline pH 10 , this result of the effect of pH on the
adsorption of metal ions by MCSAC 1 represented in Fig.3.4; It shows that alkaline medium tend to support
15
adsorption more than acidic medium. According to Low et al , at low pH values of the surface of the adsorbent
would be closely associated with hydroxium ions, by repulsion forces to the surface functional groups
consequently decreasing the percentage adsorption of metal.
Table 3.6: % Removal of the metal ions adsorbed at various pH at room temperature by MCSAC
% Removal of metals
2+
pH Pb Cd2+ Ni2+
2 53.6 65.6 42.6
4 63.4 69.8 58
6 90.4 73.8 76.2
8 93 77.6 82.4
10 88 80.2 83
27
Figure 3.4: Variation of % removal of ion by MCSAC with pH
3.4.5 Effect of degree of treatment of adsorbent (MCSAC):As the degree of treatment of adsorbent increased,
it was observed from the % removal in Figure 3.4 that the quantity of metal ions adsorbed from the solution
increased in all the three metals. This was because the adsorption capacity was increased by carbonizing the
adsorbent (improving the pore volumes and the surface structure), activating agent, CaCO3 (in addition to
increasing the surface structure, the functional groups of the adsorbent) and modification with DDPTP (which
adds more functional group like amine and hydroxyl group on the surface of the adsorbent).
Table 3.7: % Removal of the metals ions adsorbed at various levels of treatment room temperature and pH
7.5
Degree of % Removal of metal ions, Qe

2+
treatment of Pb Cd2+ Ni2+
adsorbent
A 20.4 15.5 18.5
B 35.6 28.6 33
C 79 60.2 73.6
D 98 80 92.2
Figure 3.5: Variation of % removal of ions adsorbed with different quality of adsorbents.
Table 3.7 and Fig.3.5 shows that, there was a little adsorption with non carbonized adsorbent and adsorption
increased with increase in the degree of treatment of the adsorbent due to the improvement on the adsorption site
for metals ions.
3.4.6 Effect of particle size on adsorption of Pb2+, Cd2+ and Ni2+
Particle sizes of an adsorbent tend to affect the surface area of the adsorbent in a way that, the smaller the particle
sizes of the adsorbent the more surface area 1 increased. The increased in the surface area opens up more
adsorption site for the metal ions to be adsorbed but increase in particle size decreases the surface area hence
decreasing the quantity of metal ions adsorbed.
Table 3.8: % Removal of the metal ions adsorbed at various particle sizes of MCSAC at room temperature
and pH 7.5
% Removal of metal ions
2+
Particle sizes(mm) Pb Cd2+ Ni2+
0.25 96 71.2 90
0.5 80 64 84
1.4 68.2 50 67
2.8 22 19.8 28
28
2+ 2+ 2+
3.4.7: Competitive adsorption of Pb , Cd and Ni from their mixed solution on MCSAC:
Competitive adsorption was carried out to investigated the effect of the metal ions (Pb2+, Cd2+ and Ni2+)
on each other and their influences on adsorption. Ions with higher charge and the smaller hydrated
radius have higher affinity, greater polarization facilities electrostatic exchange. The more
electronegative metals should form the strongest covalent bonds with oxygen atoms on any particular
mineral surface16.
Table 3.9: Data from the competitive adsorption of the metals with total initial concentration, Co of
the three metals ions=6mg/l (each 2mg/l), pH 9.5, contact time 30min at room temperature
METALS Ce(mg/l) Qe(mg/g) % Removal
Pb2+ 0.578 1.421 71.05
Cd2+ 1.098 0.92 45.10
Ni2+ 0.493 1.507 75.35
3.4.8: Adsorption Isotherm
3.4.8.1 The Langmuir Isotherm
The Langmuir model96 is based on the assumption that the maximum adsorption occurs when a
saturated monolayer of solute molecules is present on the adsorbent surface.
Table 3.10 a: Isotherm data for Langmuir
Pb2+ Cd2+ Ni2+
Co(mg/l) 1/Qe(mg/g) 1/Ce(mg/l) 1/Qe(mg/g) 1/Ce(mg/l) 1/Qe(mg/g) 1/Ce(mg/g)
0.5 2.2124 20.8333 3.3223 5.0251 2.2425 18.5185
1.0 1.065 16.3934 2.32 1.7575 1.2563 4.902
2.0 0.52 12.987 1.0111 0.9891 0.6203 2.5773
3.0 0.3459 9.1743 0.6254 0.7138 0.4016 1.9608
4.0 0.2701 3.3557 0.4645 0.5414 0.2933 1.6949
Table 3.10b: Langmuir isotherm constants
Ions Qm (mg/g) KL(mg/l) RL R2
Pb2+ 0.164 0.833 0.924 0.774
Cd2+ -2.740 -0.260 1.149 0.860
Ni2+ 3.086 3.028 0.398 0.910
3.4.9: The Freundlich Isotherm
Freundlich adsorption isotherm is the relationship between the amounts of metal adsorbed per unit mass of
adsorbent, Qe, and the concentration of the metal at equilibrium, Ce. The following are the Freundlich adsorption
isotherm data generated from the sorption of nickel, lead and cadmium ions.
Table 3.11a: The Freundlich isotherm
Pb2+ Cd2+ Ni2+

Co(mg/l) LogCe LogQe LogCe LogQe LogCe LogQe
0.5 -1.319 -0.344 -0.701 -0.521 -0.351 -1.267
1.0 -1.215 -0.027 -0.245 -0.366 -0.099 -0.69
2.0 -1.114 0.284 0.005 -0.005 0.207 -0.411
3.0 -0.963 0.461 0.146 0.204 0.396 -0.292
4.0 -0.526 0.568 0.266 0.333 0.533 -0.229
Table 3.11b: Freundlich isotherm constants
29
Ions n 1/n Kf (mg/g) R2
Pb2+ 0.970 1.031 17.66 0.731
Cd2+ 1.098 0.911 1.06 0.916
Ni2+ 0.891 1.122 5.38 0.916
3.5 The Pseudo-first order
The pseudo-first order rate equation (Lagergreen equation) is generally expressed as 18.
Table 3.12a: Data for pseudo-first –order
Log (Qe-Qt)
2+
Time(min.) Pb Cd2+ Ni2+
10 -0.5229 -0.6861 -0.4802
30 -0.6556 -0.7989 -0.5719
50 -1.0088 -0.9626 -0.8186
70 -1.2924 -1.1079 -1.2441
90 -2.0000 -2.6989 -1.4685
Table 3.12b: Constants of pseudo-first-order
Ions Kt (min-1) Qe (mg/g) R2
Pb2+ 0.041 0.634 0.930
Cd2+ 0.048 0.681 0.689
Ni2+ 0.030 0.557 0.960
3.6 The pseudo-second-order kinetics
The pseudo-second-order rate equation (Lagergreen equation) is generally expressed as 18.
Table 3.13a: Pseudo-second-order reaction
Time(min.) t/Qt
2+
Pb Cd2+ Ni2+
10 192.31 105.26 86.96
30 129.87 211.27 168.54
50 141.24 260.42 170.17
70 174.54 313.90 179.95
90 203.62 301.00 218.45
It was observed that cadmium with R2=0.854 followed mostly the pseudo-second-order reaction than the
pseudo-second-order reaction than the pseudo-first-order reaction.
Table 3.13b: Constants of Pseudo-Second-Order
Ions K2(mgg-1 min-1) Qe(mg/g) R2 ho(mgg-1min-1)
Pb2+ 7.46x10-4 2.976 0.111 6.61x10-3
Cd2+ 53.11x10 -3 0.405 0.854 8.7x10-3
Ni2+ 19.56x10-3 0.729 0.817 1.039x10-2
30
Conclusion and Recommendation
This research work geared towards investigating the sorption capacity of modified coconut shell activated
carbon (MCSAC) for Pb2+, Cd2+ and Ni2+ from aqeous solution conclusively and it also showed that the activated
carbon prepared from coconut shell, a local agricultural waste could be used as a potential adsorbent for the
2+ 2+ 2+
removal of Pb , Cd and Ni ions from aqueous solution and it is inexpensive materials for treating the industrial
wastewater. Kinetic studies showed that the sorption of the metal ions can best be described by pseudo-first-order
2+ 2+ 2+ 2+
( for Pb and Ni ), pseudo-second-order ( mostly for Cd and Ni ) and intra-particle diffusion models for the
three metals. Modified coconut shell activated carbon is a good adsorpbent and its adsorption capacity would be
improved by an ligand in a right proportion, probably using adsorbent; ligand ratio of (5mg:25mg). Different
adsorption temperatures using modified adsorbent should be investigated to see their effects on percentage
removal of metal ions from the aqueous solution.
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and their metal complexes 1:1, 2 –dihydroxy-1, 5-dimethyl-2-phenyl-4-[E]-(2,3,4-trihydroxyphenyl)-3H-
pyrazol-3-one(H3L) and its Co(II), Fe(III) and Os(VII) Complexes,Inter.J.Chem, 20(4):217-225.
15). Low,K.S., Lee,C.K.,Leo,A.C. (1995).Bioresource Tech.5,227.
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16). Day, J.H., P.J. Rennie,W.Stanek and G.P.Raymond, Ed.(1979). Peat testing manual.Ottawa,National
Research Council Canada.
17). Agyei, N.M.; Strydom,C.A. and Potgieter, J.H.(2000). An investigation of phosphate ion adsorption from
aqeous solution by fly ash and slag. Cem. And Concr. Res., 30 (5):823-826.
18). Lagergren,S. (1898). About the theory of so-called adsorption of soluble substances. K.Sven.
Vetenskapsakad. Handlingar Band,24,1-39.
32
Delonixregia extracts as non – toxic corrosion inhibitor for acid corrosion of mild steel in HCl solution
Olusegun K. Abiola1 *,Abdulraman O.C. Aliyu2and Suleiman Muhammed3
1
Chemistry Department, Federal University of Petroleum Resources, Effurun, Nigeria
2, 3
Chemistry Department, Kogi State University, Anyigba, Nigeria
1*
abiolaolusegun@yahoo.com
* Corresponding author.
Abstract
The corrosion inhibition properties of Delonixregia leaf extract (DLE), flower extract (DFE) and seed extract
(DSE) in 0. 5 M HClsolutions were studied using weight loss technique. Delonixregia extracts inhibited the
corrosion of mild steel in HCl solution. The inhibition efficiency increased with increasing concentration of
0
the extracts at 30 C. The seed extract (DSE) was found to be more effective than both the leaf (DLE) and
flower extracts (DFE) at the lowest concentration. The results obtained revealed that the adsorption of the
inhibitor molecule onto mild steel surface accords with Lanqmuir adsorption isotherm.
Keywords: Acid corrosion, Adsorption, Corrosion inhibitor, Mild steel, Delonixregia.
Introduction
The use of synthetic compounds as corrosion inhibitors are desirable due to their metal protecting
properties , however, the problem of toxicity, non-degradability and environmental pollution posed by
these compounds have been of serious concern [ 1, 2 ].
Consequently, attention has been focused on the need to design and develop green or non-toxic corrosion
inhibitor to replace toxic ones for a sustainable development [2 - 7]. In our previous works [5, 6], we studied
the effect of Delonixregia extract on acid corrosion of aluminium in HCl solution using chemical techniques.
The Delonixregia extract inhibited the acid corrosion of aluminium in HCl solution and the inhibitive action
was ascribed to the presence of phytochemicals in the extract.
In furtherance of our interest on the development of non- toxic corrosion inhibitors, this paper reports the
investigation on the inhibitive effects of the extracts of leaf, flower and seed of Delonixregia on the acid
o
corrosion of mild steel in 0.5 M HCl solution at 30 C.
The test specimens of dimensions 5 x 2 x 0.07 cm were mechanically press-cut from mild steel sheet of
0.04cm in thickness and 97.9% purity. The mild steel samples were prepared, degreased and cleaned as
described earlier[7, 8]. HCl was of analytical grade and 0.5 M HCl was employed as the aggressive solution
for this study. The stock solution of the plant extract was prepared as reported previously [5,6]. The stock
solution of the extract was diluted with appropriate quantity of 0.5 M HCl solution to obtain inhibitor test
solutions of 50 – 1000 mg/L concentrations. The procedure for weight loss determination was similar to
that reported earlier [7, 8].
According to this method [7, 8], previously weighed mild steel coupons were immersed in 100 ml open
beakers containing 100 ml of 0.5 M HCl (blank) and then with addition of different extract concentrations
to the 0.5 M HCl (50 – 1000 mg/L) at 30 ±0.3oC. The variation of weight loss was monitored after 4 hour
o
immersion per coupon progressively for a total of 24 hour at 30 C. The weight loss was calculated in mg as
the difference between the initial weight and the weight after the removal of the corrosion product. The
experimental readings were recorded to the nearest 0.0001 g on a Mettler digital analytical balance (digital
analytical balance with sensitivity of ±1 mg). Duplicates experiments were conducted for each
concentration of the extract.
Results and discussion
The results obtained are presented in Figures 1 – 2 and Table 1 for three different extracts of Delonixregia from
weight loss measurements.
33
Figure 1. Relation between material loss and extract concentration for mild steel in 0.5 M HCl solution for 4 h
immersion period.
-2
The amount of material loss (mg cm ) decreases significantly with increasing concentration of DLE, DFE and DSE
extracts, as presented in Figure 1. The addition of the three extracts resulted in noticeable reduction in the
o
amount of material loss from the surface of the mild steel in comparison with that of control (0.5 M HCl) at 30 C.
-
As seen in Figure 1, the weight loss decreased by a factor of 2.9, 5.8 and 8.7 over that of control at 100 mg L
1
concentration for the three extracts. This indicates that the three additives inhibit the acid corrosion of mild
steel in HCl solutions.
The values of percentage inhibition efficiency (% E) and surface coverage (è) were determined for 4 h
immersion periods from the material loss using the following equations [9]:
E% = [ wu - wb/wu] x 100 (1)
è = E % / 100 (2)
Where;wu andwbare the uninhibited and inhibited weight loss respectively.
Table 1. Percentage inhibition efficiencies of mild steel immersed for 4 hours at 30oC in 0. 5 M HCl solution
in the presence of three different extracts of Delonixregia.
Percentage inhibition efficiency at different concentrations of inhibitor

Inhibitor 50 mg/ L 100 mg/L 300 mg/L 500 mg/L 700 mg/ L 1000 mg /L
DLE 61.1 65.3 86.3 89.3 90.5 92.5

DFE 78.2 82.6 88.3 88.3 89.4 90.8
DSE 88.3 88.6 91.9 92.0 92.6 92.7
34
The values of percentage inhibition efficiency at different extract concentration are listed in Table 1. Table 1
indicates that Delonixregia extracts act as good corrosion inhibitor for the acid corrosion of mild steel in 0.5 M
HCl solution. The % inhibition efficiency increases with increasing extracts concentration and % inhibition
efficiencies were relatively high in 0.5 M HCl solution (65.3% for DLE, 82.6% for DFE and 88.6% for DSE) at 100
mg L-1 of extract concentration. The seed extract (DSE) was found to be more effective than both the flower
-1 -1
(DFE) and leaf (DLE) extracts at lower concentrations of 50 mg L and 100 mg L .
The Delonixregia extract contains 4-hydroxybenzoic acid, gallic acid, 3,4- dihydrroxycinnunmic acid, 3,5-
dinitrobenzoic acid, alkaloid, L-azetidine-2-carboxylic acid, amine base , 3,4-diydroxybenzaldehyde,
chlorogenic acid, kaempferol-3-glucoside, cyanidine-3-O-glucoside, cyanidine-3-O-rutinoside and coumarin [
10, 11].
The inhibition properties of Delonixregia is ascribed to the presence of these organic compounds in the
extracts. Organic compounds having centers for pie electrons and functional groups -OR, -NR2 and /or –SR
have been reported as corrosion inhibitors for metals in acid solutions [12- 14]. The adsorption of these
compounds on the metal surface reduces the surface area that is available for the attack of the aggressive ion
from the acid solution. The material losses decrease with increase in extract concentration due to higher
degree of surface coverage as a result of enhanced inhibitor adsorption (Figure 1). Similar view has been
expressed in our previous reports [2 – 5, 9, 12, 13] and other workers on inhibition of metals in acid solutions by
plant extracts [14,15].
Values of è were tested graphically for fit to different isotherms. As presented in Figure 2, straight line is
obtained when C/è isplotted against C and the linear correlation coefficients of the fitted data are good
(? 0.998).
35
Figure 2. Langmuir adsorption model on the mild steel surface of Delonixregia extracts in 0.5 M HCl solution for 4
o
h immersion period at 30 C.
This confirms that the inhibition is due to the adsorption of the active organic compounds onto the metal
surface and the adsorption obeys the Lanqmuir's adsorption isotherm [2 - 6] expressed as:
(3)
Where C is the inhibitor concentration and K the equilibrium constant for the adsorption/ desorption of process
of the inhibitor molecules on the metal surface.
Conclusions
§ Results obtained revealed that the three extracts of Delonixregia act as efficient corrosion inhibitors of
the mild steel in acid solution.
§ The inhibitory action of the extract is ascribed to the adsorption of the phytochemicals in the plant.
§ The adsorption of inhibitors on the mild steel surface follows Langmuir adsorption isotherm.
§ Delonixregia extract can be added to acid solution as a non-toxic corrosion inhibitor for mild steel in HCl
solution.
Acknowledgements
This research was supported by Tertiary Education Trust Fund (TETFUND), TETFUND Research
th
Projects (Batch 5 of TETFUND 2013-2014 Research Projects) of the Federal Republic of Nigeria,
through the Federal University of Petroleum Resources, Effurun, Nigeria for which OKA is highly
grateful.
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for aluminium in sulphuric acid: Correlation between inhibitive effect and electronic properties of
extracts of major constituents using density functional theory. Arabian Journal of Chemistry, 5, 351 –
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2. Babatunde, A. I., Oyelola, O., Oyelola, O. T., John, O. M., Abiola, O. K., The inhibitive effect of
Irvingagabonensis extract on the corrosion of aluminium in HCl solution, Proceedings of the 34th
international conference of the Chemical Society of Nigeria, 2011, 392 -396.
3. Abiola, O. K. and Tobun Y. (2010). Cocos nucifera L. water as green corrosion inhibitor for acid
corrosion ofaluminium in HCl solution. Chinese Chemical Letters, 21, 1449-1452.
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2methyl-5-pyrimidylmethyl)-4-methylthiazolium chloride as green corrosion inhibitor of copper in
HNO3 solution and its adsorption characteristics. Green Chemistry Letters and Reviews,4, 273 – 279.
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the inhibitive action of DelonixRegia on the corrosion of aluminium in acidic media. Anti – Corrosion
Methods and Materials, 54, 219 – 224.
6.Abiola, O. K., Aliyu, A. O. C., Muhammed, S. (In press, 2016). Eco-friendly corrosion inhibitors:
Effect of Delonixregia extract on corrosion and kinetics of corrosion process of aluminium alloy 2S in
HCl solution. African Corrosion Journal.
7. Abiola, O. K and Oforka, N. C. (2002).Inhibition of the Corrosion of mild steel in hydrochloric
36
acid by (4-amino-2-methyl-5-pyrimidinyl methylthio) acetic acid and its precursor', Journal of
Corrosion Science and Engineering 3, 1-10.
8. Abiola, O. K. and Oforka, N. C (2004).Adsorption of (4-amino-2-methyl-5-pyrimidinyl
methylthio) acetic acid on mild steel from Hydrochloric acid solution (HCl) – part 1. Materials
Chemistry and Physics 83, 315-322.
9.Abiola, O. K., Odin, E. M.,Olowoyo, D. N and Adeloye T. A (2011).Gossipiumhirsutum L. extract as
green corrosion inhibitor for aluminium in HCl solution. Bull. Chem. Soc. Ethiop., 25, 475-480.
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Chemical Ecology, 18, 2285.
11. Sharma, S., Arora, S., Phytochemicals and pharmaceutical potential of Delonixregia (Bojer ex hook).
International Journal of Pharmacy and Pharmaceutical Sciences, 7, 2015, 17-29.
12. Abiola, O. K., Aliyu, A. O. C, Elemike, E. E and Adeyemi, O. (2015). Eco-friendly corrosion inhibitor
from Tagetespatula L. for aluminium alloy (3SR) in acid fluid used in industrial operations. NISEB
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corrosion process of zinc in HCl solution. Corrosion Science, 52, 661 – 664.
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Inhibition action of some plant extracts on the corrosion of steel in acidic media. Corrosion Science,
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37
Proceedings of Annual International Conference of Chemical Society of Nigeria (CSN)
ASSESSMENT OF SOME METALS IN ROADSIDE DUST FROM DAMATURU, YOBE STATE,

NIGERIA
Zaynab Muhammed Chellube, Joseph Clement Akan and Emmanuel Mshelia
Department of Chemistry, University of Maiduguri, P.M.B 1069, Maiduguri, Borno State, Nigeria
*Corresponding Author: E-mail: joechemakan@yahoo.com
Abstract
Roadside dust samples were collected from commercial and residential areas in Damaturu, Yobe State,
Nigeria for the determination of Cr, Pb, Cu, Fe, Ni, Co, Mn, Cd, As and Zn. Sample preparation and
digestion were carried out using standard procedures. The samples were determined using flame atomic
absorption spectrophotometry after digestion with aqua regia. The levels of Cr, Pb, Cu, Fe, Ni, Co, Mn,
Cd, As and Zn were found to be significantly higher in the roadside dust samples from commercial area,
while the lowest levels of metal ions were noted in the Roadside dust samples from residential area. The
results further shows that the concentrations of all the metals were significantly higher than the control
points. From the results obtained the concentrations of all the studied metals were generally above the
WHO standard values for Roadside dust. Hence, the traffic situation in the commercial area might be
regarded as the sources of high heavy metals content in the roadside dust when compared with the
residential area.
Keywords: Heavy Metals, Traffic, FAAS, WHO
Introduction
Vehicular emission and traffic congestions are major source of road side soil pollution (World Health
Organization (WHO, 2006). Motor vehicles have directly and indirectly impacted on the metabolism of
roadside plants due to emission of trace elements (Viskari et al., 2000). There has been interest in the levels of
trace elements and their source identification associated with urban dusts in past decades (Chatterjee & Banerjee,
1999). Research has established people exposed to trace elements such as lead (Pb), Cadmium (Cd), Arsenic
(As), manganese (Mg) and nickel (Ni) develop alterations in nervous system functions, with neurophysiological
consequences constituting a health hazard (Olivero and Solano, 1998). Many studies throughout the world have
identified vehicular traffic, industrial and commercial areas as sources of trace elements in urban dusts (Al-
Chalabi and Hawker, 1997) as well as weathering or building facades (Akhter and Madany, 1993). Such toxic
elements (e.g. Pb, Cd, As, Mg, and Ni) are deposited on the roadside might be as a result of combustion,
component wear, fluid leakage and corrosion of metals. Most heavy metals are release as a result fuel burning,
tyres wearing, oils leakage and battery corrosion. Majority of heavy metals are toxic to living organisms and
even those considered essential can be toxic if present in excess (Akhter and Madany, 1993). Trace elements can
impair important biochemical processes posing a threat to human health, plant growth and animal life (Silva et
al., 2005). Research from other studies showed heavy metals can be harmful to roadside plants, animals, and
human settlements situated closer to a roads (Akbar et al., 2006).
Sampling Collection
Roadside dust samples were collected from commercial and residential areas. For each of the sampling
areas, samples were also collected 200 meters away from each sampling point to serve as a control. Roadside
dust samples were collected from the two areas and control points by using brush and plastic hand trowel to
collect settle dust along the roads.
38
Samples Preparation
Two grams of each air-dried and sieved dust samples was ashed in a muffle furnace at 500 °C for 4
hours. The ash was digested in 10 ml Aqua regia (1 part conc. HNO3 to 3 parts HCl) in a digestion tube on a
hot plate until the volume reduced to 5 ml as the heating continued. After the digestion, the digestates were
centrifuged and 10 ml HNO3 was added. Samples were cooled and then made up to the volume of 100 ml with
distilled water. The concentrations of Cr, Pb, Cu, Fe, Ni, Co, Mn, Cd, As and Zn in the Road site dust samples
were determined using Atomic Absorption Spectrophotometer (AAS, Unicam 969).
Results
The concentrations of heavy metals in Roadside dust samples from commercial area and control is as
presented in Figure 1. The concentration of Cr ranged from 1.23 to 8.22 mg/kg; 2.34 to 20.12 mg/kg Pb; 1.98
to 10.34 mg/kg Cu; 3.45 to 34.23 mg/kg Fe; 4.34 to 14.34 mg/kg Ni; 0.88 to 7.87 mg/kg Co; 1.09 to 6.34
mg/kg Mn; 1.54 to 9.23 mg/kg Cd; 0.19 to 3.98 mg/kg As and 2.34 to 32.44 mg/kg. Figure 2 present the mean
concentrations of some heavy metals in Roadside dust samples from residential area. The concentrations of
heavy metals in Roadside dust samples from residential area and control is as presented in Figure 2. The
concentration of Cr ranged from 0.11 to 3.22 mg/kg; 0.21 to 5.23 mg/kg Pb; 0.01 to 2.10 mg/kg Cu; 0.23 to
6.23 mg/kg Fe; 0.11 to 2.12 mg/kg Ni; 0.34 to 2.14 mg/kg Co; 0.02 to 1.02 mg/kg Mn; 0.13 to 1.22 mg/kg Cd;
0.21 to 2.88 mg/kg As and 0.12 to 6.33 mg/kg.
39
Discussion
Lead (Pb)
The highest Pb concentration in the road side dust was 32.44 mg/kg and was detected at the commercial
area, while the residential area shows the least value. The control points shows the lowest levels of lead as
compared to the main roads. The high concentration of Pb at commercial area when compared to other point
might be due to the differences in traffic density, metals construction work, Iron bending and welding of metals
which is a common practice along the street of the commercial area. Results also show that the levels of Pb in
dust samples were higher than the control. High values of Pb when compared to control are because all the
sample points were in the areas of high traffic density. The traffic situation in this area might be regarded as a
source of Pb in the roadside dust. Wear and corrosion of vehicle parts might also be one of the potential sources
of heavy metals. Concentrations of Pb in the dust samples from the study areas were lower than that reported
by Charlesworth et al. (2003) of 48 ìg/g, and was also higher than the range of 1.01–2.9 ìg/g reported by Al-
Khashman (2006).
Copper (Cu)
Copper may be toxic to both humans and animals when its concentration exceeds safe limits (Bakýrdere
and Yaman, 2008). The highest Cu concentration in roadside dust was found at commercial area, while
residential shows the lowest value. The high concentration of Cu at commercial area when compared to other
point might be due to the differences in traffic density and metals construction work. High values of Cu in the
sampling points when compared to the controls might be due to corrosion of metallic car parts (Al-Khashman,
2004; Al-Khashman and Shawabkeh, 2006).
Iron (Fe)
Iron is vital for almost all living organisms, participating in a wide variety of metabolic processes,
including oxygen transport, DNA synthesis, and electron transport. It is known that adequate Fe in a diet is
very important for decreasing the incidence of anemia (Lynch and Baynes, 1996). High concentration of Fe in
dust samples was observed at the commercial area, while residential area shows the least concentration. High
concentration of Fe detected at the commercial area may be attributed to differences in traffic density. (Adachi
and Tainosho, 2004).
Nickel (Ni)
Nickel pollution on a local scale is caused by emissions from vehicle engines that use gasolene which
contains Ni and by the abrasion and corrosion of Ni from vehicle parts (Al-Shayeb and Seaward, 2001). The
burning of fossil fuels as well as the refining of metals such as Cu introduces considerable amounts of Ni into
the atmosphere (Lee et al., 2005). The highest Ni concentration (6.82µg/g) in the dust samples was found at the
commercial area, while residential the least concentration. Nickel values in the dust samples from the sample
points were found to be higher than the control sites and may be the result of high traffic density within the
study area.
Manganese (Mn)
The deficiency of manganese in the human body can produce severe skeletal and reproductive
abnormalities in mammals. High doses of Mn produce adverse effects primarily on the lungs and brain. The
highest Mn concentration in the dust samples was observed at the commercial area, while the least value
detected at the residential area.
Cadmium (Cd)
It was reported that cadmium is accumulated mainly in kidneys, spleen, and liver, and its blood serum
level increases considerably following mushroom consumption (Kalac and Svoboda, 2001). Cadmium is now
most commonly encountered in cadmium–nickel battery production, although it continues to be used in paints
40
as well as in plastic production where it is an effective stabilizing agent. Highest Cd concentration in dust
samples was observed at the commercial area, while residential shows the least concentration. Cadmium values
in dust samples from the sample points were found to be higher than the control site; such variation might be
attributed to high traffic density. The concentrations of Cd in the roadside samples world-wide has been reported
to be 0.5–4.0 ìg/g (Fergusson & Kim, 1991). Cadmium levels in dust samples in the commercial area exceeds
this ranged.
Zinc (Zn)
The highest Zn concentration in dust samples was observed at the commercial area, while residential area
shows the lowest value. The traffic situation in the study area is regarded as a source of zinc in the roadside dust.
Wear and corrosion of vehicle parts (brakes, tyres, radiators, body, and engine parts) might also be one of the
potential sources of Zn in roadside dust. Zn values in the dust samples from the sample points were found to be
higher than the control sites.
CONCLUSION
In all the sampling points, Fe, Zn and Pb show the highest concentrations in roadside dust. The high
concentration of these metals in the roadside dust samples may be attributed to metals construction work, Iron
bending and welding of metals. At the same time, the traffic situation at the commercial area of study might be
regarded as a source of heavy metal content in the roadside dust. The concentrations of all the metals in the
sampling points were higher than that the control.
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Elazig, Turkey. Environmental Monitoring and Assessment, 136(1-3), 401-410.
41
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42
ANTI-OXIDANT AND ELEMENTAL ANALYSIS OF THE FRUIT OF SARCOCEPHALUS
LATIFOLIUS SMITH BRUCE (SMITH BRUCE)
*Isah Yinusa, Woli Ajoke Balkees and Hassan Adam Adeiza

Department of Chemistry Federal University, Lokoja, Kogi State, Nigeria
*Corresponding author E-mail: Isahyinusa@gmail.Com
Abstract
This work was aimed at examining the anti-oxidant activities and elemental composition of the fruit of
Sarcocephalus latifolius collected from zone eight (8) area of lokoja in Kogi state. The radical scavenging
abilities of the Sarcocephalus latifolius fruit on stable 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical were
qualitatively and quantitatively determined.The presence and concentration of pharmacologically important ten
(10) selected metallic elements which are; Cupper Cu); Manganese (Mn); Zinc (Zn); Iron (Fe); Cobalt (Co);
Nickel (Ni); Lead (Pb); Chromium (Cr); Cadmium (Cd) and Magnesium (Mg) were analyzedbythe means of
Atomic Absorption Spectrophotometer (AA240FS) Model.The methanolic extract was profound to have
reducing activity against stable freeradicals as compare to the standard (ascorbic acid).The fruit was found to
contain some valuable trace essential minerals element that could be of good health benefit. Despite that the
recommended limit for dietary consumption and average dietary daily requirement in this fruit may be lower
than expected.
Keyword: Anti-Oxidant, Sarcocephalus, Elemental, Fruit, Activity, Scavenging, Radical, Diet

Introduction
Antioxidants are substances which protect cells against the damaging effects of reactive oxygen species
otherwise called, free radicals such as singlet oxygen, super oxide, peroxyl radicals, hydroxyl radicals and
peroxynite which results in oxidative stress leading to cellular damage [1]. Natural antioxidants play a key role in
health maintenance and prevention of the chronic and degenerative diseases, such as atherosclerosis, cardiac and
cerebral ischemia, carcinogenesis, neurodegenerative disorders, diabetic pregnancy, rheumatic disorder, DNA
damage and ageing [2]. Antioxidants exert their activity by scavenging the 'free-oxygen radicals' thereby giving
rise to a fairly 'stable radical'. The free radicals are metastable chemical species, which tend to trap electrons from
the molecules in the immediate surroundings. These radicals if not scavenged effectively in time, may damage
crucial bio molecules like lipids, proteins including those present in all membranes, mitochondria and, the DNA
resulting in abnormalities leading to disease conditions [2]. Thus, free radicals are involved in a number of
diseases including: tumor inflammation, hemorrhagic shock, atherosclerosis, diabetes, infertility,
gastrointestinal ulcerogenesis, asthma, rheumatoid arthritis, cardiovascular disorders, cystic fibrosis,
neurodegenerative diseases (e.g. Parkinsonism, Alzheimer's diseases), AIDS and even early senescence [2, 3].
The human body produces insufficient amount of antioxidants which are essential for preventing oxidative
stress. Free radicals generated in the body can be removed by the body's own natural antioxidant defenses such as
glutathione or catalases [4]. Therefore this deficiency had to be compensated by making use of natural
exogenous antioxidants, such as vitamin C, vitamin E, flavones, â-carotene and natural products in plants [5, 6].
Antioxidants are often added to foods to prevent the radical chain reactions of oxidation, and they act by
inhibiting the initiation and propagation step leading to the termination of the reaction and delay the oxidation
process. Due to safety concerns of synthetic compounds, food industries have focused on finding natural
antioxidants to replace synthetic compounds. In addition, there is growing trend in consumer preferences for
43
natural antioxidants, all of which has given more impetus to explore natural sources of antioxidants.
Evaluation of elemental levels of fruits is a growing trend in nutritional studies throughout the world. Some of
these element (Trace) do not provide any calorie but they play an important role in the metabolic regulations of
the human body. Increased fruit consumption can improve the mineral regulation and reduce cardiovascular
diseases and certain cancer risks
Numerous studies have shown that fruits and vegetables are rich sources of nutrients as well as non-nutrient
molecules with antioxidant or other physiological effects, and it seems likely that given sufficient bioavailability,
these compounds may be important constituents of a healthy diet. The health promoting properties of plant-based
foods have largely been attributed to their wide range of phytochemicals, many present at relatively high
concentration [7].
Sample collection and preparation
The plant matured fruits of Sarcocephalus latifolius was collected from Crusher Area, Zone 8 in Lokoja, Lokoja
Local Government Area of Kogi State, Nigeria in November 2015. The samples were transported to the
Chemistry laboratory in an air-tight polyethylene bags. These were identified at the Herbarium of the faculty of
sciences, Federal university Lokoja, Kogi State. The voucher number (0034) of the sample was assigned. The
samples were air- dried for two to three weeks in the laboratory and afterward pulverized using mortar and pestle
to fine particles and stored in an air tight polythene bags and kept away from moisture for further analysis.
Procedure for anti-oxidant
The radical scavenging abilities of the Sarcocephalus latifolius fruit on stable 1, 1-diphenyl-2-picrylhydrazyl
(DDPH) radical were qualitatively and quantitatively determined.
Qualitative Antioxidant Assay
Dilute solutions of the Sarcocephalus latifolius root extract was prepared with methanol and spotted on pre-coat
silica gel TLCplates with capillary tube. The TLC plates were developed in solvent systems of different polarities
(polar-Methanol/Ethyl acetate; 1:2) to resolve polar and non-polar component of the extract. The plates were
allowed to dry at room temperature and sprayed with 0.02% DPPH in ethanol. Bleaching of DPPH by the
resolved spots were observed for 10 minutes and after then colour changes were observed for the spots (yellow
on purple background).
Quantitative Antioxidant Assay
Quantitative assay was done to determine the quantity of antioxidants (in percentage) in the Sarcocephalus
latifolius fruit extracts. Stock solution (1mg/ml and 0.8mg/ml) of the Sarcocephalus latifolius root extracts were
prepared in ethanol and serial dilutions were made to obtain for 50, 100, 125, 200, 250, 400 and 500µg/mL
solutions. Free radical scavenging activity of the extracts was determined by 1,1-diphenyl-2-picrylhydrazyl
(DPPH) (Aldrich, USA) using Blois method (1958), modified by [8]. Dilute extract solutions (2ml) were added
to 2ml of 0.1mM of ethanol solution of DPPH. The mixture was mixed thoroughly and kept in the dark for 30
min. The absorbance was measured at 517nm and from the values obtained, corresponding percentage of
inhibition were calculated. The percentage inhibition was plotted against log of concentration and from the graph
IC50 was calculated. Ascorbic acid was used as a standard inhibitor while DPPH solution was used as control. All
the tests were performed in triplicates and the graph was plotted with mean values. The percentage inhibition was
calculated using the formula:
44
Table 1: Anti-oxidant Activity of Methanolic Extract of the Fruit of Sarcocephalus latifolius
S. latifolius Extract Standard (Ascorbic
Concentration Acid)
(µg/ml) % Inhibition IC50(µg/ml) % Inhibition IC50(µg/ml)
100 78.012 107.08
200 76.82 107.08
400 78.947 107.08
1.52 107.08
500 71.053 107.08
800 76.535 107.08
1000 82.383 107.08
The IC50of S. latifoliuswas obtained by linear regression equations: y = 4.3145x + 43.445
The following results were obtained for each of the element so analyzed. Copper (Cu) was 0.368ppm,
Manganese (Mn) 1.185ppm, Zinc (Zn) 0.665ppm, Chromium (Cr) 0.003ppm, Iron (Fe) 2.051ppm, Cobalt (Co)
0.000ppm, Nickel (Ni) 0.091ppm, Cadmium (Cd) 0.029ppm, Lead (Pb) 1.333ppm and Magnesium (Mg)
46.213ppm. Summary of the results are as presented in table2 and figure 1. This result shows that the fruit contain
some valuable trace essential minerals that is of good health benefit. Despite that the recommended limit for
dietary consumption and average dietary daily requirement in this fruit may be lower than expected.
Table 2: Atomic Absorption Spectroscopy (AAS) Result For the Fruit Sarcocephalus Latifolius (Smith Bruce)
Element Concentration Standard Mean

(ppm) Deviation Absorbance
Cu 0.368 0.0006 0.0267
Mn 1.185 0.0026 0.0723
Zn 0.665 0.0276 0.0793
Cr 0.003 0.0004 -0.0012
Fe 2.051 0.0027 0.045
Co 0.000 -0.0002 0.0454
Ni 0.091 0.0012 0.0034
Cd 0.029 0.0003 0.0029
Pb 1.337 0.0005 0
Mg 46.213 -0.0011 0.0016
45
Figure 1: Atomic Absorption Spectroscopy (AAS) Result For the Fruit Sarcocephalus Latifolius (Smith
Bruce)
ConclusionThe antioxidant result shows that the fruit of the plant contain high level of antioxidant agent that can
be useful in health care management system. Since this fruit is believed to cure some certain ailment such as
cough and host of other ailment. This work has really shown the importance of the fruit as a good source of
natural anti-oxidant and some important macro element that are needed in the body system.
Acknowledgement: The authors wishes to thank all Staff members of Chemistry laboratory department Federal
University, Lokoja Kogi State for providing enabling environment for this research work. Our profound
gratitude goes to Professor Dosumu of department of Chemistry University of Ilorin who help in one way or the
other to see that the ant oxidant activity of the extract was carried out.
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Activity of Leaf Extracts from Different Hibiscus sabdariffaAccessions and Simultaneous
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Research, 10, 27-33.
46
PROXIMATE, MINERAL AND ANTI-NUTRIENT COMPOSITION OF Allium sativum (GARLIC)
CONSUMED IN UYO, AKWA IBOM STATE, NIGERIA
* E. I. Uwah and E. A. Moses

Department of Chemistry, University of Uyo, P.M.B. 1017 Uyo, Nigeria
* Corresponding author: emmanueliuwah@uniuyo.edu.ng; +2348028251882
Abstract
The proximate, mineral and anti-nutrient compositions of Allium sativum samples consumed in Uyo, Akwa
Ibom State, Nigeria were investigated using standard methods to ascertain the suitability of the vegetable for
human consumption for both nutritional and medicinal purposes. The results (%) for the analysis of proximate
composition revealed that moisture content was 68, crude protein 24.33, ash content 2.5, crude fat 0.5, crude
fibre 0.5 and carbohydrate 72.1. The caloric value of the vegetable samples was found to be 390.22 kcal100g.
The results (mg/100g) for the analysis of some mineral elements indicated that calcium was 180, magnesium
23.4, iron 1.7, manganese 1.54, potassium 401 and zinc 1.16. The anti-nutrient analysis showed the anti-nutrients
composition (mg/100g) to be: hydrogen cynide 0.30, oxalate 234.6, tannin 0.12, phytate 2.14 and saponin 12.60.
The appreciable levels of the proximate and mineral elements composition as well as the high caloric value of the
vegetable together with its low anti-nutrients composition (except for the high oxalate level of 234.6 mg/100g)
which could be reduced by proper processing, make the vegetable suitable for human consumption for both
nutritional and medicinal purposes.
Keywords: Allium sativum, proximate, mineral, anti-nutrient, composition, consumption, Uyo.
Introduction
Garlic (Allium sativum) is a bulb of lily like plant belonging to the same family as onion. It is commonly used as a
flavouring agent and herbal supplement and is one the vegetable substances used as spices to season foods for the
purpose of adding or imparting agreeable flavour and aroma. Garlic like other vegetable species, has the ability
to stimulate appetite and increase the flow of gastric juice, making it food assessor or adjunct [1, 2]. Garlic is
characterized by the remarkable sulphur containing compound which gives distinctive smell and pungency.
Uninjured bulb of garlic contains allinase, one of the major protein components of the plant [3]. Apart from being
used to improve sensory quality of food, it is used in perfumeries, cosmetics and toiletries. The vegetable has
long been recognized to possess medicinal properties, such as tonic, carminative and antihelminthic [1].
Being a vegetable, garlic should contain nutrients which can be absorbed by the body to be used as
energy sources, body building, regulatory and protective material [4, 5]. These nutrients are carbohydrates,
lipids, proteins, ash, fibers, vitamins and moisture. Indeed like other vegetables, garlic constitutes an important
part of the human diet [5, 6]. As vegetable, garlic is also expected to contain anti - nutritional factors such as
oxalate, cyanide, tannin and pytate that can affect the availability of the nutrients. Oxalate can bind to calcium
present in foods thereby rendering calcium unavailable for normal physiological and biochemical roles such as
the maintenance of strong bone, teeth, cofactor in enzymatic reaction, nerve impulse transmission and as clotting
factor in the blood [7]. It is reported that calcium oxalate, which is insoluble, may also precipitate around soft
tissues such as the kidney, causing kidney stones [5, 8]. It is also known that phytic acid can bind to mineral
elements such as calcium, zinc, manganese, iron and magnesium to form complexes that are indigestible, thereby
decreasing the bioavailability of these elements for absorption [5, 9]. Accordingly, Phytate is known to be
associated with nutritional diseases such as rickets and osteomalacia in children and adult respectively [5].
Proximate composition includes seven constituents, namely: moisture content, ash content, fibre,
protein, lipids, carbohydrates and caloric value. Proximate analysis is done to estimate the relative amounts
47
of protein, lipid, moisture, ash, fibre and carbohydrate in materials such as vegetables and other plants
based on the chemical properties of the materials [5].
Garlic is one of the spices vegetables widely cultivated and consumed in Nigeria including Akwa Ibom
State. This study is aimed at analysing the proximate, mineral and anti – nutrients composition of garlic (Allium
sativum) obtained in Uyo, Akwa Ibom State with a view to establishing the chemical, nutritional and
toxicological properties of the vegetable, thereby ascertaining it suitability for human consumption (both for
nutritional and medicinal purposes) or otherwise, which will go a long way in the nutritional education of the
public with a view to improving the nutritional status of the populace.
Collection and Preparation of Sample
Allium sativum (garlic) cloves were purchased from the Akpan Andem Market in Uyo, Akwa Ibom State. The
vegetable materials were procured in their unprocessed and fresh states. They were picked randomly in small
quantities from different sellers and pooled together to obtain a composite sample. The light scaly leaves on the
garlic cloves were removed and the naked cloves washed in water before being chopped into tiny pieces. The
prepared sample materials were then dried in an air-circulating oven in the laboratory and ground manually into a
fine power, using a manual grinder. The powdered sample materials were sieved through mesh (300ìm) and
stored in an air – tight cellophane bag in refrigerator prior analyses.
Analytical procedures
Proximate Analysis
The proximate composition of the vegetable sample materials was determined in accordance with the
standard methods described by [10]. Thermal drying method was used in the determination of moisture
content of the sample. Dried weight (2.0g) of sample was weighed in triplicate and placed in washed, dried
and weighed crucible. This was placed in an oven and dried at 105°C for three hours. The sample was
allowed to cool in a desiccator and then reweighed. The percentage moisture content was calculated by
computing or expressing the loss in weight on drying as a fraction of the initial weight of sample used and
multiplied by 100 as indicated below:
Moisture (% wet weight) = b – c x 100 ........................................................................................ (1)
b–a
Where: a = weight of crucible
b = weight of crucible + sample before oven drying
c = weight of crucible + sample after oven drying
The ash content was determined using the ignition method. The crucible used were thoroughly washed and pre-
heated in a muffle furnace to about 600°C. Exactly 2.0g of the oven-dried sample used in moisture determination
were weighed in triplicate and placed in the pre-heated, cooled and weighed crucible and then reweighed. The
crucible was covered with its lid and then placed in a cold muffle furnace and heated at 600°C for two hours to
burn off all the organic matter. The crucible was removed from the furnace, allowed to cool in a desiccator and
reweighed. The percentage ash content was calculated using the formula:
Ash % = weight of ash x 100 ........................................................................................... (2a)
weight of sample
OR Ash% = c – a x 100........................................................................................................ (2b)
b–a
Where: a = weight of empty crucible
b = weigh of crucible + sample before ashing
48
c = weight of crucible + ash
Determination of crude lipid content of the sample was done using Soxhlet type of the direct solvent extraction
method. The solvent used was petroleum ether (boiling range 40 - 60°C). Exactly 2.0g of the dried sample was
3
weighed in triplicate and secured in soxhlet extraction thimble. The thimble was then put into 20cm capacity
soxhlet extractor. A washed, oven-dried 100cm3 round-bottomed flask was weighed and approximately 60cm3 of
the 40-60°C boiling range petroleum ether added to it. The flask was then mounted on the heating mantle and
connected to the extractor with a condenser. The condenser and heating mantle were then activated and
extraction carried out for four hours. At the end of extraction, the solvent was evaporated and the flask dried in
the oven at 60°C. The flask was then cooled and reweighed. The percentage of crude lipid was calculated using
the formula:
% crude lipid = weight gain in flask x 100 ................................................................................ (4a)
weight of sample
OR % crude lipid = W2 – W1 x 100 ..............................................................................................(4b)
WS
Where: W2 = weight of beaker + sample
W1 = weight of empty beaker only; WS = weight of sample
Crude fibre content was determined by the Weende method described by [11]. About 2.0g of the sample
were weighed into petroleum ether for two hours and then boiled under reflux apparatus for 30 minutes with
3 3
200 cm of a solution containing 1.25% of H2SO4 per 100 cm solution. After series of filtering and washing
with boiling water until it was no longer acidic, the residue was transferred into a beaker and boiled for
another 30 minutes with 200 cm3 of solution containing 1.25g of NaOH per 100 cm3. The final residue was
filtered and washed with boiling water until it was neutral to litmus and finally washed twice with ethanol
o
and quantitatively transferred into a pre-weighed crucible and oven dried at 105 C. The sample residue was
then incinerated in a furnace at 550oC and allowed to stand at this temperature for 2 hours. The crucible was
later removed and allowed to cool in a desiccator and then reweighed. The percentage crude fibre was
estimated according to the expression below:
% Crude fibre = weight loss on ignition (g) x 100................................................................................ (5)
OR %Crude fibre =
Parameters Composition (%)

Moisture 68
Ash 2.5
Crude fibre 0.5
crude protein 24.33
Crude lipid 0.5
Carbohydrate 72.1
Caloric value 390.22 Kcal/100g
Table 2: Mineral Elements Composition (mg/100g) of Allium sativum sample obtained in Uyo
Mineral Elements Composition (mg/100g)

Calcium 180
Iron 1.7
Magnesium 23.4
Manganese 1.54
Potassium 404
Zinc 1.16
49
Table 3: Anti-nutrients Composition of Allium sativum sample obtained in Uyo.
Anti-nutrients Composition (mg/100g)

Phytate 2.14
Hydrogen cyanide 0.30
Oxalate 234.60
Saponin 12.60
References
[1] R. K. Kempaiah and K. Srinivasan (2004). Beneficial influence of dietarycurcumin capsaicin and garlic on
enythrocyte integrity in high fat fed rats. Journal of Nutrition and Biochemistry, 56(2): 150-157.
[2] N. M. Nwinuka, G. O. Ibeh and G. I. Ekeke (2005).Proximate composition and levels of some toxicants in
four commonly consumed spices .Journal of Applied Science and Environmental Management, 9(1):
150-155.
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structures of allimase from Allium sativum L. apo from and ternary complex with aminoacrylate
reaction intermediate covalently bond to PLP co-factors. Journal of Molecular Biology, 36(2): 611-625.
[4] A. N Saidu and N. G. Jideobi (2009).The Proximate and Elemental Analysis of some Leafy Vegetables
Grown in Minna and Environs. J. Appl. Sci. Environ. Manage., 13(4): 21 – 22
[5] E. I. Uwah, E. A. Moses and K. R. Okokon (2015). Proximate, minerals and Anti-nutrients composition of
two leafy vegetables commonly consumed in Idu, Uruan, AkwaIbom State, Nigeria. Int. J. Curr. Res.
Chem. Pharma. Sci., 2(10): 24-30.
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vulgaris,Cucumis sativus and Soils obtained from Gardens being Irrigated with Wastewater in Maiduguri,
Nigeria. Glo. Res. J. Agric. Biol. Scis. 3(5):373 –380.
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in Nigeria. Afr. J. Nat. Sci., 7: 71-73.
[8] O. L. Oke (1969). Chemical studies on the more commonly used vegetables in Nigeria. Afr. Sci. Ass., 11: 42-
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[9] J. N. Erdman (1979). Oily seed phytates nutritional implications. J. Am. Oil Chem. Soc.,56: 736-741.
[10] AOAC (1997). Methods of Analysis of Association of Official Analytical Chemists (16th ed).Washington,
D.C.,1: 600-792.
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[12] E. N. Onyeike, T. Olungwe and A. A. Uwakwe (1995). Effect of heat treatment and defatting on the
proximate composition of Some Nigerian local soup thickeners. Food Chem., 53:173 – 175.
[13] A. Munro and O. Bassir (1969). Oxalate in Nigerian vegetables. W. Afr. J Biol. Appl. Chem., 12: 14-18.
[14] R. B. Burns (1971). Methods of estimation of tannin in the grain Sorghum. Agronomy Jour., 63.511
[15] M. R. Kadhakrishna and J. Sivaprasad,1980. Tannins content of sorghum varieties and their roles in iron
bio-availability. J. Agric food chemistry, 28:55-57.
[16] Pearson, D, 1976. Chemical Analysis of Food, 7th edition, Churchill, London, Pp. 7-11.
[17] G. A. Otunola, O. B. Oloyede, A. T. Oladiji and A. J. Afalayan (2010). Comparative analysis of the chemical
composition of three spices – Allium sativum L.,Zingiber officinale Rose and Capsicum fruteseens L.
commonly consumed in Nigeria. African Journal of Biotechnology, 41:6927-6931.
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seeds of Monodora myristica (African Nutmeg).World Journal of Applied Science and Technology,
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[19] A. A. Akindahunsi and S. O. Salawo (2005). Phytochemical screening of nutrient and anti nutrient
composition of selected tropical green leafy vegetables. .Afr. J. Biotech., 4:497-501.
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[21] J. H Cohen, A. R. Kristal and J. L. Standford (2000). Fruit and vegetable intakes and prostate cancer risk. J.
Nat. Cancer Inst., 92:61-68.
[22] D. E. Okwu (2004). Phytochemicals and Vitamin content of indigenous species of South Eastern Nigeria. J.
Sustain Agric. Environ. 6:30-34.
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[24] FAO/WHO (1995). Evaluation of certain food Additives and Contaminants. Geneva, World Health
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Report Series, 859: 29–35.
[25] K. R. Price, I. T. Johnson and G. R Fenwick (1987). The Chemical and Biological Significance of saponins
in Foods and Feeding Stuffs. CRC Critical Reviews in Food Science and Nutrition 26:27 – 135.
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Press, New York, pp 78-113.
51
PHYTOCONSTITUENTS, PROXIMATE COMPOSITION AND MINERAL CONTENTS OF
SOYBEAN (GLYCINE MAX) FLOUR
Useh, M. U.,1* Adebiyi, A. B.,1 and Dauda, M. S.2
1
Sheda Science and Technology Complex, PMB 186, Garki, Abuja, Nigeria
2
Chemistry Department, University of Abuja, PMB 117, Abuja, Nigeria
Corresponding Author: usehmercy@gmail.com Phone: 08098239928
ABSTRACT
Soybean (Glycine max), is a species of legume widely grown for its edible bean which has numerous uses. In this
study, the phytoconstituents, proximate composition and mineral contents of soybean flour was evaluated using
standard processing techniques. The preliminary phytochemical screening indicated the presence of phenols,
cardiac glycosides, steroids, saponins, flavonoids in both the methanol and aqueous extracts. The quantitative
phytochemical screening showed that saponin had the highest content (18.4 %) followed by phenol (16.8 %) and
flavonoids (12.4 %) while phytate was the least (0.07 %). The proximate composition was in this order: protein >
carbohydrate > crude fat > moisture > crude fibre > ash content which showed that the soy flour is relatively
high in protein. The minerals analyzed for were: Ca(231.6), Fe(5.790), Mg(249.8), Zn(2.414) and Mn(0.651)
which were within the FAO/WHO standards for metals in foods.
Keywords: Soybean, phytoconstituents, protein, soy flour, saponin.
INTRODUCTION
The soybean (Glycine max (L.) Merrill) belongs to the legume family, native to Eastern Asia and introduced into
Nigeria in 1908 [1]. It has long been recognized as a plant food that, when compared with other plants, is
relatively high in protein. It is an important inexpensive food crop that contains about 40 % protein, 30%
carbohydrates, excellent amounts of dietary fiber, vitamins, minerals and 20 % oil which makes it second only to
groundnuts in terms of oil content amongst food legumes [2]. Historically it is nick-named "meat of the field"
because it is the only vegetable with a complete protein. The gold standard for measuring protein quality is the
Protein Digestibility Corrected Amino Acid Score (PDCAAS) and by this criterion soy protein is the nutritional
equivalent of meat, eggs, and casein for human growth and health [3].
Over the years, soybean consumption has been a matter of much debate. But recently researchers have taken a
very close look at the protein content of soybeans and arrived at many fascinating conclusions. Along with this
increasing interest in soy protein came the discovery of very unique proteins, typically referred to as "peptides"
which include defensins, glycinins, conglycinins and lunasin, and all are now known to provide us with health
benefits in the areas of improved blood pressure regulation, better control of blood sugar levels, and improved
immune function [4]. Soy protein has the ability to lower LDL (bad cholesterol) levels and decrease the risk of
coronary heart disease (CHD). Soybean contains isoflavones that minimizes the risk of developing certain
cancers, particularly, genistein—an isoflavone that has extensive antioxidant properties. It also serves as an
excellent source of essential fatty acids, calcium, magnesium, lecithin, riboflavin, thiamin, fiber, folate (folic
acid) and iron [5].
Malnutrition, mostly protein deficiency, is prevalent in many parts of Africa as animal protein is too expensive
for most populations. Children in most developing countries, particularly those in low-income classes, are
weaned on cheap, readily available starchy foods such as pap in which soybean flour is mainly used to
supplement this popular weaning and breakfast recipe. The beans can be processed in a variety of ways. Common
forms of soy include soy meal, soy flour, soy milk, tofu, tempeh, miso, natto and soybean oil. Soy flour is used as
an ingredient in foods, beverages, and to promote higher optimal health benefit.
Soybean seed, like other legumes, is known to contain significant amounts of all the essential amino acids,
minerals, phytate and oxalates but these anti-nutritional factors are usually removed or greatly reduced by
52
soaking, fermentation and hydrothermal treatment during processing [6]. In recent times, concerns over the
possible make-up of this wonderful legume due to its numerous health benefits made this research work
absolutely imperative.
MATERIALS AND METHODS
Sample Collection and Preparation
The soybean (Glycine max (L.) Merrill) seeds used for this work were purchased from Kubwa market in Bwari
Area Council, Abuja, Nigeria. It was transferred to the laboratory, sieved to remove stones and debris, washed
o
and soaked for 48 hours, oven dried at 40 C for 5 minutes and dehulled with the aid of hammer mill. The dehulled
beans was ground finely enough with the use of a specialized Alpine Fine Impact Mill, model A90BD to pass
through a 0.25mm screen [7]. The so made soy flour was packaged in moisture-proof, air-tight polyethylene
o
containers and kept at 4 C prior to analyses.
Methanol Extract
The methanol extract of the soybean flour was prepared by soaking 20 g of the sample in 200 ml of methanol for
12 hours. The extract was then filtered through a funnel lined with Whatman No. 42 filter paper. The filtrate was
concentrated and stored in an airtight container.
Aqueous extract
The aqueous extract of the soybean flour was prepared by soaking 20 g of the sample in 400 ml of distilled water
for 12 hours. The extract was then filtered using filter paper, concentrated and stored in an airtight container.
Qualitative Phytochemical Constituents
A small portion of the filtrate was used for the qualitative phytochemical screening using standard methods [8,
9].
Quantitative Phytochemical Constituents
Test for Phenol: This was determined using spectrophotometric method. The sample was boiled with diethyl
ether for 15 mins. 5 ml of the boiled sample was then pipette into 50 ml flask and 5 ml of distilled water was
added. Then, 2 ml of NH4OH solution and 5 ml of conc. butanol was added to the mixture. The sample was made
up to mark and left for 30 mins. to react for colour development. The absorbance was measured at 505 nm [10].
Test for Saponin: 0.5 g of the sample was added to 20 ml of 1 N HCl and was boiled for 4 hours. This was filtered
after cooling and 50 ml of petroleum ether was added to the filtrate for ether layer and evaporated to dryness. 5 ml
of acetone-ethanol was added to the residue. 0.4 ml each was taken into 3 different test tubes, 6 ml of FeSO4
reagent was added into them followed by 2 ml of concentrated H2SO4 acid. It was thoroughly mixed after 10
mins. and the absorbance was taken at 490 nm [11, 12].
Test for Flavonoid: This was done by acid hydrolysis of spectrophotometric method. 0.5 g of the sample was
mixed with 5 ml of dilute HCl acid and boiled for 10 mins. the boiled extract was allowed to cool and then
filtered. 1 ml of the filtrate was added to 3 ml of ethyl acetate and 5 ml of 1 % NH3. This was then scanned from
420 -520nm for the absorbance [13].
Taste for Phytate: This was determined by the photometric method of Latta and Esin [14]. 2 g of the test sample
was extracted with 100 ml of 2.4 % HCl acid by shaking vigorously in a vortex mixer for 1 hour and then filtered
through Whatman No. 5 filter paper. 5 ml of the filtrate was mixed with 1 ml of 0.1M Na-EDTA, 0.75 M NaOH
solution and then made up to 25 ml with distilled water before being placed on an ion-exchange (AGIX4, 100-
200mesh) column. The column was washed with 15 ml of distilled water and then 15 ml of 0.1 m NaCl before
being eluted with 15 ml of 0.7 M NaCl. The eluate was collected and wet digested in a Kjeldahl apparatus with
0.5 ml conc. H2SO4 and 3 ml HCl. The digest was cooled to room temperature, 10 ml of distilled water added and
the mixture heated again on a water bath with 2 ml of 2.5 % ammonium molybdate solution in 1 N H2SO4, 1 ml
conc. H2SO4 and then made up to 50 ml in a volumetric flask. The solution was allowed to stand for 15 mins
53
before taking the absorbance at 640 nm against a blank. This was done in triplicate [15].
Test for Oxalate: Deionized water was heated at 29oC and the sample was soaked for 1 minute. 1 ml of 5 M
NaOH, 1 ml of 5 % CaCl2 and 3 drops of phenolphthalein was added to 5 ml of extract and allowed to stand for 3
hours. This was centrifuged at 300 rpm for 15 mins. The supernatant was discarded and the precipitate was
washed with hot water and allowed to centrifuge. 2 ml of 3 M H2SO4 was pipette into it and transferred into a
conical flask. The precipitate was dissolved by warming in a water bath at 80oC. The content of each tube was
then titrated with freshly prepared 0.01M KMnO4. The titration was carried out at ordinary temperature until a
first colour change was observed. It was allowed to stand until the changed to colourless. The solution was
warmed at 70oC and titration was carried out until a pink colour persists at least for 30 seconds [16].
Proximate Composition: The proximate analysis (moisture, ash, crude protein, crude fibre, crude fat and
carbohydrate) of the sample was determined using the standard method of AOAC [17]. All determinations were
done in triplicate and reported in percentage.
Table 1: Qualitative analysis of phytochemical constituents of soy flour
Phytochemical Methanol Aqueous extract

parameters extract
Tannins - -
Phenols + +
Balsams - -
Cardenolides - -
Cardiac + +
glycosides
Phlobatannins - -
Steroids + +
Saponins + +
Resins - -
Volatile oils - -
Flavonoids + +
Terpenoids - -
Glycosides - -
Alkaloides - -
Carbohydrates - +
Key: (+) = present, (-) = absent
Table 2: Quantitative analysis of phytochemical constituents of soy flour
54
Variables Percentage (%)
Phenols 16.8
Saponins 18.4
Flavonoids 12.4
Phytates 0.07
Oxalates 0.15
Table 3: Proximate Composition of Soybean flour
Parameters Values (%) (n=3)
Moisture 6.84
Ash 4.92
Crude Protein 35.6
Crude Fibre 6.27
Crude Fat 18.5
Carbohydrate 27.87
Table 4: Mineral contents of Soybean flour (mg/kg)
Parameters Values FAO/WHO guidelines

Ca 231.6 -
Fe 5.790 48
Mg 249.8 300
Zn 2.414 60
Mn 0.651 1.0

The results of the phytoconstituents of Glycine max (L.) Merrill as determined are recorded in tables 1 and 2
while the proximate compositions and the mineral contents are presented in tables 3 and 4. The preliminary
phytochemical screening indicated the presence of phenols, cardiac glycosides, steroids, saponins, flavonoids in
both the methanol and aqueous extracts. From the active compounds indicated, it is obvious that Glycine max (L.)
Merrill is a potential source of useful drugs. From table 2, saponin had the highest percentage content (18.4 %)
followed by phenol (16.8 %) and flavonoids (12.4 %) while phytate was the least (0.07 %).
Saponins, are sterols that are present naturally in a wide variety of food-plants, including vegetables, legumes
and cereals. Legumes such as soybean and chickpeas are the major source of saponins in form of soyasaponins
and soyasapogenols in the human diet. Soybeans also contain the isoflavones genistein and daidzein, types of
phytoestrogen which are closely related to the antioxidant flavonoids. Genistein is a soy component that could
easily be singled out for its antioxidant properties! Increased activity of antioxidant enzymes—including
superoxide dismutase, glutathione peroxidase, catalase, and glutathione reductase—has now been linked to
intake of genistein from soy. Another group of antioxidant phytonutrients called phenolic acids has also been
recently investigated in soybeans. When we enjoy this antioxidant-rich legume, we also benefit from its phenolic
acids, including caffeic, coumaric, ferulic, and sinapic acid. Phytic acid in soybeans has many effects including
acting as an antioxidant and a chelating agent. It helps in reducing cancer, minimizing diabetes and reducing
inflammation. However, phytic acid is also criticized for reducing vital minerals due to its chelating effect, but
this effect is greatly reduced during processing.
From table 3, crude protein had the highest value (35.6 %) while moisture content (4.84 %) was the lowest which
showed hindrance to microorganism growth. From table 4, the minerals analyzed for were: Ca(231.6),
Fe(5.790), Mg(249.8), Zn(2.414) and Mn(0.651) which were within the FAO/WHO standards for metals in
foods.
Conclusion
55
Soybeans are perhaps best known for their fantastic blend of protein and fiber. They are a very good source of
manganese, phosphorus, and protein as well as a good source of iron, omega-3 fatty acids, dietary fiber,
magnesium, and vitamin K. There are also a wide range of unique proteins, peptides, and phytonutrients
contained in soy which include flavonoids, isoflavonoids, phenolic acids and saponins (soyasaponins from
group A and group B, and soyasapogenols) which are very useful for optimal health.
Acknowledgment
The authors wish to acknowledge the management of Sheda Science and Technology Complex (SHESTCO) for
their assistance.
References
1. Messina M. G. and Messina V. M. (2010). The Role of Soy in Vegetarian Diets Nutrients. Vol: 2 Issue: 8,
pages 855-888.
2. Ajay G. Y. and Arvind B.N. (2009) Genistein: A multipurpose isoflavone. International Journal of
Green Pharmacy. Vol: 3 Issue: 3 Pages/record No.: 176-183. 2009.
3. Amadou I. O., Yong-Hui S. J., Sun J. F and Davis M. K. (2009). Fermented Soybean Products: Some
Methods, Antioxidants Compound Extraction and their Scavenging Activity. Asian Journal of
Biochemistry. Vol: 4 Issue: 3 Pages/record No.: 68-76. 2009.
4. Anderson J.W. and Bush H.M. (2011). Soy protein effects on serum lipoproteins: a quality assessment
and meta-analysis of randomized, controlled studies. J Am Coll Nutr. 30(2):79-91.
5. Barnes S.U. (2010). The Biochemistry, Chemistry and Physiology of the Isoflavones in Soybeans and
their Food Products. Lymphat Res Biol. 8(1): 89—98.
6. Butler L.M, Wu A.H, Wang R. G. (2010). A vegetable-fruit-soy dietary pattern protects against breast
cancer among postmenopausal Singapore Chinese women. Am J Clin Nutr. 91(4): 1013—1019.
7. Chai C. B, Jul H.K, Kim S.C. (2011) .Determination of bioactive compounds in fermented soybean
products using GC/MS and further investigation of correlation of their bioactivities. Chromatogr B
Analyt Technol Biomed Life Sci. Pharmacol Rep. 63(3):643-59.
8. Harbone J.B. (2010). Phytochemical Methods. Chapman and Hall Ltd. London.
9. Klein M.A, Nahin R.L, Messina M.J. (2010). Guidance from an NIH Workshop on Designing,
Implementing, and Reporting Clinical Studies of Soy Interventions. The Journal of Nutrition 140(6):
1192S-1204S.
10. Jenkins D.J, Jones P.J, Lamarche B. (2011). Effect of a dietary portfolio of cholesterol-lowering foods
given at 2 levels of intensity of dietary advice on serum lipids in hyperlipidemia: a randomized
controlled trial. JAMA. 306(8):831-9.
11. Lammersfeld C.A, King J.U, Walker S. (2009). Prevalence, sources, and predictors of soy consumption
in breast cancer. Nutr J. 8: 2. Published online 2009 January 22. doi: 10.1186/1475-2891-8-2. 2009.
12. Lanou A.J. Soy foods: are they useful for optimal bone health? . Therapeutic Advances in
Musculoskeletal Disease vol. 3 no. 6 293-300.
13. Merritt J.C. Metabolic syndrome: soybean foods and serum lipids. J Natl Med Assoc. 96(8):
1032—1041.
14. Michelfelder A.G. (2009) Soy: A Complete Source of Protein. American Family Physician79. 1: 43-7.
15. Yimit D. F, Hoxur P.U, Amat N.I. (2011). Effects of soybean peptide on immune function, brain
function, and neurochemistry in healthy volunteers. Nutrition. 4(2): 72—82.
16. Moller N.P, Scholz-aherns K.E, Roos N.E. (2008). Bioactive peptides and proteins from foods:
indication for health effects. European Journal of Nutrition . 47(4): 171-82.
17. Association of Official Analytical Chemists (AOAC). (2009). Official Methods of Analysis. Horwitz.
18th ed.
56
Equilibrium, Kinetics and Thermodynamic Properties of Methylene Blue Adsorption by
Termite Mound
1 2 3
Anebi, P. O., Ugbe, F.A., Ikudayisi, A.V.
1
Department of Science Lab. Tech., Benue State Polytechnic, Ugbokolo
2,3
Department of Chemistry, University of Ilorin, Kwara State
Email: 2ugbefabianaudu@gmail.com
1.0 Introduction
Termites are social ants that exert significant influence on the physical and chemical properties of
tropical and sub-tropical soils (Abdus-Salam & Itiola, 2012). Mounds are widely distributed and
abundant in nature (Fufa et al., 2013). The feeding habit, the food processing and mound construction
operations introduce significant modifications to the soil on which the mound is built (Abdus-Salam
& Bello, 2015). Research carried out by Abdus-Salam & Itiola, 2012, Fufa et al., 2013 revealed that 3
dominant oxides present in TM are SiO2, Fe2O3 and Al2O3; as a result, TM is composed of Si, Fe and Al
in significant amount. The dominant functional groups of TM being Fe-OH, Fe-O and O-H as
reported elsewhere (Abdus-Salam & Bello, 2015).
Synthetic dyes are one of the main pollutant groups of water and wastewater (Shahryari et al., 2010).
Dye contamination in wastewater causes problems in several ways: the presence of dyes in water, even
in very low quantities, is highly visible and undesirable; color interferes with penetration of sunlight
into waters; retards photosynthesis; inhibits the growth of aquatic biota and interferes with gas
solubility in water bodies (Robinson et al., 2002; Shahryari et al., 2010). The highest rates of toxicity
were found amongst basic and diazo direct dyes (Shahryari et al., 2010).Therefore, it is highly
necessary to reduce dye concentration in the wastewater.
Few researchers have investigated the removal from aqueous systems of heavy metals ions such as Pb
(II) (Abdus-Salam & Itiola, 2012), Cr (VI) (Araújo et al., 2013) and Zn (II) (Abdus-Salam & Bello,
2015), using termite mound (TM) as adsorbent. Also, removal of fluoride by termite mound was
reported (fufa et al., 2013). However, in literature, there are limited or no research on the removal of
dyes using termite mound. Hence, the present study is targeted at (1) evaluating the methylene blue
(MB) sorption capacity of TM under batch adsorption setup; and (2) assessing trends of MB adsorption
with respect to variation of contact time, solution pH, adsorbent dose, initial MB concentration, and
temperature.
2.0 Materials and Methods
2.1 Collection of samples and pretreatment
The termite mound soil was collected from a farm at Opidlo, Edumoga in Benue State, Nigeria. A
sample of about 2 kg was collected and air-dried in the laboratory at room temperature for 5 days. The
sample was then pretreated according to the method reported by Abdus-Salam & Itiola, 2012. The
sample was then passed through a 0.112mm mesh sieve and used for subsequent experiment.
2.2 Adsorption Experiment
3
A 0.5 g portion of the various soil samples collected was weighed in a 100 mL conical flask and 15 cm
of working standard solutions (500, 400, 300, 250, 200, 150, 100 and 50 ppm) of MB prepared by serial
dilution of the standard solution, was added to the soil sample in the beaker. Each solution was agitated
on a flat orbital mechanical shaker for 4 hr and then filtered. The filtrate was analyzed using UV-Visible
Spectrophotometer to determine the quantity of MB remaining in the solution (Ibrahim & Jimoh,
2008).
The equilibrium concentration (concentration with the highest adsorption capacity) was then selected
57
for subsequent experiment.
Further experiments were carried out using the equilibrium concentration to examine the effects of initial
solution pH (varied from 2 to 9), variation of adsorbent (TM) dose (varied from 0.1 to 1.0g), agitation time
(varied from 5 to 120 minutes) and temperature (varied from 308 to 338K) (Abdus-Salam & Buhari, 2014,
Ugbe et al., 2014, Abdus-Salam & Bello, 2015).
3.0 Results and Discussion
3.1 Effect of Initial Dye Concentration
The result of effect of initial solution concentration on the removal of Methylene Blue (MB) by Termite
Mound (TM) is presented in figure 3.1. The amount of dye adsorbed per unit gram of Termite Mound (TM)
increased from 1.314 mg/g to 8.086 mg/g with the increasing of the initial solution concentrations from 50
mg/L up to 400 mg/L, after which the adsorption falls. This is attributed to the saturation of the adsorption
sites of TM at higher concentration (Abdus-Salam & Buhari, 2014).
3.2 Effect of Initial Solution pH

The initial pH of the dye solution is an important parameter which controls the adsorption process
particularly the adsorption capacity. The pH of the solution may change the surface charge of the
adsorbent, the degree of ionization of the adsorbate molecule and extent of dissociation of functional
groups on the active sites of the adsorbent (Ugbe et al., 2014). To observe the effect of pH on the extent of
dye adsorption, solution pH is varied from 2 to 9. The quantity of dyes removed at different pH is shown
in Figure 3.2 The quantity of dye adsorbed per unit mass of TM increased from 0.157 mg/g at pH 2 and
reaching a maximum of 11.5 mg/g at pH 8 and beyond. This is attributed to the predominance of the
negatively charged OH group on the TM surface at higher pH, resulting to increased binding of
methylene blue with the surface via the positive site (Elmorsi, 2011).
58
3.3 Effect of Variation of Adsorbent Dose
Result of the adsorption experiment using five different dosages, viz. 0.1, 0.25, 0.5, 0.75 and 1.0 g of TM
and optimum concentration of 400 mg/L MB is presented in figure 3.3 The values indicate an increase in
the adsorption capacity of MB with dosage. This is because of an increase in the number of available
adsorption sites of the adsorbent which results in enhanced removal of the dye (Eman et al., 2014).
3.4 Effect of Agitation Time

The effect of agitation time for the adsorption of MB on TM was studied between 5 and 120 minutes as
illustrated in figure 3.4. Result shows that the uptake increased from 8.514 mg/g at 5 minutes and
reaching equilibrium at 90minutes with 11.514 mg/g uptake. This is as result of saturation of adsorption
sites of TM at higher contact time (Eman et al., 2014). However, the variation of agitation time does not
produce a remarkable difference in the quantity adsorbed due to high affinity of TM for the dye (Eman et
al., 2014).
3.5 Adsorption isotherms

The adsorption data obtained from the adsorption of methylene blue experiment were tested for fitness
of data against three common adsorption equations, Langmuir, Freundlich and Temkin adsorption
isotherms, respectively. The isotherm parameters are presented in table 3.1. The observed R2 = 0.9807
shows that the Langmuir isotherm fitted well the adsorption data for MB. This indicates that adsorption
of MB onto TM may follow a monolayer coverage, uniform energies of adsorption onto the surface and
no transmigration of MB in the plane of the surface.
The favourability of this adsorption process was subjected to the equation of separation factor RL:
(Shahryari et al., 2010).
The RL value obtained from this adsorption process is 0.048 which fall into the range 0 < RL < 1
indicating that the adsorption process is favourable. Similar result was obtained elsewhere (Abdus-
59
Salam & Itiola, 2012). The data obtained did not fit into Freundlich and Temkin isotherm appropriately
and therefore cannot be analysed further.
Table. 3.1. Isotherm constants and correlation coefficients
Isotherm Model
Langmuir Freundlich Temkin
KL qm
R2 (L/g) (mg/g) R2 KF 1/n R2 A B
0.9807 0.053 8.628 0.3537 2.6212 0.1803 0.507 23.59 0.8362
3.6 Adsorption Kinetics
The adsorption kinetic models are important in the process of removal of toxic heavy metals from the
environment because it provides information on pollution flux (Abdus-Salam & Itiola, 2012). Pseudo-
first order, pseudo-second order and intra-particle diffusion models were tested with the data obtained to
establish the model that best fits the data (table 3) using equations reported elsewhere (Lagergren, 1898,
Ho & McKay, 2000, Weber and Morris, 1963).
2
As shown in table 3.2 pseudo-second order model, yields the best plot with a regression coefficient R =
0.9982, pseudo-second order rate constant k2 = 0.0204 g/mg/min and qe = 11.862 mg/g. Therefore, the
kinetics is best described by the pseudo-second order kinetics, implying that the rate-limiting step could
be chemical adsorption involving valence forces through the sharing or exchange of electrons between
adsorbent and adsorbate. This trend was the same as reported elsewhere (Abdus-Salam & Itiola, 2012,
Fufa et al., 2014). The linear plot of qt versus t1/2 shows a good fit but does not pass through the origin.
This indicates that the intraparticle diffusion is not the only rate controlling step (Hameed et al.,
2008).
Table 3.2. Kinetics constants and correlation coefficients
Adsorption Kinetics
Pseudo First Order Pseudo Second Order Intra-particle Diffusion
k1 qe
2
R (min-1) (mg/g) R2 k2 (g/mg/min) qe (mg/g) R2 Ki C
0.9798 0.0436 2.359 0.9982 0.0204 11.862 0.9578 0.3619 7.8795
3.7 Thermodynamics of Adsorption

In order to examine in greater detail the effect of temperature on the sorption of MB by TM,
thermodynamic parameters such as Gibbs free energy change (G), enthalpy change (H), entropy change
(S) and isosteric heat of adsorption (Hr) were estimated using equations reported elsewhere (Ugbe et al.,
2014, Saha & Chowdhury, 2011). The estimated thermodynamic parameters are shown in table 3.3.
As shown in table 3.3, the values of the free energy change (G) are negative above 308K,
indicating the feasibility and spontaneity of the process at temperature above 308K. Also, G became
increasingly negative with temperature; an indicative of feasibility of the process at higher temperature.
Similar resulted have being reported elsewhere (Noshin et al., 2009). The positive value of enthalpy
change (H) showed that the process is endothermic; as evident in enhanced removal at increased
-1
temperature. The magnitude of H falls into the range of 80–200 kJ mol , an indication that binding of
MB onto TM follows chemisorption mechanism (Muhammad et al., 2014, Saha & Chowdhury, 2011).
Positive value of the entropy change (S) showed greater affinity of the TM towards the dye. In addition,
it showed increased randomness at the solid/solution interface with some structural changes in the MB
60
and the TM. The adsorbed solvent molecules, which are displaced by the MB, gain more translational
entropy than is lost by the MB, thus allowing for the prevalence of randomness in the system (Saha &
Chowdhury, 2011). The value of the isosteric heat of adsorption is > 40 kJ/mol, illustrating a
chemisorption mechanism (Muhammad et al., 2014).
Table 3.3 . Thermodynamic Parameters for sorption of MB on TM
Temperature Parameters
qe (mg/g) ? G ?S ?H ? Hr
(K)
(kJ/mol) (J/mol/K) (kJ/mol) (kJ/mol)
308 4.0714 +1.7067 +461.73 +142.68 -113.952
318 10.957 -6.2184 +461.73 +142.68 -113.952
328 11.514 -8.6329 +461.73 +142.68 -113.952
338 11.871 -12.717 +461.73 +142.68 -113.952
Conclusion
From the adsorption data, the sorptive property of the mound was found to be dependent on
initial solution concentration, agitation time, pH, dosage and temperature. The equilibrium adsorption
data showed satisfactory correlation with the Langmuir adsorption data and was also found to fit the
pseudo-second order and intra-particle diffusion kinetics. The favorable adsorption of MB on TM was
signified by the obtained value of the separation factor, RL and the Gibbs free energy change, G. The
equilibrium removal of MB increased as the temperature of solution increases, because the adsorption
was chemical as evident in the magnitude of enthalpy change and isosteric heat of adsorption. The
removal of MB by TM is an exothermic and a spontaneous process.
Therefore, the results of the study could provide fundamental information to evaluate TM for the
practical removal of MB from waste water.
References
Abdus-Salam N. & Bello, M.O. (2015). Kinetics, thermodynamics and competitive adsorption of lead and
Zinc ions onto termite mound. International journal of environmental Science and Technology, 12 (11),
3417-3426.
Abdus-Salam, N. & Itiola, A.D. (2012). Potential application of termite mound for adsorption and
removal of Pb (II) from aqueous solutions. Journal of the Iranian Chemical Society. DOI 10.1007/s13738-
011-0047-2
Abdus-Salam, N. & Buhari, M. (2014). Adsorption of Alizarin and Fluorescein Dyes on A d s o r b e n t
prepared from Mango Seeds. Pacific Journal of Science and Technology, 15(1), 232-244.
Araújo, B.R., Reis, J.O.M., Rezende, E.I.P., Mangrich, A.S., Wisniewski, A., Dick, D.P. & Romão,
L.P.C (2013). Application of termite nest for adsorption of Cr (VI). Journal of E n v i r o n m e n t a l
Management, 129, 216-223
Elmorsi, Taha M. (2011). Equilibrium Isotherms and Kinetic Studies of Removal of Methylene Blue
Dye by Adsorption onto Miswak Leaves as a Natural Adsorbent. Journal of Environmental
Protection, 2, 817-827.
Eman, A. Ashour, Maha, A. Tony& Patrick, J. Purcell (2014). Use of Agriculture-Based Waste for Basic
Dye Sorption from Aqueous Solution: Kinetics and Isotherm Studies. American Journal of
Chemical Engineering 2 (6), 92-98.
Fufa, F., Alemayehu, E., Lennartz, B. (2013). Defluoridation of Groundwater using Termite M o u n d .
Water Air Soil Pollution, 224, 1552.
61
Fufa, F., Alemayehu, E., Lennartz, B. (2014). Sorptive removal of arsenate using termite mo Journal of
Environmental Management, 132, 188-196
Ibrahim, M.B. & Jimoh, W.L.O. (2008). Adsorption Studies for the Removal of Cr (VI) Ion F r o m A q u e o u s
Solution. Bayero Journal of Pure and Applied Sciences, 1(1), 99-103.
Muhammad, A.A., Pam, A.A., Audu, U.F. & Onakpa, S.A. (2014). Thermodynamic Study of the Competitive
Adsorption of Chromium (III) Ions and Halides onto Sweet Orange (Citrus Sinensis) Peels as Adsorbent.
Journal Environmental Analytical Chemistry, 1 (2), 1 – 7.
Noshin, H., Somaieh, K. & Hossein, A. (2009). Equilibrium and Thermodynamic Studies of Cesium Adsorption
on Natural Vermiculite and Optimization of Operation Conditions. Iran. j. chem. Chem.. eng. 28 (4), 2009.
Robinson, T., Chandran, B. & Nigam, P. (2002). From an artificial textile dye effluent by two agricultural
waste residues, corn corb and barley husk. Environ. Int. 28: 29-33.
Saha, P. & Chowdhury, S. (2011). Insight into Adsorption Thermodynamics. InTech, 16, 350-364.
Shahryari, Z., Soltani A.G. & Azadi, M. (2010). Experimental study of methylene blue adsorption from
aqueous solutions onto carbon nano tubes. International Journal of Wa t e r R e s o u rc e s a n d
Environmental Engineering, 2 (2), 016-028.
Ugbe, F.A., Pam, A.A. & Ikudayisi, V.A. (2014). Thermodynamic study of chromium (III) ion Adsorption
by sweet orange (citrus sinensis) peels adsorbent. American Journal of Analytical Chemistry, 5, 666-
673.
62
PETROCHEMICALS: UPSTREAM TO PLASTIC INDUSTRY USING IDEAS, METHODS,
DEVELOPMENTS AND PROCEDURES IN DEVELOPING COUNTRIES.
OGODO AEMURERHIMEN DICKSON (Snr.) (Chartered Chemist)(A2652)

(HND, PGD, M.Sc., Ph.D (In- View), AISLT, C.CHEM., MICCON, MCSN, MNES, MSPE)
School of Applied Science, Department of Science Laboratory Technology, Chemistry Laboratory, Delta State
Polytechnic, P.M.B. 1030, Ogwashi –Uku, Delta State, Nigeria: Tel: 08030738401, 08094921724, E-mail:
dicksonogodo@yahoo.com. Formerly, Pioneer Co-ordinating Chairman, Research Institute for Tropical
Agriculture, School of Science and Technology, Department of Science Laboratory Technology, Chemistry
Laboratory, Delta State Polytechnic, P.M.B.5,Ozoro, Delta State, Nigeria.
Abstract
Plastic still remains today the real driving force of the petrochemical industry. Petrochemical is a branch of
Chemistry that employs hydrocarbons, Oil and Gas as raw materials. This scientific research work clearly
reveals that the petrochemical industry produces polymers (Plastics, Synthetic Fibres and Rubber),
Detergents, Fertilizers, Fine Chemicals as well as alternative fuels. It dwells extensively and specifically on
Petrochemicals and Petrochemical Products which are in great world over. It enumerates the CHEMISTRY
OF PLASTIC FORMATION and various characterization tests and typical feed characterization as
evaluated in the CARBON BLACK PROCESSING UNIT and the routine test conducted. The research
concludes that Polymers is the product that account for the bulk of petrochemical output and technology
breakthrough, economies of scale and competition between producers have sustained this trend of which the
reversal does not appear to compromise the future of plastic, while growth prospects remain good. The
research recommend that since some saturation could appear above consumption, a level at which problems
of waste, pollution and recycling becomes important and should be taken into consideration for the better
future of our environment.
Keywords: Petrochemicals, Upstream, Plastic/Petrochemical industry, Polymers, Plastic Formation.
Introduction
The term “Petrochemicals” is generally applied to the branch of chemistry that employs hydrocarbons, oil and
gas as raw materials. It occupies a leading position today in the field of chemistry, but an even greater one in the
history of material progress in the twentieth century.
The petrochemical industry produces polymers (plastics, synthetic fibers and rubbers), detergents,
fertilizers and fine chemicals, as well as alternative fuels. This enormous range of products and the degree of
domestic comfort that they offer could never have been achieved with natural products alone1.
63
Petroleum Cuts Natural Gas
Basic units
Major Intermediates
Ammonia/Methanol Olefins Aromatics
Alternative fuels Fertilizers Plastics Synthetic fibers Detergents
and rubbers Fine chemicals
Chemistry and the Petrochemical industry

The petrochemical industry is concerned with large scale production of petrochemicals and petrochemical
products: which includes Rubber, Solvents, Paints, Dye, Drugs, Agricultural input such as (Fertilizers,
2
Insecticide, Herbicides and Pesticides), Pharmaceuticals etc . The Products of Petrochemicals are used in
several Companies such as: Paper Industry: where the naturally occurring cellulose in plants gives only a low
grade news print papers, consequently, some appropriate products are incorporated into the cellulose to produce
the well desired high quality glossy paper
• Solvent Industry: Where indispensable solvents such as Ethanol, Methanol, Ether, Petrol, acetone as
well as Benzene (all Petroleum Products) are used as raw materials.
• Production of Plastic containers of various shapes, dimension and sizes.
• Several Products generated from the Petrochemical Industry are involved in the manufacture of drugs
.
• Production of Fertilizer is heavily dependent on refinery products and several other Industry3,4,5.
EXPLORATION AND
DRILLING
MARKETING
AND TRANSPORTATION GAS ANALYSIS
LUBE PRODUCTION AND REFINING

LUBRICATION THE CHEMIST AND THE OIL
INDUSTRY PETROCHEMICALS
RESEARCH AND DEVELOPMENT
POWER GENERATION
ENVIRONMENT AND
POLLUTION CONTROL
SAFETY AND WASTES
MANAGEMENT
64
Source: 7
Fig. 1: Shows an illustration of the Chemist Reaching at to the Various Division of the Oil
Industry
The Nigerian Petroleum and Petrochemicals/Chemical Industry
At present the Petrochemical Industry in Nigeria can be said to be virtually at an infant state of development. What
we have is an assortment of small to medium scale downstream Plants that process about 80% imported plastics or
Petrochemical to produce a variety of consumer products, and three Petrochemical Plants(namely Polypropylene
and Carbon Black in Ekpan near Warri and Linear AlkylBenzene Plant in Kaduna) producing less than 20%
basic raw materials for the downstream industries6.
Between 1970 and 1978, the number of plastic processing factories in Nigeria increased from 12 to120. Plastics
consumption rose from 43,400 tonnes in 1972 to 124, 300tonnes in 1978. Most of the factories were engaged in
technically non – exacting operations such as simple injection and compression moulding, film and pipe extrusion
to fabricate household wares such as jerry cans, buckets, film for packaging, floor tiles, footwear and synthetic
rubber products etc.
The most obvious unwholesome features of these polymer processing factories is not only their high dependence
on imported raw materials but also the total absence of facilities for the manufacture of technical components and
sophisticated machinery and automobiles. Such products are invariably imported into this country at high cost.
These high levels of foreign exchange depletion have in no small way led to our present economic difficulties and
the rather precarious position of our local manufacturing industries, especially those dependent on petrochemical
raw materials6.
Ideas and Methods
Upstream
Exploration and Drilling
In the exploitation or production of crude Oil, the optimum use and interplay of chemicals and additives is a
desideratum for success of production activities especially in the modern techniques of Enhance Oil Recovery
(EOR). The Chemist is thus well integrated in the art of Petroleum Engineering, usually called the Production
7
Chemist (e.g. Seplat Petroleum). He is expected to find solution to the myriads of chemicals oriented problems
encountered in the course of drilling. These include well stimulation which is concerned with evaluation and
identification of chemicals (acids, toluene and additives) that will enhance oil recovery from reservoirs. In drilling
fluid engineering, the Chemists has to design the correct drilling mud composition that will ensure the balance and
control of formation pressure to avoid blow-outs. Table 1 shows typical composition of drilling mud's as used in
the industry. The Chemists is also involved in the design of well casing and additives for use in the cement mixture
thus further reducing the risk of blow-outs5,7,4.
Table 1
Chemicals Functions
Barite, Calcium Carbonate Weighing agents
Bentonite, Xanthan, Polyacryl Viscosifiers
Amides
Lignosulphonate, Tanin Dispersants (Deflocculating Agent)
Carboxynethylcellulose, Starch Fluid Loss Additives
Polyacrylates
Emulsifiers, Sodium Dichromate Specialty Chemicals
65
Gas Analysis
In furtherance of Upstream activities, Chemists also determine various gaseous composition. With the creation
of the Nigerian Gas Company Ltd as a subsidiary of NNPC and the construction of the Escravos - Lagos Pipeline,
the need for stringent monitoring (gas transmission) becomes more paramount. Gas sent as feedstock to the
Egbin NEPA power Station must meet certain specification with respect to Methane, Ethane, Propane and
Butane composition. This analysis is performed by Chemists using the Gas Chromatograph. The hydrocarbon
composition enables such significant parameters as the Calorific value to be determined. Besides, Chemists
determine the Moisture Content and Dew Point (which is the temperature at which moisture condenses into
water droplets7.
The dew point is a very important parameter for gas transmission as the moisture contained in gases can together
with the hydrocarbons form solid gas hydrates which may end up blocking pipes and filters. The moisture
content of gases can also be directly linked to corrosion problems. The control of these phenomena which can
negatively impact on operational activities is achieved through Chemists7.
Development and Procedures
Petrochemicals
Petrochemicals as the name implies are chemicals derived from petroleum and at the bench scale were originally
developed by Chemists2. The contribution of the petrochemicals industry to enhance living standards worldwide
and the multiplier effects so created are positive attributes, which can rightly be claimed by Chemists. In the
Nigerian petrochemical scene, the Chemists have equally proved their mettle in their various roles in Poly-
6,7,8
propylene, Carbon Black and Linear, Alkyl Benzene Plants .
Polypropylene
In Propylene Polymerization the Chemists first ensures that the Quality of the raw materials are in conformity
with the standard specification as stipulated by Plant licensors. These raw materials include Propylene (main raw
material) and other additives and catalyst materials. A typical Propylene specification is shown in the Table 2
below 7.
Typical Propylene Specification
Component Specification
Propylene 94% mole (min.)
Propane 6% “(max)
Hydrogen 10 -12 ppm wt.
Nitrogen 100 pp Max
Methane 20 – 50 ppm
Moisture 1 ppm wt. Max
Ethane 25 ppm max
Ethylene 30 – 50 ppm max
Butylenes 5 ppm wt. max
Butanes & Heavier 100 ppm wt. max
1, 3 Butaliene 10 ppm wt. Max
Cyclopropane 10 ppm wt. Mx
Carbondioxide 1 ppm wt max
Carbon monoxide 1 ppm wt. Max
Carbonyl Sulphide 25 ppb
Sulphur 1 ppm.
66
Raw propylene from the refinery is passed through the purification unit to meet the above specification which is
the polymerization grade quality. Chemists thus monitor the efficiency of the purification unit. consequently,
7
evaluating the extent of Hydrogen Sulphide, and Carbonyl Sulphide removal, among others .
Carbon Black
Chemist in Carbon Black processing unit ensure that the main feedstock Decant Oil meets design specification
and also control the quality of the intermediate and final products in order to obtain marketable products with
desired and specified quality. A number of tests are thus performed at various stages of manufacture and storage.
Typical feedstock characteristics as evaluated by the Chemist is as shown.
Typical Decant Oil or Carbon Black Feedstock Characterization
Table 3.
Propane 6% “(max)
Hydrogen 10 - 20 ppm wt.
Nitrogen 100 pp Max
Ethane 25 ppm max
Cyclopropane 10 ppm wt. Max
Sulphur 1 ppm7.
For purpose of production control, samples of Carbon black are taken from various Plant outlets and routine test
conducted. These include:-
Iodine Adsorption No: For surface area control
§ Sieve Residue: To evaluate particle size and presence of foreign matter e.g gift.
§ Dibutylphthalate Adsorption: For evaluation of Carbon Black internal structure.
§ In addition to the above tests, Rubber compounding tests are also performed prior to shipment or
evacuation of final product.
Plastics
The plastics “revolution” only truly began immediately after the Second World War. Production grew from 1950
to 1973 at a steady and sustained rate of 15% per year, increasing from 1 Mt in 1950 to over 40 Mt in 1973.
Plastics still remain today the real driving force of the petrochemical industry, with a growth rate that is likely to
remain higher than 4% per year. Low weight, constantly improving mechanical (rigidity or flexibility), chemical
(corrosion resistance) and thermal properties, adjustable to the user's needs, reinforcement feasible by glass or
carbon fiber, these attributes have all helped to open up many markets for these new materials, including
67
building, packaging, home appliances, electronics and the automobile, in which they have partly replaced
conventional materials such as steel, glass and wood.Technological breakthroughs, economics of scale and
competition between producers have sustained these trends and caused a steady drop in prices (PVC cost 440 $/t
in 1960, 255 $/t in 1970, and them then 300$/t in mid-1989), of which the reversal does not appear to compromise
1
the future of plastics .
Wide Variety of Materials

The wide variety of sources, structures and properties make it difficult to classify plastics. A distinction is drawn
between thermoplastics, which lend themselves to extrusion and hot molding, and thermoset resins, which
harden irreversibly under the action of heat. The former, with two-thirds of total tonnage, include polymers such
as polyethylenes and polypropylene (for sheets and moldings), PVC (flexible or rigid for pipes, films, anti-
corrosion linings of storage tanks), polystyrene and its derivatives (for household objects), and polyamides for
many industrial uses. Thermosets include alkyd, epoxy, urea formaldehyde and phenolic resins (for paints,
adhesives, reinforced materials), silicones and polycarbonates1. The polyurethane family offers many varied
materials, ranging from glasses to foams. The sectors using plastics also vary widely according to country, and
can be summarized as follows in 1986/1987:
Developments in Developing Countries

Highly Concentrated Production and Consumption
Per capita plastics consumption figures vary considerably from one country to another, without any right link
with the level of development. It may be observed that the two biggest consumers are Germany and Belgium,
with about 100kg/year. The world average is very low, with 16kg/year per capita, despite the rapid growth of
consumption in a number of developing countries (including Brazil). Production and consumption are hence
very concentrated in the industrialized countries, including Western Europe, which has been in first place since
1967, with about 70% of world consumption, Total world consumption of plastics reached 86 million tons in
19881.
Uses
An Evolving Market
In the absence of new uses, the market for any plastic gradually becomes saturated. Thus the growth in the
quantities of PVC employed in electrical engineering is barely more than that of the user sector. A new
1
breakthrough in 1965, PVC for food .
68
121 (kg / year / inhabitant
103
98
96
72
59
20 20
16
3.55
FRG Belgium U.S.A Sweden Japan France USSR Latin Africa World
America
Fig.3: Plastics – Consuming Countries in 1987 (kg/year per capital)
WORLD PRODUCTION
14.5Mt1 30.0Mt1 45.0Mt1 60.0Mt1 82.0Mt1
31% Western Europe
32% North America
11% Japan
12% Eastern Europe
14% Miscellaneous
1985 1970 1973 1980 1967

Period
Fig.4: Plastics – Producing Areas

Source: Japan Plastic Industries Federation and Miscellaneous
Packaging temporarily reversed the trend, with a market that became saturated but which is expanding in the area
of pipes, for instance. Plastics also compare with each other, with polystyrene and propylene sharing the
household products field, for example.
Among the industries not yet saturated which are likely to use large amounts of plastics in the future is the
automobile industry, which, in addition to upholstery, inside fixates and the dashboard, offers outlets for plastics,
especially reinforced ones, in the structural components and the body. The trend is impressive. From 20 kg in
1965 to 95kg in 1980, the American car could incorporate 130 to 150kg of plastics, i.e. more than 12% of its total
weight. Even in France, this proportion has already reached 9 to 10%, and even 11% in 1995, for certain car
models (Result 25, Citroen BX). In other areas, telecommunications, aerospace, the petroleum industry, and
69
biomedical applications demand increasingly high performance synthetic materials with very high added value,
opening up markets for “technical” plastics, certain elastomers/thermoplastics and composite materials.
Growth prospects for plastics remain good, with an annual rate 4% in the industrialized countries (1988/1993).
Some saturation could appear above consumption of 100 to 120kg per capita, a level at which problems of waste,
pollution and recycling also become important. The developing countries are likely to witness steadier growth
which could raise African composition to 16kg/year per capita and South American composition to over
30kg/year in year 2000. World production of plastics was accordingly around 130 to 140 Mt in year 2000. The
share of the western Countries (North America, Western Europe and Japan) in this production could drop by 3 to
1
5 percentage points in year 2000, in favor of the Middle East, Latin America and Asia/Oceania .
Some Properties of Plastics

Plastics are complex compounds produced by either polymerization or condensation of various chemicals. It
includes Polyalkene such as Polythene, Polyvinyl Chloride (PVC) and Polystyrenes. Plastics have various
qualities, which make them useful to man. These include, high resistance to oils, greases, and fatty acids, having
good tear strength …. Fold endurance, 'excellent resistance to weak mineral acids, unaffected by inorganic salt
solutions, … and do not corrode in the electrical sense9.
Certainly, these are valuable qualities needed in any durable utility material and it is these that have made plastics
the ubiquitous choice material in applications in all spheres of our life. Ironically, these same desired qualities
have turned out to be properties that present problems when its usefulness loses value and needs to be disposed.
Plastic in Solid Waste: Implications for Solid Waste Disposal in Nigeria
Some types of synthetic materials began to appear in solid waste collections. One of such synthetic materials is
plastic, a product of Petrochemical origin (Akumagba,2001). Thus, plastic remain and accumulate where they
are when other components of solid wastes have long decomposed9.
Plastic Wastes
By 1915, record shows that in Nigeria, the consumption of plastic had reached 1kg /head (NNPC, 1981) in9at the
time, Nigeria was only muting the idea of a Petrochemical Industry and all plastics were imported when
Petrochemicals came on stream in the U.S.A, it was predicted that plastics apparently will be the fastest growing
outlets for some Petrochemicals. This prediction holds true for Nigerian now that the Petrochemical Industry
concerned with the production of raw materials for plastic production has come on stream. However, even from
imported raw materials presently used in plastic production today, plastics are already everywhere in Nigeria.
This can be seen from the application of large quantities of plastic in all sections of the economy: Agriculture,
Building, Fashion, Automotive, Pharmaceuticals, Home items and so on with a consistent increase in plastic
production and application, comes a commensurate increase in the quality of plastics discarded as refuse into the
environment by man. Nigeria at the moment has no such data on plastic waste but the enormous quantity being
discarded into the environment annually can only be best appreciated after a visit to a few solid waste dump in
our towns and cities where large quantities of plastic wastes may be
identified in a single large refuse dump or in the various back street where low income families resides9.
Implications of the properties of Plastics to Current Solid Wastes Disposal Method in Nigeria
The current disposal method of solid waste in common practice in Nigeria is such that depends entirely on
elements of nature to eliminate it from the environment.
Such natural agents depended upon to do this task include:
70
1. Macro-decomposers and Micro-decomposers to bring about biodegradation of solid wastes over a
period of time.
2. Erosion and /or corrosion process or action of natural acids which will bring about the gradual
disintegration of any waste not subject to biodegradation and its eventual disappearance from the
environment.
However, to be able to decompose a material present in solid waste, a micro-organism (Fungi and bacteria),
besides the required external conditions must have the capability of utilizing that material as a carbon source.
Plastics have proved unassailable to micro-organisms as a source of nutritional carbon making it not
biodegradable by micro-organism. Also because plastic are resistant to weak acids and inorganic salt solution
and does not corrode, they do not undergo swift degradation from the action of these agents.
This amount to one fact that plastics remained (even when buried) in the environment, long after other solid
waste components have disappeared under natural conditions. The higher the molecular weight of the polymer
the less the likelihood of biodegradation …… (and) the lack of rapid breakdown of most plastics is a problem in
9
refuse disposal .
This authorities in Nigeria applying the present method of solid wastes disposal, which depend completely on
nature to eliminate it from the environment have to contend with this fact that the plastic materials components of
the waste is not vulnerable to their chosen agent of disposal term. Here in lies the major implication of the
presence of plastic in solid wastes disposal in current practice in Nigeria. In addition since plastics in solid wastes
do not readily degrade, but accumulate, their presence in large Quantities have some other environment and
9
aesthetic implications .
Fire –Spread Hazard

Reduce the quality of tie-up land resources that may be used for farming or recreation.
1. Constitute an eye –sore as litters in the streets.
2. Possible blockage of water drainage routes that may lead to flooding
3. Retard percolation of water through the soil thereby affection soil fertility
4. Reduce soil microbial activity and microbial nutrient recycling and therefore tie-up available carbon in
nature.
Suggestions for Tracking the Problems

The overall thrust here is to
a. Reduce the quality of plastics in the solid wastes to be disposed or
b. Make plastic items that are easily degradable or
c. Develop effective and efficient methods of solid wastes disposal that can take care of plastics along with
other components of the waste.
The following are plausible suggestions:

1. Reducing the quantity of plastic wastes: this can be achieved through reducing at user source of the
quantity released into the environment. For example a polythene film bag for conveying commodities
from the market may be re-used several times after washing instead of discarding it or in some cases the
container can be returned to the producer for a taken price (e.g. Bottle crates)
2. Making plastic items that are easily degradable. This may be achieved by impregnating carbohydrates or
other such biodegradable substance into plastic items during production (as fillers). This will serve as
71
spots that will be easily biodegraded and thereby quicken the disintegration of the plastics in the
environment. Another alternative is the production of plastics that could be partly chemically
disintegrated, say, ultra violet light, into smaller molecules that are then more easily decomposed by
micro -organism.
3. Recycling of plastic in the waste. Possibilities now exist for recycling discarded Plastic. Plastics can
therefore be scavenged from waste dump and recycled to produce fresh plastic product.
4. Applying an effective and efficient Solid Waste Disposal Methods. Whatever the method chosen, it must
be such that will rapidly get rid of plastics along with other components of the solid waste. For example
burning in a well designed incinerator may provide the answer. An alternative may be to convert the
solid waste along with plastics containers into Reused Derived Fuel (RDF) which can then be used to
generate useful heat. This may seem far-fetched considering the level of our present technology but
records shows that it has been in use successfully since 1974 in U.S.A9.
Environmental Waste Managements

In addition to hiring services to dispose of waste properly, many Local Governments and Industries also employ
Environmental Waste Consultants to create innovative, financially sound ways to reduce the waste produced by
their community or operation. These consultants create environmental waste solutions, such as recycling, and
re-use efforts, to reduce the amount of waste that is sent to landfills or otherwise disposed of consultants also
work with waste management services to determine proper disposal methods and quantities to meet the Local
Communities Waste disposal needs without damaging the environment. In an effort to control waste and
environmental damage, many unique environmental waste solutions have been created landfills area common
14
waste management solutions in many parts of the world .
Plastic /Pure Water Sachet Recycling Technology

This Technology present the best form of managing plastic waste products including pure water sachets that
typified the entire Nigerian hands cape. The Machine is capable of recycling several tones of pure water sachets
and other plastic waste that are in abundance in the state into a peletized plastic products which is in very high
demand, the pellets is used in several other applications for the production of new plastic products like plastic
hangers, shopping bags, conduct pipe, also an exportable commodity for international market for instance; this
machine is capable of providing direct employment opportunities for about Nine Thousand (9,000) people in
Abia State alone, if at least 2 machines are installed in each of the 17 Local Government Areas of the State. It will
interest you to know that, the cost implication of the entire technology is very low, while it has very high return on
the investment. Plastic recycling plant at Olushosun for the conversion of water sachet into garbage bags – the
15
Government introduced a buyback programme for water sachet, cartons .
Glass Recycling
A U.S based company – container Recycling Alliance, has a facility that can sort and process approximately
800,000 tons of glass annually. It services all major glass bottle container manufactured in U.S. The company
reported a net income of 503 Million Us Dollars from revenue of 11.3 billion U.S Dollars in year 2001. This feat
can also be achieved this past of the world. In Carro Egypt, recycling of various types of waste has led to the
establishment of very vast small and medium industries enterprises that employed several thousands of workers.
It is equally pertinent to mention here that most of the equipment that can recycle most waste generated in this
part of the world is available in this country 15.
72
Conclusion
The problems of plastics in solid wastes accumulation due to non-degradation when other components have long
disappeared is here with us and quite noticeable therefore the earlier we begin to recognized and attend to the
problem of plastic in our solid waste, vis –a- vis the method of solid waste disposal impracticable in Nigeria, the
9
better it shall be for us, our future generation and the environment .
Chart 1. Different Plastic items (from Warri Market Surveys).

1. Plates
2. Tea cup and saucer
3. Spoon
4. Cups and tumblers
5. Jugs
6. Trays
7. Bottles
8. Jerry cans and barrels
9. Bucket
10. Bag
11. Shoes and footwear
12. Table covering and mats
13. Rain Coat
14. Pipes
15. Canopy-Covering
16. Wall clocks
17. Radio and Television Casings
18. Video and Audio Tapes
19. Chairs
20. Toys
21. Baskets
22. Ceiling Tiles
23. Machine Parts
24. Car Upholstery
25. Car Battery Casing
26. Packaging for Drugs
27. Polystyrene Packaging Casings
28. Water Tanks
29. Electrical Insulation
30. Plastic Film for Green House
31. Mattress
All aspect of human endeavours are associated with waste generation10.

Sustainable development, cannot happens without an efficient waste management system as affirmed by Section
I Paragraph 12 (g) of the General Assembly Resolution 44228 of the United States incorporated into Agenda 21
which States that environmentally sound waste management must go beyond the mere safe disposal or recovery
of wastes that are generated11. There is need to issue waste Management guidelines that is implemented and
73
practicable and waste management technologies adopted in Nigeria to reflect Nigeria's present level of
development and specific requirement and standards that are determined by technological, environmental and
12
economic criteria . A look at many of our major cities would reveal the abundance of refuse and waste that line
the roads, river banks, markets, and motor parks, in addition to those in the traditional refuse dumps. To
effectively keep our streets clean, there is the urgent need to convert most of these refuse into useful products
such as fertilizers, cooking gas etc. This is a big challenge to environmentalist, other scientists as well as other
13
stakeholders .
The first Phase of the Petrochemical Plant are in Warri and Kaduna in Addition to the Commissioning of the Four
Refineries. With the coming on stream of the National Fertilizer Company (NAFCON) at Onne there is massive
Financial input into Agricultural Section aimed at achieving self sufficiency in Food Production. It is generally
recognized that in any modern economy the three sectors; Agriculture, Iron and Steel and Petrochemical /
Chemicals interlock to effect rapid industrial developments and raise the standards of living of the citizens of the
Nation, hence economic independence6
Problemsand Prospects
The Polymer and Carbon Black Plants for the Phase I Petrochemicals have been commissioned. Also the Phase
II (Eleme Petrochemicals) comprising the Olefins Complex and another Polypropylene Plant have been
commissioned. Substantial funds have to be sourced and made available for PhaseIII comprising (BTX)
Benzene Toluene and Xylenes Complex. This will be the precursor for the birth of true Petrochemical
downstream industries.
The development of this Country's Petrochemical industrial base has suffered setback since the seventies mainly
because successive governments have pursed that laudable objective with less than total commitment. The
financial and opportunity losses to this nation arising from indecision of the executive are simply staggering.
The estimated cost of the entire Petrochemical Complex in 1970-1974 development plan has escalated to more
than 800% in the current plan. It is realistic to expect even more outrageous upwards revisions project costs so
long as precious time is lost. If the basic Petrochemicals Industry had been established here in the seventies, it is
most likely that a reasonable industrial base would have been achieved in this Country by now6.
In forecasting the prospects of Petrochemicals in the Industrial development of Nigeria towards the beginning of
this century, one only needs to draw attention to the impressive records of achievements of other developing
countries similarly endowed with indigenous petroleum. Notable example is Mexico that started a total
production more than 100,000 tonnes of five different basic Petrochemicalin the early sixties. The Mexican
Petrochemical industry increased total productions to nearly 2 Million tones in 26 different products by 1970 and
by 1981 a total production of 9million tons of 40 different products had been achieved. What has been achieved
in Mexico is also possible in Nigeria if the requisite political will and motivation is present especially in the light
of the present democracy6.
The Old Port Harcourt Refinery was Commissioned in 1965. The Warri Refining and Petrochemicals Company
was Commissioned in 1978. To optimize it's gaseous and Fluid Catalytic Cracking (FCC) bottom Products
Polypropylene Plant and the Carbon Black Plant respectively were Commissioned in 1988. The Kaduna
Refining and Petrochemicals Company was Commissioned in 1980. It has the Linear Alkyl Benzene (LAB)
Plant among others units. The New Port Harcourt Refinery is a modern Plant, Commissioned in 1989 with
8
Polypropylene Plants . NNPC had (Until recently) the Monopoly of Petroleum Products Supply to over 7000
retail outlets mostly operated by the Seven Sisters – Agip, ElF, Mobil, Chevron, Shell, Exxon and Texaco8. In
74
conclusion, economic independence implies ability and capacity to shun foreign powers working through IMF
or World Bank. It implies ability to be self sufficient in food and Agriculture. Ability to support transportation.
Ability to defend ourselves from foreign aggression. This was Hilter's strategy before he commenced the 2nd
Word war. Nigeria can be in that position and a sound and carefully planned chemically industrialized
6
development strategy is the only way to complete economic independence .
Recommendations
1) People should recycle as much as possible to reduce waste. Many major cities offer recycling for plastic,
14
paper and glass products .
2) Plastic/Pure water sachet Recycling Technology present the best form of Managing water p l a s t i c
waste products including pure water sachets that typified the entire Nigerian Landscape.
3) Plastic recycling plant at Olushosun for the conversion of water sachet into garbage bags …. the
15
Government introduced a buy back programme for waste sachet, cartons, paper and glass .
In Cairo Egypt, recycling of various types of wastes has led to the establishment of very vast small and medium
industries/enterprises that employed several thousands of workers15.
References
1. J. Masseron (1990), Petroleum Economics Edited. Part 5: Petrochemicals by J. Cheli. 1990 Edition
Technip, Paris and Institute Francais du Petrole Publications. Rueil – Malmaison. 4th Edition Updated
and Expanded. Pages 387 – 431.
2. A.D. Ogodo (2008), Oil-field Chemistry, Petroleum and Petrochemical Industries: The Role and
Contribution of the Chemist. Conference on: General Studies: Goal to a Balanced Education. Ozoro
Journal of General Studies (OJOGS), Delta State Polytechnic, Ozoro, Nigeria, Volume 1 (2008)
Published by School of General Studies, Delta State Polytechnic, Ozoro. 1(353-362).
3. A. O. Igbuku (1999), Chemistry and Nigeria's Industrialization. “The Delta Chemist” (A Biannual
Magazine of the Delta State Chapter of the Chemical Society of Nigeria Delta CSN). 1 (1). October
1999. Pages 35 – 38.
4. A.D Ogodo (2008), Oil Field Chemistry, Petroleum and Petrochemical Industries: The Role and
Contribution of the Chemist. Ozoro Journal of General Studies (OJOGS) Delta State Polytechnic,
Ozoro, Nigeria. Vol. 1, 2008. Published by School of General Studies, Delta State Polytechnic, Ozoro.
Pages 353 – 362.
5. A.D Ogodo and H. I. Esemuede (2011), Career Prospects and Opportunities for Scientists in Science
Laboratory Technology for the Service of Man Through the Study of Science. Conference on: Prospects
th th
and Challenges of Science Laboratory Technology (SLT) Professions in Nigeria. 26 – 29 October.
Oduduwa Hall, Obafemi Awolowo University, Ile-ife, Osun State, Nigeria. 27th Annual National
75
Conference/Scientific
6. E.E Nkang (1999), Chemical Industries: The Sure Way to Nigeria Economic Independence.The Delta
Chemist. A Bi-Annual Magazine of the Delta State Chapter of The Chemical Society of Nigeria.
October.1 (1). Pages 23 – 27.
7. E.E. Nkang (2001), Chemists in the Oil Industry. “The Delta Chemist” (A Biannual Magazine of the
Delta State Chapter of the Chemical Society of Nigeria (Delta CSN). Volume 3. September 2001. Pages
27 – 31.
8. E. E. Nkang (1999), Refining Imperatives for the Next Millennium: The Nigerian Challenges. “The
Delta Chemist” Magazine. A Biannual Magazine of the Delta State Chapter of the Chemical Society of
Nigeria (Delta CSN) Publication No 1. October 1999. Pages 39–42.
9. P.E. Akumagba (2001), Plastics in Solid Waste: Implications for Solid Waste Disposal in Nigeria.
September, 2001. The Delta Chemist, Publication No.3. A Bi-annual Magazine of the Delta State
Chapter of the Chemical Society of Nigeria. Page 20 – 22.
10. A.D Ogodo (2004), Modern Trends and Strategies for Effective Waste Management in Nigeria.
th th
Conference on: Effective Waste Management: A key to National Development. 25 -27 November. Obi
th
Wali Integrated Cultural Centre, Port Harcourt, Rivers State, Nigeria. 34 Annual Conference and
General Meeting of the Nigerian Society of Chemical Engineers (A Division of Nigerian Society of
Engineers (NSE) (Port Harcourt 2004).
B
11. A.D. Ogodo (2006 ), Effective Waste Management Issues in Nigeria. Edited Text of a Paper Originally
meant for Presentation at the Delta State Stakeholders Forum on Effective Waste Management, Slated
for Unity Hall, Government House, Asaba, Delta State, Nigeria on the 28th of June, 2006 Published in
Chemistry in Nigeria Magazine, A National Magazine of the Chemical Society of Nigeria Vol. 3 No. 2:
September, 2008. Pages 16-19.
12. A.D. Ogodo (2007), Developing Waste Management Strategies for A Systematic Approach to
th
Management of Waste in the Oil and Gas Industries. Proceedings of the 30 Annual International
Conference of the Chemical Society of Nigeria, Abuja, Nigeria. Pages 169 – 174.
A
13. A.D. Ogodo and Q. O. Idehen (2006 ). The Need for Harmonization, Effective Enforcement and
Implementation of Existing Environmental Laws and Guidelines on Waste Management Problems in
Nigeria: The Future Direction. ChemTech Journal (An Official Publication of the Chemical Society of
Nigeria, Edo State Chapter) Volume 2:386 – 399.
14 A.S. Daboner (2014), Environmental Sanitation, Waste Management and Management Systems. A
Paper Presented on the Occasion of Uvwie L.G.A Environmental Sanitation Awareness Campaign flag-
off. 28th August, 2014. Uvwie L.G.A. Secretariat, G.R.A., Effurun, Delta State, Nigeria.
15. L.E. Tudararo – Aherobo (2014), A New Era in Waste Management in Uvwie Local Government Area.
A Seminar Paper Presented on the Occasion of Uvwie Local Government Council Environmental
76
Sanitation Week Tagged “Operation Keep Uvwie Clean” Official flag-Off by Hon. Sir. Bright
Edejewhro, Chairman, Uvwie Local Government Council, Effurun, Under the Distinguished
Chairmanship of Chief Sunny Daboner, JP, The Atetcte I of Uvwie Kingdom. 28th August. Uvwie Local
Government Council Secretariat, Agric Road, G.R.A. Effurun, Delta State, Nigeria.
77
Kinetics of the Photocatalytic Degradation of Chlorendic Acid (Flame Retardant) in Aqueous
TiO2 Suspension
*Boisa N., and Obunwo C.C.

Department of Chemistry, Rivers State University of Science and Technology, Port Harcourt,
Nigeria
*-Corresponding author's email: boisa.ndokiari@ust.edu.ng
Kinetics of the Photocatalytic degradation of chlorendic acid [1, 4, 5, 6, 7, 7-hexachlorobicyclo-(2, 2, 1)-

hept-5-ene-2, 3-dicarboxylic acid], a flame retardant found in the environment, has been investigated in
aqueous heterogeneous solutions using TiO2 (Degussa P-25) powder in the presence of artificial
ultraviolet radiation and oxygen. High-performance liquid chromatography (HPLC) with detector set at
221 nm was used to monitor the concentration of chlorendic acid during the degradation. The degradation
kinetics followed a Langmiur-Hinshelwood type equation. The apparent photocatalytic rate constant k,
-1 -1
was calculated as 12.0 ± 1.38 mg L min . The influence temperature and catalyst loading were studied.
Results showed that initial degradation rates increased with reaction temperature. The optimal TiO2
concentration for the photocatalytic process was determined as of 2 g/L. Kinetic data on the degradation of
persistent organic pollutants will assist in the design of efficient protocols for their removal from the
environment.
Keywords: Chlorendic acid, TiO2, HPLC and degradation
Introduction
Chlorendic acid (Figure 1) is used mainly as a flame-retardant monomer in polyester resins, epoxy resins
and polyurethane foams [1]. It is also serves as a hardening agent for epoxy resins used in printed circuit
boards [1]. In 1981 chlorendic acid was listed as a priority chemical and thus recommended for acute and
chronic toxicity tests to aquatic invertebrates, macrophytes, algae, and fish [2]. The release of chlorendic
acid to the environment may occur during preparation of flame retardant polymers, and subsequent
hydrolysis of chlorendic anhydride [2, 3]. The solubility of chlorendic acid in water is 0.35 g / 100 g at 22.8
o
C [4] and based on this, in the environment favours the aqueous compartments [2]. Its degradation in
the environment is slow [2]. The desire to find faster methods of degrading known environmental
pollutants has lead researchers to try catalyst, including photocatalyst like titanium dioxide (TiO2).
Photocatalytic degradation with TiO2 is an evolving method for the removal of organic pollutants from
water [4]. The investigation of the kinetics of degradation of chlorendic acid with TiO2 is the focus of this
study.
Chlorendic acid bears the hexachloronorborene unit in its molecular structure. The
hexachloronorborene unit is degraded only slowly after application in the environment. The
hexachloronorborene structure renders it inert to chemical and biological degradation. The inertness
can be explained by steric and inductive effects due to the chloride atoms [5]. They bioaccumulate and
biomagnify in animals that are higher in the food chain [6]. In a feed study conducted for a period of 24
months, results indicated clear evidence that chlorendic acid is carcinogenic in male and female rats and
male mice [7]. It has been suggested to be a human carcinogen [7, 8].
78
Cl Cl
Cl
Cl H
COOH
Cl
Cl
H COOH
Figure 1: Molecular structures of chlorendic acid.
Several techniques have been attempted for the removal of chlorendic acid from water. The methods included,
enhancement of the SBRs process with activated carbon [4, 9], enhancement with different solar conditions
[10] and ozonation [11] Of the methods listed only ozonation achieved close to 100 % degradation but this
method requires the use of environmentally toxic organic solvents. The heterogeneous catalysis with TiO2 has
been a promising method for the removal of organic pollutants from water. Titanium dioxide (TiO2) is metal
oxide that undergoes photo-excitation in the presence of light; displaying a relatively high efficiency when
applied as a photocatalyst. One of the unique features exhibited by TiO2 is that it absorbs irradiation that falls
within the near UV- visible region.
The titanium dioxide used for this experiment is the Degussa P-25; a binary mixture of the rutile (20%) and the
anatase (80%) form. The Degussa P-25 has a specific BET area of 50 m2g-1 and a mean particle size of 21nm. At
the molecular level the photocatalysis process involves; diffusion towards the catalyst surface, adsorption,
surface reactions, product desorption and diffusion away from the catalyst surface. The aim of this study is to
investigate the kinetics of the photodegradation of chlorendic acid using titanium dioxide (TiO2) as the
photocatalyst.
Experimental
Titanium dioxide (TiO2) photocatalyst was used as obtained from the laboratory. Chlorendic acid (99% purity)
was purchased from Sigma-Aldrich Logistik GmbH, Germany, and used as received. 6.0 M nitric acid solution,
0.1M potassium thiocyanate solution and 0.5% tartrazine indicator were freshly prepared from analytical grade
chemicals available in the Laboratory. 0.1 M silver nitrate solution and HCl solutions were prepared from
available stock solutions.
A laboratory built photoreactor [12] was employed for this study. The UV-Visible absorption spectra were
recorded using UV Evolution 300 (Thermo Electron Corporation) spectrophotometer. The high-performance
liquid chromatography-diode array detector (HPLC-DAD) with a Varian 9050 HPLC system interfaced with a
Jasco PU-980 pump and a SP 4270 integrator was used to analyze chlorendic acid. A Centaur MSE centrifuge
instrument was used to precipitate the titanium dioxide in aliquots collected for analyses. A Jenway 3010 pH
meter with a 662-1759 international probe was used for the measurement of aliquots extracted during
photodegradation reaction.
All chlorendic acid solutions used during this were systematically obtained from a 1000 mg / L stock solution. A
100 mg / L solution of chlorendic acid was scanned with the UV-visible spectrophotometer to obtain the
absorption spectrum. Nine working solutions of concentrations ranging from 10 to 500 mg / L were prepared
for calibration plot with the HPLC instrument set at 221 nm. For the Kinetic study duplicate aqueous
suspensions of chlorendic acid (200 mL, at fixed concentrations) and TiO2 (1.0 g) were placed in 400 mL Pyrex
beaker and left in the dark for 20 minutes to permit the adsorption / desorption equilibrium to be reached, then
subsequently irradiated with the UV lamp, under oxygen-equilibrated conditions in the photoreactor. To
investigation the influence of UV radiation, the degradation process was also conducted with the UV lamps
79
switched on but without TiO2. Also the experiment was conducted using varying concentrations of the catalyst
and at different temperatures. At designated times samples were extracted from the reacting systems with
pipette, centrifuged, filtered, and analysed quantitatively with the HPLC instrument. Artificial sea water of 600
mg / L chlorendic acid was prepared and subjected to photodegradation at optimal conditions identified during
this study.
The degradation of chlorendic acid with UV radiation alone and with UV radiation and TiO2 photocatalyst were
investigated. The equilibrium concentrations between 0 – 120 mins suggested slow degradation with UV
radiation when compared with UV radiation and TiO2 (Fig 2). Based on these data it can be suggested the
photocatalyst facilitates the degradation of chlorendic acid in water. To assess the efficiency of TiO2 as a
photocatalyst varying initial concentrations of the acid were subjected to photodegration. The results suggest
that the photocalyst was effective in degrading the acid at concentrations between 105 – 254 mg/L (Fig 3). For
the studied concentrations after 60 min of TiO2 aided photodegradation about 50 % of the initial equilibrium
concentrations were degraded (Fig 2).
Variation in the initial rate of decomposition of chlorendic acid as a function of catalyst loading was
investigated. It can be seen that the initial rate of degradation increases proportionally to the concentration of
the catalyst until it gets to ~ 2 g / L, then it starts decreasing as the concentration of the catalyst is increased (Fig
4). Above the value of 2 g / L the initial rate is affected negatively by increasing TiO2 loading. The decrease in
reaction rate occurred because the rate became dependent on the screening effect of the excess particles. The
excess particles mask part of the photosensitized surfaces, this happens when the solution layer thickness
exceeds the optical penetration length at the applied illumination [13]. The optimal concentration of TiO2 (2
g/L) obtained for chlorendic acid during this study fall within the range (0.15 to 8 g/L) reported [46] for several
slurry photoreactors employing TiO2.
The rate of degradation of the chlorendic acid increases proportionally with temperature within temperature
o
range (20 – 40 C) selected for this study. This suggests that the photodegradation process is enhanced by
increased temperature (Fig 5). The effect of temperature on degradation may be dominated by the rapid
desorption of substrates and intermediates from the surface of the TiO2 catalyst. The dependence of
desorption of substrates and intermediates from the catalyst at higher temperatures, leading to larger effective
surface area for the reaction have previously been reported [13, 14].
The decrease in the concentration of the chlorendic acid could be accounted for by a Langmuir-Hinshelwood
Kinetic model:
R = kè = kKCe/(1+KCe), Linear form: 1/R = 1/k + (1/kK)1/Ce. Where: R is the oxidation rate of the acid,
è the surface coverage, k the apparent rate constant. The value of the rate constant k, 12.0 ± 1.38
mg/L.min was estimated from Figure 6 with a correlation coefficient of 0.9621.
Figure 2: Photolysis and photocatalysis of chlorendic acid at 25 oC
80
Figure 3: Chlorendic acid photocatalytic degradation trends for varying equilibrium concentrations
1.2
Initial rate of Chlorendic acid
Dissappearance (mg/min)
0.8
0.6
0.4
0.2
0
0 1 2 3 4 5 6
TiO 2 (g/L)
Figure 4: Variation of the initial rate of chlorendic acid disappearance as a function of

concentration of catalyst.
3.5
Initial rate of degradation (mg/min)
2.5
1.5
0.5
0
0 10 20 30 40 50
Temperature ( o C)
Figure 5: Initial rate of degradation of chlorendic acid as a function of temperature
81
Linear transform of reciprocal initial rate vs. reciprocal equilibruim
concentration
0.7 y = (53.3 + 6.10)x + ( 0.0832 + 0.0378)

0.6 R2 = 0.9621
1/Ro ( L/mg.min)
0.5
0.4
0.3
0.2
0.1
0
0 0.002 0.004 0.006 0.008 0.01 0.012
1/Ce (L/mg)
Figure 6: Plot of inverse of initial rate against inverse of equilibrium concentration. Experiment conducted at
o
25 C using TiO2 (5 g/L) in 200 solution. Equilibrium concentration used: 105, 143, 199, 240 and 740 mg /L
Conclusion
Heterogeneous photocatalytic degradation of chlorendic acid using UV-irradiation and TiO2 (Degussa P-25)
have been achieved.
The quantity of TiO2 that produced highest rate of removal was obtained at a concentration of 2 g / L.
Temperature increase enhances the photocatalytic degradation of chlorendic acid TiO2.
Degradation kinetics of chlorendic acid in water is first-order and it follows the Langmiur-Hinshelwood model
with kinetic constant k, 12.02 ± 1.38 mg / L.min.
References
1. http://ntp.niehs.nih.gov/ntp/roc/eleventh/profiles/s033chlo.pdf seen 20/07/05 seen 20/07/2016.
2. Federal Register / Vol. 47, No. 25, 1982/ Notices
3. Segev O., Kushmara A., and Brenner A., Int Environ Res Public Health, 2009, 6, 478
4. Honda K., J. Photochem. Photobiol. A: Chem, 2004, 166, 63.
5. Schulte-Hostede S, Gab S. And Kort F., Chem. Ber., 1978, 111, 2646.
6. Cochrane W.P. and Fobres M.A., Chemosphere, 1974, 3, 41.
7. http://www.inchem.org/document/iarc/vol48/48-01.html seen 20/07/2016
8. http://www.inchem.org/document/ehc/ehc/ehc185.htm#SubSectionnumber:1.16 seen
17/07/2016
9. He Z., Yang S., Ju Y., and Sun C., J of Environmental Sciences, 2009, 21, 268
10. Rashed M.N., and El-Amin A.A., Int J Phys Sci, 2007, 2, 073
11. Stowell J.P. and Jensen J.N., Wat. Res. 1991, 25, 83.
12. Boisa N., J.Appl Sci Environ Manage, 2013, 17, 335
13. Carp O., Huisman C. L. and Reller A., Progress in Solid State Chemistry, 2004, 32, 33
14. Al-Rasheed R., and Cardin D.J., Chemosphere, 2003, 51,925
82
GAS CHROMATOGRAPHY-MASS SPECTROMETRIC ANALYSIS OF METHANOLIC LEAF
EXTRACT OF BLIGHIA SAPIDA
THERESA I EDEWOR AND NIMOTALAI O KAZEEM
DEPARTMENT OF PURE AND APPLIED CHEMISTRY, LADOKE AKINTOLA UNIVERSITY OF
TECHNOLOGY, OGBOMOSO, OYO STATE, NIGERIA
EMAIL: tiedewor@lautech.edu.ng
ABSTRACT
Blighia sapida is a medicinal plant whose fruits and leaves possess therapeutic properties. This research is aimed
at identifying the phytochemical constituents present in the leaves. The plant leaves were extracted with n-
hexane and methanol. Phytochemical screening was carried out on both extracts while only the methanolic
extract was subjected to GC-MS analysis. The phytochemical screening revealed presence of saponins and
steroids in the n-hexane extract while tannins, steroids and flavonoids were present in the methanolic extract.
Twenty one compounds were separated and identified using GC-MS. The predominant compounds were
Ethanol, 2-butoxy- (23.283%), n-hexadecanoic acid (12.214%) and Phytol (9.803%). Some of the identified
compounds possess biological properties. The presence of these compounds in the leaves of Blighia sapida
confirms its ethnomedical uses.
Keywords: Blighia sapida; medicinal; Leaves; ethnobotanical; Phytochemicals; GC-MS
INTRODUCTION
Health is as important to man as is his food. The maintenance of man's health has gone beyond the use of only
synthetic drugs to the use of plant materials which could serve as both food and drug. Synthetic drugs either have
side effects that affect the consumer negatively or are too expensive. A safe alternative is to resort to natural
products such as plant materials whose potency in alleviation of ailments have been proved by numerous
researches. There are several medicinal plants around the world and 15% of these have been screened for their
therapeutic values.1 Herbal drugs can either cure or act as supplement. Some people consume herbal drugs so as
to aid the body system in its function.2 Medicinal plants can serve as raw material base for the production of semi-
synthetic drugs. The growing awareness of the medicinal importance of plants has led scientist to find
3, 4
correlations between the phytochemicals present in a plant and its pharmacological activity.
Blighia sapida also known as Ackee is a tree that grows across West African countries. It grows to a height of
about 10 m. The leaves are 15-30 cm in length and 5-8 cm wide. The fruit is pear shaped and green but turns red
when ripe. It contains three shiny seeds. Blighia sapida is a member of the Sapindaceae family of plants. Much
research has been carried out on the fruit which is eaten as food in some African countries, Jamaica and other
South American countries.5, 6 The unripe fruit is known to contain some poisonous substances known as
7, 8
hypoglycins. The aqueous extract of the seed is used for the treatment of headache, cold and expulsion of
9 10 11
parasites while the leaf extract is used to treat diarrhea diabetics, dysentery, cold and pains. Other uses
include ascaricidal and insecticidal,12 the juice from the leaves is used as eye drops in opthalmia and
conjunctivitis,13 The aim of this research work is to determine the phytochemicals that are present in the leaves of
Blighia sapida and thereby ascertain its ethnobotanical use.
EXPERIMENTAL
Sample collection and preparation
The plant leaves were collected from a medicinal farm in Egbe, Kogi State, Nigeria. The plant was shown to Prof.
S. O. Owa of Landmark University, Omu aran, Kwara State, Nigeria; Who got the local name and the picture of
83
14
the plant and identified the plant from the book by Z. O. Gbile and M. O. Soladoye . The leaves were air dried in
the laboratory for two weeks and pulverized using a sterilized food blender. All solvents used were obtained from
Sigma Chemical Co. (St. Louis, MO. USA) and were distilled before use.
Extraction
The plant leaves were extracted with n-hexane and later with methanol using a Soxhlet extractor. The extract was
concentrated by distillation and later evaporated to dryness.
Phytochemical analysis
Harborne, 199315 method for determination of the presence of phytochemicals in a crude extract was used to
analyze the crude extracts for the presence of flavonoids, alkaloids, saponins, tannins, terpenoids, steroids and
glycosides.
Gas Chromatographic-mass spectrometric analysis
The crude extract was subjected to GC-MS. Model 7890A; Agilent Technologies interfaced with mass selector
detector (MSD) model 5975C was used. 5% diphenyl and 95% dimethyl siloxane were used to pack the column.
The sample was dissolved in methanol and 1µl of the sample was taken and separately used for the analysis. The
samples were injected in the split mode with ratio 50: 1. Helium was used as the carrier gas which had a constant
flow rate of 1.5 ml/min. The injection temperature was 300oC while the total running time was 49 mins. The
o o
ionization voltage was kept at 70eV. The oven temperature was kept at 80 C held for 4 mins and ramped to 270 C
o
at the rate of 3.5 C/min holding for 6 min.
RESULTS AND DISCUSSION
The identification of phytochemicals present in plant materials which are used either for food, cosmetics and
medicine is very important. This can serve as a source of information in the discovery of therapeutic drugs and the
disclosure of new sources of economic phytochemicals which can be modified through synthesis to produce
complex chemical substances that will be beneficial to human health. Blighia sapida is a medicinal plant and it is
important to have knowledge of the phytochemicals that are present in the plant. Phytochemical analysis of the
leaf extracts identified saponins and steroids in the n-hexane extract while tannins, steroids and flavonoids were
present in the methanol extract as presented in Table 1. These phytochemicals have been reported to possess
antidiarrhoeal, antihemorrhagic,16 anti-inflammatory,17 antioxidant, antimicrobial and health promoting
18
properties.
Table 1: Phytochemical screening of the crude extracts of Blighia sapida
Extracts Alkaloids Saponins steroids terpenoids Tannins flavonoids glycosides
n-hexane + - + - - - -
methanol - - + + + + -
The total ion chromatogram of the methanolic leaf extract of Blighia sapida showing the separated
compounds with their peak areas and retention times is depicted in Fig. 1.
84
Fig. 1: Total ion chromatogram of the methanolic leaf extract of Blighia sapida
The GC-MS analysis of the methanolic extract revealed presence of twenty one compounds as shown in Table 2.
The first compound to emerge from the GC was o-xylene with retention time
Table 2: GC-MS analytical report on the methanolic crude extract of Blighia sapida
S/N Retention Name of compound Molecular Molecular
time weight formula
1 5.291 o-xylene 106 C8H10
2 7.310 Ethanol, 2-butoxy- 118 C6H14O2
3 9.117 Benzene, 1-ethyl-3-methyl- 120 C9H12
4 10.389 Benzene, 1, 2, 3-trimethyl- 120 C9H12
5 28.959 Naphthalene, 1, 2, 4a, 5, 8, 8a-hexahydro-4, 204 C13H24
7-dimethyl-1-(1-methylethyl)-, [1S-(1a,
4aa, 8aa)]-
6 36.202 Tetradecanoic acid 228 C14H28O2
7 37.474 10-heptadecen-8-ynoic acid, methyl ester 278 C18H30O2
8 37.734 1, 2-benzenedicarboxylic acid, bis(2- 278 C18H26O4
methylpropyl)
9 37.961 3, 7, 11, 15-teramethyl-2-hexadecen-1-ol 296 C20H40O
85
10 38.221 Phthalic acid, butyl-2-ethylbutyl ester 306 C18H26O4
11 38.401 Hexadecanoic acid, methyl ester 270 C17H34O2
12 38.833 n-hexadecanoic acid 256 C16H32O2
13 39.611 8, 11-octadecadienoic acid, methyl ester 294 C19H34O2
14 39.650 9, 12-octadecadienoyl chloride, (Z, Z)- 298 C18H31ClO
15 39.752 Phytol 296 C20H40O
16 39.815 Heptadecanoic acid, 16-methyl- methyl 298 C19H38O2
ester
17 39. 925 17-octadecynoic acid 280 C18H32O2
of 5.291min while the last to emerge was squalene with retention time of 44.780 min. The predominant
phytochemicals are Ethanol, 2-butoxy- (23.283%), n-hexadecanoic acid (12.214%) and Phytol (9.803%). These
compounds were identified based on their peak area, retention time, molecular formula, molecular structure,
molecular mass and calculated fragments. The interpretation of the mass spectra of the unknown compounds was
carried out by comparing the mass spectrum of the unknown compound with a matching spectrum obtained from
NIST 2011 database incorporated into the computer system of the GC-MS equipment. Identified bioactive
phytoconstituents such as phytol and n-hexadecanoic acid have also been identified in the leaves of Woodfordia
19 20 21
fruticosa, Kedrostis foetidissima and Aloe vera. The presence of phytol in the plant leaves confirms its use as
an antimicrobial and anti-inflammatory agent. Squalene has been reported to possess antioxidant, antitumor and
anticancer properties.17 Some of the identified compounds have been reported to exhibit biological properties as
shown in Table 3.
Table 3: Biological properties of Blighia sapida methanolic leaf extract
S/N Name of compound Type Properties
1 Squalene Triterpene Antibacterial, antioxidant, antitumor,
immunostimulant, lipooxygenase inhibitor.
Pesticide.
2 Phytol Diterpene Antimicrobial, anticancer, diuretic, anti -
inflammatory.
86
3 9, 12 -octadecadienoyl Linoleic acid Antiinflammatory, nematicide, insectifuge,
chloride, (Z, Z)- hypocholesterolemic, cancer preventive,
hepatoprotective, antihistaminic, antiacne,
anti arthritic, antieczemic, 5á -reductase
inhibitor, antiandrogenic.
4 n-hexadecanoic acid Fatty acid Antioxidant, nematicide, insectifuge,
hypocholesterolemic, 5á-reductase inhibitor.
5 Hexadecanoic acid, Fatty acid Antioxidant, hypocholesterolemic.
methyl ester ester
6 3, 7, 11, 15 -teramethyl- Terpene Antimicrobial
2-hexadecen-1-ol alcohol
Source: Dukes. Phytochemical and Ethnobotanical Databases. Phytochemical and Ethnobotanical Databases.
22
www.ars-gov/cgi-bin/duke/. 2013
The presence of these compounds in the plant leaves confirm its use against dysentery, diarrhea, insects and
nematodes.
CONCLUSION
The methanolic leaf extract of Blighia sapida contains terpenes, Fatty acids, fatty acid esters, other esters,
terpene alcohol and aromatic compounds. The biological properties of the plant leaves are in agreement with its
tradomedical use. Research is on-going to isolate and determine biological and pharmacological parameters of
the phytoconstituents.
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of variation and preferences in ackee (Blighia sapida KD Koenig) fruit traits in Benin: implications for
domestication and conservation. Journal of ethnobiology and ethnomedicine 6 (12): 1-14.
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Indies, Jamaica, pp 254.
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Phytochemistry (Oxford), 19, 19, 199-203.
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13. Gbile Z. O, Soladoye M. O. (2002).Vernacular Names of Nigerian plants (Yoruba). 2 ed. Forestry
Research Institute of Nigeria, Ibadan, pp.72.
14. Harborne J. B. (1993). Phytochemical methods, A guide to modern techniques in plant analysis.
Chapman and and Hall, New York, pp.60-66.
15. Dukes. Phytochemical and Ethnobotanical Databases. Phytochemical and Ethnobotanical Databases.
www.ars-gov/cgi-bin/duke/. 2014.
16. Dhivya R, Manimegalai K. (2013). Preliminary Phytochemical Screening and GC- MS Profiling of
Ethanolic Flower Extract of Calotropis gigantea Linn. (Apocynaceae). J Pharma. Phyto. 2(3): 28-32.
17. Salini S. T and Shankar S. (2014). Phytochemical and GCMS Analysis of Canthium coromandelicum
Leaves Extract Int. J. PharmTech Res. 6 (5): 1731-1735.
18. Priya D, Rajaram K and Suresh KP. (2011). Phytochemical studies and GC-MS analysis of Caralluma
fimbriata wall. Int. J. Pharm. Res. Dev. 3: 105-110.
19. Grover N, Patni V. (2013). Phytochemical characterization using various solvent extracts and GC-MS
analysis of methanolic extract of Woodfordia fruticosa (L) Kurz. Leaves. Int. J. Phar. & Pharma. Sci.
5(4): 291-295.
20. Kalaisezhiyen P, Sasikumar V. (2012). GC-MS evaluation of chemical constituents from methanolic
extract of Kedrosti foetidissima (Jacq.) Cogn. Asian J Pharm Clin Res 5(4): 77-81.
21. Arunkumar, S. and Muthuselvam, M. (2009). Analysis of phytochemical constituents and antimicrobial
activities of Aloe vera L. against clinical pathogens. World J. Agric. Sci. 5: 572-576.
88
CHARACTERIZATION AND DISTRIBUTIONS OF ALIPHATIC AND POLYAROMATIC
HYDROCARBONS IN SOILS OF OIL SAND DEPOSITS AREA OF ONDO STATE, NIGERIA
T.A .Adedosu1*, O.K. Adeniyi2, O.H. Adedosu3
1
Department of Pure and Applied Chemistry, PMB 4000, Ladoke Akintola University of Technology
Ogbomoso, Nigeria
2
Department of Chemistry, Rhodes University Grahamstown, South Africa
3
Department of Science Laboratory Technology, PMB 4000, Ladoke Akintola University of Technology,
Ogbomoso, Nigeria
*Corresponding Author: Dr. Adedosu Taofik Adewale (Associate Professor), Department of Pure and
Applied Chemistry, PMB 4000, Ladoke Akintola University of Technology, Ogbomoso Nigeria.
Mobile No: +234- 07055302824; +234-8146683211
Email address: taadedosu@lautech.edu.ng; terryad2001@yahoo.com
ABSTRACT
The concentration, distribution profile and possible sources of n-alkanes and polycyclic aromatic hydrocarbons
in the soil of oil sand deposits in Ondo Southwest Nigeria were investigated to determine the present level of
contamination prior to exploitation of the oil sand deposits. Surface soil samples (0-15 cm) were collected from
Ilubinrin, Agbabu and Oloda communities. The level and distribution of polyaromatic hydrocarbons and n-
alkanes in the soil samples was analyzed using Gas chromatography coupled with a flame ionization detector.
The mean concentrations of PAHs in the soil of Agbabu, Ilubinrin and Oloda were 36.07±1.61 ìg/kg, 16.09±0.76
ìg/kg and 10.04±0.55 ìg/kg and the mean concentrations of identified, resolved n-alkanes were 14.03±0.55
mg/kg, 22.38±0.99 mg/kg and 30.37±1.20 mg/kg respectively. The odd-numbered alkanes were dominant in
Agbabu and Ilubinrin while the even numbered alkanes are dominant in Oloda. The distribution of total
combustion specific PAHs was higher in most of the area than the non-combustion specific PAHs and the highest
percentage of total carcinogenic PAHs were 28%, 33% and 45% in Oloda, Ilubinrin, and Agbabu respectively.
The studied area was not polluted with PAHs, but there was evidence of PAH's accumulation in the soil. Various
n-alkanes and PAHs ratios were obtained to identify the possible sources of PAHs and n-alkanes. Mixed sources
of pyrogenic and petrogenic PAHs were discovered with predominant pyrolytic sources due to combustion of
wood, biomass, and bitumen seepage.
Keyword: Polycyclic Aromatic Hydrocarbons, Oil sand, Diagnostic ratio, Aliphatic Hydrocarbons,
Bitumen.
1.0. INTRODUCTION
Mining and exploitation of natural resources have posed environmental, social and economic problems to many
communities in developing countries. Nigeria has abundant mineral resources, including solid minerals and
fossil fuel. The fossil fuel consists principally of crude oil, coal, and bituminous sand. The Nigerian oil sands
region is one of the largest natural bitumen deposits in the world with an estimated 30 to 40 million barrels and
potential bitumen recovery of about four billion barrels (Adegoke et al., 1991). Despite the economic importance
of these resources, there is growing concern over the environmental and health risks that could result from
contaminants released during mining, exploration, refining, and transportation of these resources (Ite et al.
2013). PAHs and n-alkanes are ubiquitous in the environment and released through different anthropogenic
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activities and natural sources (Wilckle 2000). PAHs and n-alkanes accumulate in soil from atmospheric
deposition, seepage from natural sources, biogenic sources and discharge of wastes. Petrogenic sources and
pyrogenic sources are the main anthropogenic sources of PAHs (Inengite et al., 2010). Identifying the impact of
oil sand industrial activities on the ecosystem requires knowledge of the background level and pre-development
concentration of contaminants in environmental media (Wilkund et al., 2012). Previous studies on the Nigerian
Oil sand regions have focused on the geology, mineralogy (Egunyomi and Olatumile, 2010) and methods of
processing with limited studies on the pre-development level of potential contaminants in the area. Therefore,
this study describes the level, distribution pattern and sources of hydrocarbons (n-alkanes and PAHs) in soil of
selected communities within the Nigerian oil sand deposit belt.
2.0. MATERIALS AND METHODS

Site Description
The Nigerian oil sands region stretches across Edo, Ondo to Lagos state covering about 140 km. The area falls
within the Afowo formation and the Araromi shale of the Eastern part of the Dahomey basin. The area is
composed of sandstone impregnated with bitumen. Okosun, (1990) described the stratigraphic settings of the
basin. Three different communities with the largest reported oil sand deposits, namely Oloda, Agbabu and
Ilubinrin were selected for the study. These communities are small villages surrounded by scattered farm
settlements and forest. Oil sand outcrops and seepage of liquid bitumen from the subsurface are seen in the area,
contaminating soil and farmland, surface and groundwater. Figure 1 is the map of Ondo state showing the
selected communities.
Sampling and Sample preparation
Soil samples were collected from locations close to the oil sand deposits from three selected communities.
Samples were collected between February and March 2015. Soil sampling stations were established at 100 m X
100 m away from the deposit and each other. Ten soil samples were randomly collected from different points
within each sampling station and composite. Soil samples (0-15 cm) were collected into pre-cleaned glass
containers with a calibrated soil auger, wrapped with aluminum foil, covered and composite. The composite
sample was homogenized and air-dried. Four composite soil samples were collected at the study areas. The air
dried samples were ground with a pestle and mortar, and sieved through a 2 mm sieve and kept in brown bottles
before analyses
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Fig. 1 Map of Ondo State showing sampling Area
Extraction and Instrumental Analysis
The air-dried soil samples were extracted according to USEPA method 3550C-ultrasonic extraction. The
concentrated extract was fractionated into aliphatic, aromatic (PAHs) and polar fractions by eluting with of n-
hexane, n-hexane/dichloromethane (2:3 in v/v) and methanol respectively. The identification and quantification
of PAHs and n-alkanes were performed with an Agilent 7890A Gas chromatograph (GC) with a flame ionization
detector (FID). The GC was equipped with an HP5 fused silica capillary column (30 m x 0.25 mm internal
diameter with 0.25 µm film thickness). Helium gas was used as the carrier gas at a flow rate of 1 ml/ min and
nitrogen was used as the makeup gas. 1 ìl of the fractioned extract was injected in a splitless mode. The GC oven
temperature was programmed as follows: For the aliphatic fraction, the oven temperature was programmed from
60 ºC held for 2 min and ramped at 6 ºC/min to 300 ºC and held for 20 min. For PAHs analysis the oven
temperature was programmed from 60 ºC (held for 2 min) to 180 ºC at a rate of 30 ºC min-1, from 180 ºC to 250 ºC
at a rate of 5 ºC min-1 and from 250 ºC to 330 ºC at a rate of 15 ºC min-1 (held for 8 min). The USEPA 16 priority
PAHs and C8-C30 standard mix were analyzed using the same instrumental conditions. Identification of n-
alkanes and PAHs compounds was based on a comparison of samples retention times with that of the reference
standard mix and quantification was performed with five points external calibration method.
3.1. Characterization and Spatial Distribution of n-alkanes
Table 1 shows the distributions of n-alkanes in soil samples from the oil sand deposit region of Ondo state,
Nigeria at different sampling locations. The spatial distribution of n-alkane varies significantly between
the sampling locations. The total mean concentrations of n-alkanes in Oloda (14.03±0.55 mg/kg dw),
Ilubinrin (22.38±0.99 mg/kg dw) and Agbabu (30.37±1.20 mg/kg dw) were lower than the Nigerian
EGASPIN intervention concentration (50.00mg/kg dw) (DPR 2012) and the Oklahoma Department of
environmental quality limit (50.00mg/kg) (Fagbote and Olanipekun 2010). This is comparable to the total
concentration of n-alkanes recorded in soil of other communities; outskirt of Beijing, China (0.60-39.92
mg/kg: Zhu et al., 2005); Port Harcourt, Nigeria (16.74-2025 mg/kg; Olajire et al., (2007) observed lower
concentration of total n-alkane in soil within the vicinity of the Agbabu oil sand deposit (0.422-2.289
mg/kg), lower concentrations were also recorded around a petroleum industrial park in China (0.576-
1.202 mg/kg; Teng et al., 2015). The soil is characterized by n-alkanes ranging from C12-C30 with the highest
concentrations of n-C15 and the absence of light chains n-alkanes between C8- C11 (Table 1). The highest n-
alkane concentrations were between C15 to C25 in the entire sampling locations (Table 1) suggested
pollution from biogenic or heavy hydrocarbon sources. The total sum of odd numbered alkane; 7.034
mg/kg, 12.52 mg/kg and 16.31 mg/kg were higher than the even number alkanes; 6.995 mg/kg, 9.861 mg/kg
and 14.06 mg/kg in Oloda, Ilubinrin and Agbabu. Odd numbered alkanes were dominant at all the
sampling stations in Ilubinrin and Agbabu. However, there was spatial variation in the spatial distribution
of the odd and even numbered alkane within the sampling station in Oloda (Figure 2). The dominance of
even numbered alkane is commonly observed in bitumen and oil derived from carbonate or evaporate
rock (Ficken et al., 2000; Fagbote and Olanipekun 2012), even-numbered alkanes dominant the n-alkanes
in Oloda 1, 2 and 4 while odd number alkanes were predominant in Oloda 3. This suggested a significant
petrogenic input of n-alkanes in the soils of Oloda. The nC17 n-alkane is considered as a typical indicator of
hydrocarbons from algae (Zegouagh 1998). The nC17 is high in all the sampled soil, this indicates a
significant input of petrogenic source of n-alkanes in the soil. The carbon preference index, CPI show the
relative contribution of n-alkanes from biogenic and anthropogenic sources. A CPI ? 1 indicates the
relative contribution of n-alkane from natural (biogenic) sources and a CPI ? 1 show anthropogenic
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(petroleum) sources. The values of CPI for the whole range of hydrocarbon, CPI1 (∑(C9-C29)/ (∑ (C8-C30)
in the soil of Oloda (1.01), Ilubinrin (1.26) and Agbabu (1.16) were about 1 (Table 1). This indicated that
the main sources of n-alkane in the soil were from anthropogenic activities and not from natural biogenic
sources. The CPI in the range of nC13- nC25 ? 1.0 indicated petrogenic sources of normal alkane (Wang et
al., 2003; Vladimir et al., 2012). The CPI2 (∑ (C13-C25)/ (∑ (C12-C24) in the soil of the studied areas were
shown in Table 1. The CPI2 values in the soil of the study areas ranged from 1.1 to 1.4. The ratio of total n-
alkanes to n-C16 is less than 15 for petroleum-contaminated soil (Ou et al., 2004). The value of ∑(THC)/n-
C16) in the area was greater than 15. The ratios showed that petroleum input of n-alkane was evident in the
areas. The ratio of short chain to long chain alkanes (SCC/LCC) less than 1 usually represent n-alkanes
produced by higher plants, marine animals, and sedimentary bacteria. SCC/LCC values close to 1 suggest
that n-alkanes are mainly from petroleum and plankton sources and SCC/LCC values greater than 2
often show the presence of fresh oil (Wang et al., 2006). Petroleum distillates have the SCC/LCC value
smaller than one (Wang and Fingas, 2005). The ratio of SCC/LCC (Table 1) in the soil samples were
greater than one suggesting an anthropogenic source of n-alkanes.
Table 1 Mean concentration of n-alkanes in the soil of Oloda, Ilubinrin and Agbabu (mg/kg, dwt)
Sampling Site
n-alkane
Oloda Ilubinrin Agbabu
C8 ND ND ND
C9 ND ND ND
C10 ND ND ND
C11 ND ND 0.045
C12 0.001 0.001 0.114
C13 0.505 0.610 2.027
C14 0.587 1.031 1.81
C15 1.300 2.968 3.679
C16 1.393 2.308 3.245
C17 0.912 2.023 2.814
C18 1.157 1.487 2.191
C19 0.912 2.295 2.139
C20 1.032 2.069 2.088
C21 1.202 0.617 1.116
C22 1.605 1.861 2.513
C23 0.988 2.023 2.225
C24 0.704 0.364 0.988
C25 1.136 1.894 2.112
C26 0.409 0.635 0.904
C27 0.046 0.059 0.131
C28 0.084 0.093 0.193
C29 0.033 0.033 0.026
C30 0.023 0.012 0.011
THC 14.029 22.383 30.371
SD 0.552 0.99 1.20
∑even HC 6.995 9.861 14.057
∑Odd HC 7.034 12.522 16.314
MH C22 C15 C15
CPI1 1.01 1.26 1.16
CPI2 1.07 1.36 1.24
∑( TCH)/C16 10.07 9.70 9.36
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THC: n-alkane total concentration; CPI1: carbon preference index calculated as ∑C9–C29)/∑C8–C30); CP2:
carbon preference index calculated as ∑C13–C25)/∑C12–C24); MH: Major hydrocarbon; ∑Odd HC: sum of
odd numbered alkane; ∑even HC: sum of even numbered alkane; SCL/LCL: ratio of short chain to long
chain alkanes. ND: Not detected.
Fig. 2 Distribution of even and odd-numbered alkanes in soil of Oloda, Ilubinrin, and Agbabu
3.2. Distribution and Concentration of PAH in soil
Table 2 show the average concentration and distribution of the US EPA priority PAHs in the soil of Oloda,
Ilubinrin and Agbabu. The average concentration of PAHs (?16PAHs) in the soil of Agbabu, Ilubinrin and Oloda
were 36.069±1.61 ìg/kg, 16.087±0.76 ìg/kg and 10.043±0.55 ìg/kg respectively. The total average
concentration of PAHs in the soil were higher than the average concentration of PAHs (0.07±0.036 ìg/kg) earlier
reported in the soil of Agbabu (Fagbote and Olanipekun,2010) and the endogenous concentration (1-10 ìg/kg)
resulting from plant synthesis, anthropogenic input results in value of total PAHs greater than 10 ìg/kg (Olajire
and Brack, 2005). The total PAHs concentration in the soil samples was comparable to the concentration of PAHs
in some of the communities (Odiabidi (367 ìg/kg) and Ahoada (168 ìg/kg);) hosting crude oil deposits in the
Nigeria Niger Delta reported by Ana et al. (2009). Maliszewska-Kordybach (1996) classified soil contamination
based on the total concentration of PAHs (?16PAHs) as non-contaminated (? 200 ìg/kg), weakly contaminated
(200-600 ìg/kg), contaminated (600-100 ìg/kg) and heavily contaminated (? 1000 ìg/kg). The soil of Agbabu,
Ilubinrin and Oloda were non-contaminated but there is increasing accumulation of PAHs in the soil of the area
from anthropogenic activities. The proportion of combustion specific PAHs in Agbabu, Oloda and Ilubinrin were
55%, 44% and 45% of the total mean PAHs concentrations respectively. The ring size distribution of PAHs in the
soil was shown in Figure 3. The results showed that all ring sizes of PAHs were present in the soil samples with
an increasing order of 3>4>5>2>6- ring PAHs in all the studied sites.
Table 2 Mean Concentration of PAHs in soil of Agbabu, Oloda and Ilubinrin (Mean±SD µg/kg, dwt)
?TPHA: Total sum of 16EPA priority PAHs; ?LPAH: Low molecular weight PAHs; ?HPAH: High molecular
93
PAHs Abbreviation
Oloda Ilubinrin Agbabu
Naphthalene NA 0.607±0.21 2.552±0.81 3.032±0.92
Acenaphthylene Ant 0.903±0.26 1.335±0.34 1.722±0.61
Acenaphthene Ace 0.924±0.27 1.815±0.60 2.497±0.51
Fluorene Fl 1.585±0.80 1.736±0.34 2.949±0.54
Phenanthrene Ph 0.880±0.81 0.655±0.70 1.962±0.58
Anthracene Anth 1.612±0.18 1.584±0.42 4.845±1.74
Fluoranthene Flu 0.639±0.37 0.723±0.20 1.787±0.67
Pyrene Py 0.072±0.02 0.342±0.31 1.166±0.51
Benzo(a)anthracene BaA 0.095±0.04 0.560±1.05 2.099±1.07
Chrysene Chr 1.128±0.33 1.375±0.57 5.133±2.04
Benzo(b)fluoranthene B(b)F 1.076±0.50 1.964±0.54 0.798±0.46
Benzo(k)fluoranthene B(k)F 0.056±0.02 0.404±0.16 5.141±1.68
Benzo(a)pyrene B(a)p 0.333±0.10 0.428±0.54 1.395±0.69
Indeno(1,2,3cd)pyrene Icdp 0.080±0.03 0.450±0.54 1.051±0.38
Dibenzo(a,h)anthracene DahA 0.047±0.02 0.153±0.22 0.426±0.41
Benzo(g,h,i)perylene Bghip 0.009±0.01 0.013±0.00 0.063±0.34
? TPHA 10.043±0.55 16.087±0.76 36.069±1.61
?LPAH 6.510±0.41 9.676±0.62 17.010±1.11
?HPAH 3.530±0.44 16.410±0.59 19.060±1.80
?COMPAH 4.432, (44%) 7.271, (45%) 19.798, (55%)
?CPAHs 2.814, (28%) 5.334, (33%) 16.045, (45%)
weight PAHs; ?COMPAH: sum of combustion specific PAHs; ?CPAHs; sum of carcinogenic PAHs.
Fig 3: Ring size distribution of PAHs in soil of Agbabu, Oloda and Ilubinrin
The International Agency for research on cancer, IARC have identified seven PAHs as priority carcinogens. The
sum of the seven identified carcinogenic PAHs (benzo(a)anthracene, chrysene, benzo(b)fluoranthene,
benzo(a)pyrene, dibenzo(a,h)anthracene, indeno(1,2,3-cd)pyrene and benzo (k)fluoranthene) were 2.814
µg/kg, 5.333 µg/kg and 16.045 µg/kg in Oloda, Ilubinrin and Agbabu accounting for 28%, 33% and 45% of the
total mean PAHs concentration in the soil (Table 2). Highly toxic and class 1 carcinogen benzo(a)pyrene was
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detected in all the samples. The mean concentration of benzo(a)pyrene in soil of Agbabu, Ilubinrin and Oloda
were 1.395±0.69 ìg/kg, 0.428±0.50 ìg/kg and 0.333±0.10 ìg/kg respectively, thus posing high risk to the
community despite the low concentration of PAHs recorded in the area when compared to other crude oil host
communities in the Niger Delta.
3.3. Identification of sources of PAHs in the soil
Anthropogenic sources of PAHs are due to petrogenic and combustion input, PAHs derived from petrogenic
sources are characterized by the predominance of low molecular weight PAH (LPAHs) (Emoyan et al., 2015),
while PAHs from combustion sources are characterized by a higher proportion of high molecular weight PAHs
(HPAHs). Low-temperature combustion of biomass also results in the formation of LPAHs (Kumar and Kothiyal
2012). Low molecular weight PAHs are classified as 2-3 ring PAHs and high molecular weight PAHs as 4 to 6
rings PAHs (Mathieu and Friese 2012). The total sum of low molecular weight PAHs (LPAH) were higher than
the high molecular weight PAHs (HPAH) in Oloda and Ilubinrin accounting for 65% and 60% of the total PAHs
concentrations (Table 2), the abundance of LPAH could have originated from the outcrop of bitumen sand
deposits and from the combustion of biomass. The PAHs pattern at Agbabu was dominated by high molecular
weight PAHs (52% of the total PAHs concentration in Agbabu). The ratio of LPAH/ HPAH can be used to
differentiate different PAHs sources, the values of LPAH/HPAH were 1.85, 0.59 and 0.89 in Oloda, Ilubinrin and
Agbabu respectively (Table 4). LPAH/HPAH less than 1 is an indication that PAHs in Ilubinrin and Oloda are
mostly from combustion sources (Kumar and Kothiyal, 2012).
The ratio of phenanthrene to anthracene (Phe/An) and fluoranthene to pyrene (Flu/Pyr) are used to
identify PAHs sources. Combustion of organic matter at high temperature generates PAHs characterized by
Phe/An ? 10 and slow decomposition of organic matter lead to higher Phe/An ? 10 (Wang et al., 2014). The
values of Phe/An in Oloda, Iluninrin and Agbabu (Table 4) were less than 10 indicating pyrolytic sources of
PAHs in the soil. The ratio of Flu/Pyr less than 1 are characteristics of petrogenic origin and greater than 1 are
characteristics of pyrolytic origin, the values of Flu/Pyr in the entire study area were greater than 1, indicating
pyrolytic sources of PAHs. The ratio of anthracene to the sum of anthracene and phenanthrene (Ant/(Ant+Phe)),
can also be used to identify PAHs sources. The ratio of Ant/(Ant+Phe) ? 0.1 indicates PAHs sources to be of
petroleum sources while Ant/(Ant+Phe) ? 0.1 indicates combustion sources of PAHs (Budzinski et al., 1997),
the Ant/(Ant+Phe) ratios in Agbabu, Oloda and Ilubinrin were greater than 0.1(Table 3), confirming that
combustion products were the Predominant PAHs sources in the studied areas.
Table 3 Diagnostic Ratios of PAHs in Agbabu, Ilubinrin and Oloda soil

Diagnostic Ratio
Sampling
Point Flu/ An/Phen Inp/Inp+
Phen/An Bap/chr Na/Ace Flu/Flu+pyr BaA/BaA+Chr
Pyr +An Bghip
Oloda 1 0.37 12.1 0.14 0.00 0.73 0.92 0.13 0.90
Oloda 2 0.68 10.48 0.38 0.84 0.59 0.91 0.34 0.86
Oloda 3 0.58 5.97 0.37 1.21 0.63 0.85 0.35 0.92
Oloda 4 0.78 6.39 0.38 0.88 0.56 0.86 0.35 0.91
Ilubinrin 1 0.26 0.99 0.23 1.15 0.79 0.50 0.19 0.99
Ilubinrin 2 0.42 1.86 0.34 1.34 0.71 0.65 0.18 0.95
Ilubinrin 3 0.44 7.68 0.37 2.00 0.69 0.88 0.29 0.87
Ilubinrin 4 0.45 1.41 0.61 1.33 0.68 0.58 0.32 0.91
Agbabu 1 0.37 1.40 0.36 1.49 0.72 0.58 0.25 0.94
Agbabu 2 0.38 1.51 0.25 1.23 0.72 0.60 0.18 0.97
Agbabu 3 0.47 1.82 0.18 0.91 0.68 0.65 0.14 0.89
95
Also, the ratio of benzo(a)anthracene to benzo(a)anthracene+chyresene (B(a)A/(B(a)A+Chyr) in Ilubinrin and
Agbabu showed a mixed source of pyrogenic and petrogenic origin. The ratio of (B(a)A/(B(a)A+Chyr) greater
than 0.2 indicates petroleum sources and between 0.20 - 0.35 indicate mixed sources while values greater than
0.35 indicates pyrolytic sources (Yunker et al., 2002; Hwang et al., 2003). The ratio of fluoranthene to the sum of
fluoranthene and pyrene (Flu/(Flu+Pyr)) could differentiate petroleum combustion sources from other types of
combustion sources. Olajire and Brack, 2005 and Inengite et al., (2010) suggested that the ratio of
Flu/(Flu+Pyr)? 0.4 indicates petroleum combustion sources, values between 0.4-0.5 indicates pyrogenic, while
? 0.5 indicates combustion of coal, grasses, and woods. The Flu/(Flu+Pyr) ratios in Ilubinrin (0.50-0.88), Agbabu
(0.58-0.65) and Oloda (0.85-0.92) were greater than 0.5 thus suggesting that the PAHs in the soil samples likely
originated from pyrogenic sources derived from the combustion of wood and grasses. Also, it is presumed that
the ratio of indeno(1,2,3cd)-pyrene (Ind/Ind+BghiP) less than 0.2 indicate petroleum sources. (Ind/Ind+BghiP)
in the range ≥ 0.2 and ≤ 0.5 indicates petroleum combustion sources and >0.5 as grass, wood and coal
combustion sources. The ratios of Ind/Ind+BghiP were greater than 0.5 in the entire study area confirming that
burning of wood and grasses are the major sources of pyrogenic PAHs in the entire study area.
2.0. Conclusion
The concentrations and origin of PAHs and n-alkanes in the soil of selected communities in the oil sand region of
Nigeria were reported. The level of PAHs are below the regulatory standard value and the area is classified as
unpolluted by PAHs, but the accumulation of PAHs and n-alkane in the soil were observed. Analysis of
diagnostic ratios and molecular indices reveals that anthropogenic activities and seepage of bitumen from the
bitumen deposit are the major sources of n-alkane and PAHs in the soil of the area. The n-alkanes in the soil are
predominately from petrogenic sources from the oil sand deposit sites while the PAHs are from mixed petrogenic
and pyrogenic sources with predominant input from the combustion of wood, grass. The observed values of n-
alkanes and PAHs will serve as useful baseline data for monitoring and assessment when exploration begins.
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diagenetic alterations in Arctic surface sediments (Lena River delta and Laptev Sea, Eastern Siberia), II.
Molecular and isotopic studies of hydrocarbons. Org Geochem 28: 571-583.
97
Comparative Assessment of the Environmental Dynamics of Dissolved Organic Carbon (DOC),
Nitrogen (DON), Phosphorus (DOP) and Characterization of DOM in Subtropical Wetlands of Some
Northern Nigerian States.
1
Department of Chemistry, Sule Lamido University, P. M. B. 048, Kafin Hausa, Jigawa State, Nigeria
2
Department of Pure and Industrial Chemistry, Bayero University, P. M. B. 3011, Kano, Nigeria
Email: sayzay37@yahoo.com; aaaudu.chm@buk.edu.ng
ABSTRACT
This study focuses on the comparative assessment of the environmental dynamics of Dissolved organic carbon
(DOC), nitrogen (DON) and phosphorus (DOP) using the persulphate wet-oxidation method. The dissolved
organic matter (DOM) was characterised using FTIR. The averaged values for DOC, DON and DOP during the
sampling period ranged from 7.454±0.73 to 13.686±1.50 mg/l, 9.485±1.83 mg/l to 21.796±14.73 mg/l and
1.651±0.38 mg/l to 6.109±0.58 mg/l respectively in the wetlands studied. These values are above the 4 mg/l, 0.03
mg/l and 1 mg/l limits set for DOC, DOP and DON respectively. The infrared spectrum showed the structural
composition of the DOM present in the wetlands. FTIR results indicated that the DOM of these wetlands contain
the phenolic hydroxyl groups, hydroxyl group, conjugated double bond of aromatic family (C=C), and free
carboxyl groups. Analysis of variance (ANOVA) indicated that there is a significant difference (p<0.05) between
the DOD, DOP and DON concentrations in both the wet and the dry seasons. High concentrations of DOC, DOP
and DON in water have being implicated for trihalomethane formation, eutrophication and, methemoglobinemia
in young infants and cancer respectively.
Keywords: Dissolved organic carbon, Dissolved organic matter, FTIR, wetlands
INTRODUCTION
Dissolved organic matter (DOM) are formed from the incomplete decomposition of organic materials, which
include soil organic matter, plant residues and soluble particles released by living organisms, including bacteria,
algae and plants (Reddy et al. 1999).
DOM is the key component in the global carbon cycle, the storage pool of carbon (C), nitrogen (N), and
phosphorous (P), the important supplier for heterotrophic microbes, a regulator of pH of the lake water, and the
complexant and sorbent of the inorganic and organic pollutants, affecting their mobility, transformation, and
toxicity (Wu et al. 2008).
Sources and compositions of DOM are key topics in DOM research (Wu et al. 2008). Chemical properties of
DOM determine their fates and roles in environments, such as reactivity, degradation rates, and photochemical
behaviour. Chemical characteristics and environmental behaviour of DOM from environments have not been
given much attention in the northern wetlands of Nigeria.
Wetlands are areas of marsh, fen, peatland or water whether natural or artificial, permanent or temporary, with
water that is static or flowing, fresh brackish or salt, including areas of marine water the depth of which at low tide
does not exceed 6 meters (Ramsar 1994).
The ecological significance of DOM is not very well understood and has not been clearly defined. This is due to
lack of a clear understanding of DOM composition and its cycling, biodegradability and relationships with
nutrients.
Natural wetlands, as significant carbon pools, have been increasingly recognized as important DOC sources for
adjacent waters (Baker et al., 2008). Much work has been completed concerning the concentration and
bioavailability of soil dissolved organic carbon from wetland ecosystems (Zhang et al., 2005; Zhang et al., 2008;
Guo et al., 2010), yet rare information is available about how the dynamics of DOC are derived from surface
ponds in wetland ecosystems (Wang et al., 2010; Song et al., 2011).
A study of seasonally flooded, freshwater cypress-tupelo wetlands in South-eastern United States showed high
98
concentrations of DOC, DON and DOP in the summer season than in the spring season (Alex, et al. 2012).
Seasonal changes in DOC and DON concentrations have been previously documented in both wetland (Fraser, et
al. 2001) and forested landscapes (Qualls and Haines 1991; Yano, et al. 2004).
Previous studies found that urban areas contribute to DOC, DON and DOP concentrations whereas agricultural
land contributes predominantly to DON and DOP (Harrison et al., 2005). Additionally, wetlands are intensively
cultivated with extensive use of nitrogen-based fertilizers and organophosphoric insecticides. More so, Pittal et
al. (2014) reported that intensive industrial activities, extensive use of nitrogen-based fertilisers and
organophosphoric insecticides in the Evros basin resulted in waters enriched in DOC, DON and DOP
concentrations.
The aim of the research is to investigate the dynamics of DON, DOP, DOC and characterize DOM in some
wetlands in Northern Nigeria
The Study Area
The Hadejia-Nguru wetlands are located between latitudes 12o15'N and 12o55'N, and between longitudes 10oE
o
and 11 E in the sudan savanna of Nigeria. Baturiya wetland occupies an area of approximately 101,095 ha and is
located between latitude 12°31'N and longitudes 010°29'E.
The Lower Kaduna-Middle Niger wetlands

The wetlands are located on the extensive floodplain of the mid-section of River Niger (in Nigeria) and the lower
? ' ? ' ? '
course of one of its main tributary, River Kaduna. The wetlands extend from Jebba (9 00 N 4 50 E) to Baro (8 35 N
6?25'E) along the Niger, and from Wuya Bridge on the River Kaduna, along the Jebba – Bida (9?00'N 6?00'E) road
downstream to Pategi (8?45'N 5?37'E) on the south bank of the Niger. Jebba and Lokoja wetlands were studied in
this area.
Material and methods
All chemicals that were used in this study are analytical reagent grade. Solutions were prepared using distilled-
deionised water.
Sampling and sample treatment
3
Surface-water samples were collected in 1000 cm white, low-density polyethylene Carboy bottles from 35 sites
spread across four sampling locations during the early part of the dry season (i.e 5 Oct. 2014 to 28 Jan 2015) and
the wet season (i.e May to September, 2015). Composite samples were collected 200 m apart from each sampling
?
location. The samples were kept at about 4 C and taken to the laboratory for analysis. The bottles were washed by
-1 -1
soaking in 0.5 molL HCl followed by 0.1 molL NaOH for 24 h each. Water samples were filtered through
precombusted (470?C for 4 h) 0.7-µm GF/F glass fiber filters (Whatman International) rotary evaporated and
freeze-dried for FTIR.
Determination of Dissolved Organic Phosphorus and Nitrogen
Sample preparation
To 50 cm3 of the sample in a 125 cm3 Erlenmeyer flask 1 cm3 of 5.5 M H2SO4 solution and 25 cm3 of alkaline
persulphate digestion reagent were added. This was boiled gently on a preheated hot plate until a final
3 3
volume of 10 cm is reached. Cooled and diluted to 30 cm with water. 1 drop of phenolphthalein indicator
solution was added and neutralized to a faint pink colour with NaOH solution.
S2O82- + H2O 2 HSO4- + ½ O2
Procedure for phosphate determination
30 cm3 of the sample was Pipetted into a 50 cm3 calibrated flask, 8 cm3 of the mixed reagent (125 cm3 of 2.5
M tetraoxosulphate (VI) acid, 37.5 cm3 of 0.0324 M ammonium molybdate, 75 cm3 of 0.1 M of ascorbic acid
3
and 12.5 cm of 0.008214 M potassium antimony tartrate solution) was added and dilute to volume with
water. This was mixed thoroughly. After at least 10 minutes but not more than 30 minutes, the absorbance
99
of each sample was measured at 880 nm, using reagent blank as the reference solution (Charles, et al.
2003). DOP was determined as the difference between total phosphorus and hydrolysable phosphorus.
Procedure for nitrate determination
3 3 3
5 cm of digested sample was pipetted into a volumetric flask, 20 cm of water was added and 10 cm of the
reducing reagent (hydrazine sulphate, copper (II) sulphate, zinc sulphate) was added and heated for 15
minutes at 52 oC. It was allowed to cool to room temperature to form nitrite. 1 0 cm3 of 0.0581 M acidic
sulphanilamide (10:1) solution was added, shaken thoroughly for 5 minutes for the diazotization reaction to go to
3
completion. Then, 10 cm N-(1-Naphthyl) ethylenediamine dihydrochloride solution was added to form an
azo dye and the contents were diluted to 50 cm3 with water. After dilution to 50 cm3 with water, the absorbance of
the pink coloured dye was measured at 540 nm against the corresponding reagent blank (Johnes, et al. 1992).
- -
DON was determined as the difference between total nitrogen and inorganic nitrogen (NO3 , NO2 and
+
NH4 )
+
Procedure for NH4 -N Determination
3 3 3 3
25 cm of the sample was transferred into a 50 cm Erlenmeyer flask, then 1 cm phenol solution, 1 cm sodium
nitroprusside solution and 2.5 cm3 oxidising solution (a mixture of 100 cm3 alkaline citrate solution and 25 cm3
sodium hypochlorite) were added with thorough mixing after each addition. The samples were covered with
o
plastic wrap and kept in the dark at room temperature (25 C) for 1 h (Nikolaos et al., 2010)
The absorbance was measured at 640 nm. Six standards were used to prepare the calibration graph. The blank
was treated like the standards.
Determination of Dissolved Organic Carbon
Dissolved organic carbon was determined according to the method described by (HACH, 2013).
S2O82- heat 2 SO4- + e-
H 2O heat H + OH
- 2-
SO4 + H2O SO4 + H+ OH
2- -
S 2 O 8 + H 2O 2 HSO4 + ½ O2
(Liang, C., Wang, Z., Bruell, C., 2007)
Instrumental analysis
A 755S UV-VIS spectrophotometer equipped with Deuterium Lamp and Halogen-Tungsten Lamp was used for
the determination of DOC, DON and DOP concentrations. Working standards were prepared by further dilution
of 1000 ppm stock solution (of KHC8H4O4, KNO2 and KH2PO4 respectively) and a calibration curve was
constructed by plotting absorbance versus concentration. By interpolation, the concentrations of the DOC, DON
and DOP in samples were determined. Fourier Transform Infrared (FTIR) spectrometer (SHIMADZU KN S72-
120, Japan) KBr. pellet technique was used for the characterization of DOM. The spectra resolution was 2 cm-1
and 10 scans were averaged.
Data Analysis
All analyses were performed in triplicates. Results were expressed by means of ±SD. In addition, the SPSS 11.5
software was used in the statistical analysis. The difference in DOC, DON and DOP concentrations among the
different sites were tested by analysis of variance tests (ANOVA). In analysis where P<0.05, the comparisons
were considered statistically significant.
RESULTS AND DISSCUTION
Table 1: Mean Concentration of DOC
100
Wetlands Mean conc. of DOC (mg/l)
Dry season Wet season
Jebba 10.786±0.43 8.240±1.03
M-Uku 12.943±1.23 9.616±1.82
Lokoja 13.686±1.50 11.140±1.01
Baturiya 13.071±2.34 10.300±2.43
Nguru 8.271±1.64 7.454±0.73
Table 2: Mean Concentration of DOP in Dry & Wet Seasons
Wetlands Mean Conc. of DOP (mg/l)

Dry Season Wet season
Jebba 1.846±0.07 3.696±0.16
Matara-Uku 2.602±0.55 5.548±0.24
Lokoja 2.650±0.31 6.109±0.58
Baturiya 2.523±0.46 4.927±0.9
Nguru 1.651±0.38 3.650±0.37
Table 3: Mean Concentration of DON in Dry & Wet Seasons
Wetlands Mean Conc. of DON (mg/l)

Dry Season Wet season
Jebba 14.071±3.92 13.883±1.97
Matara-Uku 13.655±3.43 11.257±2.62
Lokoja 21.045±5.04 9.587±1.46
Baturiya 21.796±14.73 9.485±1.83
Nguru 7.135±2.40 11.12±1.78
Figure 1: Concentration of Dissolved Organic Phosphorus in Wetlands
101
Figure 2: Concentration of Dissolved Organic Carbon in Wetlands
Figure 3: Concentration of Dissolved Organic Nitrogen in Wetlands
102
a = Jebba, b = Baturiya, c = Lokoja, d = Nguru wetlands
Fig. 4: FT-IR spectra of the DOM from Wetlands
Marked inter-seasonal variations in DOP, DOC and DON concentrations were observed among the thirty five
sites during the study period. Variations in DOP, DOC and DON concentrations in both dry and wet seasons are
illustrated in Figure 1, 2 and 3 respectively.
The DOC concentrations showed significant difference (p<0.05) among the wetlands studied except for Matara-
Uku, Baturiya and Lokoja wetlands where the DOC concentration were not significantly different.
DISCUSSION
The mean DOC concentrations recorded in this study are 10.786, 12.943, 13.686, 13.071 mg/l and 7.454 mg/l
(Table 1) for Jebba, Matara-Uku, Lokoja, Baturiya and Hadejia-Nguru wetlands respectively during the dry
season and 8.271, 8.24, 9.616, 11.14, 10.3 mg/l and 7.454 mg/l (Table 1) are obtained for Jebba, Matara-Uku,
103
Lokoja, Baturiya and Nguru wetlands respectively during the wet season for the thirty five sites analysed.
The mean DOC concentration in River Dee system in north-east Scotland ranges from 1.2±0.2 to 10.6±1.7 mg/l
(Aitenhead, et al. 1999). This is in agreement with the concentration of DOC analysed in sites J1, J2, J3, J4, J5, J6, J7,
BR1, Ng1, Ng2, Ng3, Ng4, Ng5, Ng6 and site Ng7, which ranges from 6.42 mg/l to 11.2 mg/l but lower than those
values recorded in sites M1, M2, M3, M4, M5, M6 and site M7, which ranges from 11.6 mg/l to 16.8 mg/l. However,
the DOC concentration in a coastal freshwater forested wetland in Winyah Bay, South Carolina ranges from 15 mg/l
to 55 mg/l (Alex, et al., 2012). This is higher than the values we obtained in sites J1, J2, J3, J4, J5, J6, J7, BR1, Ng1,
Ng2, Ng3, Ng4, Ng5, Ng6, Ng7, M1, M2, M4, M5, M7, LA1, LA2, LA3, LA7, BR1, BR2, BR3, BR4 and site BR7,
which ranges from 6.42 mg/l to 13.7 mg/l but conformed with the DOC concentrations we obtained in sites M3, M6,
LA4, LA5, BR4 and site BR5, which ranges from 14.7 mg/l to 16.8 mg/l. The high values obtained are attributed to
high litter fall, land use for agriculture and decomposition of saw grasses observed in these wetlands. In contrast, the
minimum concentration of stream water DOC is 2 mg/l and the highest DOC concentration obtained in sleepers river
research watershed is 5 mg/l (Stephen, 2007). These values are lower when compared with the DOC
concentration recorded in this study. During the study period, inter-seasonal variations of DOC concentrations
were identified in different sites of the wetlands (Fig. 2). During a one year study undertaken by Wang, 2013, in
different sites of a wetland in Sanjiang plain, he obtained average values ranging from 7.08±1.38 to 48.73±3.26
-1
mg l . This range of values agrees with what we obtained in this study.
In contrast, a mean value of 23.25 mg/l was recorded for DOC concentration in the surface ponds at five natural
wetlands sites in Sanjiang plain (Wang, et al. 2013), this value is higher than what we obtained in this study. Nagao et
al. (2007) observed that DOC concentrations in river waters from tributaries of the Amur River in the Sanjiang Plain
were generally lower than 10 mg/l. The high concentrations we recorded suggest that the natural wetlands in this
study could serve as alternative DOC sources for tributaries along these wetlands. Similarly, Waiser and Robarts
(2004) reported that natural wetlands across the Canadian prairies exhibited high DOC concentrations (>10 mg/l),
acting as significant DOC pools for adjacent waters. Raphael et al. (1996) and Arrigoni et al. (2008) pointed out that
the Tivoli Bay's freshwater tidal wetlands exported DOC to the main channel of the Hudson River, but the magnitude
of DOC concentrations (<7 mg/l) in their studies were much lower than the values recorded in this study. These
variations might result from different rates of DOC production, solubility and transport under freshwater and tidal
wetlands.
In this study, we found that DOC concentrations (Table 2) in the wetlands showed the decreasing sequence of
Lokoja>Baturiya>Matara Uku>Jebba>Nguru wetlands during the study period. The greater DOC values from the
Lokoja, Baturiya, and Matara Uku wetlands compared to Jebba and Nguru wetlands might result from the varying
hydrological conditions, different rate of decomposition of litter fall and grasses in these wetlands. Nguru wetlands
were flooded with higher water levels which would lead to more anaerobic conditions, lower soil carbon
decomposition rates, and thus less release of DOC from soil and litter decomposition. Another potential driver of the
observed differences might be the contrasting nutrient contents of the wetlands. DOC is a product of SOM, soil total
nitrogen and soil total phosphorus, this could control the DOC production/consumption processes (Chapin et al.,
2002).
A great dynamics was observed in the DOC concentrations in Jebba and Lokoja wetlands in both seasons even though
they are in the same belt. This could be due to differences in the hydroperiod, production rate, adsorption and
solubility of the dissolved organic carbon. Similarly, seasonal dynamics was observed in DOC concentration in the
various sites in hadejia-Nguru (Matara-Uku, Nguru and Baturiya) wetlands. There is a significant difference in the
DOC Matara-Uku, Baturiya and Nguru wetlands. This could be attributed to the differences in soil organic carbon
content along with different hydrologic characteristics.
The Jebba and Lokoja wetlands have a slightly higher DOC concentration (13.201±0.97 mg/l) in the dry season than
104
the Hadejia-Nguru wetlands which have a value of 13.165±1.74 mg/l. There is no significant difference in the
DOC concentrations between the two wetlands location, despite their varying climatic conditions. Although the
more litter fall observed in the Hadejia-Nguru wetlands may release DOC to the water column, photolysis and
microbial utilization may reduce DOC concentrations. In contrast, in the wet season, the DOC concentration
(10.710±1.02 mg/l) in the Hadejia-Nguru wetlands is slightly higher than the value recorded (10.540±1.66 mg/l)
in the Jebba and Lokoja wetlands, though there is no significant difference in the DOC concentration obtained in
both wetlands. This may be attributed to the difference in temperature, precipitation, adsorption, solubility and
different organic carbon content of the soil as well as the hydrologic characteristics of the area.
Studies in UK have also reported that the DOC releases from different wetland types depended on nutritional
grounds (Freeman et al., 2004). The significant differences in DOC concentrations between the wetlands might
come from the fact that they held different contents of soil organic carbon along with different hydrologic
characteristics (Wang et al., 2010).
When Total Organic Carbon (TOC) is greater than 4.0 mg/l, THM levels may exceed 100ìg/l if the residence
time in the network is 2-3 days. (Ryan 2012).
As shown in fig. 1, the highest DOP concentration was found during the wet season. The high DOP concentration
was probably attributed to the accumulation of nutrients during the dry period. A large fluctuation was observed
in DOP concentrations from the different sites. Such irregular pattern could be due to the solubility and
adsorption of DOM.
The mean DOP concentration (2.259±0.46 mg/l) in Hadejia-Nguru wetlands is higher than the mean
(2.248±0.19 mg/l) obtained in Jebba and Lokoja wetlands. The extensive use of organophosphorus insecticides
in Hadejia-Nguru wetlands may have led to this dynamics observed in the dry season. In contrast, Hadejia-Nguru
wetlands recorded a low mean DOP concentration (4.708±0.503 mg/l) in the wet season when compared to the
mean value (4.903±0.37 mg/l) obtained in the Jebba and Lokoja wetlands. This may be due to the dilution of
Hadejia-Nguru wetlands and wastewaters from residential homes enter into the Lokoja wetlands.
Furthermore, the mean DON concentration (17.558±4.48 mg/l) in the Jebba and Lokoja wetlands is higher than
the mean DON concentration (14.195±6.85 mg/l) observed in the Hadejia-Nguru wetlands in the dry season.
This dynamics is also attributed to the use of nitrogen-based fertilizers and the inflow of wastewater into the
wetlands. Similarly, the mean DON concentration (11.735±1.72 mg/l) in the Jebba and Lokoja wetlands in the
wet season is also higher than the mean DON concentration (10.621±2.08 mg/l) obtained in the Hadejia-Nguru
wetlands.
Baturiya and Lokoja wetlands have significantly high DON concentration (Table 3); these could have been
caused by runoff from rainstorms. Residential buildings are also around this area of the wetlands and could also
impact the nitrate levels and cause them to rise.
Kennedy, et al. 2004 recorded average of 14.07 mg/l NO3 in wetland surface and soil-water. This value is in
agreement with what we observed in this study. DON Concentrations ranging from 2.0 to 4.0 mg/l were observed
within the Boxford marsh riparian wetland on the river Lambourin in southern England (Prior and Johnes, 2002).
This range of values is lower than what we observed in this study (9.485±1.83 to 21.796±14.73).
Compounds such as phosphate and nitrate are considered nutrients to organisms such as algae. The wetlands
contain high levels of phosphate and nitrate which both promote algae growth; however, phosphorus appears to
be the limiting factor. As the algae grow, they use up the phosphorus causing the phosphate levels to drop. The
phosphate runs out, resulting in the lower concentrations that were observed during this research.
Differences in nutrients dynamics between wet and dry seasons, suggest that litter fall, fertilizers, use of
organophosphoric insecticides and run off are the major source of DOC, DON and DOP in these wetlands but
precipitation and temperature controlled the DOC, DON and DOP concentrations in wetlands (Alex, et al. 2012)
105
FT-IR spectra
For the characterization of the DOM, the spectrum of the FTIR was recorded. The FTIR spectrums are presented
in the (Fig. 4). FT-IR spectra of the four wetlands locations were quite similar. The profile of every spectrum was
dominated by five prominent bands at 3,408, 1,645, 1,510, 1,420, and 1,145 cm- 1. The broad band around 3,408
cm- 1 was attributed to N–H or H bonded O–H stretching of phenols, alcohols, carboxylic acids, and/or other
-1
substances which contains hydroxyl groups (Francioso et al. 1996). The sharp band at about 1,645 cm
-
corresponds to aromatic C = C vibration, olefinic carbon–carbon bonds, symmetric stretching of COO groups,
and H-bonded C = O of conjugated ketones (Francioso et al. 1996; Senesi et al. 2003).
The band at 1,420 cm- 1 was assigned to phenolic O–H and symmetric bending vibrations of aliphatic C–H
-1
groups (Agnelli et al. 2000; He et al. 2009). The sharp peak near 1,145 cm can be attributed to alcoholic,
polysaccharide, polysaccharide-like C–O stretching (Olk et al. 2000; He et al. 2009). More so, the peak at 1,145
cm- 1 was contributed by C–O stretching of ethers, esters, phenols and carboxylic acids (Senesi et al. 2003; Chang
Chien et al. 2007). In spectra of all the wetlands studied, there were also minor peaks at 2,362 and 2,940 cm- 1,
which was attributed to C–H stretching of methyl and methylene groups of aliphatic chains (Agnelli et al. 2000;
-1
He et al. 2006). The weaker band at 1,510 cm in the spectra was ascribed to stretching vibrations of aromatic C =
C bond or C = O vibrations of amide groups (Duarte et al. 2005).
These results indicate that the DOM of these wetlands might contain the phenolic hydroxyl groups, hydroxyl
group, conjugated double bond of aromatic family (C=C), and free carboxyl groups.
CONCLUSION
During the study period, inter-seasonal dynamics of DOC, DON and DOP concentrations were identified in
different sites of the wetlands. The averaged values for DOC, DON and DOP during the sampling period ranged
-1
from 7.454±0.73 to 13.686±1.50 mg L , 9.485±1.83 mg/l to 21.796±14.73 mg/l and 1.651±0.38 mg/l to
6.109±0.58 mg/l respectively at the thirty five sites studied. The infrared spectrum showed the structural
composition of the DOM present in the wetlands.
The results from the DON, DOP and the DOC analysis suggest that the wetlands contained high concentrations
of these substances, which may be carried by flood water to the surrounding rivers and lakes.
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108
SYNTHESIS, CHARACTERIZATION AND KINETIC STUDIES OF Fe (II) and Cu (II) COMPLEXES OF NICOTINIC
ACID HYDRAZIDE.
*1
O.W. Salawu, 2M.S. IORUNGWA and 3M.U. Adaji.
*1
Department of Chemistry, Federal University of Lokoja Nigeria
2
Chemistry Department, University of Agriculture, P.M.B. 2373 Makurdi
3
Isa Memorial Islamic Academy, Ajiolo - Ojaji, P.O Box 324, Anyigba, Kogi State.
*
Email: olalekansalawu1@gmail.com
ABSTRACT
Complexes of nicotinic acid hydrazide [NAH] with M (II) sulphates (M = Fe, and Cu) have been synthesized by
elemental analysis, conductance, magnetic susceptibility and electronic spectral studies. The infrared data
suggest that the ligand act as neutral bidentate donors ligating through the carbonyl O and the azomethine N.
the magnetic and spectra data indicate a distorted octahedral geometry in all the complexes.The temperature
effect showed that the reaction rate increased slightly with increase in temperature. The plot of the natural
logarithm of the concentration of complexes obtained over time (sec) showed that the reaction was second
order. The activation entropy obtained from plots of ln(kobs/T) vs. 1/T of the complexes were negatives agreeing
with an associative mechanism of reaction. Also, the negative activation enthalpy for the complexes was an
indication that the reaction was rather exothermic. The lower activation energy Ea is an indication that the
complexes required lower energy before it crossed over to the product formation state at lower temperature.
Keywords: Synthesis, hydrazine hydrates, Nicotinic acid hydrazide [NAH], metal complexes, characterization,
kinetics
INTRODUCTION: The term “Kinetics” has become a popular term in several field of research such as organic
chemistry, industrial chemistry, microbiology, engineering, geochemistry etc. The term has been studied
extensively with several ground breaking and findings. While some are useful, others require the contributions
from basic inorganic chemical reactions which could serve as a bed rock guiding these findings and more so, the
kinetic study of complexes is an aspect beckoning for intensive attention. It is worthwhile to note that works on
kinetics mainly describe the correlations between time of a reaction and its concentration. The rate constant,
reaction order, and activation energy are mostly determined from every kinetic investigation I. It is impossible to
draw any conclusions about the reaction mechanism when these parameters are absent. Apart from the
unique nature and the different donating sites of nicotinic acid hydrazides, little information is available
regarding the kinetic studies of its metal complexes thus, it is pertinent to synthesize a wide variety of Fe (II) and
Cu (II) complexes of nicotinic acid hydrazide and carry out the kinetic studies on them using gravimetric method
if the information on these complexes will be made more available.
Reagents / Solvents and Instrumentation: FeSO4.7H2O, CuSO4.5H2O and Methylnicotinate were of analytical
grade reagents and obtained from commercial sources (Aldrich Chemical Co. and BDH) and used without
further purification. Elemental analyses were performed on an Elemental Vario EL III Carlo Erba 1108 analyzer.
FT–IR spectra were recorded in KBr medium on a Perkin Elmer RX1 spectrophotometer in wave number 4000 –
400 cm – 1. Electronic spectra (in DMSO) were recorded using perkin Elmer Lambda – 2B – spectrophotometer.
Molar conductance measurements were conducted using 10 – 3 M solutions of the complexes in DMSO on Elico
– CM 82 Conductivity Bridge at room temperature. Melting point /decomposition temperatures of the ligand
and complexes were determined using a Gallenkamp melting point apparatus. The magnetic susceptibilities
measurements were taken on a Gouy balance at room temperature using mercuric tetrathiocyanatocobaltate
(II) as the calibrant.
II
Synthesis of the hydrazide: The hydrazide was prepared using standard method . 14.59g, [0.29 moles] of
109
hydrazine hydrate was added to 44.01g, [0.29 moles] of methyl nicotinate in 100 mL of absolute ethanol.
Antibumping granules were added to prevent bumping. The mixture was refluxed for six hours, in a 250 mL
round bottom flask. It was poured into a beaker which was left overnight to crystallize. The resulting crystals
were re-crystallized with minimum quantity of ethanol. The final crystals obtained was allowed to dry over
CaCl2 in a vacuum desiccator and weighed. [% Yield = 73]
Preparation of metal complexes: The metal complexes have been prepared by mixing the solution of
CuSO4.5H2O and FeSO4.7H2O to the ethanolic solution of the hydrazide [NAH] in 1:2 molar ratios. The resulting
mixture was then refluxed on water bath for about 10 – 15 minutes. A coloured product appeared on standing
and cooling the above solution. The complexes were filtered, washed with ether and dried under reduced
pressure over anhydrous CaCl2 in a desiccator. They were further dried in an electric oven.
Kinetic measurements: The kinetic studies were conducted in accordance with the method described by III in
order to study the rate of formation of the complexes at varied time intervals. The complexes formed at each
180, 360, 540, 720 and 900 seconds were allowed to cool in an ice bath and the crystals obtained at each time
intervals were dried over CaCl2 in the desiccator and weighed. The reaction was carried out at 328, 333, 338,
343, 348 and 353K respectively.
Results and Discussion : The preparation of the Hydrazide as ligand from the Ester and hydrazine hydrate is
giving in the equation below.
O O
C OCH 3
C NHNH 2
6hrsReflux
+ NH 2NH 2.H 2O
ethanol + CH 3OH + H 2O
N
Hydrazine hydrate N
Ethyl nicotinate
Nicotinic Acid Hydrazide [NAH]
Figure 5: Schematic Diagram for the preparation of Nicotinic Acid H ydrazide
The reaction of the metal (II) salts and the hydrazide from equations the above to form metal complexes can be
represented in the equation below:
MSO4.nH2O + RCONHNH2 [M(RCONHNH2)2.SO4.nH2O + nH2O

2+ 2+
Where M = Cu and Fe
Table 1: Elemental Analysis of the Ligand and Metal Complexes

Molecular Mol.wg Elemental analysis
formula t
%C %H %N %M %SO42-
Calcd Found Cald Found Cald Found cald Found Calcd Found
C6H7N3O 137.14 52.50 52.11 5.10 5.25 30.63 30.23 - - - -
C12H20N6O9SCu 487.82 29.53 29.30 4.10 4.01 17.21 17.14 13.03 12.79 19.68 19.52
C12H20N6O9SFe 480.13 29.99 29.77 4.17 4.11 17.50 17.69 11.63 11.53 19.99 19.85
110
Table 2: Physical Constants for Ligand and complexes
Complexes Molecular form. Color Form. M.p Mol. Cond.
0 2 -1
Wgt C Scm Mol
NAH C6H7N3O White 137.14 161 0.01
Cu(NAH)2SO4.3H2O C12H20N6O9SCu Blue 487.823 219 0.19
Fe(NAH)2SO4.3H2O C12H20N6O9SFe Brown 480.125 146 0.80
FT-IR Spectra: The data of the FT - IR spectra of the ligand and it metal complexes are listed in Table 3. The
IR spectra of the complexes were compared with those of the free ligand in order to determine the
involvement of coordination sites in the complexes. Characteristic peaks in the spectra of the ligand and
-1
complexes were considered and compared. The band in the spectra of the ligand at 1655 cm have been
assigned to v(C=O). This band was lowered by 7 to 17 cm-1 on the complexes, indicating the involvement of
the carbonyl group in the bonding to metal VIII. The imino group i.e ?(C=N) band at 1549 cm – 1 for the free
–1
ligand and 1431 - 1555 cm for the metal complexes which are found to be shifted to lower frequency
region to 6±118 cm on complexion which suggest the involvement of the nitrogen atom of the ?(C=N)
–1
azomathine nitrogen in the bonding with the metal (II) ions IV. The stretching vibrations of the sulphate ion
v(SO42-) are observed to have a sharp singlet band at 1102 - 1032 cm-1 indicating that SO42- was involved in
the inner sphere coordination since the stretching vibrations of the sulphate ion are usually observed as a
–1 –1 V
sharp singlet around 1100 cm and a medium band around 680 cm . The appearance of broad band at
3368 – 3404 cm – 1 in the complexes have been assigned to ?(OH) which was associated with coordinated or
solvent water molecules which is indicative of the presence of water molecules in the complexes and
supporting band in the range of 611 – 951 cm attest for the distorted vibration of ?(OH) band and this
-1
VI
implies that water molecules held by a crystal lattice is present in the complexes . Hence it is concluded
that the compound (NAH) act as a neutral bidentate ligand VII. The v(M-N) stretching bands are not observed
in the spectrum of the ligsnd but are observed in the spectrum of the complexes probably because, there is
VIII
no (M-N) coordinated in the ligand . Similarly, there is no v(M-O) stretching band in the ligand because,
IX
there is no metal coordinated to oxygen .
-1
Molecular formulae FTIR in KBr pellets (cm )
S/No
2-
C=N N-N N-H C=O M-O M-N SO4 H2O
1 C6H7N3O 1549 1337 3219 1655 - - - -
2 [Cu(C6H7N3O)2]SO4.3H2O 1555 1368 3258 1638 483 398 1032 3404
3 [Fe(C6H7N3O)2]SO4.3H2O 1431 1351 3233 1643 458 407 1102 3390
111
Table 3: FTIR Spectra Bands of Ligand and Complexes
Electronic absorption spectra: Electronic absorption spectra of the ligand and there transition metal
complexes in the UV-Visible range 200–800 nm were recorded on ultraviolet visible spectrophotometer in
ethanol using a 1 cm quartz cell. The absorption bands for the complexes will help to give an idea of their
structure. The absorption spectra and magnetic moments of the metal complex are given in Table-4. The
2+
diffuse electronic spectrum of Cu complex shows two broad bands around 305 nm and 311 nm. The first band
may be due to a 2B1g → 1A1g transition while the second band may be due to charge transfer. The first band
shows structures suggesting a distorted octahedral structure for the Cu2+ metal complex. The higher value of
2+ X
the magnetic moment of the Cu complex supports the same . When compared the complexes with the free
ligand, there was a shift in frequency.
Table 4: Electronic and Magnetic data for the complexes
Compounds µeff (B.M) ?max (nm) Assignment Geometry
Cu(NAH)2SO43H2O 1.87 311
305 2B g → 1A g Octahedral
1 1
Fe(NAH)2SO43H2O 2.867 287 5T g → 5Eg Octahedral
2
Kinetic Measurements: After monitoring the reaction at the temperature of 328 K, the reaction were further
consider at higher initial temperatures of 333, 338, 343, 348 and 353 K. The reactions were observed to be
faster as temperature of the reaction was increased. Most of the reactions were observed to be completed at a
temperature of about 353 K and at a time of 360 seconds. A plot of the natural logarithm of the concentration of
the complexes against time was made (figures 1 – 2) and it showed that the reaction had a second order
dependence on the complexes formed. The observed rate constants (kobs) were obtained from the slopes of the
plots. Figure 1 - 2 show that, the rate of formation of the complexes increases with increase in time. These
XI
results agreed with the earlier studies . These regular variations in the observed rate constants as the
temperatures were increased are suggestive of the fact that the rate of formation of the complexes depended
highly on the temperatures. This fact is in agreement with research findings made by XII. For each of the
XVIII
complexes another plots of ln(kobs/T) against 1/T (Fig 3) was made, this was in accordance with . The graph
was found to be linear with very nice regression coefficient.
Fig 1: A plot of ln[Cu(NAH)2]SO4.3H2O vs time (secs) for various temperatures
112
Fig 2: A plot of ln[Fe(NAH)2]SO4.3H2O VS time (secs) for various temperatures
The activation enthalpies and entropies (? H‡ and ? S‡) was obtained from the slopes and intercepts of another
plot of ln(kobs/T against 1/T (fig5). This is in accordance with the XIII equation.
ln(kobs T) = ln(kb h) + ? S R - ? H RT
-23 -14
Where, T is the temperature in Kelvin, Kb is Boltzman's constant (1.38 X 10 J/k), h is plank constant (6.63 X 10
Js), and R is the molar gas constant (8.314 J/K/mol). The activation energy was therefore computed from the
relationship Ea = ? H - T? S
Fig 3 show the plot of ln(kobs/T) vs 1/T
Table 5: Activation data for the complexes formed at 328 K
Complexes -? H‡(KJ/mol) -? S‡(J/K/mol) -Ea(KJ/mol)

Cu(NAH)2SO4 2.403 327.314 107.361
Fe(NAH)2SO4 2.619 320.247 105.038
113
Conclusion: The complexes were recognized to be non electrolyte. The FT -IR spectra of the ligand and the
metal complexes show a clear evidence of formation of the complexes, the Uv spectra of the complexes
2+
recognized that the complexes were octahedral complexes. The magnetic moment for Fe indicates that the
2+
complex is a high spin type paramagnetic while the magnetic moment for Cu complex was also found to be
within the expected value. The kinetic results show that the rate of formation of the complexes increases with
increase in time and temperature for all the complexes formed. The thermodynamic parameters show a lower
activation complex Ea for all the complexes formed which is an indication that the complexes required lower
energy before it crossed over to the product formation state at lower temperature. Also the negative value of
the entropies is a suggestive that the reactions follow associative mechanisms. The negatives values of the
enthalpy implying that the reaction is rather exothermic.
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114
ELEMENTAL ANALYSIS AND CYANIC ACID CONTENTS OF SELECTED CASSAVA BASED
FOODS IN NIGERIA
Orishadipe Abayomi*, Afolayan Michael, Odukomaiya Doyinsola, Aguzue Onyinye & Bwai Macham David
Chemistry Advanced Laboratory, Sheda Science & Technology Complex (SHESTCO), P.M.B 186, Garki –
Abuja, Nigeria.
*Corresponding author email: oorisat@yahoo.com
ABSTRACT
This study was carried out to evaluate the hydrocyanic acid and the heavy metal contents of some foods produced
from cassava (white and yellow garri, lafun, fufu, wet and dry abacha) as obtained from the market in
Gwagwalada, Abuja - F.C.T., Nigeria. The hydrocyanic acid content was analyzed based on the method provided
by method 915 of A.O.A.C., while the heavy metal level was determined using the atomic absorption
spectrophotometer (AAS-Shimadzu Japan). The cyanic acid content reveals both wet and dry abacha as having
the highest value of 0.022mg/g while fufu had the lowest value of 0.005mg/g. White garri, lafun and yellow garri
had cyanic acid contents of 0.010mg/g, 0.015mg/g and 0.016mg/g respectively. The heavy metals analysis of all
the food samples showed that Cobalt was present only in white garri at 0.00002mg/g. Cadmium was present in
white garri, lafun and fufu while Nickel was absent only in Lafun. Lead and zinc were however present in all the
food samples analysed. From all the results obtained, it was discovered that the hydrocyanic content of processed
foods from cassava compared to cassava in its unprocessed form is lower and in a minute level depending on the
adequacy of the processing and the heavy metals level were low which makes them safe for consumption.
Keywords – Hydrocyanic, Heavy metals, Cassava, Spectrophotometer, Garri
INTRODUCTION
Cassava (Manihot esculenta, Crantz) is the most staple food crop in tropical Africa, it has a unique adaptivity to
various ecological conditions thereby earning the status of the most important famine reserve crop. (Onwueme,
1978). It is an all-season crop of the tropics and it is one of the top food crops in the world (FAO, 2002). The
cassava tuber contains 30% to 40% dry matter with about 90% starch or sugar, while the crude protein content is
2 to 4% (Adeshinwa et al., 2011). The tubers contain significant amounts of vitamins, particularly vitamin C,
thiamine, riboflavin, niacin, calcium and phosphorous (FAO, 2002). The proximate nutritive value of cassava
tuber was expressed as: moisture (59.4g.kg-1), total carbohydrate (38.1g.kg-1), lipid (0.2 g.kg-1), protein (0.7
g.kg-1), Ca+ (50 mg.kg-1), P (40 mg.kg-1), Fe+ (0.9 mg.kg-1), niacin (0.3 mg.kg-1), vitamin C (25 mg.kg-1),
thamin (0.05 mg. kg-1) and riboflavin (0.1 mg.kg-1) (Perera et al, 1989).
Two of the major constraints to the development of cassava post-harvest systems include perishability of the
fresh tubers and presence of cyanogenic compounds. Hydrogen cyanide is a highly volatile, colorless and
extremely poisonous liquid, its solution in water is called hydrocyanic acid or prussic acid.
Cassava naturally contains cyanogenic glycosides (cyanogens, mainly composed of linmarin, acetone
cyanohydrin and hydrocyanic acid). If the cassava is not adequately pressed to reduce the level of cyanogens
prior to consumption, these compounds can lead to formation of hydrocyanic acid in the gut. Exposure to this
acid can cause symptoms ranging from vomiting and abdominal pain to coma and death. (Miles, 2011)
This study is aimed at assessing the hydrocyanic acid and heavy metal levels in various processed foods from
cassava as compared with that found in unprocessed cassava.
Collection of Samples
Samples of cassava based foods (white and yellow garri, lafun, fufu, wet and dry abacha) and raw cassava were
115
purchased from Gwagwalada market, Abuja. They were stored in air-tight containers and kept in a refrigerator
until needed for analysis.
Chemicals and reagents
All the chemicals and reagents used in this study were of analytical grade and obtained from the Chemistry
Advanced Laboratory of Sheda Science and Technology Complex, SHESTCO.
Determination of hydrocyanic acid
10-20g of the sample was kept in the distillation flask and 200ml of water was added and allowed to stand for
4hrs. Distillate of about 150-200ml was collected in a solution of 0.5g NaOH in 20ml of water, 100ml of the
distillate was diluted to 250ml and an aliquot of 100ml in which 8ml of 5% potassium iodide has been added was
titrated with 0.02N silver nitrate using a microburet, (AOAC, 1990).
Elemental analysis
The samples were prepared for mineral analysis by adding 10ml of Conc. HNO3 to 2g sample and digested until a
clear solution was obtained. The digest was allowed to cool and then transferred into a 100ml standard flask and
made up to mark with de-ionized water. The mineral elements were analyzed with atomic absorption
spectrophotometer (GBC Avanta Ver 2.02 Model, Australia) equipped with air-acetylene flame. The following
metals were determined after the samples were digested: cadmium (Cd), cobalt (Co), nickel (Ni), lead (Pb) and
zinc (Zn) The concentration of the samples were reported in milligram per gram (mg/g).
The result of the heavy metal contents and hydrocyanic acid contents of raw cassava and other processed foods
from cassava is represented in Table 1. A bar chart representation of the results is also shown in Figure 1.
TABLE 1: Hydrocyanic acid and Heavy metal contents of cassava and cassava – based foods
samples Hydrocyanic acid Heavy Metal Contents (mg/g)

content (mg/g) Cd Co Ni Pb Zn
White 0.010 0.00002 0.00002 0.0061 0.0037 0.0097
Garri
Yellow 0.016 - - 0.0475 0.0007 0.0091
Garri
Wet 0.022 - - 0.0006 0.0006 0.0106
Abacha
Dry 0.022 - - 0.0663 0.0021 0.0121
Abacha
Lafun 0.015 0.00005 - - 0.0008 0.0071
Fufu 0.005 0.00008 - 0.0009 0.0013 0.0035
Cassava 0.054 0.00006 - - 0.0038 0.0089
116
Figure 1: Chart showing concentrations of HCN and Heavy metals in samples
The level of HCN in some cassava based food in Nigeria as shown ranges between 0.005 to 0.022 mg/g while it is
0.054 mg/g in raw cassava. Hydrogen cyanide is a colourless, very poisonous and highly volatile liquid that boils
o
slightly above room temperature at 26 C. Hydrogen cyanide is weakly acidic and partly ionizes in solution to
- -
give the cyanide anions CN (Blum et al., 1962). Cyanide ion (CN ) concentration of 300ppm in air will kill a
human being within a few minutes. The toxicity is caused by the cyanide ion, which prevents cellular respiration.
Blum et al (1962) report the lethal dose of HCN for adult man as 50-60mg/kg. The least concentration of HCN in
fufu is due to fermentation that is responsible for the removal of residual HCN (Ukhun et al., 1989; Otuga et al.,
1989; O'Brien et al., 1992; Milingi et al., 1993 and Oboh et al., 2007). While the finished products from cassava
may be safe for human consumption depending on how adequately it has been processed and also the time frame
for the processing as this was indicated in the value of the hydrocyanic acid content obtained for fufu (which had
the lowest value), white garri, yellow garri, lafu due to the way they were processed as they are washed more in
water than the wet and dry abacha.
Cadmium is non-essential element in foods and natural waters and it accumulates principally in the kidney and
liver (Divirikli et al., 2006). The presence of cadmium in white garri, lafu, fufu and cassava is very low when
compared with WHO standard.
One of the most important metals for normal growth and development in human beings is zinc (Divrikli et al.,
2006). Concentration of Zn in the samples reported in this study varied between 0.0035 to 0.0121mg/g with
lowest in fufu and highest in dry abacha. Therefore these cassava based foods if taken can contribute zinc to the
body.
Cobalt has little direct activity on its own in the body as it is an integral component of vitamin B12 and as such its
effects, sources and uses are very similar to that of vitamin B12. Very little information has been reported on its
concentrations in food materials. However, for this study, it's only presence in white garri at low concentration of
0.00002mg/g.
Nickel also plays some role in body functions including enzymes functions. It activates some enzyme systems in
trace amount but its toxicity at higher levels is more prominent (Divrikli et al., 2006). The Ni levels in the samples
tested varied between 0.0006 to 0.0663 mg/g. The lowest was observed in wet abacha and highest in dry abacha.
The results indicate that the heavy metals in the cassava based foods were below the safe limits of 0.0004, 0.06,
0.005 and 0.0002 mg/g for cobalt (Co), Zinc (Zn), lead (pb) and cadmium (Cd) respectively set by WHO/FAO.
The heavy metal content in each samples were quite low ranging from 0.00002mg/g-0.0663mg/g and cobalt's
level is very negligible and it was only found in white garri at a very minute level.
117
CONCLUSION
The hydrocyanic acid content of cassava processed food compared to cassava in its unprocessed form is lower
and in a minute level depending on the adequacy of the processing and the heavy metals level were low which
makes them safe for human consumption.
REFERENCES
1. Onwueme IC. The tropical tuber crops: yams, cassava, sweet potato and coco yams, Wiley, New York.
1978.
2. FAO (Food and Agriculture Organization) (2002). FAOSTAT Statistics Database. Available at:
http://aps.fao.org [Accessed 6 September 2012].
3. Perera WDA, Jayasekera PM, Thaha SZ. Table of Food Composition as Used in Sri Lanka Medical
Research Institute. 1989; Sri Lanka (UNICEF)
4. Miles D, Jansson E, Mai MC, Azer M, Day P, Shadbolt C, Stitt V, Kiermeier A, Szabo E. A survey of total
hydrocyanic acid content in ready to eat cassava-based ships obtained in the Australian market in 2008.
J Food Prot. 2011; 74, 6, 980-5,
5. Association of Official Analytical Chemist (AOAC). Official methods of analysis. 15th Ed, Association
of Official Analytical chemists. Washington D.C. 1990; pp. 808:831-835
6. Blum MS, Woodering JP. Secretion of Benzaldehyde and hydrogen cyanide by the millipede
pachyesmus crasicutis 1962.
7. Ukhun ME, Nkowocha FO. The HCN Content of Garri flour made from Cassava, (manihot spp) and the
Influence of Length of Fermentation and Location of Source, Food Chemistry, 1989; 33, 107-113.
8. Otuya CO, Ukpong E, Adesina AA. Modelling of the Rate Date from the Fermentation of Cassava
Slurry, Letters in Applied Microbiology, 1989; 9, 13-16
9. O'Brien GM, Mbome L. Variation in Cyanogen Content of Cassava during Village Processing in
Camerouns. Food Chemistry, 1992; 44, 131-136.
10. Milingi NV, Assey VD, Swai ABM, Mclaren DG, Karlen H, Rosling H. Determinants of Cyanide
Exposure from Cassava in a known Afflicted Population in Northern Tanzania, Int. J. of food Sci. and
Nutri. 1993; 44, 137-144
11. Oboh G, Oladunmoye MK. Biochemical Changes in Micro-Fungi Fermented Cassava Flour Produced
from Low and medium-Cyanide Variety of Cassava Tubers. Nutr. Health, 2007; 18, 4, 355-367
12. Divrikli U, Horzum N, Soylak M, Elci L. Trace heavy metal contents of some spices and herbal plants
from western Anatolia, Turkey. Int. J. Food Sci. Technol. 2006; 41, 712-716.
st
13. WHO. Evaluation of certain food additives and contaminants (41 Report of the Joint FAO/WHO Expert
Committee on Food Additives). World Health Organization Technical Report Series 1993; No. 837,
1993. Sydney: WHO.
118
PRODUCTION OF METHYL ESTER (BIODIESEL) USING MELON SEED OIL
AS A RAW MATERIAL
1 2
AGBO, I.U. and ONYEZEKA, E.G. (M.Sc. in progress)
1.Department of Pure and Industrial Chemistry
Faculty of physical Sciences
University of Nigeria, Nsukka, Enugu State.
2.Department of Chemistry
College of Physical and Applied Sciences
Michael Okpara University of Agriculture, Umudike, Abia State.
E-mail: ginikanwaonyezeka@gmail.com
ABSTRACT:
The production of the methyl ester was carried out using raw melon seed oil as a raw material through the process
of transesterification. The most important variable which influences the transesterification reaction time and
conversion were put into consideration.
INTRODUCTION
Biodiesel has been defined as the mono alkyl esters of long chain fatty acids derived from renewable feedstocks,
such as vegetable oils or animal fats, for use in compression ignition (diesel) engines. The biodiesel is commonly
composed of fatty acid methyl esters that are prepared from the triglycerides in vegetable oils by
transesterification with methanol. The resulting biodiesel is quite similar to conventional diesel fuel in its main
characteristics.
Biodiesel production consists of the reaction of triglycerides with methanol in the presence of a catalyst usually
NaOH or KOH. In this reaction, glycerol is obtained as by-product. Glycerol is also a useful starting material for
1
other chemicals and when purified, it is a valuable supplement for pharmaceutical products.
Materials
Material preparation
Melon seeds was purchased from Obollo Afor market, Nsukka of Enugu State on the 20th Feb. 2006. The raw
material was prepared by shelling and removing the seeds manually (hands). The raw shelled melon seeds was
sun-dried. The sun-dried melon seeds were put inside an air-tight polyethylene container, neatly labeled and kept
in the refrigerator pending analysis.
METHODS
Analytical procedures
The melon seeds was grinded using an electric grinder. The powdered sample was stored in an air-tight plastic
o o
container. The oil was extracted with petroleum ether (40 C-60 C) for 12 hours using a soxhlet apparatus. After
drying the solution with anhydrous sodium sulphate, the solvent was removed by vacuum distillation at 30oC and
the oil was analysed for specific gravity, acid value, saponification value, iodine value and pH by standard
2 o
methods as described by Winton , The refractive index was obtained using an Abbe refractometer at 40 C, while
viscosity measurement was performed using a universal torsion viscometer in a temperature controlled
circulatory bath.
Preparation of the methyl ester (biodiesel)
The reaction was carried out in a closed vessel reactor, equipped with a stirrer and a thermometer. To shift the
reaction to the right and to obtain a maximum ester yield, the molar ratio was increased to 6:1 of alcohol to oil.
Thus the required molar volume of the methanol and the quantity of the NaOH was obtained to be 243cm3mol-1 of
methanol and 3.4g of NaOH. The molar mass of the oil (884gmol-1) was obtained from the fatty acid composition
-1
of the melon seed oil and with the density measured to be 0.911gcm , the molar volume of the oil was obtained to
119
3 -1
be 970.36cm mol .
Based on the standard methods described by Lang3 whereby 1000cm3 of the oil requires 200cm3 of methanol and
3.5g of NaOH. 970.36cm3 of the oil requires 243cm3 of methanol and 3.4g of NaOH. In the same vein,100cm3 of
3
the oil was transesterified using 25.04cm of methanol and 0.350g of NaOH. The reaction was timed as soon as
the reactants were mixed. The reactor was made slightly air-tight to avoid the evaporation of the methanol. The
stirring was carried out at a temperature of 30oC for 30mins and stopped. The reactor was allowed to stand for 24
hours. The two layers of methyl ester (upper layer) and glycerol (lower layer) were separated. The methyl ester
was washed with 10ml of hot/warm water for 3 times to remove unreacted methanol, alkali, small glycerol and
soap. This was done by pouring the water into the separating funnel, shaking vigorously and releasing pressure
from time to time and allowed to stand until a clear separation of ester at the upper layer and water at the lower
layer was obtained. The layers were then separated. The methyl ester was dried for 24 hours in a desiccator over
anhydrous sodium carbonate. The fuel properties of the raw oil and its methyl ester product was determined
according to the methods of the Institute of Petroleum, London.4
The results are presented in the table below;
Parameters Raw melon seed oil Methyl ester
Refractive index at 30oC 1.480 1.448
Specific gravity 0.911 0.875
Viscosity (Cst) 33.0 6.40
Flash point (oC) 186 162
pH 5.80 7.23
Iodine value (mg/g) 128.80 119.29
Saponification value 189.30 98.18
(mgKOH/g)
Acid value (mg/g) 4.61 0.92
DISCUSSION
The chemical characteristics of the extracted oil are shown in the table above. The iodine value of the melon seed
oil classifies the oil as semi-drying oil with high degree of unsaturation. Thus judging from the iodine value of the
5
oil, the seed appears to be a viable source of oil for paint formulation and resin.
The iodine value of biodiesel is within the hydrocarbon –based diesel oil range. The specific gravity of the raw oil
is above the diesel oil range while the specific gravity of the biodiesel is within the diesel oil range. The flash
point of the oil and its methyl ester were very much higher than the limits for diesel fuel grades which is from 100-
o
130 C. The viscosity of the biodiesel is very low which implies that its combustion will not cause cold engine
start up, misfire and ignition delay.
Therefore, the fuel parameters of melon seed methyl ester has a great industrial potential as substitute for diesel
fuel.
REFERENCES
1. Vincent , et al., (1998). Industrial crops and production.
2. Winton,L.A.and B.K., (1945)., The analysis of foods, John Wiley, New York, p482.
3.Lang,X. et al. (2001). Preparation and characterization of biodiesel from various bio oils, Biores Technol
4. Institute of Petroleum, London (1993), Standard Methods for Analysis and Testing of Petroleum and Related
products,Vol.1 Willey,New York,pp1-280.
5. I. C. Eromosele, C.O. Eromosele, P. Innazo and P. Njerim (1998), Studies on some seeds and seed oils. Biores.
Technol., 64, 245-247.
120
2+ 3+ 2+
Studies on Biosorption of Co , Cr and Cd from Aqueous Solutions by Piliostigma Malabaricum seed
pod.
*Onwu, F. K and Amadi, O. K.
Department of Chemistry, Michael Okpara University of Agriculture Umudike, P.M.B 7267 Umuahia,
Abia State.
Email: frank4kalu2007@yahoo.com OR frank4kalu2013@gmail.com
ABSTRACT
2+ 3+ 2+
The potential of acid treated orchid Malabar seed pod for adsorption of Co , Cr and Cd was investigated in a
batch process. The effects of various reaction conditions such as pH, initial metal ion concentration and
temperature were studied. The adsorption process was found to be dependent on pH and the results indicate that
the optimum pH for adsorption of Cr3+, Co2+ and Cd2+ occurred at pHs 4.0, pH 7.0 and pH 4.0 respectively.
Studies on the effect of temperature revealed that the optimum temperature for the removal of the metal ions is 40
o
C. Various thermodynamic parameters calculated viz; Gibbs free energy of adsorption (G), enthalpy of
adsorption (H) and entropy of adsorption (S) showed that the process is spontaneous and endothermic. The
results obtained show that biosorption process could serve as a promising alternative for the removal of these
metal ions from aqueous solution.
INTRODUCTION
The awful rise in the use of heavy metals and their compounds in our industries presents a possible human health
risk. This is because of heavy metals toxicity, accumulation in the food chain and persistence in nature. Toxic
metal compounds coming from the earth's surface not only reach the earth's waters (seas, lakes, ponds and
reservoirs), but also contaminate underground water in trace amounts by leaking through the soil. Therefore, the
earth's waters may contain various toxic metals [1[. Heavy metals contamination of water sources is of great
concern because of their non-biodegradability and toxic effects on human beings, animals and plants in the
environment even at very low concentrations [2]. The main source of heavy metals contamination is from
various industrial, agricultural and domestic activities, such as mining operations, metal smelting and plating,
electric device manufacturing, power plants, waste incinerators, battery manufacturing, domestic effluents,
agricultural run-off etc. The effects of heavy metals in water and wastewater range from beneficial to toxic,
depending on concentration [3].
Since majority of the heavy metals are non-biodegradable, effective removal of heavy metal ions from aqueous
solutions and waste effluents through various technologies (physical or chemical) is important in the protection
of environmental quality and public health. Various chemical and physical methods have been used to remove
metal ions from solutions in the past few decades. These include: chemical precipitation, solvent extraction, ion
exchange, evaporation, reverse osmosis, electrolysis and adsorption. Among these methods, chemical
precipitation, solvent extraction, ion exchange and adsorption are more commonly used [2]. Chemical
precipitation has traditionally been used to remove heavy metal ions from wastewater with relatively high
concentrations of metals. The operation of chemical precipitation is simple but generates large quantities of
sludge, which require further disposal. In addition, chemical precipitation is usually not effective in removing
trace levels of metal ions from aqueous solutions. Solvent extraction has been widely used in organometallic
removal. Although the process may have fast kinetics and high capacity, solvent extraction is often costly due to
the quantity and specific types of solvents needed. Ion exchange method has commonly been used to remove
metal ions from water or wastewater, but the process has slow kinetics, consumes additional chemicals,
generates hazardous streams and is not well applied to heavy metal ions due to the possible problem of resin
pollution [2, 4]. Biosorption as a sub branch of adsorption, aims at the use of cheaper materials of biological
origin as adsorbents in the removal of metal ions and particulates from solutions [5]. The present study aims at
121
assessing the potential of acid-treated seed pod of Orchid malabar, a plant waste in the removal of some metal
ions from aqueous solution.
Preparation of the adsorbent
The adsorbent, Orchid malabar seed pod was obtained at Michael Okpara University of Agriculture, Umudike,
Abia state Nigeria. The uncooked shells were carefully removed, washed with deionized water, dried and
crushed with a blender. The crushed samples were sieved through to obtain 60 ìm mesh size. The sieved samples
were soaked in 0.3 M HNO3, stirred for 30 min and left undisturbed for 24 h. They were then filtered through
Whatman no. 41 filter paper and rinsed thoroughly with de ionized water until a pH of 7.4 was obtained. The
adsorbent was kept in an oven at 100 oC for 12 h and finally stored in a tight plastic container. The treatment of the
adsorbent with 0.3 M HNO3 helps to oxidize the adhering organic materials and the removal of any debris or
soluble bio-molecules that might interact with the metal ions during sorption.
Preparation of the metal ion solutions
All the reagents used were of analytical grades and were used without further purification. De-ionised water was
used in the preparation of all sample solutions. Stock solutions of 1000 mg/L of each of chromium, cobalt and
cadmium were prepared from their salts, Cr(NO3)3.9H2O, Co(NO3)2 and CdSO4.8H2O respectively. This was
3
done by dissolving appropriate amount of each salt in 1000 cm of de-ionized water, and made up to the mark of
volumetric flask. From the stock solutions, working concentrations of 10 to 50 mg/L of each of the metal ions
were prepared by serial dilution.
Batch adsorption experiments
Equilibrium sorption of Co2+, Cr3+ and Cd2+ onto Orchid malabar was studied as a function of the following: pH,
temperature and initial metal ion concentration. The effect of pH on the adsorption of the metal ion was studied at
o
a temperature of 30 C and at a fixed metal ion concentration of 50 mg/L. The pH of the solutions was adjusted to
2, 4, 6, 7 and 8 using 0.1M HCl or 0.1M NaOH solution as the case may be. Exactly 50 ml of each metal ion
solution of different pH were introduced into different 250 ml Erlenmeyer flasks containing 0.25 g of the
adsorbent. The mixture was agitated intermittently for 2 h in a thermostated shaker and then filtered. The metal
ion content of each filtrate was finally determined using UNICAM Atomic Absorption Spectrophotometer
(Solar AAS 500).
The effect of temperature on the sorption process was investigated at temperatures ranging from 30 oC to 70 oC.
The adsorption experiments were performed in a constant speed thermostated shaker. The adsorbent (0.25 g of
particle size 60 ìm) was put in different 250 ml Erlenmeyer flasks containing 50 ml of 50 mg/L of the different
metal ion solutions at a pH 6. After shaking for 2 h, each reaction mixture was filtered into sample bottles and the
metal content of the filtrate determined using UNICAM Atomic Absorption Spectrophotometer (AAS).
The influence of initial metal ion concentrations on the equilibrium sorption of the metal ions by Orchid malabar
was carried out using 50 ml of various concentrations (10, 20, 30, 40, and 50 mg/L) of the metal ions at a contact
o
time of 2 h, temperature of 30 C and at pH 6. The adsorbent (0.25 g of particle size 60 ìm) was put into 50 ml of
each of the metal ion solutions and the mixture agitated intermittently. At the end of the contact time (2 h), the
mixtures were filtered rapidly into separate sample bottles. The metal ion concentration of the filtrate was
determined using Atomic Absorption Spectrophotometer (Solar AAS 500).
Data analysis
The amounts of Cr3+, Co2+ and Cd2+ adsorbed by the Orchid malabar during the batch investigations were
determined using a simplified mass balance equation:
qe = (Co – Ce ) V/m (1)
Where qe is the equilibrium sorption capacity (mg/g), Co and Ce are respectively the initial and residual
(equilibrium) concentrations (mg/L), V is the volume of metal ion solution in L while m is the dry mass of the
122
adsorbent in g.
Effect of pH on the adsorption of the metal ions by the adsorbent
3+ 2+ 2+
The amounts of Cr , Co and Cd adsorbed as a function of pH are shown in Figure 1. The pH of an aqueous
solution is an important factor affecting both the solution chemistry and the availability of binding sites on the
adsorbent [6, 7]. The range of pH values 2.0 to 8.0 selected in our study were chosen to avoid the precipitation of
the metal ions in solution at higher pH. Results show that the amount of metal ions adsorbed from the solutions by
the adsorbent increased as the pH of the metal ion solution increased from pH 2.0to pH 7.0 and then decreased as
the pH is increased beyond pH 7. The decrease in the amount adsorbed at low pH may be attributed to the fact that
at low pH values the protons compete with the metal ions for the binding sites on the adsorbent surface leading to
reduced adsorption of the metal ions by the adsorbent as a result of electrostatic repulsion. At higher pH, the
proton concentration decreases thereby creating more binding sites on the adsorbent leading to enhanced metal
Fig 1: Effect of pH on the adsorption of metal ions by orchid Malabar

Effect of initial metal ion concentration
3+ 2+ 2+
The effect of initial metal ion concentration on the amounts of Cr , Co and Cd adsorbed are shown in Figure 2.
From Figure 2, it can be observed that with increase in metal ion concentrations from 10 mg/L to 50 mg/L, the
adsorption capacity of orchid malabar increased for all the metal ions used in the study. The amounts of Cr3+, Co2+
and Cd2+ adsorbed onto orchid malabar increased from 2.07566 mg g-1, 2.9019 mg g-1 and 0.8098 mg g-1 to
-1 -1 -1
10.0816 mg g , 10.145 mg g and 6.2802 mg g at the initial concentrations of 10 mg/L and 50 mg/L employed
respectively. This result showed that for all initial concentrations used, there was initial rapid uptake of the metal
ions from solution onto the adsorbents. Results further showed that the trend of removal is in the order Co2+ > Cr3>
Cd2+ from the solutions. This trend of removal could be attributed to the affinity of the active groups of the
biomass for the metal ions.
123
Fig 2: Effect of metal ion concentration on the adsorption capacity of Orchid malabar
Effect of temperature on the adsorption process
The influence of temperature on the equilibrium adsorption capacity for the metal ions by the adsorbent was
determined at temperatures ranging from 30oC to 70oC, with initial metal ion concentration of 50 mg/g and pH 4
and the results presented in Figure 3. The results obtained showed that adsorption increased with increase in
temperature up to a maximum at 40 oC and beyond 40 oC there was a decrease in amount adsorbed. It had been
stated that physisorption usually decreases with increase in temperature while chemisorption increases as the
temperature is increased [8]. This result suggests that the bonding of the metal ions to the adsorbents in our study
could have involved a combination of chemical interaction and physical adsorption. However, an increase in the
amount adsorbed at temperature 40 oC may have resulted from enlargement of the pores at higher temperature
leading to a corresponding increase in the surface area available for adsorption, diffusion and penetration of metal
ions within the pores of the adsorbent and this is in line with result of previous work [9]. Also, increasing the
temperature is known to increase the diffusion rate of adsorbate molecules within the pores as a result of
decreasing solution viscosity and this in turn will alter the equilibrium capacity of the adsorbent for the metal ions.
The decrease in adsorption capacity (or percent metal adsorption) with increasing temperature above 40 oC could
have resulted from the weakening of attractive forces between the active sites of the sorbent and the metal ions and
also between the adjacent molecules of the adsorbed phase [10]. This weakening of attractive forces may have
caused the metal ions to detach from the adsorbent surface into the solution phase.
Fig. 3: Plot of sorption capacity (qe) vs temperature for adsorption of the metal ions onto orchid malabar.
Thermodynamic study
In order to study the feasibility of the adsorption process, the thermodynamic parameters viz: Gibbs free energy of
adsorption kj/ mol), enthalpy of adsorption kj/mol) and entropy of adsorption (J/K/ mol) were obtained from the
following equations:
K = Cad/Ced. (2)
In (K/T) = In R/Nah + S/R - H/RT (3)
124
23
Where R is gas constant (8.314 J/K/mol), Na is Avogadro's number (6.02214 x 10 / mol), A is Arrehnius
-34
constant (or pre-exponential factor) and h is the Planks constant (6.626 x 10 Js). From equation 3, a
plot of In(K/T) versus 1/T should produce a straight line with slope equal to – H/R and intercept equal
to InR/Nah + S/R respectively. The values of H and S calculated from the slope of the transition
state plots are shown in Table 1. The free energy G) values were also calculated from equation 4 and
presented in Table 1.
G= H-T S (4)
The positive values of H indicate the endothermic nature of the process while the positive values of
. S indicate increased disorder and randomness at the solid-solution interface. The negatives values of
. G at the temperature of 303 K indicate that the adsorption process was spontaneous and that the
Orchid malabar surface had good affinity for the metal ions.
Figure 5: Variation of ln (K/T) with 1/T for the adsorption of the metal ions by the adsorbent.
Table 1: Thermodynamic values as calculated from the transition state plots at 303 K
Metal ions H (kJ/mol) S (J/mol) G (kJ/mol)

Cd(II) 33.14 189.80 -57.48
Cr(III) 32.55 134.80 -40.81
Co(II) 12.25 60.01 -18.17
CONCLUSION
This study investigates the potential of Orchid malabar as an adsorbent in removing Cr3+, Co2+ and Cd2+
from aqueous solutions. The influences of pH, temperature, and initial concentration were examined.
The affinity of the adsorbent for the metal ions follows the trend: Cr3+ > Cd2+ > Co2+. The adsorption
process was found to be dependent on pH and the results indicate that the optimum pH for adsorption of
Cr3+, Co2+ and Cd2+ occurred at pHs 4.0, pH 7.0 and pH 4.0 respectively. Studies on the effect of
o
temperature show that the optimum temperature for adsorption of the metal ions is 40 C. The various
thermodynamic parameters, viz; S, H and G evaluated suggest that the sorption process is
125
spontaneous and endothermic. The results from this study indicate that the biosorbent studied could be
employed as a low cost adsorbent for the removal of Co(II), Cd(II) and Cr(III) ions from aqueous
solutions.
REFERENCES
1. U, IIhan and G. Fuat (2000). Adsorption of some Heavy Metal lons from Aqueous Solution by Activated
Carbon and Comparison of Percent Adsorption Results of Activated Carbon with those of some Other
Adsorbents. Turk J. chem. 24: 291-297.
2, N. Li, (2006). Study of Chitosan-Based Biopolymer Adsorptions and their Applications in Heavy metal
Removal. Ph. D Thesis, National University of Singapore, 239 pages.
3. S. Chintana (2002). Removal of Heavy Metals from Waste Water by Adsorption Using Chitosan. M.
Eng Thesis, Suranaree University of Technology, p. 96.
4. M. F. Cybelle, K. Chi-chuan, L. P. Maria and W. W. Meng (2011). Nickel Removal from Aqueous
Solution in Fixed Bed Using Chitosan-Coated Bentonite. Sustain Environ. Res, 21(6): 361-367.
5. F.K. Onwu, S.O Ngele and N.I.Elom (2014). Biosorption of Ni(ll) and Pb(ll) ions from aqueous
solutions by bush mango (Irvingia gabonensis) husks. Int. J. Chem. Appli., 6(1): 69-80.
6. A.A. Farghali, M. Bahgat, A. A. Enaiet and M. H. Khedr (2013). Adsorptionof Pb(II) ions from Aqueous
Solutions Using Copper Oxide Nanostructures. Journal of Basic and Applied Sciences, 2:61-71.
7. F. K. Onwu and S.P.I. Ogah (2010). Studies on the effect of pH on the Sorption of cadmium(ll),
nickel(ll), lead(ll) and Chromium(VI) from aqueous solutions by African white star apple
(Chrysophyllum albidium) shell. Afri. J. Biotech., 9(42): 7086-7093.
8. R. Houwing and G. Salomon (1965). Adhesion and Adhesives, 2nd edn. Elsevier. pp. 50-75.
9. M. Saleem, T. Pirzada, and R. Qadeer (2007). Sorption of acid violet 17 and direct red 80 on cotton fibre
from aqueous solutions. Colloids Surf, A Physicochem Eng. Asp 292: 246-250.
10. S. Ahmet and T. Mustafa. (2008). Biosorption of cadmium (II) from aqueous solution by red algae
(Ceramium virgatum); equilibrium, kinetic and thermodynamic studies. J. Hazard Mater.157:448-454.
126
CORROSION INHIBITION PROPERTIES OF COMMIPHORA AFRICANA (A. RICH.) ENGL. GUM
EXUDATES ON MILD STEEL IN ALKALINE MEDIUM.
1
Odoemelam, S.A., 2Onukwube, N.D. and 3
Eddy, N.O.
1
Department of Chemistry, Michael Okpara University of Agriculture, Umudike Abia State, Nigeria.
2
Department of Chemistry, Abia State Polytechnic, Aba. Abia State, Nigeria.
3
Department of Chemistry, Ahmadu Bello University, Zaria. Kaduna State, Nigeria.
Corresponding Author:
decon4real@yahoo.com
08032976594
Abstract.
The effect of Commiphora africana (CA) gum exudates on the corrosion of mild steel in 2.5 M Na2CO3 has been
studied using weight loss (gravimetric) and thermometric methods at 303 and 333 K. Results obtained suggest
that Commiphora africana acts as a good corrosion inhibitor as the inhibition efficiency increased with increase
in the concentration of the inhibitor. A decrease in the %I was observed with increase in temperature from 303 to
333 K. Values of inhibition efficiency from weight loss method were found to be significantly higher (p ≤ 0.05)
than values obtained from gasometric method, even as the (I%) values from the two methods correlated strongly.
Also values of ? Gads, Ea and Qads suggest physical mechanism for the adsorption of the inhibitor molecules on the
surface of mild steel even as Temkin adsorption isotherm was found to best suit the adsorption mechanism within
the temperature range under study.
Keywords: Mild Steel, Adsorption Isotherm, Corrosion Inhibition, Activation Energy.
Introduction
Corrosion scientists and engineers have continued to grapple with the manifestation of corrosion and corrosion
products on steel structures [1-2]. One of the most practical methods of protection of metals against corrosion in
various media is the use of inhibitor which helps to effectively isolate the metal from the corrosive agents.
Inhibitors are important in corrosion monitoring because they prevent or reduce corrosion without significant
reaction with the components of the environment. Compounds containing heteroatoms are known to be effective
and efficient organic inhibitors [3]. An effective inhibitor should be able to not only displace water from the
surface of a metal, but should interact with the anodic or cathodic reaction sites to retard the oxidation and
reduction corrosion reactions, as well as prevent transportation of water and corrosion active species on the
surface of metal. Synthetic organic inhibitors have been continually replaced with naturally occurring
substances which are cheap, readily available, ecologically and environmentally friendly. Most importantly,
they are biodegradable and renewable sources of materials. Recent studies have shown plant materials as
effective good inhibitors for metals in aggressive media [4-8]. Also reports abound on the corrosion inhibitive
effectiveness of metals by Gum arabic [9], Raphia hookeri [10], Acacia seyal var seyal [11], Guar gum [12],
Anogessus leocarpus [13], Pachylobus edulis [14]. The growing interest in environmentally friendly corrosion
inhibitors has necessitated this work which seeks to investigate the inhibiting effect of gum exudates from
Commiphora africana (CA) on mild steel corrosion in alkaline medium using gravimetric and thermometric
techniques at 303 - 333 K. Also considered were the thermodynamic, activation and adsorption parameters that
govern metal corrosion.
Materials and Methods.
127
Materials.
Mild steel sheets of composition (wt%) Mn (0.6), P (0.36), C (0.15), Si (0.03) and the rest iron were used in
this study. The sheet was mechanically cut into different coupons, each of dimensions 4 x 3cm. Each coupon
was degreased by washing with ethanol, rinsed with acetone and allowed to dry in air before they were
preserved in desiccators. All reagents used for the study were Analar grade and double distilled water was used
for the preparations. The samples were purified using the procedure earlier described by [13].
Corrosion Inhibition Studies.
Gravimetric method.
Gravimetric study using Commiphora africana gum was done by dipping a previously weighed metal (mild
steel) coupon into 20 ml of the test solution maintained at 303 and 333 K in a thermo stated bath. The weight
loss was determined by retrieving the coupons at 1 hour intervals progressively for 7 hours. Prior to
measurement, each coupon was immersed in a solution of 20 % sodium hydroxide containing 200 g/l of zinc
dust to terminate the corrosion reaction and then rinsed in acetone before drying. The difference in weight was
taken as the weight loss of the mild steel. From the average weight loss (mean of three replicate analyses)
results, the degree of surface coverage (è), the inhibition efficiency (%I) of the inhibitor, and the corrosion rate
of mild steel (CR) were calculated using the following equations.
where W0 and W1 are the weight losses (g) for mild steel in the absence and presence of the inhibitor in Na2CO3
2
solution, ? is the degree of surface coverage of the inhibitor, A is the area of the mild steel coupon (in cm ), t is the
period of immersion (in hours) and W is the weight loss of mild steel after time, t.
Thermometric method.
Thermometric analysis was also carried out as reported elsewhere [15]. From the rise in temperature of the
reaction system per minute, the reaction number (RN) and the percentage inhibition efficiency were calculated
using the following equations.
Where
Tm = Maximum temperature attained by the system.
Ti = the initial temperature.
t = the time (min) taken to reach the maximum temperature.
RNaq = Reaction number in the absence of inhibitor
RNwi = Reaction number in the presence of inhibitor.
Results and Discussion.

The variation of weight loss with time for the corrosion of mild steel in 0.1 M Na2CO3 containing various
concentrations of Commiphora africana gum exudates at 303 and 333 K are as shown in figures. 1 and 2
respectively. From these figures, it was observed that the rate of corrosion of mild steel increased as the
128
period of immersion increased, but decreased with increase in the concentration of Commiphora
africana gum exudates, confirming the inhibition of corrosion of mild steel by gum exudates of CA
in Na2CO3 within the temperature range under review. However, the inhibition efficiency of the gum
exudates was found to have a direct proportional relationship with the concentration of the exudates
gum, but varies inversely with the temperature, suggesting physical adsorption on the surface of mild
steel. Similar findings were equally deduced from thermometric study.
Fig 1: Variations of weight loss of mild steel with time for the corrosion of mild steel in 0.1 M
Na2CO3 containing various concentrations of Commiphora africana at 303 K.
Fig 2: Variations of weight loss of mild steel with time for the corrosion of mild steel in 0.1 M Na2CO3
containing various concentrations of Commiphora africana at 333 K.
129
Effect of temperature.
The effect of temperature on the rate of corrosion of metal cannot be overemphasized. In corrosion involving
basic medium, there is an exponential increase in corrosion rate with increase in temperature and so the
integrated Arrhenius type equation which is experimental dependent is observed between the corrosion rate
and temperature.
Where Ea is the activation energy, CR1 and CR2 are the corrosion rates of mild steel at the temperatures T1 (303 K)
and T2 (333 K) respectively.
The heat of adsorption of Commiphora africana on the surface of mild steel was calculated using equation (7),
since the corrosion inhibition was carried out at constant pressure, the heat adsorbed should approximate the
enthalpy change [9, 15].
Where ? 1 and ? 2 are the degrees of surface coverage of the inhibitor at temperatures, T1 (303 K) and T2 (333 K)
respectively and R is the gas constant. Calculated values of Qads are negative as recorded in Table 1, indicating that
the adsorption of Commiphora africana gum on mild steel surface is exothermic.
Table 1: Calculated values of activation energy and heat of adsorption for mild steel corrosion in 0.1 M
Na2CO3 with various concentrations of Commiphora africana gum exudates.
Conc. of (CA) (g/l) Ea Kjmol-1 Qa (Kjmol-1

Blank 11.97
0.1 13.71 -18.19
0.2 16.84 -20.16
0.3 20.13 -22.01
0.4 22.27 -22.01
0.5 22.71 -32.18
In the present study, it could be seen that the values of Ea in the presence of different concentrations of CA were
progressively higher than that in its absence. This could be attributed to the formation of an adsorptive
electrostatic film of physical character [16]. Thus, a physical barrier to charge and mass transfer is created by the
adsorbed molecule. The degree of surface coverage decreased with increase in temperature as signified by the
negative values of Qads. This could be attributed to the fact that attainment of physical adsorption equilibrium is
usually rapid and the process is readily reversible and exothermic, whereas in chemical adsorption, the
occurrence of chemical reactions at the metal surface makes the process relatively slow and not readily
reversible. This is in line with earlier suggestion [5].
Adsorption and Inhibition Efficiency.

The adsorption capacity of molecules determines their protective ability in metal corrosion [17]. The adsorption
film that results isolates the metal surface from the corrosive medium such that the corrosion rate indicates the
number of free corrosion sites remaining after some sites have been effectively shut out by the adsorbed inhibitor.
Table 2 indicates that the %I increased with increase in the concentration of Commiphora africana, but decreased
130
with increase in temperature. This is attributed to the decrease in formation of the protective film on the metal
surface (or desorption of the inhibitor molecule from the metal surface) at elevated temperatures [18]. Physical
adsorption mechanism is thus suggested. Values of inhibition efficiency obtained from thermometric method
were found to be significantly higher than values from weight loss method (p ≤ 0.05). Thus the inhibitor could
be said to be more efficient in instantaneous inhibition than inhibition over a length of time.
Table 2: Corrosion Rates, Inhibition Efficiencies and Reaction Numbers of various concentrations of
Commiphora africana for the corrosion of mild steel in Na2CO3
Conc. of Weight Loss Thermometric

(CA) g/l 303 K 333 K %I RN
CR X 10-3 %I CR X 10-3 %I
Blank 2.50 3.85
0.1 2.17 13.33 3.56 7.43 66.67 0.075
0.2 1.71 31.49 3.14 18.27 77.78 0.055
0.3 1.33 46.67 2.75 28.48 82.22 0.040
0.4 1.07 57.14 2.39 37.78 86.67 0.030
0.5 1.04 58.57 2.36 38.69 91.11 0.020
Attempts were made to fit the degree of surface coverage values (? ) to various adsorption isotherms. Temkin
adsorption isotherm was found to best suit the experimental data using equation (8) [19].
exp(-2f? ) = Kads C. … (8)
Where Kads, C, f and ? represent the equilibrium constant of adsorption process, additive concentration,
molecules interaction parameter and degree of surface coverage respectively. Taking the logarithm of
equation (8) with rearrangement results to equation (9).
A plot of ? against Log C would give a straight line with intercept Log K. Fig. 3 is an indication that Temkin
adsorption isotherm is obeyed.
Fig 3: Temkin isotherm plots for the adsorption of Commiphora africana on mild steel surface in 0.1 M
Na2CO3.
Calculated values of equilibrium constant and standard free energy of adsorption are as shown in table 3 as
obtained from equation (10).
? Gads = -2.303RTlog (55.5K) ... (10)
131
Where R is the gas constant, 55.5 is the concentration of water in the solution in Mol/L.
The positive value of f shows the existence of attractive force at the neighbouring adsorption sites in the
adsorption layer [14].
Table 3: Thermodynamic Parameters for adsorption of Commiphora africana (CA) on mild steel
surface at 303 and 333 K
Temp (K) Log K ?G0ads f R2
303 -1.18 -3.27 2.912 0.990
333 -0.05 -10.80 1.045 0.981
Generally, ? G0ads values up to -20 Kjmol-1 are consistent with physical adsorption [20]. The values of standard
free energy from table 3 are in consonance with physical adsorption mechanism and also show that the
adsorption of Commiphora africana (CA) on the surface of mild steel in an alkaline medium is spontaneous. Also
higher value of (f) at 303 K shows a better interaction of the molecules at lower temperature confirming the
mechanism of physical adsorption.
Conclusion.
The following conclusions were made from the results of this study:-
(i) Gum exudates of Commiphora africana is a good inhibitor for the corrosion of mild steel in alkaline
medium within the temperature range under study.
(ii) The inhibitor follows the physical mechanism of adsorption onto the surface of mild steel as the
inhibition efficiency decreased with increase in temperature.
(iii) The efficiency of the inhibitor increased with increase in the concentration of the inhibitor.
(iv) The adsorption behaviour of the inhibitor is consistent with Temkin adsorption model.
References.
1. Tretchewey, K.R. and Chamberlain, J.(1995). Corrosion for Science and Engineering,1st edition,Longman,
United Kingdom, 31 – 40.
2. Ita, B. I. and Offiong, O.E. (2000). Inhibition of mild steel corrosion in hydrochloric acid by 2
–Ammopyridine and 2 – Aminomethyl pyridine, Global J. Pure and Appl. Sci. 6 (1): 51 –55.
3. Fekry, A.M., Ameer, M.A.,(2010). Corrosion inhibition of mild steel in acidic media using newly synthesized
heterocyclic organic molecules. International Journal of Hydrogen Energy 35: 764 - 771.
4. Avwiri, G.O. and Igbo, F.O. (2003). Inhibition action of Vernonia amygdalina on the corrosion of aluminium
alloys in acidic media. Materials Letters 57: 370 - 375.
5. Oguzie E.E. (2007). Corrosion inhibition of aluminium in acidic and alkaline media by Sansevieria trifasciata
extract, Corros. Sci., 49: 1527-1539.
6. Emregul K.C. and Abbas Aksut A.,(2008). The behavior of aluminum in alkaline media, Corros. Sci., 42:
2051-2067.
7. Abdel-Gaber A.M., Khamis E., Abo-ElDahab H. and Adeel Sh.,(2008). Inhibition of aluminium corrosion in
alkaline solutions using natural compound, Mater. Chem. Phys.,109: 297-305.
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8. Singh A., Ahamad I., Singh V.K. and Quraishi M.A.,(2011). Inhibition effect of environmentally benign
Karanj (Pongamia pinnata) seed extract on corrosion of mild steel in hydrochloric acid solution, J. Solid
State Electrochem., 15: 1087-1097.
9. Umuoren, S.A., Ebenso, E.E., Okafor,P.C., Ekpe, U.J. and Ogbobe, O., (2006). Effect of halides ions on the
corrosion inhibition of aluminum in alkaline medium using polyvinyl alcohol. J. of Appl. Polymer Sci., 103:
2810-2816.
10. Ebenso, E.E., Obot, I.B., Obi-Egbedi, N.O and Umoren, S.A., (2009) ,The Inhibition of aluminium
corrosion in hydrochloric acid solution by exudate gum from Raphia hookeri, Desalination, 250 : 225-236.
11. Buchweishaija ,J. and Mhinzi, G.S.,( 2008). Natural products as a source of environmentally friendly
corrosion inhibitors: The case of natural tree gum exudates from Acacia s e y a l var s e y a l , Portugaliae
Electrochimica Acta, 26: 257-265.
12. Abdallah, M., (2004). Guar gum as corrosion inhibitor for carbon steel in sulphuric acid solutions”
Portugaliae Electrochemica Acta, 22: 161-175.
13. Eddy, N.O., Ameh, P., Gimba, C.E. and Ebenso, E.E., (2011). Corrosion Inhibition potential of Anogessus
leocarpus gum exudates for the corrosion of mild steel in solutions of HCl. Int. J.of Electrochem. Sci., 6 :
581-585.
14. Umoren, S.A., Obot, I.B., Ebenso, E.E. and Okafor, P.C., (2008). Eco-friendly Inhibitors from
Naturally Occurring Exudate Gums for Aluminium Corrosion Inhibition in Acidic
Medium.Portugaliae Electrochimica Acta, 26: 267-282.
15. Umuoren, S.A., Ebenso, E.E., Okafor,P.C., Ekpe, U.J. and Ogbobe, O., (2006). Water soluble polymers as
corrosion inhibitors. Pigment and Resin Technol., 35(6): 346-352.
16. Obot, I.B., Obi-Egbedi N.O and Umoren, S.A. (2009)” Antifungal drugs as corrosion inhibitors for
aluminium in 0.1 M HCl . “Corrosion Science, 51 (8): 1868-1875.
17. Haider, A.A. (2011). The study of the inhibitory properties of Omeprazole on the corrosion of
Aluminum 6063 in alkaline media. Basrah Journal of Scie. .28(1): 74-93.
18. Doche L. M., Rameau J.J., Durand R., Novel-Cattin F., (1999) “Electrochemical behaviour
of aluminium in concentrated NaOH solutions”, Corros. Sci. 41: 805 -810.
19. Okafor P.C. , Liu X. and Zheng Y.G. ,(2009)." Corrosion inhibition of mild steel by
ethylamino imidazolline derivative in CO2 –saturated solution" Corrosion Science, 51:
761-768.
20. Scendo M. (2008)”The influence of adenine on corrosion of copper in chloride solutions “

Corrosion Science, 50: 2070–2077.
133
OGODOMETRICS:ISO APPROVAL/CERTIFICATIONS/PRODUCT DESIGN, INNOVATION USING
GLOBALLY HARMONIZED INTERNATIONAL REFERENCE STANDARDS IN SONGHAI DELTA
AMUKPE SAPELE AS TARGETED WORLD COLLABORATING CENTRE.
OGODO AEMURERHIMEN DICKSON (Snr.) (Chartered Chemist)

(HND, PGD, M.Sc., Ph.D (In- View), AISLT, MCSN, MICCON, C.CHEM., MNES, MSPE)
School of Applied Science, Department of Science Laboratory Technology, Chemistry Laboratory, Delta
State Polytechnic, P.M.B 1030, Ogwashi – Uku, Delta State, Nigeria: Tel: 08030738401, 08094921724, E-
mail:dicksonogodo@yahoo.com
Abstract
This Research Work shows that Innovation involves a series of Scientific, Technological, Organizational,
Financial and Commercial activities. It clearly reveals actions taken to Design or Redesign and Innovate or Re-
innovate more and new materials substances and ideas. It dwells specifically with collaboration for
commercialization as a National Project through World-Wide Franchise/ Sole Concession Rights in Member
Countries after endorsement as Nigerian Official Internationally RecognizedPackaging Materials for storage
of products using Songhai Delta Amukpe Sapele Innovative Technology Transfer by signing a Global
Memorandum of Understanding (GMOU)for the Packaging Container Law(PCL)using the Packaging Added
Value(PAV) being regulated in Nigeria in collaboration with the 154 ISO Countries, The Study concludes that
ISO Certifications requires that the manufacturing facilities are certified for the Design, Development,
Manufacturing, Assembly, Sterilization by Irradiation and meets the US Pharmacopoea Light Transmission Test
of Ogodometrics Superior Material produced in a Plant awarded ISO Quality Management System and
Environmental System and complied to ISO for permanency of material. The Study recommends that a Co-
ordinating Office under a Presidential Scientific or Technical Adviser is the only option for Nigeria using
Science Governance. Secondly, there should be effective Global Technical Partnership/Collaboration/Synergy
for National Development through Technical and Commercial Scientific Licensing with Proposed Member
Countries to generate Nigeria Vision 20:2020 target for Global Competiveness. Finally, there should be no Sole
Concession/Patent /Franchise Rights in Proposed Member Countries without Ogodo (Nigeria) Standards
International (OSI) Permission except as Permitted by Law through Technology Transferred Agreement of
Intellectual Property Rights (IPR).
Keywords: ISO Approval/Certifications, Product Design/Innovation, Globally Harmonized International

Reference Standards (Songhai Delta Amukpe Sapele Targeted World Collaborating Centre), Ogodo
(Nigeria) Standards International (OSI).
Introduction
The chemical industry is in business of manufacturing products to sell to customers locally and overseas – The
Market- As manufacturing, distribution, communication and technology expand and develop, purchasers are
faced with a wide variety of choices. They can obtain what they want from several sources and at any time they
want it. This creates competition and the factor of choice is Quality Products. Poor-quality products become
rejected in favour of what satisfies the desire of the purchaser. Therefore any Company whose management
places little or no value on quality becomes unprofitable and impossible to sustain. Thus, it is no longer good
enough to simplify make products according to specification or some standard. Companies must actively
identify their customers requirements and use this information to develop better and better products or services.
(1)
.
134
ISO Aprroval
The Packaging Materials should be produced in a Plant awarded the ISO 9001:2008 for Quality Management
System, ISO 14001:2004 for Environmental Management System and complies to ISO 9706 for Permanency of
Packaging Materials.
ISO Certifications
ISO Certifications requiress that the manufacturing facilities are certified to ISO 13485: 2003 and ISO 9001:
2000 for the design, development, manufacturing,surface treatment, printing, assembly, sterilization by
irradiation to customer specifications per ISO11137 and to meet the US Pharmaeopoea Light Transmission Test
of Ogodometrics Superior Material produced in a Plant awarded the ISO 9001:2008 for Quality Management
System, and ISO 14001:2004 for Environmental Management System and complies to ISO 9706 for permancey
of Packaging Materials and closures meets the requirment of 21CFR177:1520 for Food and Beverage use.
The NIS ISO 9001: 2000 Standard

The standard is useful as a tool for the implementation of which gives confidence that the system will
consistently guarantee customer satisfaction and operation within legal requirements. It is usually a
strategic decision for an Organization to make for the installation of a Quality Management System
(QMS). The design and implementation of the QMS will vary depending on the needs, specific
objectives, products provided, the processes used and the size and structure of the Organization.
This implies that any Organization can use the Standard to demonstrate its ability to meet customers',
regulators', and the Organizations' own internal requirements. Internal or external parties to assess
the Organization's ability to meet customer requirements may also use the standard. It has in-built
flexibility to accommodate the documentation structure of any Organization without necessarily
(2)
changing it .
The International Standard does not include requirements specific to other management systems,
such as financial management or risk management. However, it enables an Organization to align or
integrate it own quality management system with related management system requirements. It is
possible for an Organization to adapt its existing management system(s) in order to establish a QMS
that complies with the requirements of the standard.
135
Basic Concepts of ISO 9001: 2000
Quality Management
All the methods used to achieve, sustain and improve quality which are quality control, quality
assurance and quality improvement form quality management. It can be seen as management of
success or elimination of failure. The ISO 9001: 2000 aims to enhance customer satisfaction through
the effective implementation of QMS and assurance of conformity to customer and applicable
regulatory requirements. This pre-supposes that you plan to succeed and so it is also implied that you
will prevent failure. Since quality cannot be repeated through change, it has to be built into the product
or service deliberately. Even when we inspect or test it, it is only to gather data for decision making.
Then we may make changes but if we underestimate the effect of the change, it can become
unmanageable. Thus we need a tool that we will practice to enable us achieve our objectives while
(2)
preventing failures from occurring. .
In the United States, a standard was set for quality with the introduction of the Malcolm Baldrige
National Quality Awards in 1987. The new global measures for quality are called ISO 9000 standards.
ISO 9000 is the common name given to Quality Management and Assurance standard published by
the International Organization for Standardization (ISO). Prior to the establishment of such standards
in 1987, there is no international standard of quality against which to measure Companies. ISO
Standards, established in Europe, provide a “common denomination” of business quality accepted
(2)
around the world .
Pigmented Silica-Based Packaging Materials (Ranking Order)

st
Silica-Based 1 Ranking – Best Superior (AGB)
Packaging Materials 2nd Ranking – Better Superior (GGB)
RD
3 Ranking – Good Superior (CGB)
Polyurethane
Packaging Materials 4th Ranking – Poor Superior (CPB)
Songhai - Delta Amukpe, Sapele (SDAS) Background Information

This Delta State Owned Songhai-Delta Amukpe, Sapele, is a replica of Songhai Port-Novau in the
Benin Public. It was established in the year 2001 for the development of Livestock and Crop
Production as well as a Training Centre for Farmers to acquired new techniques in modern agricultures
as a profitable venture. The most significant landmark in this research work was the visit/study tour to
Songhai-Delta Amukpe Sapele on the 7th of July, 2003 in discovering the best superior Silica-based
packaging materials that will protect the Shelf-life of the various juices, Beverages and Vegetables Oils
being processed by Songhai Delta, Amukpe, Sapele. The Delta State Owned Songhai-Delta situated
rd
at Amukpe, Sapele uses the 3 ranking order in protecting the Shelf life of its products. The thrust of this
innovative Technology Transfer Centered on the use of the 1st Ranking Order-Best Superior Packaging
Materials which is more effective in improving the Shelf-life of it products(3).
Songhai Amukpe Will Ensure Vision 20-2020 Target

The Management of Songhai – Delta, Amukpe Farm, a special seed and Animal Development center
located in Amukpe, Sapele, Delta State, Nigeria rested its commitment in ensuring that the Nation's
vision 2020 and the recently launched Delta Vision 2020 meets its target. The Management of
136
Songhai, Delta Amukpe brought a new vision into farming “as it makes people now identify with the
occupation of generality of Africans, Delta inclusive. The programme and vision in Songhai Delta
Amukpe farm was one that the nation could rely on “because the implementation is favoured to
achieved maximal Agricultural Productivity in positioning Nigeria and indeed Delta State, as among
the top 20 economies in 2020, leading to self sufficiency in food production(3).
The History of the Quality Movement

Japan, Germany and the United States of America have always accorded Science and Technology high priority
so much so that 2-3% of their enormous Gross Natural Product (GNP) are Spent on Science and Technology.
Japan was defeated in World War II and her industrial and financial strength collapsed. She had no natural
resource and limited sources of foods for her people. There was an urgent need to completely rebuild their
economy to be able to develop products that were good enough to sell abroad to generate revenues in order to
import food. This was the starting point of the Quality Movement whereby the Union of Japanese Scientists and
Engineers invited a Dr. Deming to Japan in 1947 to help the Japanese on how to improve on the quality of
manufactured good.
Deming taught them how they could manufacture high quality goods and how they could do it faster and cheaper.
In 1954, Dr. Joseph Duran came to add his knowledge to the quality improvement train and succeeded in raising
the level of quality management from the factory to the totality of a whole Organization (1).
He stressed the importance of evolving quality from the product design stage, prototype testing, proper
equipment operation and accurate process feedback. As part of his educational campaign in Japan, Duran
introduced the Total Quality Control (TQC) concept involving company-wide activities in quality control,
quality circles, quality audits and promotion of quality management principles.
By 1968, the educational campaign has caught on and the Japanese have assimilated the quality concept such that
one of their own, Kaoru Ishikawa has become an expert on his being the pioneers of Japanese quality movement.
This formed the foundation of Japan's industrial revolution that eventually came to challenge the African might
and system.
Thus, hitherto apparently “lazy” Japan woke up from her deep slumber, and swore never to allow herself to suffer
such a national humiliation any longer. SHE THEREFORE SIGNIFICATLY REVIEWED THE EDUCTIONAL
CURRICULUM OF THE CHILDREN TO MAKE SCIENCE & TECHNOLOGY COMPULSORY FOR
EVERY CHILD. Today, Japan is a world power to recon with in anything Science and Technology. The products
of Japanese technology, including cars, electronic gadgets, communication systems etc flood al countries in the
world today, including the USA and Africa. The result of Japan's decision to make Science and Technology
compulsory for every child is clear for all to see today. Today Japan is one of the most highly developed and
respected countries in the world. This status is certainly not based on Japan's prowess in football, but on the
expertise of its citizens in Science & Technology.
However, it is pertinent to note that Science & Technology, as a way of life, is not a field for short-term
engagement. It is a full-life engagement. Scholars whose names top the list of contributors to Science &
Technology in the world did only start as children, but actually devoted their lifetime to it. Examples
137
abound in such great man as Isaac Newton of the Mathematics and Physics fame; Thomas Edison the
Incandescent Lamp Inventor; Lawrence Oppenheimer (Atomic Energy), Mendeleev, the Periodic Table
Inventor, Michael Faraday (Electromagnetic Forces and Gregor Mendel (Genetics). Today, in the USA, young
Bill Gates is a leading world Computer “guru”. These are but a few examples of world's worthy citizens who
have found it worthwhile to pursue & Technology, and had changed the history of the world for good. But the
truth is that Nigeria is not short of people endowed with brains to do better. But do we have the political
will to encourage them? What lessons did Nigeria learn from the Biafran war? Can Nigeria emulate Japan in
making Science compulsory for every Nigeria child? These questions constitute good food for our thought menu
(4)
.
The hope we all have for a better tomorrow and indeed for a better future depend on those Inventors and
Innovators who have and are striving to make the world a better place through their creative energies and
sustained hard work (5).
Nigeria's desire to be one of the 20 leading economies in the world by the year 2020 would be a mere dream
except Science and Technology takes the leadership role as the chief driver of the nation's vision Agenda, if
Ogodometries is challenged in using Innovative Technology Transfer for common regulation of Intellectual
Property Rights (IPR) policy through International Reference Standards for Regulation in Nigeria through
(6)
Synergy/Collaboration .
Vision 20/2020 cannot succeed without Science and Technology (7). Nigeria is endowed with so many Chemists
and it painful for her not to be among the industrialized nations of the world due to our inability to generate a
national vision (such as Projects Ogodometrics International) Vision 20:2020 Target for Songhai - Delta,
Amukpe Sapele Innovative Technology Transfer and implementation with distinct strategies to accomplish set
goals through documentation and modalities for it implementation in research and development and create a
desire in citizens to patronize indigenous efforts, and avoidance of importation of items that can be manufactured
within Nigeria and promulgate the Packaging Container Law (PCL) by signing a Global Memorandum of
understanding (GMOU) with other Countries/ ISO/SON and declare that without a Packaging Added Value
(PAV) of Ten Naira (N10.00) (Ogodometrics) per every bottle of finished product imported into Nigeria, other
Countries cannot bring their finished products to Nigeria or Africa (8). Also effective management of existing
manufacturing or allied Companies in Nigeria with improved maintenance culture, with virtues that we lack in
this country (Nigeria) is necessary to achieve the greatness. These and other virtues are very important for our
nation Nigeria to transit to an industrial nation. Chemistry is truly fundamental to industrialization (9).
Innovation
Knowledge-idea and the people who generate them are the new coin of the currency. The innovative stage
rewards new competencies and strengths. CSN can transfer ideas into a new or improved products through
entrepreneurship used in industry or commerce (10). Innovation is the application of knowledge in production
through learning, research or experience. To innovate is to produce something that is novel in a given setting
(Silica-based Packaging Materials) for International Reference Standard for Regulation in Nigeria. Innovation is
the technical and commercial activities involved in the marking of a new improved product or the first
commercial use of a new/improved process or equipment. A technological innovation cannot be said to have
been fully implemented until it has been introduced on the market (product innovation). Innovations therefore
involves a series of scientific, technological, organizational, financial and commercial activities.
138
Collaboration with National and International Agencies
We have to think of ways to partner with International bodies to move what we are doing forward. We will take a
look at foreign Technologies exhibited and compare with our own so as to enable us train the young scientist to
11
the quality of the foreign scientists ( ). Many countries will be attracted to establish their collaborative efforts
with us hawing seen how possible it was will us. Our capacity building through Technology Transfer can
compete to some extent with the imported ones and even better than the ones from Asia which are not as good as
12
was taught. With time, our Technology Transfer will compete Favourably with other parts of the world ( ).
We require exchange of Experience and input between communities, Agencies and Nation. The collaboration
can be done at two levels.
1. Technical Support and Capacity Building.

This area of collaboration is accessed through Bilateral arrangements between Nigeria and other
Countries/Agencies within the frame work of such arrangements.
2. Collaboration in Programme Delivery. Project delivery should be facilitated through a network of

collaborative activities with local communities, local and state Governments, and Agencies (12).
Collaborative Research and National Development

Collaborative research, development and deployment (RDD) is positively influencing development in nations of
the world. A great number of nations have remarkably improved their socio-economics problems through huge
investment in collaborative research where individuals in the state of harmony bring their expertise together to
solve a typical problem beneficial to the society. Such synergy results in breaking down barriers in the way of
inventions, discoveries or innovations which otherwise would not have been possible when one individual is
working on the project all alone. Instance bound where such an approach has worked wonders. (13).
Collaborative research, is also yielding huge investments in India, China, Malayia, Singapore. India has
developed to the extent that her software exports were about Six billion dollars in 2000.
India and the Philippines are home to call centres of Africa's largest companies. Microsoft, American Express,
IBM, Hewlet, etc. The large-scale off-shoring of factories and plants to places like China, Malaysis and
Singapore is fast redefining the manufacturing sector of the American economy (14). In 2004, about 2500
13
Americans' tax returns were prepared by graduate accountants in India ( ). Even our fathers of science
collaborated in their works to achieve greater results, impacted on the universe as a whole and won highly Prized
Nobel Prizes.
The husband-and-wife team of Jean Frederic Joliot-Curie (1900-1958) and Irene Joliot Curie (1897-1956) won
15
the Nobel Prize in Chemistry in 1935 for the discovery of new radioactive elements prepared artificially ( ). One
could go on and on. The products of such collaborations are what we are enjoying today in the shade of modern
life (13). So, our society will be much better as we develop interest in collaborative works, identify relevance areas
and work as a team of experts to achieve a goal. International Organizations (UNDP, UNESCO, UNICEF,
Common Wealth, World Bank, etc) prefer giving grants to projects carried out as a team.
139
COUNTRY A COUNTRY B
(TECHNOLOGY SUPPLIER) (TECHNOLOGY USER)
Management & executionSTOCK

of OF STOCK OF Management & execution of
TECHNICAL
Research & development TECHNICAL Research & development
TRANSFER OF TECHNICAL
KNOWLEDGE
TRANSFER OF PRE-INVESTMENT
140
CONSULATANCY SERVICES
Management and execution
Part of pre- Management of fall, and
TRANSFER OF DESIGN AND of design and engineering
Management and execution
investment and ENGINEERING SERVICES
execution of some pre-
of capital goods production STOCK OF
STOCK feasibility studies TRANSFER OF CAPITAL GOODS investment and feasibil ity TECHNOLOGI
OF
Management and execution of CAL
TECHNOLOGIC studies
installation and commission etc.
AL RESOURCES
TRANSFER OF INSTALLATION
RESOURCES
Some design and
AND COMMISSIONING SERVICES
Fig 1. Technology Transfer Process: Specialized Complementarity

engineering services
OPERATIONAL PRODUCTION Sequence of techn-managerial
Sequence of technology
transforming tasks SYSTEM Decision-making
Technology Transfer
Technology transfer may be conceptualized as an active process during which technology is carried
across the border of two entities. The entities could be Companies, Countries or even individuals
(Fig.1). Thus, technology transfer could be defined as positive goal – oriented process of interaction
between two or more social actors. The interaction may be in the form of formal training, printed
matters or mere discussions. In this time of interaction, the pool of technological resources remains
stable or increase through the transplantation of one or more components of technology(16).
Technology transfer could be in tangible (e.g. capital goods) or intangible forms such as brand names
or proprietary designs. It is thus recognized that technology transfer plays a central role in the process
of economic development of both developed and developing Countries.
At this stage, it is pertinent to make a further elaboration on the process of technology transformation
herein referred as the flow of technology (Fig. 2). The three categories of the overall flow of technology
have been recognized and this can be seen as Flow A, B and C.
EXPORTING IMPORTING
FIRMS FIRMS
TECHNOLOGY TRANSFERRED
Flow A Engineering services,

New
Management service
Technology Production
and capacity goods Subsequent
capacity Capacity
Flow B Skills and know-how for investment
operation and maintenance projects to
create new units

Flow C Knowledge, expertise and
(16)
Fig 2. Types and patterns of technology transfer flows .
141
OGODO (NIGERIA) REFERENCE STANDARD OS EN ISO
INTERNATIONAL/PROJECT OGODOMETRICS INTERNATIONAL
14001:2004
Implementation of ISO 14001:2004
Songhai Delta Technology Transfer
Specification With Guidance For Use

The European Standard EN ISO 14001: 2004 will have the Status
of Ogodo (Nigeria) Reference Standard International
NO SOLE CONCESSION/PATENT RIGHTS WITHOUT OGODO

(NIGERIA) REFERENCE STANDARD INTERNATIONAL OSI
PERMISSION EXCEPT AS PERMITTED BY LAW(17).
OSi/POi
Fig 3.Ogodo (Nigeria) Reference Standard International/Project
Ogodometrics International
142
Technology Development and Transfer
(18)
According to the report of G. Arthur Brown (1991) , technology development has been held
responsible for creating entirely new processes, products and services, and improving existing ones.
In addition, technology has been contributing towards raising educational and social productivity,
which increase standards of living, and improves the fractional terms of trade, which enables
Countries to protect or enhance their international competitiveness in particular products or services.
In addition, the report pointed out that no one country could expect to be completely autonomous
technologically, and because technologies could be transferred internationally, a Country would not be
using its resources optimally if it tried to do so. But all Countries should aim to enhance their skills in
acquired technologies, and disseminating, absorbing and adapting them efficiently. It has been
strongly recommended that the developing Countries should recognize the crucial need to enhance
their ability to absorb, adapt and utilize technology by reorienting their human resource development
policies so that workforces can contribute effectively to development and ensure that their Countries'
International competitive positions are maintained or improved in view of the widening gap in scientific
and technological capacity(18).
The commonwealth Group of Experts (1991) has enumerated the main proven means of technology
transfer. These were listed as through;
i. Direct foreign investment (equity participation, joint ventures, and subsidiaries of
transnational corporations);
ii. Direct transfer of technology (Patenting, Licensing Agreements, Technical Consultancies,
Management Services Contracts, On-And Off-The-Job Training);
iii. Imports of capital goods (often in association with other transfer elements such as
information on Technical Specifications, Training, Consultancy);
iv. Turn key projects (sometimes involving an entire enterprise);
v. Reverse Engineering (i.e. Unpackaging Technologies; and
vi. Non-negotiated transfer, such as screening Foreign journals, direct observation by
Scientists and Technicians at Universities and Research and Development Institutions
Abroad, e.t.c.
As would be expected, these methods of transfer involve different costs and benefits to the
receiving country in terms of price, control and ability to absorb or adapt the technology concerned,
or develop totally new processes or products. Any one method may not be appropriate in particular
circumstances, but the aim of the developing countries is to acquire technology at a fair price, on
reasonable terms and conditions, and with maximum effectiveness in developing their indigenous
assimilative and innovate capacities.
It has also been recommended that the developing Countries would need to strengthen their
human capacities in order to evolve a co-ordinated technology development strategy. These could
be through:
i. Technical Education, Vocational Training.
ii. Institutional systems (Research Centers, Consultancies, Diffusion and Extension
Services;
143
iii. Organization Arrangements (Data-Banks, Patenting, Rule-and Standard-Setting); and
iv. Administrative Capabilities (Technology Forecasting and Assessment, Technology and
Related Industrial and trade Policy-Marking, Negotiating Capacity, e.t.c)(18).
Intellectual Property Rights (IPR)

Intellectual Property Rights (IPR) enhances a Nation's development, business image and integrity.
Such protections stimulate advances that imparts positively on the entire world be it technology,
medicine, chemical or biochemical processes. The protection of IP right is crucial to a Nation's public
(6) th
Health and safety. Why in any case should IP right be entrenched in Nigeria? . At the close of the 20
Century and at the dawn of the 21st Century, Global Economic output was generated and maintained
by services that are largely dependent and sustained on the wings of New and emerging technologies.
An increase in GDP during the period was traceable to innovation deliberately and conscientiously
pursued by those nations that had the ambition to lead others. The hope we all have for a better
tomorrow and indeed for a better future depend on those inventors and innovators who have and are
striving to make the world a better place through their creative energies and sustained hard work(5).
Among them are the following famous Inventors and Achievers:
Antoine Lavoisier, the father of modern Chemistry

Francis Bacon, the father of experimental Philosophy
Hippocrates, the father of Medicine
Copernicus, the father of modern Astronomy
Dr. AlfredBernhard Nobel, who invented the Dynamite
Roger Bacon, who invented gun powder
Michael Faraday, who invented the dynamo and Electric Transformer
Alexander Graham Bell, who invented the telephone
Professor David Edward Hughes, who invented the Microphone
James Watt, who invented the Steam Engine
George Stephenson, who invented the Locomotive
Orville and Wilbur Wright, who invented the Aeroplane
(5)
Sir Charles Algernon Parsons, who invented the Steam Engine .
Galileo Galilei, who Invented the Thermometer and the Telescope and
Ogodo Aemurerhimen Dickson, the father of international Reference Standards of Silica-based
Packaging Materials for Standardization for Regulation in Nigeria.
Innovations, Intellectual Property, Its Value and Its Uses

A new kind of society is taking shape in the modern world, one that is based on knowledge and
information and one that is oriented toward not so much on the creation of information as its
management. The wealth of nations is also changing. While prior centuries were dominated by nations
with superior industrial or agricultural capabilities, the innovation stage rewards new competencies
and strengths(10). Knowledge – ideas and the people who generate them– is the new coin of the realm.
Innovation capacity will be the key driver of future economic prosperity with emerging technologies,
such as standardization of Silica - based packaging materials using International Standards
144
Organization (ISO) Standards
Patent
· What is a Patent? A Patent is a document that gives the Patentee/Assignee the right to exclude
others from making, using, and selling what is claimed as the invention for a limited period time.
· What is useful? Something that benefits the society. Utility must be specific, substantial, and
credible.
·Patentable? Patentable things include the following:

·New and useful process
·Manufacturing method
·A machine
·An unknown combination of known parts
·New and useful improvement on a process, machine, etc
·Patents are granted in many parts of the world on a 'first to file' basis. It is, however, granted on
(10)
a 'first to invent' basis in the United States of America .
Innovation Versus Invention

Invention and innovation, though closely related, are not the same. Innovation is often confused with
research and measured in terms of scientific or technological outputs. It is rather the application of
knowledge in production. This knowledge might be acquired through learning, research or experience,
but until it is applied in the production of goods or services it cannot be considered innovation. It is a
distinctly different activity from invention. To invent is to create or design something new and preciously
non-existent. An invention is thus anything created that is novel. To innovate is to introduce something
new and previously non-existent in a particular setting. An innovation is thus anything that is novel in a
given setting(10). Innovation is the technical and commercial activities involved in the marketing of a
new/improved product or the first commercial use of a new/improved process or equipment. It includes
continuous improvement in product design and quality, changes in Organization and Management
routines, creativity in market and modifications to production processes that brings cost down,
increase efficiency and ensure environmental sustainability. For an invention, the focus is on novelty
and this is a key criterion for patenting. In other words, it must be new to its field all over the world.
Innovation on the other hand does not have to be new except in a specific situation. Thus, innovation
consists of the 'process by which Firms master and implement the design and production of goods and
services that are new to them, irrespective of whether they are new to their competitors, their Countries
(10)
or the World . An innovation system is conceptualized as a network of Firms and other economic
agents who, together with the Institutions and policies that influence their innovative behavior and
performance, bring new products, new processes and new forms of Organization into economic use. It
is an economic concept. A technological innovation cannot be said to have been fully implemented
until it has been introduced on the market (product innovation). Innovations therefore involve a series
of Scientific, Technological, Organizational, financial, and commercial activities. A key ingredient in the
transformation of an innovative idea to an innovative product is entrepreneurship.
Fostering economic democracy and entrepreneurship is one tool for self-sufficiency.

· Entrepreneurship is the core organizing principle for the continued building of a Company. This
145
is because Entrepreneurs:
·Change the business and civic landscape
·Open and lead markets
·Innovate
·Take risks(10).
Repositioning Science, Technology & Innovation System to Meet the Challenges of Vision 20:2020.
X-rayed National Consensus, Shared Vision & Values

v The extent of the development of endogenous capacity of nation depend on its ability to construct a
19
consensus to define its national goal ( ).
v Vision 20: 2020 has provide the national goal.
v There is need for elites to subscribe to the same vision; to believe that Nigeria can be catapulted within a
given short period of time.
CHALLENGES OF VISION 20: 2020

v Nigeria's current rankings in the world makes vision 20: 2020 look bleak or totally impossible:
st
- CIA world fact book ranked Nigeria as the 41 on the list of development economics based on GDP for
2007.
- For the same year, world bank ranked Nigeria 40th
- On the group description of knowledge index, Nigeria ranked 66th amongst the Lagging Followers
Group
th
- On the global competitiveness ranking, Nigeria ranked 95
Vision 20: 2020 is Achievable

19
v Daunting as these challenges may seem, vision 20: 2020 is achievable ( ).
v Our way of thinking and doing things must however be differently
v Though it took Britain 58 years to double its per capital output, China has been doing it every 7 years in
the last 2 decades
v Compared with their predecessors, they did things differently.
v They invested in manufacturing; Advanced Manufacturing Technology (AMT)
v Using AMT, the Chinese have become the fourth growth zone along with the US, Japan and Germany
(19).
On the Way Forward, Science Governance is Important:

v The professional, intellectual apparatus and ideological commitment of policy makers and science
administration are very crucial to the attainment of building and utilizing SETI capacity for national
development.
v In Nigeria's economic management at the top echelon, no single scientist of note sits on any of the two
bodies.
v There is need for correction here in order to facilitate the required synergy to realize the nexus between
science and the national economy.
v The prevailing national dictum of buying & not making which export our jobs and competencies needs
to change (19).
146
Innovation, Invention & Intellectual Property
v Knowledge, ideals and people who generate them - is the new currency.
v Innovative capacity will be the key driver of future economic prosperity.
v Competition is not for control of raw material, but for the most dynamic strategic asset: “productive
knowledge”
National Innovation System Links S & T to Economic Growth

v A technological innovation cannot be said to have been fully implemented until it has been introduced on
the market (Product Innovation).
v Innovations therefore involves a series of Scientific, Technological, Organization, Financial, and
Commercial activities.
v A key ingredient in the transformation of an innovative ideal to an innovative product is
entrepreneurship.
v Fostering economic democracy and entrepreneurship is one tool for self – sufficiency(19).
Harnessing Science & Technology Research Results for Sustainable National Development
The Nigerian Vision 20:2020
v Be one of world's 20 largest economies by 2020 based on remarkable potentials:
§Population of 1 50million
§Median age of 18.63 years
§Proven resourceful people
§8th largest oil exporter
§6th largest producer of gas
§34 different minerals
§Rich arable land and friendly climate
v Assumptions underlying the Vision:
§GDP growth from 3.5% (2009) to 7% (2015) to 13.5% (2020)
§Poverty level will reduce from 70% to 30% in 2020 (using $2/day).
§Power supply increase from 3 000MW to 5 0,000MW in 2020 (20).
v However, significant investments in S&T education leads to innovations.
v Need for leadership 'Yes! We can attitude and work together.
v Need to strengthen STEM (Science, Technology, Engineering and Maths) education.
v Vision 20:2020 education free at primary and secondary levels by 2003 and 2009 respectively, 26% or
more of national budget to education.
v More than visions, time to implement e.g. Science, Technology and innovation (STI).
v STI more than traditional sciences but includes humanities.
v “In the final analysis, it is basically the mastery and utilization of modern S&T that distinguishes the south
from the north- AbdulSalam (l979 Nobel Laureate in (20)).
v Need to develop capacity for processing Mineral/Agricultural Products
20
v Metrics for determining knowledge economies GDP, R & D expenditure, patents, electricity etc ( ).
v For example need to achieve l,000Wper capita
v STI results Japanese post World War II economic expansion Teknion University & Weizmann Institute in
Israel, China etc.
v Sound scientific knowledge fundamental to addressing micro-economic transformation, bio-fuels,
147
embryonic stem cells, GM crops etc.
v
Knowledge accumulation accelerated by ICT.
v
Targeted R&D needed to fast-track developing economies.
v
Reverse engineering research institutes provide road-map to circumvent roadblocks to indigenous
technology enhancement(NASENI).
v
National S&T strategy consult
v
Change from consumer to producer nation WTO, GATT Agreements, WMO Montreal protocols
reviewed.
Strategies and Challenges

v STI vision: “to build an STI system that will drive competitive knowledge economy towards 20-2020.
st
v 1 strategy build science based workforce
- Share of manufacturing foreign and 50%by 2020 to increase from 4% (2009) to 20% (2015).
- Productivity improvement 25% by 2015 and 50% by 2020
nd
v 2 strategy forge national innovation system encompassing existing & new STI institutions:
- 5% of R&D patentable 2015 (50,000 apps) and by 2020 (100,000 apps).
- 30% of patentable R & D commercialized by 2015 and 50% by 2020
- Expected increase in investments (public / private) in activities to 5.6% by 2020.
v 3rd Strategy engender culture of STI in society:
- Improve emoluments of S&T professionals.
- Pre-1980 policy of enhanced scholarships to S&T students.
v 4th Strategy for Government: Commitment through creation of Nations Foundation for Science,
Innovation and Competitiveness.
- Also national awareness of S&T in populace.
- Reinforce strong academic background (20).
Other Strategies:
- Emerging technologies (biotech, nanotech, Project Ogodometrics International).
- Develop renewable energy sources.
- Space capabilities for socio-econ purpose.
- Promote value addition to agro, mineral & petroleum resources
- Optimize use of research facilities & humans.
- Enhanced IT capability
- Development of indigenous & orthodox medicine
v CHALLENGES AGAINST NIGERIAN VISION (NV) 20:2020 INCLUDE:

Lack of political will, policy inconsistency.
- No STI-supporting infrastructure
- Under-funding of research ETF
- Activities/Scope of R&D institutions limited
- Non-identification of areas of comparative advantages
- No linkages between R&D, & manufacturing
- Inhibiting cultural practices & superstition Professor Obe summarized by stating that:
148
Wealth of nation measured by inherent knowledge
Research institutes, Universities & Academy key role
Research institutions to develop research policy with an agenda
Challenges to Nigeria Vision (NV ) 20:2020 can be overcome through S&T
Build capacity for appropriate technologies merit-based System
Train Scientists, Engineers etc.
Promote grass-roots inclusive innovation
Develop technology transfer know how (Project Ogodometrics International for International
21,22,6,23& 24
Reference Standards for Technology Transfer) ( ).
Strength capacity of local science institutions to conduct research
Collation and marketing of S&T research results by NAS (Science) and NAE (Engineering)(20).
- Open and lead markets
- Innovate
- Take risks
v Entrepreneurship in the curriculum of Science and Technology studies is crucial.
v This needs to be done for science and engineering curriculum in the universities nationwide.
Table 1
COUNTRIES THAT HAVE HEAVILY INVESTED IN R&D HAVE REAPED SUBSTANTIAL
ECONOMIC BENEFIT (24).
SOUTH SOUTH NIGERIA
KOREA CHINA INDIA AFRICA
Population in million
(2007 estimate) 48.0 1329.1 1164.7 49.2 147.7
R&D Expenditure as %
GDP (2000 - 2005) 2.6 1.4 0.8 0.8 0.20*
Researchers in R&D per
million people (1990 - 3187 708 119 307 125.9*
2005)
Royalties and license fee
in USD per person 38.2 0.1 - 0.9 ?
(2005)
Patents to residents per
million people(2000 - 1113 16 1 - ?
2005)
Source: Human Development Report, 2007/2008/2009.

v Developing nations can no longer compete based only on their natural resource endowments
andlocation advantages.
v For a nation to withstand in this era of globalization there is need for such to identify its niche areas and
build on it by to application of scientific methods (e.g. Brazil: SUGARCANE; Malaysia: OIL PALM,
Ethiopia: Cut flower, Now Nigeria:
v International Reference Standards through Project Ogodometrics International (which way?
High - tech? Low - tech?).
v What kind of R & D do we need? High - tech? Low - tech?
v No economic gains from R & D unless the outputs (including patents) are exploited (25).
149
v Translation to innovation occurs through entrepreneurial activities that take care of marketing.
v When R & D is demand drive and solves real life problems.
v No economic gain from R & D unless the outputs (including patents) are exploited.
v Translation to innovation occurs trough entrepreneurial activates that take care of marketing
v When R & D is demand driven and solves real life problems model for the relationship among Research,
Development and Diffusion
a = Research and development;
b = Commercialization;
c = Publication, etc;
d = Research, development and Diffusion,
e = the Nation Innovation System (NIS), the domain in which all activities take place.
How Technology Can Diversify Economy
In the present Globalized World, Science and Technology is the bench mark on which national progress and
development is measured. No nation can be competitive in the modern world without properly harnessing and
developing its science and technology potentials. Socio-economic development is in most pact dictated by the
techno scientific advancement a global trends that is glaring.
The strategy on how technology can diversify the nation's economy is focused on four (4) key elements.
(1) Re-orienting values

(2) Reducing poverty
(3) Creating wealth
(4) Generating employment in realization of national goals to improve the quality of life in Nigeria for the
realization of the goals of MDGS and NEEDS (26).
The requirement or agents are;

(a) Talents.
(b) Infrastructures.
(c) Material Resources
(d) The Embedding of Science and Technology (S & T) innovation.
(5) Chemists in Nigeria corridors of power should obtain the necessary connections for political
appointment so that our interest is represented when taking major decisions on Science and Technology.
(6) We should articulate a policy that will harness science and technology in practicable and functional ways
for sustainable development.
26
(7) We should be more productive in terms of patents as well as research and publications ( ).
(8) We should improve productivity in accordance with government policies by propagating the results of
scientific and technological research to benefit the country in sectors such as Agriculture, Industry,
Commerce and Health.
(9) The drive towards advancement in technology ought to move in three interrelated directions.
a. Improvement of indigenous technologies to revive self confidences in our potentials.
b. Understanding the imported technology transferred on our needs and resources.
c. Development of new technologies based on needs and resources and the competitive advantage
which its offer.
150
(10) These technologies should reach the market as quickly as possible.
(11) For higher institution in this country to progress, detail attention must be paid to infrastructural facilities,
equipments, laboratories, libraries, human capital development, quality of research and publications.
(12) For any national economy to compete internationally there is need for manpower with the right skills and
productivity through the National System of Innovation (NSI).
(13) There should be an institutional arrangement between Universities, the State and Industries aimed at
generating University spin – off firms, internal initiatives for knowledge – based economic development
and strategic alliance among firms ( large, medium and small operating firms in different areas with
different levels of technology for the national system of innovations to succeed.
(14) A straight forward benefit of science and technology for the demand side of the economy lies in the
provision of quality control, standardization and measurement techniques, international standards and
measurability are important since they will allow Nigeria products to benchmark themselves against,
26
and conform to global standards ( ).
(15) We should therefore provide adequate funding for science and technology sector, ensure a stable polity
and a conducive science and technology environment that encourage local researchers and lure home
those abroad, while ensuring a well co – ordinated national system of innovation under the aegis of the
Federal government (26).
Recommendations
The Study recommends that
1) A Co-ordinating Office under a Presidential Scientific or Technical Adviser is the only option for Nigeria
using Science Governance.
2) There should be effective Global Technical Partnership/Collaboration/Synergy for National
Development through Technical and Commercial Scientific Licensing with Proposed Member
Countries to generate Nigeria Vision 20:2020 target for Global Competiveness.
3) There should be no Sole Concession/Patent /Franchise Rights in Proposed Member Countries without
Ogodo (Nigeria) Standards International (OSI) Permission except as Permitted by Law through
Technology Transferred Agreement of Intellectual Property Rights (IPR).
4) Standardization of Mr. Ogodo A. Dickson Certificated Firms (Songhai –Delta, Amukpe-Sapele)

for Award Winning Technology can be used as a tool for enhancing trade through Technology
Transfer across the boarders of two entities in the process of economic development of both
Developed and Developing Countries. The OGODO INTERNATIONAL REFERENCE
STANDARDS for Silica-based Packaging Materials should be Documented, Recommended ,
Accepted and Implemented around the World and approved by the International Organization
for Standardization (ISO) in Geneva in collaboration with Standard Organization of Nigeria
(SON), National Agency for Food, Drug, Administration and Control (NAFDAC) etc after the
Global Memorandum of Understanding (GMOU) is entered into between the Federal Ministry
of Science and Technology (FMST), Ministry of Industries, SON, NAFDAC, HAVARD
151
UNIVERSITY U.S.A., AMERICAN OIL CHEMISTS SOCIETY, (AOCS) U.S.A., WORLD
TRADE CENTRE, WORLD BANK ETC. AND OGODO REFERENCE STANDARDS
INTERNATIONAL.
5) The Songhai-Delta (Amukpe-Sapele) Innovative Technology Transfer for Silica-based

Packaging Materials will be transferred to the 253 Countries of the World, the Seven
Continents of Africa, North America, South America, Asia, Antartica, Europe and Australia
should be backed with the signing of a Global Memorandum of Understanding (GMOU) for
Sole Concession/Worldwide Patents/Franchise Rights in Members Countries for Product
Specifications and applications with qualified professional Chemists / Scientists to attract
Global Participation of Stakeholders in Industries, Academia, Scientists bringing together
the industries professionals, Researchers, investors, Government and Corporate
Establishments to examine and Share ideas in ways to improve the Songhai-Delta (Amukpe,
Sapele) Technology Transfer to Develop local Expertise and Optimize integration of Industries
with other vital sectors of the Economies of the World.
6) This Vision 20-2020 Document should be gazetted in the Corporate Affairs Commission in
Abuja.
7) A Position Paper/White Paper should be presented in White House / Nigerian House in London
for implementation of the International Reference Standard Regulation in Nigeria.
8) The Silica-based Packaging Materials should be Registered with the Standards Organization
of Nigeria, Nigerian Patent Office and United States Patent Office.
9) The Songhai-Delta, Amukpe Sapele Innovative Technology Transfer and Implementation

Project should be adequately funded.
10) The Scientific Work should be patented in the United States Patents Office.
11) Songhai-Delta (Amukpe-Sapele) in Nigeria is the Technology Supplier (Country A) of the Silica
based Packaging Materials while all other Countries (Country B) are the Technology users for
the Transfer of the Stock of Technical Knowledge. (Skills and know-how, knowledge, Expertise
for generating and Managing Technical Change)
12) The Federal and State Law-Makers should give Songhai-Delta, Amukpe Sapele the required
Constitutional backing after Submission of the position papers to the President, His
Excellency, Mohamadu Buhari, and the Senate President for approval to make provision for
its regulation and usage with its functions and powers for the effective implementation of such
functions.
13) A bill for a law to make provisions for the Control, Regulation and Implementation of Superior
Pigmented Silica-based Packaging Materials should be enacted by the National Assembly of
the Federal Republic of Nigeria and by Authority of same through the member Representing
Sapele Constituency in the Delta State House of Assembly/Hon. Speaker of the Delta State
152
House of Assembly , Hon. (Chief) M. O. Igbuya and Commissioner for Science and
Technology, Delta State, Hon. Joyce Overahto recommend this acceptable Nigerian officially
Internationally Recognized Standard that will be documented as the best quality Superior
Pigmented Silica-based Packaging Materials.
14) There should be no Patent Rights/Franchise Rights/Sole Concession Rights in Proposed

Members Countries such as Austria, Belgium, Denmark, Finland, France, German, Greece,
Spain, Sweden, Switzerland and the United Kingdom without OGODO STANDARDS
(NIGERIA) INTERNATIONAL (OSI) PERMISSION EXCEPT AS PERMITTED BY LAW.
15) The OGODO STANDARDS (NIGERIA) INTERNATIONAL (OSI) should be given the status of
a National Standard by endorsement at the Latest by March 2017 and Conflicting Standards on
Superior Pigmented Silica-based Packaging Materials shall be withdrawn at the latest by
March 2017.
16) The OGODO STANDARDS (NIGERIA) INTERNATIONAL (OSI) should be approved by the
International Organization for Standardization (ISO) in collaboration with Standards
Organization of Nigeria (SON) and National Agency for Food, Drug, Administration and Control
(NAFDAC) before March 2017. OGODO STANDARDS (NIGERIA) INTERNATIONAL (OSI)
Proposed Members Countries should comply with OGODO Standards (Nigeria)
International/National Agency for Food, Drug, Administration and Control/European
Committee on Standardization (OSI/SON/NAFDAC/ISO/CEN) International regulations which
stipulates the conditions for giving the OGODO STANDARDS (NIGERIA) INTERNATIONAL
(OSI) the status of a National Standard without any alteration and this should be completed in
36 Months.
17) The bill for the Implementation of this Innovative Technology Transfer should be passed into
Law by the approval of the State and National Assembly in its first and Second hearing after
President, His Excellency Mohamadu Buhari has gone through it for approval to the House for
Implementation.
18) A panel for the implementation of this Innovative Technology Transfer should be constituted
and to submit their report while a Blue Print for the establishment and reactivation of existing
Companies in Sapele, Delta State, Nigeria will be submitted to the Federal Government for
Consideration.
19) Worldwide Patent Rights through domains in Professionalism can be got from America through
the Assistance of Delta State Governor, Senator Dr. Arthur Ifeanyi Okowa.
20) A Three-Year Action Plan should be set up by the United Nations Agency for International
Development and a reasonable amount of fund should be approved for the Implementation in
Nigeria.
21) The Delta States and Federal Government should do everything within its power to see to the
realization of this innovative technology transfers since it will bring revolving funds from other
parts of the World to Nigeria.
153
22) The Federal Government via the Federal Ministry of Science and Technology should help to
tackle this matter by making Companies to participate in this programme.
23) One of the functions of Standards Organization of Nigeria (SON) is to ensure Reference
Standards, for calibration and verification of measures and measuring instrument to compile
an inventory of products requiring Standardization. This study is one of such requiring
Standardization of Silica based Packaging Materials.
24) The support of the Standards Organization of Nigeria (SON), International Standards
Organization (ISO), National Agency for Food, Drug, Administration and Control (NAFDAC),
Federal Ministry of Science and Technology (FMST), Manufacturers Association of Nigeria
(MAN)etc, should be sought in the Implementation of this Innovation Technology Transfer and
gazetting the position paper in the Corporate Affairs Commission in Abuja.
25) Ogodometrics will join in the fight to kick Polio and other killers diseases out of Nigeria from the
funds made available through the International Reference Standards from the Innovative
Technology Transfer from Nigeria to other nations through the Vision 20-2020 documents.
26) Some Orphans will benefits from Ogodometrics every 15th of February of every year.
27) Ogodometrics will award Scholarship to brilliant but indigent students through a written test
every 15th of February of every year.
28) The proceeds from the Packaging Added Value (Ogodometrics) will be use to collaborate with
the Federal and States Government to establish FREE EDUCATION UNIVERSITY IN
AMUO-OGODO TOWN, SAPELE, DELTA STATE, NIGERIA whereby every State in Nigeria
will contribute 51% of its yearly declared profits to the funding of this University.
29) The Delta State Government should establish a Packaging Trust Society International in
Amuo-Ogodo Community, Sapele, Delta State, Nigeria and a bill should be proposed and
Titled “Development of Standards, Research on Standardization and Publication of
Standards, Certification, Delta State Bureau of Standards, Delta State, Nigeria. The bill
should be immediately passed to the relevant House Committee to study and recommend to
the general House for legislation after which it will be passed into law in the tenure of the current
legislature.
30) This research work should serve as the National and International Reference Standards for
Silica-based Packaging Materials, through national and International collaboration to cause
rapid development of the Nation through Science and Technology by using OGODOMETRICS
in the Context, of the National Science Technology and Innovation System in the Federal
Ministries of Education, Science and Technology and Finance.
154
31) The Delta State Government should work in collaboration with the National Board for Technical
Education (NBTE) and National Universities Commission (NUC) in the Areas of National and
International Collaboration with the reservoir of skills and expertise available among its team of
28
resource persons ( ).
32) The African Network of Scientific and Technological Institutions (ANSTI) should assess major
areas of Constraint that requires urgent attention using Science and Technology Education in
Nigeria and provide financial support for this institution.
33) To start with, the Delta State Higher Education should be determined to set the pace for
Nigerians and make research activities a top priority program. This it can do by setting up an
outlet of government bureau in every tertiary institution in the state. This is to make a grass root
contact with the potential researchers dispersed from the government office in Asaba. In
consonant with the above, the steps should be adopted to achieve a vibrant research
atmosphere in education system.
34) A reasonable proportion of the State budget should be allocated to fund research.Maintenance
and Purchase of Research Equipment should be given top priority under the direct supervision
of the Research Committee set up by the institution with a representative from the government.
35) Sharing of limited resources including exchange of equipment, data and information:
stimulating and energizing Higher Institution and Research Institute to respond to community
need and repositioning their curricular for better relevance to industry and national
development.
36) Provision of free access to internet facilities for all Academics Staff as a means of accessing
current information as well as cross-fertilizing ideas locally and internationally.
37) Establishment of viable and well equipped research centers in all Tertiary Institutions. The
nature of these centers should be geared to reflect and meet the set goals and aspiration of the
institutions.
38) Research and travel grants should be made available to prospective researchers without
politicizing the scheme.
39) Reward should be periodically given to the best researcher. This will stimulate and motivate
others to venture into researches(37).
40) Government Should Adequately Fund Chemistry Programme in Higher Institutions/Research

Institutes/Agency Because the Chemical Industry is a Crucial one in the Nation's Economy. The
Nigerian Government should make it mandatory for the Industrial sector to join International Bodies to
facilitate development and a good example of such Organization in Scientific and Industrial
28 29)
Development is the World Association of Industrial and Technological Research Organization ( ), ( .
155
41) Non - Commercialisation of Research Results
Nigeria is yet to realize the critical linkage between science, technology innovations and industries for
the benefit of the citizenry. The knock – out effect of this is the over reliance on the foreign sources of
even the basic commodities and services with the manufacturing sectors contributing only about 3% of
30
the nations, Gross Domestic Product (G D P) ( ). In Nigeria, unfortunately the critical linkage between
Science Technology Innovations (S T I) and industries has not been realized. The knock out effect of this
reliance on foreign sources of even the most basic commodities and services with the industrial
manufacturing sector contributing a mere 3% to the G D P. Some of the critical challenges facing
Nigeria's STI system include
(a) Non – commercialization of successful research results
(b) Non - demand driven research and development (R & D) activities
(c) Duplication of functions among research institutions and competitions from foreign import,
among others.
There is a need to reverse these unwholesome trends in order to entrench the culture of good S T I practices and
30
how the best outcome of the S T I can facilitate industrial growth and production ( ).
Research result should be designed to achieve the following:

(a) Identifying and promoting strategies and procedures that would accelerate the commercialization of R
& D result in Nigeria.
(b) Forging a national innovation system that encompasses all existing and new S T I institutions towards
the achievement of vision 20-2020
(c) Engendering the activities as well as stimulate the active interest of Nigeria in science and technology
and innovation' among others
(d) Even though all the parastatals under the Ministry of Science and Technology are doing a lot in the areas
of research and development, these achievements has never been translated to the desired benefits of the
average Nigeria.
(e) It is time for the country to step up in the area of research and development from the laboratories and pilot
stages to full commercialization, so that Nigerians could begin to enjoy the result of research and
development in Science and Technology (30).
(f) Most of the outcomes of research and development in Nigeria have unfortunately remained unutilized, a
development which has obviously stunted industrial development in the country.
(g) We should proffer a framework that would make the outcomes of R & D easily accessible, especially to
investors and entrepreneurs and to identify and promote strategies and procedures that would accelerate
the commercialization of these results and forge a NATIONAL INNOVATION SYSTEM (NIS), that
31
would drive the achievement of vision 20 -2020 ( ).
Conclusion
The Research Study concludes that:
1. ISO Certifications requires that the manufacturing facilities are certified for the Design, Development,
Manufacturing, Assembly, Sterilization by Irradiation and meets the US Pharmacopoea Light
Transmission Test of Ogodometrics Superior Material produced in a Plant awarded ISO Quality
Management System and Environmental System and complied to ISO for permanency of material.
156
2. Adequate Funds should be made available to provide the necessary systematic Technology Transfer to
Nigeria on new challenges of Vision 20:2020 Agenda.
3. Expand, Collaborate with other countries with appropriate human and material resource through
PROJECT OGODOMETRICS INTERNATIONAL for Vision 20:2020 Agenda of Technology Transfer
Arrangement/ Agreement. The scope of the Intellectual Property Rights (IPR) should be Via synergy
with Harvard University, USA; International Standards Organization Geneva and American Oil
Chemist's Society, USA etc. United States of America (USA), South African Government, National
Office for Technology Acquisition and Promotion (NOTAP) and NIGERIAN RESEARCHERS
SHOULD TEAM UP TO DEVELOP THE NEW REVENUE FORMULA FOR NIGERIA WEALTH
23
CREATION ( ).
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160
CHEMICAL CONSTITUENTS OF SANDBOX TREE (HURA CREPITANS LINN.) AND ANTI-
HEPATOTOXIC ACTIVITY OF THE LEAVES AND STEM BARK EXTRACTS
Ganiyat K. Oloyede1*, Oluwatosin A. Adaramoye2 and Mutairu B. Olatinwo1

1
Natural products/Medicinal Chemistry Unit, Department of Chemistry, University of Ibadan, Nigeria.
2
Department of Biochemistry, University of Ibadan, Nigeria.
* Author to whom correspondence should be addressed.
Ganiyat Kehinde OLOYEDE (Ph.D)
E-mail: gk.oloyede@ui.edu.ng/oloyedegk@gmail.com Telephone: +234 803 562 2238
ABSTRACT
Objectives: The use of natural products derived from plants for therapeutic purpose is as ancient as human
civilization. Sandbox tree, Hura crepitans L. (Euphorbiaceae) is one of such plant and has been reported to have
many ethnomedicinal applications especially as antimicrobial, anti-inflammatory and antihepatotoxic effects.
This recent study was designed to determine the anti-hepatotoxic activity of the ethylacetate soluble fraction of
the leaves and stem bark of H. crepitans and to isolate secondary metabolites.
Methods: Chromatographic technique was used for isolation and Ultraviolet-Visible (UV), Infra-red (IR) and
Nuclear Magnetic Resonance (NMR) spectroscopies were used for structural elucidation. Anti-hepatotoxicity
study was carried out using carbon tetrachloride (CCl4) induced rat model and biochemical parameters: alanine
aminotransferase (ALT), aspartate aminotransferase (AST), L-ã-glutamyltransferase (GGT), urea and creatinine
(CREA) were assayed on the serum. Phytomicrographs of the liver samples were also taken and analyzed.
Results: Our present study showed that biochemical studies of blood samples of CCl4 treated rats with value
105.0±0.001 AU inALT showed significant increase in the level of serum enzyme activities reflecting liver injury
but, 69.0±13.23 AU for leaves and 53.3±2.52 AU for bark (p<0.05) indicated protection of hepatic cells. AST,
GGT, urea and CREA also reduced significantly. Daphnane diterpenes, daphnetoxin acid and huratoxin were
isolated from H. crepitans in this recent study along with apocynin and methylpentadecanoate.
Conclusion: H. crepitans significantly reduced the level of biochemical parameters indicating protection against
hepatocellular injury. Isolates obtained from this plant could also serve as lead compounds in therapy of diseases
involving hepatic injury.
Keywords: Daphnane diterpenes, apocynin, methylpentadecanoate, hepatoprotective, histopathology.
INTRODUCTION
Mineral, plant and animal products were the main sources of drugs for a long time and the use of these natural
products with therapeutic properties is as ancient as human civilization (1-2). But recently, interest in therapeutic
use of natural products, especially those derived from plants deepened because report has it that certain
percentage of drugs prescribed worldwide and those considered as basic and essential by the World Health
Organization (WHO) are exclusively of plant origin or many are synthetic drugs obtained from lead compounds
(1-3). Examples of important drugs obtained from medicinal plants are morphine and codeine from Papaver
somniferum, artemisinin from Artemesia annua, vincristrine and vinblastine from Catharanthus roseus, digoxin
from Digitalis spp., quinine and quinidine from Cinchona spp., and atropine from Atropa belladona. The
importance of these natural products cannot be overemphasized as these secondary plant metabolites can be lead
compounds, allowing for the design and rational planning of new drugs, development and the discovery of new
therapeutic properties not yet attributed to known compounds. In addition, compounds such as physostigmine,
161
cannabinoids and yohimbine obtained from plants, are important tools used in pharmacological, physiological
and biochemical studies (1-8).
Sandbox tree, Hura crepitans L. (Euphorbiaceae), a plant common to the tropics has been reported to have many
ethnomedicinal applications especially as antimicrobial, anti-inflammatory and antihepatotoxic effects (9).
Adedire and Ajayi (2003) (10) reported the insecticidal activity of the seed oil. Huratoxin was isolated from the
milky sap of H. crepitans along with hexahydrohuratoxin and keto-enal. Crepitin is also a toxalbumin derived
from H. crepitans and has been shown to be toxic (11-12). Hurain, the proteolytic enzyme of the sap of H.
2+ 2+ 2+ 2+ -
crepitans, has also been reported to be activated by Fe and inhibited by Ni , Zn , Cd and Cl ions; its activity
diminished in the presence of trypsin inhibitors from soy bean, black bean and Ascaris lumbricoides (13-14).
Lectins which are carbohydrate-binding proteins, specific for sugar moieties were isolated from the latex of H.
crepitans L. and Euphorbia characias L. (Mediterranean spurge) (15). Recently, phytochemicals such as
glycosides, alkaloids, flavonoids, tannins was reported in the seed oil (16-18). The presence of some fatty acids,
sterols and acyclic aliphatic ketones in the root of H. crepitans has also been reported (19) while antimicrobial
activity of the extracts from leaves and stem bark; chemical composition and antimicrobial activity of
essential oil of H. crepitans leaf (20) have also been investigated. Not much report of the nature of secondary
metabolites and pharmacological activity of the aerial parts of H. crepitans L. (leaf, stem or stem bark) was found
in literature. The objective of this research work therefore was to isolate, purify and characterize the chemical
constituents in H. crepitans leaves and stem bark and to screen the ethylacetate soluble fraction for
hepatoprotective activity against carbontetrachloride-induced liver damage in rats since it has been established
in a separate assay that H. crepitans ethylacetate fraction has antioxidant activity when compared to other
fractions (21). Plants with antioxidant activity have been observed to have protective effect against hepatic injury
(22). Hepatotoxicity implies chemical-driven liver damage. Liver plays a central role in transforming and
clearing chemicals and is susceptible to the toxicity from these agents. There are different kinds of disease
conditions that affect the liver, some of which are liver cirrhosis, hepatitis, liver cancer amongst others.
Chemicals that cause liver injury or damage are called hepatotoxins and more than 900 drugs have been
implicated in causing liver damage. Hepatotoxic substances that are commonly used for in vivo and in vitro
assays are CCl4, vinyl chloride, bromobenzene and ethanol (23-24).
Plant collection and identification
Fresh leaves and stem bark of H. crepitans were identified and authenticated by Mr E. Donatus of the Herbarium
unit of Department of Botany and Microbiology of the Faculty of Science University of Ibadan and were
collected in September 2009 at the Botanical Gardens, University of Ibadan, Ibadan, Oyo state, Nigeria. This
plant was also compared and found to be the same with a Herbarium specimen labelled FRIN 10565 at Forestry
Research Institute of Nigeria, Ibadan, Oyo state, Nigeria.
Animals: Adult Wistar (albino) rats of average weight 160 g (14 weeks old) bred in the animal house of the
Department of Biochemistry, University of Ibadan were used. The animals were divided into 8 groups. Five
animals were used per group. The rats were kept at 27 ± 20C for 1 week before and during the experiments and
were fed with balanced livestock feeds from Pfizer, Plc and water, ad libitum. The animal experimental protocol
was approved by the Institutional Animal Ethics Committee.
Chemicals and Reagents: Hexane, ethyl acetate (EtOAc), methanol (MeOH), butanol (BuOH), chloroform,
hydrochloric acid, naphthol, bismuth nitrate, potassium iodide, sodium hydroxide, copper acetate, NaOH,
sodium chloride, copper sulphate pentahydrate, ferric chloride, conc. tetraoxosulphate (VI) acid, conc. HCl,
ammonia solution, sodium potassium tartarate, potassium chloride, glacial acetic acid, Molisch reagent,
Dragendroff's reagent, disodium hydrogen phosphate, dihydrogen potassium phosphate, iodine crystals,
162
copper acetate, carbon tetrachloride were all BDH (England) chemicals and solvents (distilled prior to use),
silica gel 70-230 microns (Merck, Germany), and Randox Laboratory kits for biochemical assays (Randox,
UK).
Equipment and Apparatus: Soxhlet apparatus and condenser (Pyrex, UK), Mettler analytical balance H80
(Mettler, UK), Water Bath (Gallenkamp, UK), Rotavapor RII0 (Buchi, England), silica gel GF254 (precoated
aluminium sheets - Merck Germany), Centrifuge (Gallenkamp, UK), pH meter (Jenway model, UK), Astel
Hearson Oven (Gallenkamp, UK), Teflon Homogenizer (Gallenkamp, UK),, UV-Visible spectrophotometer
(UVD-2960 model equipped with a UVWIN software version LABOMED INC, USA), IR Spectrophotometer
(Perkin Elmer FT-IR system, Spectrum BX model of IR Spectrophotometer, England), Varian-Mercury NMR
Spectrophotometer (England) operating at 200 MHz for H and 50 MHz for C nuclei.
Sample preparation, extraction and fractionation procedure
The leaves and stem bark of H. crepitans were weighed separately and air-dried for 4 and 6 weeks respectively
until the weight was constant and then pulverized using mill machine at the Wood extraction laboratory,
Department of Chemistry, University of Ibadan. The pulverized dried leaves (3.5 kg) and bark (2.5 kg) were
extracted with 6.0 L and 5.0 L of distilled methanol respectively. Methanol was used to mimic local gin (alcohol)
used in preparation of the plant H. crepitans in folklore medicine. Soxhlet apparatus was used for extraction since
it has the tendency of extracting large amount of plants giving high yield within the shortest possible time. The
extracts were collected and concentrated with the aid of Bucchi rotavapor at 37 0C and partitioned into various
fractions with distilled water, hexane, ethylacetate and butanol in other to separate plant constituents based on
polarity. The EtOAc and BuOH fractions of leaves and EtOAc fraction of bark obtained above were subjected to
0
column chromatography using silica gel of mesh size 70-230 microns (activated in an oven at 105 C for 1hr) as
adsorbent. Slurry of adsorbent was prepared by mixing activated silica gel with n-hexane in optimum proportion
(ratio of silica gel to extract was 30:1) and wet-packed into the column gently to avoid air-bubbles and cracking
which could affect the resolution. Ethylacetate and butanol fractions (15 g each) of the leaves and bark were pre-
adsorbed in silica gel and dried before loading onto the column. Hexane was used to wash the packed column
prior to analysis. The loaded sample was then covered with pure white sand and small piece of cotton wool. The
top of the column was usually filled with enough solvent/mixture of solvents to avoid cracking. The elution
started with 100% non-polar hexane after which the polarity of the solvent(s) was increased gradually with
EtOAc and methanol in a gradient elution chromatographic technique. 25 ml of each of the eluents was collected.
The purity of the fractions was assessed by thin layer chromatographic analysis (TLC) using pre-coated
aluminium TLC plates. The spotted plate was developed in different solvent systems depending on the degree of
polarity of each constituent at room temperature. If not coloured after development, it was visualized in the
iodine tank. Solvent systems used were Hexane/EtOAc (1:1), (3:1), (1:3), (4:1); Hexane/MeOH (1:1), (1:3),
(3:1), (1:4); Hexane/Ethanol (1:1), (1:2), (2:1), (1:3), (4:1), (1:4) and Hexane/EtOAc/MeOH (1:1:1), (3:1:1),
(4:1:1), (2:1:4).
Elution of fractions: A total of 126 ethylacetate fractions of leaves were collected, analyzed using TLC and
pooled according to Rf values; 1-5 (H1), 6-11 (H2),12-19 (H3), 20-22 (H4), 23-33 (H5), 34-45 (H6), 46-52 (H7),
53-58 (H8), 59-74 (H9), 75-81 (H10), 82-91 (H11), 92-95 (H12), 96-101 (H13), 102-103 (H14), 104-126 (H15).
Fraction 53-58 obtained as solid crystals was recrystallized to obtain a dark greenish substance tagged Ola 3.
Butanol fraction of leaves gave 117 fractions, analyzed using TLC and pooled accordingly; 1-6 (C1), 7-16 (C2),
17-30 (C3), 31-40 (C4), 41-53 (C5), 54-69 (C6), 70-85 (C7), 86-101 (C8), 102-117 (C9). Fraction 17-30
obtained as solid crystals was recrystallized to give a dark brownish solid (Ola 5). While elution of ethylacetate
fraction of H. crepitans stem bark gave 153 fractions and analyzed using TLC. Fractions having the same Rf value
were bulked accordingly: 1-11 (HC1), 12-16 (HC2), 25-31 (HC3), 32-40 (HC4), 41-53 (HC5), 54-69 (HC6), 70-
85 (HC7), 86 (HC8), 87 (HC9), 88-100 (HC10), 101-119 (HC11), 120-153 (HC12). Fractions 25-31 and 87
163
which gave single spots when analyzed by TLC were purified to obtain dark greenish solids named Ola 2 and Ola
1 respectively while fraction 88-100 (brown solid; Ola 4) was obtained in small yield.
Analysis of the isolates: Spectroscopic techniques; UV, IR and NMR were used for structural elucidation.
Samples (0.01% w/v) were scanned between 190 nm and 400 nm with the aid of UVD-2960 model UV/Visible
spectrophotometer. Data from chart/recorder gave a graph of Absorbance against wavelength (nm). Vmax (cm-1)
from IR data obtained from a Perkin Elmer FT-IR system, Spectrum BX model of IR Spectrophotometer using
1 13
the KBr disc method also confirmed the structures. The H NMR and C NMR spectra of the pure compounds
were determined using a 200 MHz machine for 10% (w/v) solutions in deuterioacetone or
deuteriodimethylsulphoxide using trimethylsilane (TMS) as internal standard. Pulse irradiation technique
employed was FT NMR at ambient temperature. Chemical shifts are reported in ppm. Also anti-hepatotoxicity
study was carried out on the ethylacetate soluble fractions of the leaves and stem bark using carbon tetrachloride
induced rats.
Experimental design for Antihepatotoxicity Study
Rats which were bred to mature body mass were randomly assigned to eight groups with each group containing
five rats (n=5). Animal experimental conditions are as described above. The animals were administered 100
mg/kg body mass of extracts (vehicle for the extracts was corn oil) and toxicant (CCl4) intraperitonially. Group 1
served as control (corn oil), Group 2 (CCl4 Only), Group 3 (CCl4 + HCEt (leaves) pre-treatment), Group 4 (CCl4 +
HCEt (bark), pre-treatment), Group 5 (HCEt (leaves)), Group 6 (HCEt (bark)), Group 7 (CCl4 + KV pre-treatment)
and Group 8 (KV) where HCEt is ethylacetate fraction of H. crepitans and KV is Kola viron (standard). CCl4 was
administered 3 times in a week for two weeks. Extracts and control were administered 5 times in a week for two
weeks for those without pre-treatment while those pre-treated were administered 5 times in a week for three
weeks. The dose regimen and animal experimental conditions are in line with standard procedure for
antihepatotoxicity screening (25-27).
Biochemical Assay
Determination of haematological parameters
The male albino rats were sacrificed by cervical dislocation after an overnight fasting. Blood samples were
collected by cardiac puncture technique into clean dry centrifuge tubes and allowed to clot by standing for 30
minutes and later centrifuged at 3000 g for 15 minutes. The supernatant (serum) was collected and stored in the
refrigerator for preservation prior to estimation of various biochemical parameters. Also collected was the liver,
rinsed in ice-cold 1.15% potassium chloride (KCl), blotted on filter paper and weighed appropriately. The liver
tissues were then macerated, rinsed and homogenized in 4 volumes of homogenizing buffer (pH 7.4) using a
Teflon homogenizer. The homogenates were spun for 10 minutes at 10000r/min using cold centrifuge at 4oC.
Aliquots of the supernatant obtained (the post-mitochondrial fraction) was stored at 20oC while the liver and
serum for biochemical assay were kept at ice-cold temperature.
Histopathology studies
The liver for histopathology was stored in 10% formalin and then fixed in bovine solution; processed for paraffin
embedding following the standard micro technique. Sections of liver were stained with haematoxylin-eosin and
were observed microscopically for any histopathological changes. Phytomicrographs of the liver samples were
taken and analyzed (25-29). The following biochemical parameters: Alanine Aminotransferase (ALT), Aspartate
Aminotransferase (AST), L-ã-Glutamyltransferase (GGT), Urea and Creatinine (CREA) were assayed on the
serum according to the methods described in the kits manufactured by Randox Laboratories Ltd, United
Kingdom. In ALT determination, á- oxoglutarate reacted with L-alanine to give L-glutamate and pyruvate and
ALT was measured by monitoring the concentration of pyruvate hydrazone formed with the 2, 4-
dinitrophenylhydrazine. Transaminase activities in some sera were stimulated by high concentrations of
164
aldehydes, ketones or oxo acids. Measurement against a serum blank instead of a reagent blank eliminated this
effect. Also for AST determination, á- oxoglutarate reacted with L-aspartate to give L-glutamate and
oxaloacetate, AST was measured by monitoring the concentration of oxaloacetate hydrazone formed with 2, 4-
dinitrophenylhydrazine. L-ã-Glutamyltransferase (ã-GT/GGT) however, involved a reaction of L- ã-
glutamyl-3-carboxy-4-nitroanilide with glycylglycine in the presence of ã-GT to give L-ã-
glutamylglycylglycine and 5-amino-2-nitrobenzoate. CREA, in alkaline solution reacted with picric acid to
form a colored complex. The amount of the complex formed was directly proportional to the creatinine
concentration and was so determined. Lastly, Urea in serum was hydrolyzed to ammonia in the presence of
urease. Ammonia was then measured photometrically by the Berthelot's reaction (30).
Determination of protein
Protein concentration of the various samples was determined by means of the Biuret method as described by
Gornal et al., (1949) (31) with some modification. Biuret reagent contained CuSO4, KI and potassium sodium
2+
tartarate (KNaC4H4O6.4H20). Proteins form a coloured complex with cupric ions (Cu ) in an alkaline solution.
Potassium iodide (KI) was added to the reagent in order to prevent the precipitation of Cu2+ ions as cuprous oxide.
Procedure for Protein Determination in the samples
The post mitochondrial fractions of the supernatants were diluted 5 times with distilled water so as to increase the
sensitivity range of Biuret method. Diluted sample (1 ml) was added to 3 ml of Biuret reagent. The mixture was
incubated at room temperature for 30 min after which the absorbance was read at 540 nm using distilled water as
blank. The protein content of the samples was extrapolated from the standard Bovine Serum Albumin (BSA)
curve (29-31).
Data analysis
Graphs were plotted from the experiment carried out and Absorbance Vs U/I values given in the kit served as the
calibration curve. ALT, AST, GGT, Urea and CREA activities in the serum were then obtained by extrapolation
from the graph. The mean value ± SEM was calculated for each parameter and analyzed separately using
ANOVA followed by Dunnets 't' test.
Secondary plant metabolites
Secondary metabolites found in H. crepitans leaves and stem bark extracts were alkaloids, flavonoids, tannins,
steroids and phenolic compounds. Five pure compounds were obtained and recrystallized but four were fully
characterized. They are daphnetoxin acid, huratoxin, apocynin and methylpentadecanoate. Daphnetoxin; a
daphnane diterpene and apocynin though are known compounds are newly reported in H. crepitans. These
1 13
compounds were identified by their UV, IR, H and C - NMR. The spectra data obtained are in complete
agreement with literature values. Structures of the compounds reported here are shown in Fig. 1.
1-(4-hydroxy-3-methoxyphenyl) ethanone (Apocynin; Acetovanillone) (Ola 1): UV [EtOH]nm (log å): 264
-1 1
(0.423), 301 (0.162); IR (KBr) Vmax cm : 3440.21 (O-H), 1750.14 (C=O), 1601-1487(C=C aromatic); H NMR
(200 MHz; Acetone): ppm: 9.6 (O-H), 6.75 - 7.9 (3H, m, aromatic), 3.2 - 4.6 (3H, OCH3), 2.6 (3H, CH3), 13C NMR
(50 MHz, Acetone) ppm: 206.2 (C=O), 104.0 (C-unsaturated ring system), 58.6 (O-CH3), 28 (CH3); Molecular
0
formulae: C9H10O3, Mpt 112-115 C.
Huratoxin (Ola 2): UV [EtOH]nm (log å): 290.65 (0.257), IR (KBr) Vmax cm-1 : 3433.26 (O-H), 2928.72 (C-H),
1653.97 (C=O), 1508.13 (C=C alkene), 1458.02 (C-H bending), 1261.68 (C-O alcohol), 1024.37 (C-O ether),
1
995.46 (C-H, ring), 827.00 - 632.26 and 383.39-370.49 (fingerprint region, C-H aliphatic , C – O, C – C); H
NMR (200 MHz; DMSO): ppm: 8.8 (1H, m, cycloalkene,H-1), 6.6-7.3 (3H, m, OH), 5.2-5.5 (6H, m, alkene) 3.3-
165
4.2 (10H, m, cyclic), 0.4-2.8 (28H, m, polymethylene CH2/terminal CH3); 13C NMR (50 MHz, DMSO) ppm:
207.190 (C-3, C=O), 177.275(C-1), 174.933 (C-15), 172.473 (C-2),145.984 (C-3'), 145.823 (C-4'),138.839 (C-
5'), 109.173(C-12), 108.968 (C-16), 79.259 (C-8), 77.472(C-2'), 56.387(C9,13,14, 3 C-O, bridged), 53.356 (C-
4,5,20, 3C-O, (OH)), 51.921 (C-6, 7), 36.649(C-10), 32.974(C-11), 30.602(C-18), 30.206(C-17), 29.825-24.935
(C-6' - 13'), 19.019(C-1'), 18.814(C-19), 14.451-14.012 (C-14') Molecular formulae C34H48O8, Mpt 217-2190C.
-1
Daphnetoxin acid (Ola 3): UV [EtOH]nm (log å): 295.07 (0.257); IR (KBr) Vmax cm : 3400.14 (O-H stretch,
broad), 2907.06 (C-H, saturated), 1712.97(C=O), and 1508.13 (C=C), 1475.02 (C-H bending), 1361.68 (C-O,
alcohol), 1234.37 (C-O, ether), 995.46, 827.00, 752.26 (C-H, aromatic); 1H NMR (200 MHz; DMSO): C9,13,14,
3 C-O, bridged ppm, J/Hz: 12.85 (OH, carboxylic acid),, 7.763 (5H, cycloalkene), 6.104 (CH2, ethylene,
cyclopentene), 4.6 - 3.8 (OH, alcohol), 3.653- 3.362 (H, methine, cyclopentene, cyclohexane), 2.913 - 2.491(H,
oxiran, saturated ring system), 1.589-1.137 (H, methyl); 13C NMR (50 MHz, DMSO) ppm:, 173.146 (C=O, C-
21), 171.301(C-2'), 162.530 (C-15), 150.143(C-2), 144.872 (C-3'), 136.994(C-7'), 136.291(C-5'), 135.398(C-
4'), 135.427(C-6'), 132.859(C-1), 128.984(C-1'), 128.736(C-16), 123.054(C-13), 104.429 (C-9), 97.152(C-14),
94.252(C-6), 89.069(C-7), 52.639(C-20), 51.277 (C-5), 40.763(C-4), 39.928(C-11), 39.519(C-12), 38.584(C-
3), 38.260(C-8), 22.534(C-10), 17.305(C-19), 11.903(C-17), 10.556 (C-18) Molecular formulae: C28H32O9, Mpt
193-1960C.
Ola 4 is a dark brownish solid substance; UV [EtOH]nm (log å): 290.65 (0.38125); IR (KBr) Vmax cm-1 : 3399.29
(O-H stretch, broad), 1700.18 (C=O),1653.82 - 1507.35 (C=C), 1457.81 (C-H, bend), 1030.85 (C-O), 665.89,
432.89, 373.41, 361.91(fingerprint region, C-H aliphatic , C – O, C – C). No enough data to characterize it but
chemical test confirmed it was a flavonoid.
Methylpentadecanoate (Ola 5): dark brownish solid; UV [EtOH]nm (log å): 300.34 (0.374); IR (KBr) Vmax
-1 1
cm : 2914.08 (C-H), 1710.02 (C=O), 1076.23 (C-O), H NMR (200 MHz; DMSO): ppm, J/Hz: 3.2-3.6 (3H,s, -
13
CH3-O), 2.5 (2H,s, -COCH2-), 0.8-1.6 ( -CH3-, Aliphatic -CH2- unresolved sharp singlet). C NMR; ATP
(50MHz, DMSO) ppm: 191.303 (C=O), 53.4 (OCH3), 29.474 - 38.750 (-CH2-), 15.7 (CH3); Molecular formulae
O O
O
HH
o OHOH
O O OH
O
HOH
O O HO O H
O
(CH2) HH O
HO 4
OH Me
HO
(i) (ii) (iii)
O
H3C
(iv) O CH3
Figure 1: Structures of (i) Apocynin, (ii) Huratoxin, (iii) Daphnetoxin acid and (iv) methylpentadecanoate.
Spectroscopic studies showed that the compounds were unsaturated; The UV absorption values above 260 nm
obtained for these compounds falls into the range for cyclic compounds. Transition in 270-350 nm is n-ð*
transition of ketones. IR spectrum displayed characteristic absorptions for signals due to OH, C=C, C=O, C-O
functional groups. Oxygen function group with signals of aromatic hydrogen in the 1H NMR was observed
typical of aryl hydrogen adjacent to oxygen functions. These data are in complete agreement with those reported
166
for these compounds in literature (32-33).
Apocynin or acetovanillone is an organic compound found in natural source and has been reported to have
pharmacological activities (33). It is however newly reported in H. crepitans and was obtained as a dark green
solid. Absorption peaks for OH at 3440.21 cm-1 and 1750.14 cm-1 for C=O were prominent in the IR region. The
1
H NMR revealed that it has aromatic group (6.7-7.9 ppm), ether/hydroxy/ketone group (3.2-4.6 ppm) and
13
methylene group (2.6 ppm). The C NMR revealed the presence of carbonyl function at 206 ppm.
Huratoxin was also obtained as a dark greenish solid. It is a piscicidal compound (widely used to catch fish in
different parts of the world) and had been isolated from the milky sap of H. crepitans. Prominent peaks
-1 -1 -1 -1
corresponding to OH (3433.26 cm ), C-H (2928.72 cm ), C=C ring (2129.23 cm ), C=O (1653.97 cm ) were
observed but absorption for C=O was unusually low because C=C bond adjacent to a carbonyl group results in
the delocalization of the ð electrons in the C=O and C=C bonds resulting in the lowering of the frequencies. The
1
H NMR revealed absorption peaks at (ppm) 6.6-7.3 and 0.4-2.8 due to free OH and polymethylene
13
(CH2/terminal CH3) respectively; C NMR showed prominent peaks due to C=O at (ppm) 207.190 and C-O at
56.387- 51.921. About fourteen isomers of this compound are known to exist (11-13).
Daphnetoxin is a benzoic acid-ortho ester of a diterpenoid and contained carbonyl, aromatic, hydroxy/ether and
methylene/alkyl groups. Acid displacement of the cyclopentenone functional group was observed in the IR Vmax
-1 1
cm : 3400.14 for O-H stretch, 2907.06 due to C-H (saturated) and 1712.97(C=O). Signals in the H NMR at ppm:
12.85 due to OH of a carboxylic acid and 2.913 - 2.491 due to proton of a saturated ring system, oxiran; data
from 13C NMR (ppm) further confirmed the structure, C=O at 173.146 and C-O at 52.639 - 51.277.
Methylpentadecanoate is an ester. Prominent peaks due to C-H at 2914.08 and C=O at 1710.02 were observed in
1
the IR spectrum. The H NMR displayed characteristics resonance peaks in the aliphatic region. Protons in three
different environments were observed. The presence of methyl proton signal next to an ester group at 2.9-3.6
ppm, methylene protons next to a carbonyl was observed at 2.1-2.7 ppm while the terminal protons of a methyl
group and other methylene protons were accounted for by signals observed at 0.8-1.2 ppm. The attached proton
test experiment (ATP) revealed the presence of quaternary carbonyl carbon at 191.303 ppm and saturated
carbon; methylene and methyl groups without electronegative element at 29.474 while peak at 41.247 ppm was
assigned to saturated carbon attached to an electronegative atom (C-O). The presence of this ester was accounted
for as a break down product from the side chain of the daphnane diterpene isolated in the compound. Daphnane
diterpenes with a tricycle [9.3.0] tetradecane ring system have been isolated from the extracts of sap and seeds of
Euphorbiaceae plants. Some of these compounds have been shown to exhibit cytotoxic, irritant, and analgesic
activity (15-18, 23, 33-34).
Hepatoprotective activity
Carbon tetrachloride (CCl4) is a hepatotoxin commonly used for the production of experimental liver toxicity or
damage (34-37). Assessment of liver function is done by estimating the activities of CREA and Urea as well as
that of serum marker enzymes like ALT, AST, GGT (22, 25-27). When liver cell plasma membrane is damaged, a
variety of enzymes normally located in the cytosol are released into the blood stream. Their estimation in the
serum is a useful quantitative marker of the extent and type of hepatocellular damage (38-40). In this present
study, it was revealed that the ethylacetate extracts induced suppression of increased activities of the enzymes
ALT, AST, GGT, as well as Urea and CREA (which are nitrogenous waste products excreted from the body)
when compared to the CCl4 group (P<0.05). Table 1 shows the effect of ethylacetate fractions of leaves and
stem bark of H. crepitans on different biochemical parameters in the serum of rats. There was a significant
increase in ALT and AST levels (P<0.05) in group 2 (CCl4 only) when compared to other groups (control, CCl4 +
leaves extract, CCl4 + bark extract, leaves extract only, bark extract only, CCl4 + kolaviron and kolaviron only).
The observed increase can be attributed to the release of these enzymes from the liver into the serum indicating
hepatotoxin effect of CCl4. The co-administration of CCl4 and the extracts as well as the standard kolaviron at 200
mg extract/kg animal body weight in groups 3, 4 and 7 brought about a decrease in the ALT and AST activity
167
indicating that the extracts reduced the effect of tetrachloride exposure on both biochemical parameters. There
was also significant decrease in ALT level (P<0.05) in group 1 (control) when compared to group 6 (bark extract),
group 7 (CCl4 + kolaviron) and group 8 (kolaviron) indicating that there was no kidney impairment. Gamma-
glutamyltransferase (GGT) is elevated in all forms of liver disease like liver cancer or cirrhosis and originates
primarily from the hepatobiliary system. There was significant increase in GGT level (P<0.05) in group 2 (CCl4
only) when compared to other groups indicating that the toxicant was able to induce liver diseases. GGT level
however decreased in the other groups indicating protection (Table 1). The toxic metabolite CC13 radical was
produced by cytochrome P450 which further reacts with oxygen to produce trichloromethyl peroxy radicals.
These radicals bind covalently with the macromolecule and cause peroxidative degradation of lipid membrane of
the liver. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are members of transaminase
(amino-transferase) family of enzymes and facilitate certain chemical reactions within the cells. ALT is majorly
found in the liver while AST is found in the liver, kidney and brain. When cells are injured or necrosis occurs,
enzymes such as ALT, AST, GGT are released from the cells into the blood stream to protect the cell organs
against the damage. Urea and CREA (nitrogenous waste products excreted from the body in the urine) levels also
increased. The determination of these transaminases in the serum is therefore often used as one of the essential
makers of the cells functionality. Thus, the reduction of ALT, AST, GGT, Urea and CREA activities by the
extracts is an indication of repair of hepatic tissue damage induced by CC14 and that serum transaminases
returned to normal activities with the healing of hepatic parenchyma and regeneration of hepatocytes (41).
Table 1: Effect of Ethyl acetate fractions of leaves and stem bark of Hura crepitans on different
biochemical parameters in the serum of rats measured in absorbance units (AU)
Serum parameters
Group Treatment ALT AST GGT Urea Creatinine Protein
1. Control 83.3±7.37** 37.3±9.45** 3.09±3.34** 8.85±3.07** 49.71±8.28 10.17±1.04
(corn oil)
2. CCl4 105.0±0.001* 100.0±15.0* 8.69±7.47* 13.38±2.89* 53.85±7.18 9.87±1.44
3. CCl4 + HC Et 78.3±6.56** 45.3±9.87** 8.42±6.55** 3.52±0.81*,** 52.2±4.39 9.00±0.01
(leaves)
4. CCl4 + HC Et 70.0±6.56** 60.0±19.08*,** 8.39±7.47*,** 5.79±1.39** 65.25±21.97 8.88±0.63
(bark)
5. HCEt 69.0±13.23** 37.0±8.89** 6.66±7.64** 6.06±0.48** 54.38±20.53 8.67±0.29
(leaves)
6. HCEt (bark) 53.3±2.52*,** 47.5±0.71** 3.47±1.16** 8.29±0.11** 64.21±9.49 9.17±0.29
7. CCl4 + KV 63.7±4.73*,** 49.0±1.41** 2.32±0.001** 6.48±0.40** 43.50±10.76 9.00±0.50
8. KV 60.0±8.19*,** 27.5±9.19** 2.89±0.81** 5.87±2.38** 43.50±0.001 8.00±2.02
* = The mean difference is significant at P<0.05 when compared with the control.
** = The mean difference is significant at P<0.05 when compared with the toxicant (CCl4).
KV = Kola viron, Et = ethylacetate, CCl4 = carbon tetrachloride.
Creatinine and urea which are nitrogenous waste products excreted from the body in the urine via the kidney are
used exclusively in the assessment of the kidney function. Elevation of creatinine and urea is indicative of kidney
impairment. There was no significant increase in creatinine (P>0.05) when group 2 (CCl4) and group 1 (control)
were compared with other groups but there was significant increase in urea level (P<0.05) in group 2 (CCl4 only)
when compared to other groups. The observed increase can be attributed to the release of urea from the kidney
into the blood stream indicating nephrotoxin effect of the toxicant. The effect was reduced by co-administration
with the extracts. Thus, administration of ethylacetate fractions of H. crepitans leaves and stems bark gave
168
hepatoprotective activity against the toxic effect of CCI4, which is also supported by histological studies. Also
lack of variation in protein concentration of the various samples as determined by the Biuret method showed that
there was no significant increase (P>0.05) in group 2 (toxicant only) when compared with other groups (Table 1).
Comparative histopathology studies of livers from different groups
A comparative histopatological study of liver from the groups further corroborated the hepatoprotective
effect of H. crepitans. Histopathology of liver from normal control group shows prominent central vein,
normal arrangement of hepatic cells. Microscopical examination of carbon tetrachloride treated liver section
showed various degrees of pathological changes starting from centrilobular necrosis of hepatic cells and
central lobular fatty regeneration. Photomicrographs of the liver samples are shown Fig 2.1 – 2.8.
Figure 2.1 Liver section from group 1 (control/corn oil) showing normal histology, prominent
central vein with normal arrangement of hepatic cells (Mag x 400).
Figure 2.2 Liver section from group 2 (CCl4) showing periportal hepatic necrosis/cellular
infiltration, bile duct hyperplasia and central lobular fatty regeneration (Mag x 400).
169
Figure 2.3 Liver section from group 3 (CCl4 + ethylacetate fraction of leaves) showing
periportal hepatic necrosis/cellular infiltration (mild), bile duct hyperplasia (Mag x 400).
Figure 2.4 Liver section from group 4 (CCl4 + ethylacetate fraction of bark) showing portal
congestion (mild), moderate bile duct hyperplasia (Mag x 400).
Figure 2.5 Liver section from group 5 (ethylacetate fraction of leaves) showing no visible lesions
(Mag x 400).
Figure 2.6 Liver section from group 6 (ethylacetate fraction of stem bark) showing portal
congestion (mild) and moderate normal arrangement of hepatocytes around the central vein
and absence of necrosis (Mag x 400).
170
Figure 2.7 Liver section from group 7 (CCl4 + kola viron) showing no visible lesions (Mag x
400).
Figure 2.8 Liver section from group 8 (kola viron only) showing moderate normal arrangement of
hepatocytes around the central vein (Mag x 400).
Histopathological studies revealed that group 1 control animals showed normal hepatocytes (Fig 2.1), group 2
(CCl4 treated) showed periportal hepatic necrosis /cellular infiltration (mild), bile duct hyperplasia (Fig 2.2) and
also exhibited intense centrilobular necrosis, vacuolization and macrovesicular fatty change, group 3 (CCl4 +
leaves extract) and group 4 (CCl4 + bark extract) showed mild periportal hepatic necrosis/cellular infiltration and
moderate bile duct hyperplasia (Fig 2.3 and 2.4), group 5 (leaves extract only) and group 6 (bark extract only)
showed no visible lesions (Fig 2.5 and 2.6), group 7 (CCl4 + kolaviron) (Fig 2.7) had normal arrangement of
hepatic cells while group 8 (kolaviron only) showed very mild ventral venous congestion (Fig 2.8). Thus, the
animals treated with ethylacetate fractions of leaves and stem bark of H. crepitans exhibited significant liver
protection against the toxicant as evident by the presence of normal hepatic cords, absence of necrosis and lesser
fatty infiltration. The present histopathological and biochemical studies of the ethylacetate fraction of H.
crepitans confirm its medicinal application as hepatoprotective agent in traditional herbal practise. There were
also indications that the presence of daphnane diterpenes in H. crepitans may have played a major role in the
observed activity however the pure compounds must be subjected to hepatoxicity study.
CONCLUSION
Spectroscopic analysis of the pure compounds obtained from ethyl acetate and butanol fractions of H. crepitans
1 13
leaves and stem bark using UV-Visible, Infrared, H and C-NMR revealed the presence of carbonyl, hydroxyl,
aromatic alkene and methylene functional groups in the pure compounds. These compounds namely
daphnetoxin acid, huratoxin, apocynin and methylpentadecanoate were reported. The result of the
hepatoprotective activity revealed that the ethyl acetate soluble fractions of H. crepitans leaves and stem bark
171
showed significant hepatoprotective activity by reducing elevated levels of serum enzymes, ALT, AST, GGT, as
well as Urea and CREA levels. A comparative histopathological study of the liver from different group further
corroborated the hepatoprotective efficacy of H. crepitans as it showed a significant regeneration of hepatocytes.
This present study has therefore been able to establish the hepatoprotective influence of the leaves and stem bark
of H. crepitans and secondary plant metabolites of medicinal importance have been isolated. However, more in
vivo assays are also essential to characterize them as biological hepatoprotective agents. Additional work is also
necessary to subject the pure isolated compounds from ethylacetate soluble fractions to hepatoprotective activity
to know the isolates that are responsible for the observed activity in this promising plant.
Acknowledgements: The authors would like to thank the MacArthur Foundation for the award of a Start-up
Research Grant (2010 Grants) tenable at the University of Ibadan, Nigeria to G.K. Oloyede in respect of this
work. The staff of Multi Research Central Science Laboratory, University of Ibadan and Central Science
Laboratory Obafemi Awolowo University Ile-Ife, Osun State are also gladly acknowledged for the use of
spectroscopic equipment. Our appreciation also goes to the staff of the Bio-membrane Research Laboratory,
Department of Biochemistry, University of Ibadan, Nigeria for assisting in carrying out the hepatoprotective
assay.
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174
STUDIES ON THE MOLAR RATIOS OF OPTIMAL INTERACTIONS OF NR-PVC BLEND USING
IR SPECTROSCOPY
1 2
*Okoye, O. N. N., Eboatu, A.N
1
Department of Chemical Sciences, Evangel University Akaeze
2
Department of Pure and Industrial Chemistry, NnamdiAzikiwe University, Awka
*Corresponding Author: OkoyeOkechukwuNnaemeka N.
Email: ookoye94@yahoo.com,Phone Number: +234(0) 7038378186
ABSTRACT
Blends of Natural Rubber(NR) and poly (vinyl chloride) (PVC) were made by the liquid-liquid technique. The
analytical study was carried out by the means of Infra- Red spectroscopy. The aim of the research is to locate
blend compositions of optimal interaction, between the components. Using frequency method the functional
groups of optimal interactions were; C-Cl, C=C, and CH3. The result obtained for establishing the base mole
ratio of optimal interaction by means of frequency average deviation for the polymer pair was 0.395. It was noted
from this study that there was interaction between the polymer pair and points of the degrees of the polymer pair
interactions were indicated.
Key words: Blend, PVC, NR, Molar Ratios, FTIR
INTRODUCTION
1
Polymer blends, by definition, are physical mixtures of structurally different homopolymers or copolymers. By
1976 blending was being extensively carried out with a wider range of polymers, though the earliest significant
use of blends as plastic material dates from the late 1940's.2Polymer blending provides a powerful route to
engineering new properties in materials using available polymers. By this technique it is possible to produce a
3
range of materials with properties that are superior to that of each individual component polymers. That is, from
an industrial point of view, polymer blends fill the deficiency in price/performance of existing homopolymers
with a relatively minor capital investment. A shorter period of time and effort is required to develop a new
4
product via blending existing materials in comparison with that needed to develop a new polymer. Techniques in
resin blending are simple and efficient method for improving the properties of polymers, and have been used
widely in polymer modification field.5Blending methods usually involve solid - to - liquid, solid - to -solid or
liquid - to - liquid mixing.6As an important type of polymer material, rubber is widelyused due to its high and
reversible deformability. Since theessential modulus and strength of neat rubber are low, anadditional
7 8
reinforcing phase is necessary for the practicaluses of rubber materials. According to Hafeziet al , blending
plasticand rubber helps to improve the physical, thermal, and mechanicalproperties as well as to modify the
processing characteristics and costreduction of the final product. To this effect this study seeks to find out the
molar ratios of optimal interaction of NR-PVC components of the blend.
EXPERIMENTAL
Chemicals
THF (Tetra hydro furan) is Qualikems make. Toluene solvent is BDH make, procured from theBridge-Head,
Onitsha.Natural Rubber (NR) crepe, obtained from the Rubber Research Institute, Iyalamo, Benin, Nigeria,
0
while Poly (Vinyl Chloride) (PVC) resins, which has the temperature of melting, 88 C, was obtained from LG
Chemical Ltd, Korea.
Blend preparation and Analysis
2.0g NR crepe was dissolved in 200ml Toluene solvent while 6.0g of PVC resin was dissolved in 100ml THF
solvent, after which the two were blended together in different proportions as follows:
Table 1: Volume Ratio of the Different Blends
175
Polymer Solvent Volumes of the blended solutions (ml)
NR Toluene 10.0 10.0 10.0 10.0 10.0 10.0 10.0
PVC THF 0.0 1.0 2.0 5.0 7.0 10.0 15.0
The Solution mixtures were then poured on glass petri-dishes to producethin films by evaporation. FTIR of
these thin films were obtained using the Model_Shimadzu FTIR 84005, after which seven results were
obtained. The NRhomopolymer, which was not mixed with the PVC solution was used as the control; from
which the frequency, absorbance, and energy deviation tables were deduced, as compared with others (the
different blends).The base mole ratios of each homopolymer, and the different blends were calculated using
the formulae:
Table 2: Base Mole Ratios of the Blends

Polymer Solvents Base Mole Ratios
NR Toluene 1.00 0.605 0.480 0.234 0.179 0.133 0.093
PVC THF 0.00 0.395 0.566 0.766 0.821 0.867 0.907
176
Studies Using IR Spectroscopy
When two or more polymers interact, there is perturbation and the vibration frequencies of these polymers shift
from their normal frequencies. The vibrational spectrum of a molecule is considered to be a unique physical
property and is characteristic of the molecule. As such, the infrared spectrum can be used as a fingerprint for
identification by the comparison of the spectrum from an ''unknown'' with previously recorded reference
spectra.Also it is possibleto deduce whether specific functional groups are present. If detected, one is also able to
9
determine local orientation of the group and its local environment and/or location in the structure.
Tables and Graphs
Based on the spectra obtained from the FTIR, the difference in volumes between pure NR and the different NR-
PVC mixtures (as seen in table 2) were calculated and tabulated. This is based only on the frequency of
interaction of the spectra, as follows:
Table 3: Establishing the functional group of optimal interaction of NR: PVC blend using Frequency
Frequen cy Change (? F)
Peak
Numbers C1 C2 C3 C4 C5 C6
1 148.52 78.12 80.05 1.93 25.08 160.1
2 4.82 142.74 144.67 0.96 1.93 57.86
3 94.52 100.31 126.34 0 2.89 141.77
4 124.41 129.9 132.13 0.97 2.9 226.64
5 132.12 152.39 155.28 0.96 2.89 260.39
6 219.89 67.51 69.44 0.96 16.4 298.01
7 238.21 106.09 102.23 0.96 5.78 232.43
8 301.87 231.46 233.39 0 8.68 171.77
9 412.78 1259.56 73.3 0.97 3.86 331.77
10 407.96 1183.36 991.44 2.9 305.73
11 901.75 649.07 454.25 613.38
12 1111.99 740.69 361.67 401.121
13 1065.71 569.98
14 183.25
15 379.03
177
Using table 3, a graph was plotted (fig. 1), which indicates the points where the NR-PVC mixtures interacted.
Using the correlation table data, the functional groups responsible for such interactions were detected. Below are
the discussions:
Fig 1: The functional group of optimal interaction of NR: PVC blend by means of Frequency
C1, C2, C3, C4, C5, C6, are base mole fractions PVC of one homopolymer.
From Fig 1, the functional group of optimal interaction of NR: PVC blend are C-Cl, C=C, and CH3; with peak
numbers 0-1, 2-3 and 7-8, which have 608.56, 836.17-693.43, and 1428.34-1329.96, frequencies, respectively.
The vibrations may have shifted the frequencies from 1680-1600 of alkenes to 836.17cm-1.This could be as a
result of the electron withdrawing Cl attached to the PVC carbon or the double bond present in the poly isoprene
of the Natural Rubber molecule.
Table 4: The base mole ratio of optimal interaction of NR: PVC blends using Frequency average deviation.
PVC base mole fractions 0.395 0.566 0.766 0.821 0.867 0.907
Frequency Average Deviation 381.789 252.009 230.019 199.076 112.25 266.739
Fig 2: The base-mole ratio of optimal interaction of NR: PVC

From Fig 2 the highest point of interaction is at 0.395 base mole fractions. This means that at this point there is the
highest level of miscibility and bonding of the participating polymers.
Conclusions
The polymers used in this work are NR (hydrocarbon) and PVC (halogen containing polymer). It was noted from
this study that there was interaction between the polymer pair, and points of interactions and the functional
groups responsible for them were established. This, of course, will aid polymer chemists in knowing at what
points to get NR-PVC blends of optimal interaction.
Reference
178
1. J.M.R.C.A. Santos, and J.T.Gutherie, (2006), Polymer Blends: PC-PBT case, Journal of Material
Chemistry, Vol. 16, p. 237.
2. T.U. Onuegbu, A.N. Eboatu, R.U. Arinze and P.N.Ekemezie (2008), Further Studies on Polystyrene-
Polyisoprene Rubber Blend, Anachem Journal, 3(1): 393.
3. M.S. Khan, R.A.Qazi and M.S. Wahid (2008), Miscibility studies of PVC/PMMA and PS/PPMA blends
by dilute solution viscometry and FTIR, African Journal of Pure and Applied Chemistry, 2(4): 041.
4. I.S. Miles, and S.Rostani(1992), Multicomponent Polymer Systems, Polymer Science and Technology
Series, Longman Scientific & Technical, Longman Group UK Limited, p. 63.
5. X. Liu, X. Fan, M. Tang, and Y.Nie(2008), Synthesis and Characterization of Core- Shell Acrylate Based
Latex and Study of its Reactive Blends, International Journal of Molecular Science, vol. 9, p. 342.
6. A.N.Eboatu, B.Animashaun, S.T. Spiff, and, F.M.Abdulrahman(1990), Studies on the Thermal
Characteristics of Polymer Alloys, Journal of Thermal Analysis, Vol. 36, p. 85.
7. G. Sui, W.H.Zhong, X.P. Yang, Y.H. Yu and S.H. Zhao (2008), Preparations and Properties of Natural
Rubber Composites Reinforced with Pretreated Carbon Nanotubes, Polym. Adv. Technol., Wiley
Interscience, p. 1.
8. M.Hafezi, S.N.Khorasani, F.Ziaei and H.R.Azim (2007), Comparison of Physicochemical Properties of
NBR-PVC Blend Cured by Sulphur and Electron Beam, Elastomers and Plastics, Vol 39, p. 152
9. J. Coates (2000), Interpretation of Infrared Spectra, A Practical Approach,Encyclopedia of
Analytical Chemistry, R.A. Meyers (Ed.), Ó John Wiley & Sons Ltd, Chichester, p. 1.
179
Antibacterial And Antioxidant Activity of Justicia Spicigera Extracts: Activity Enhancement by
addition of Metal Salts.
1 2 3
Ohenhen O. N ., Njoku P. C ., Igara C. E .
1
Department of Chemistry, Michael Okpara University of Agriculture, Umudike.
2
Department of Chemistry, Alvan Ikoku Federal College of Education, Owerri.
3
Department of Science Laboratory Tech., Akanu Ibiam Federal Polytechnic, Unwana, Afikpo.
E-mail: offinel2012@gmail.com
ABSTRACT
Justicia spicigera was screened for its phytochemicals. The results showed the presence of alkaloids, flavonoids,
tannins, phenols, saponins and steroids. Extracts of Justicia spicigera prepared with solvents of different
polarities (n-Hexane, Chloroform, Methanol, and Ethyl acetate) was used to analyze the antibacterial properties
in the absence and in the presence of metal salts. The antioxidant activity was analyzed using 2, 2-diphenyl-1-
picrylhydrazyl reagent (DPPH). Quantitative determination of total phenolics was carried out using
spectrophotometric method. The results indicate that the tested bacterial strains were most sensitive to the n-
Hexane extract but moderately sensitive to the chloroform and methanol extracts. The chloroform, methanol,
and ethyl acetate extracts showed potent radical scavenging activities. The ethyl acetate and methanol extracts
3+
showed higher total phenolic content than the other extracts. Antioxidant activity was found in the extract-Fe
2+
ion combination. The extract-Zn ion combination showed enhanced antibacterial activity against tested
bacterial strains compared to the extract alone. The results scientifically establish the efficacy of using the plant
extracts and its metal salt combination as antibacterial and antioxidant agents.
Key words: Justicia spicigera, total phenolic content, antibacterial activity, antioxidant.
INTRODUCTION:
Justicia spicigera a shrub from the Acanthaceae family has been found to produce biologically active species
that are responsible for the plant therapeutic properties (Dominiguez et al., 1990; Euler et al., 1982). It is
commonly referred to as firecracker plant due to their narrow red flowers, mutile in Mexico and Iyip abasi by the
Efiks in Nigeria. J. spicigera has been used in traditional medicine for many years as a stimulant, for
gastrointestinal disorders, diabetes, inflammations, blood depurative treatment, uterine cancer, skin and kidney
infections (Marquez et al., 1999). The infusion of Justicia spicigera has characteristic red color.
Currently there is much interest in the protection of low density lipoprotein, important cells and organs as well as
food systems against oxidative damage caused by superoxide, hydroxyl and peroxyl radicals. This has led to an
increasing demand to evaluate the antioxidant properties of direct plant extracts or isolated products from plant
origin rather than looking for synthetic ones. Antioxidants may guard against reactive oxygen specie (ROS)
toxicities through the prevention of ROS construction by the prevention of ROS attack, scavenging reactive
metabolites and converting them to less reactive molecules or by enhancing the resistance of sensitive biological
targets to ROS attack (Sen, 1995). Free radicals, reactive oxygen species (ROS), and reactive nitrogen species
(RNS) are associated with many pathological conditions such as atherosclerosis, arthritis, ischemia, reperfusion,
injury of many tissues, central nervous system, gastritis, cancer and AIDS (Cook et al., 1996; Kumpluainen et al.,
1999). The anti-oxidative properties of phenolics are due to mechanisms such as scavenging of free radicals,
chelation of metal ions, and inhibition of enzymes responsible for free radical generation (Akinmoladun et al.,
2007).
Several studies have been made in order to determine the antioxidant properties of plants, especially those used
in traditional medicine (Jang et al., 2007; Surveswaran et al., 2007). This study is in support of other studies that
have been made to replace synthetic antioxidants that are carcinogenic with natural antioxidants from natural
180
sources.
MATERIALS AND METHODS.
The plant samples were collected and identified by comparing with the leaves inflorescence in standard text
(Akobondu and Agyakwa, 1998); it was further authenticated by Prof. Nmeregini of Forestry Dept., MOUAU.
Extraction: The extraction was performed according to standard method of (AOAC, 1995). The Methanol
extract was extracted using 80% methanol and labeled ME, the Ethyl acetate extract was prepared with Ethyl
acetate and labeled (EAE), Chloroform extract labeled CE and n-Hexane extract labeled (n-HE).
Phytochemical Determination: The Phytochemicals determination was done according to the standard method
by Harborne et al. (1998).
Determination of Total Phenolic content: The total phenolic content was measured by spectrophotometric
determination with a modified Folin-Ciocalteau method Tuberoso et al. (2007) using Gallic acid as a standard
phenolic compound. The total phenolic content was expressed as Gallic acid equivalent (GAE) in g/100g of dry
wt. of extract. Mixture of water and reagent was used as blank. Sample measurement was done in triplicates and
the mean and standard deviations calculated in each case.
DPPH Free radical scavenging activity: The free radical scavenging activity of the plant extracts and extracts-
metal ions was evaluated by the stable radical DPPH (2, 2-diphenyl-1picrylhydrazyl) reagent described by
Miliauskas et al. (2004). Radical scavenging potential was expressed as inhibiting conc. IC50 value. This
represents the sample conc. at 50% of the DPPH radicals scavenged. The assays were done in triplicate and the
results were expressed as mean values ± standard deviations.
Antibacterial Screening: Extracts of the leaves 1g was dissolved in 1ml of dimethyl sulphoxide (DMSO) to get
a concentration of 100mg/ml. This was used as stock solutions for the antimicrobial assay (Okigbo and
Omodamiro, 2006).
Four representative test microorganisms were used in testing the antimicrobial activity of the leaves extracts of
Justicia spicigera– n-Hexane (n-HE), Chloroform (CE), Methanol (ME) and Ethyl Acetate (EAE). The test
microorganisms used were Streptococcus pneumonia, Salmonella typhi,Escherichia coli and Aspergilus niger.
The test cultures were originally obtained from Federal Medical Centre (FMC) Umuahia, Abia State, Nigeria and
were maintained in nutrient agar.
The antimicrobial test used the filter paper disc diffusion method (Anderson, 1974). Ciprofloxacin drug 0.1g
dissolved in 1ml of distilled water was used on a separate disc. This served as a standard while the disc with
DMSO (100mg/ml) served as the control. The mean was taken as the diameter zone of inhibition. To increase the
synergistic effect the metal salts Fe(III) and Zn(II) ions was combined with the extracts for the antibacterial tests.
Aliquot containing solutions with a constant concentration of extract 10equivalent and freshly prepared metal
salts ZnSO4, FeCl3 solution with a stoichiometric equivalent (1-5eq) were added to give a final extract-metal ion
ratio of 10:01, 10:02, 10:03, 10:04, and 10:05. The obtained product was filtered and collected. The experiment
was repeated three times for each extract and the means of these values were recorded.
RESULTS:
Table 1: Total Phenolic content of Justicia spicigera
Extrac t Total Phenolic content (g GAE per 100g) dry wt. of extract
n - HE 3.28±0.01
EAE. 5.65±0.02
CE 3 99±0.02
ME 4.54±0.04
- Data are means± standard deviations from two extraction replicates

Table 2: Antioxidant activity of J. spicigera with and without metal ion
181
Sampl e DPPH (IC 50 )µg/m l
n - Hexane 34.18±0.50
3+
n-Hexane-Fe 34.01±0.15
2+ 36.73±0.80
n-Hexane-Zn
EAE 13.73±0.21
3+ 11.64±0.18
EAE-Fe
EAE-Zn
2+ 12.68±0.39
ME 18.71±0.74
ME-Fe
3+ 18.01±0.12
2+ 18.50±0.19
ME- Zn
14.94±0.16
CE 13.28±0.51
CE-Fe3+ 14.57±0.13
CE-Zn2+
14.94±0.30
AA (Ascorbic acid)
Data are means± standard deviations from two extraction replicates
Fig 1: Inhibition zone (mm) of extracts against four bacterial strains with standard drug (S= Ciprofloxacin).
182
3+
Fig 2: Inhibition Zone (mm) of extract-Fe combination against four bacterial strains at concentrations 10:04
and 10:05.
183
Figure 3:Inhibition Zone (mm) of extract-Zn2+ combination against four bacterial strains at concentrations 10:04
and 10:05.
Discussion:
The phytochemical screening done on all the plant extracts revealed the presence of alkaloids, phenolics,
flavonoids in all the samples except n- Hexane. Steroids were present in all the extracts but more in the ethyl
acetate extract. Tannins and saponins were found in methanol and ethyl acetate extracts. The results of the total
phenolic content of Justicia spicigera is shown in table 1.There is a positive correlation between the total
phenolic content and antioxidant activity of Justicia spicigera with that reported for other plant species of
Acanthaceae (Sawadogo et al., 2006; Jang et al., 2007). The total phenolic content of Justicia spicigera ranged
from 3.28±0.01 to 5.65±0.02g GAE per 100g dry wt. of extractwith methanol extract richer in phenols than the
other extracts. The results of CE, ME, EAE were found to be significant at P<0.05 but not significant for n-
Hexane extract at P>0.05 when compared to the blank.
The results of the DPPH radical scavenging activity of the extracts and ascorbic acid are presented in Table 2. The
results show that the extracts exhibited a concentration dependent anti radical activity and the lower IC50 values
are indicative of a higher anti radical activity. At low concentrations (10:01,10:02) the extract-metal ion
combination showed results comparable to the extract alone but a discernable effect was observed of the free
radical scavenging activity at concentration 10:03. ME-Iron(III) exhibited highest percent inhibition of DPPH
radical as compared to other fractions. CE-Iron(III) ion and EAE-Iron(III) ion exhibited modest scavenging
activity than the extracts alone. The results of the radical scavenging activity for the chloroform extract,
methanol extract and ethyl acetate extract were found to be significant at P<0.05 but not significant for n-Hexane
at P>0.05 when compared to the reference standard ascorbic acid. The extracts-Zn(II) ions did not show
significant improvement in radical scavenging activity.
The antibacterial result of the extracts in the absence of metal salts is shown in figure 1. The results indicate that
the tested bacterial strains were sensitive to all the extracts. In comparison the extracts were most sensitive to the
n-Hexane extract, moderately sensitive to the chloroform and methanol extracts least sensitive to the ethyl
3+
acetate extract. The antibacterial result of extracts with metal salts at different concentrations with Fe is shown
in figure 2,the antibacterialresults for extracts with metal salts at different concentrations with Zn2+is shown
infigure 3. The results indicate that the extract-metal salt combination enhanced the antibacterial activity than the
crude extracts alone. The extract-metal salt combination possessed stronger antibacterial activity against Gram
+ve bacteria as compared to the Gram -ve bacteria. This may be because gram -ve bacteria have been reported to
be more resistant than gram +ve bacteria (Russel, 1991). At lower conc. of extract-metal salt combination (10:01;
10:02, 10:03 ratio) no recordable growth was observed. Further increase in metal ion salt conc. to (10:04, 10:05)
had significant effect but further increase above that resulted in almost same growth for all tested bacteria. This
confirms the study by Simon et al., (2009) which showed that antibacterial properties of natural products can be
enhanced by the addition of metal ions due to positive participation of extracts in coordination with the metal
ions.
Conclusion:
Many plant species are currently used as sources of nutritional additives because of their wide antibacterial and
antioxidant properties that increase immunity to some diseases. This study has looked at the biological
evaluation of the various extracts of Justicia spicigera and the enhanced synergistic activity by the addition of
metal salts. The results suggest that combination of Fe(III) ion with EAE, CE and ME could be considered as a
new potential source of natural phenolic antioxidant for food, pharmaceutical, and cosmetic industries. The
184
considerable antibacterial activity of n-HE-Zn(II) ion suggests that Justicia spicigeracan serve as selective agent
for the maintenance of human health.
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186
KINETICS AND MECHANISM OF THE OXIDATION OF POTASSIUM
TRISOXALATOFERRATE(III) BY PERMANGANATE ION IN AQUEOUS HYDROCHLORIC
ACID MEDIUM
1
B.O. Ogori, Y.N Lohdip and 2J.N Egila
2
1
Department of Chemistry, F.C.E. Pankshin, Plateau state.
2
Department of Chemistry, University of Jos, Jos Nigeria
Corresponding author: ogorib@yahoo.com
Phone: 07036820097
ABSTRACT
The kinetics and mechanisms of the oxidation k3[Fe(C2O4)3].3H2O by permanganate ion in acid solution has been
studied. The data are consistent with the rate law:
3-
d [ Fe(C 2 O4 ) 3 ]
- 3- -
= b[ H +
(a + ]) [Fe(C 2 O 4 ) 3 ] [ MnO4 ]
dt
-1 6 2 -1 0 + -3 -3
with a = 0.12 s and b = 1.35 dm mol s , at T = 27.0±1.0 C,[H ]= 0.2 mol dm , I= 0.2mol dm (NaCl),
and ëmax = 420nm. There is evidence for the formation of an intermediate complex of significant stability and free
radicals are important in the reaction. The reaction is rationalised in terms of an inner-sphere mechanism.
INTRODUCTION
There has been a lot of interest in the study of the oxidation-reduction reactions involving Trisoxalato
complexes with a variety of substrates [1-3]. This interest is due to the fact that these complexes have played a
leading role in many aspects of co-ordination chemistry especially in their spectroscopic behavior, kinetic
behavior in solution and in some cases their resolution into enantiomers [4]. These complexes have found useful
applications in photography, medicine, analytical chemistry and in actinometry.
K3[Fe(C2O4)3].3H2O is most abundantly used in platinum/palladium photographic printing, where its
aqueous solution is first converted to the acidic hydrogen ferric oxalate, after which the resulting acid is then
used as a sensitizer in developing a photographic prints [5]. The compound has been successfully used in
volumetric analysis to estimate iodine in iodized salt [6]. In medicine the complex is used as a desensitizer in
patients that exhibit a hypersensitive dental condition [7]. K3[Fe(C2O4)3].3H2O is also used in chemical
actinometers because of its sensitivity over a wide range of wavelengths [8].
2-
Recently, we have investigated the redox reaction of K3[Fe(C2O4)3].3H2O with SO4 [9]. In continuation
-
of our effort in this direction, we now report the results of the redox reaction of K3[Fe(C2O4)3].3H2O with MnO4 in
aqueous hydrochloric acid.
EXPERIMENTAL
Materials and reagents
The complex, K3[Fe(C2O4)3].3H2O was synthesized and characterized according to a known procedure
involving the direct reaction of potassium oxalate monohydrate and iron(III)chloride hexahydrate as reported in
the literature [10]. Sodium chloride (BDH, analar) stock solution was prepared without further purification or
-1
standardization and used to maintain the ionic strength at 0.2 mol dm while HCl was used to investigate the
effect of hydrogen ions on the rate of the reaction. All other chemicals were used as supplied.
Stoichiometric studies
The stoichiometry of the reaction was determined by spectrophotometric titration using the mole ratio
- 3-
method. Solutions containing known concentrations of MnO4 and varying concentrations of [Fe(C2O4)3 ] (0.01-
0.040 mol dm3) were reacted at [H+] = 0.05 mol dm-3, [MnO4-] = 0.001 mol dm-3 and I = 0.2 mol dm-3 (NaCl) and
allowed to stand for 24 hours for the reaction to go to completion. The absorbance of the solutions was measured
187
at 420nm and the stoichiometry was evaluated from the plot of absorbance versus mole ratio.
Kinetic measurements
All kinetic runs were performed under pseudo-first order conditions with the concentration of
[Fe(C2O4)33-] in at least 10 fold excess over that of MnO4-. The rate of reaction was monitored by measuring the
rate of decrease in absorbance of the mixture at 420nm using a Seaward photoelectric colorimeter [AE-11D] at
0 + -1 -1 -1
27.0±1.0 C, [H ] = 0.2 mol dm and I = 2.0 ×10 mol dm (NaCl) unless otherwise stated. Pseudo-first order rate
constants were obtained from plots of log (At-A∞) versus time.
Stoichiometry
The results of the spectrophotometric titration indicated a 1:2 stoichiometry and the overall reaction is
given by equation (1).
[Fe(C2O4)3]3- + 2MnO4- + 8H+ [Fe(C2O4)3-] + 2Mn2+ + Other products ……… (1)
Product analysis
At completion of reaction, the presence of Mn(II) ions was confirmed by adding a solution of KIO4 to the
colourless reaction mixture and warming gently. The characteristic purple colour reappeared, owing to
oxidation of Mn(II) back to Mn(VII).
Determination of order
3-
Kinetic measurements were made under Pseudo-first order conditions with [Fe(C2O4)3 ] = 0.01-0.045
-3 -3 -3
moldm , [Mn] = 0.001 mol dm and I=0.2 moldm (NaCl). Under this conditions, plots of log (At-A∞) versus
time were found to be linear to more 80% of the extent of the reaction, indicating that the reaction is first order in
-
[MnO4 ] (At and A∞are absorbance at time t and infinity respectively). Values of the observed rate constants are
3-
given in Table 1. A plot of log k1 versus log [Fe(C2O4)3 ] was linear. The slope of 1.2 (r = 0.939) signifies first order
in [Fe(C2O4)33-] (figure 1). The rate law for the reaction is therefore represented as:
3-
- d [ Fe(C 2 O4 ) 3 ] 3- -
K 2 [Fe(C 2 O 4 ) 3 ] [ MnO4 ] ………………........ (2)
=
dt
The second order rate constants calculated from k2= were fairly constant, confirming
-
first order in [MnO4 ]. The observed rate constants, k1 and second – order rate constants are presented in table
1.
3- -
Table 1: Second order rate constants for the oxidation of [Fe(C2O4)3 ] by MnO4 .
[Fe(C2O4)33-]= (0.01-0.040) moldm-3, [Mn] = 0.001 mol dm-3 I =2.0× 10-1 moldm-3(NaCl), T= 27.0±1.00C
and ë = 42 nm.
[Fe(C2O4)33-] 10[H+] I, (NaCl), mol dm-3 10[k2]
Mol dm-3 mol dm-3 dm3 mol-1s-1
0.010 2.0 0.2 1.68
0.015 2.0 0.2 2.14
0.020 2.0 0.2 1.80
0.025 2.0 0.2 1.55
0.030 2.0 0.2 1.41
0.035 2.0 0.2 1.25
0.040 2.0 0.2 1.15
0.020 2.0 0.2 1.56
0.020 2.4 0.2 1.66
0.020 2.8 0.2 1.69
188
0.020 3.2 0.2 1.78
0.020 3.6 0.2 1.80
0.020 4.0 0.2 1.89
0.020 4.4 0.2 1.91
0.020 2.0 0.2 1.02
0.020 2.0 0.3 1.05
0.020 2.0 0.4 1.11
0.020 2.0 0.5 1.21
0.020 2.0 0.6 1.24
0.020 2.0 0.7 1.32
0.020 2.0 0.8 1.34
0.020 2.0 0.9 1.50
0.020 2.0 1.0 1.63
Acid dependence
The reaction rate was observed to increase in the presence of acid (0.02-0.048) moldm-3. The reaction
showed a first order dependence on [H+], (Table 1). A least square plot of k2 versus [H+] was found to be linear (r
=0.977) with a positive intercept, and the relationship is given by equation (3).
+
k2 = a + b [H ] ................................... (3)
-1 6 2 -1
a = 0.12 s and b = 1.35 dm mol s .
Substituting the expression for k2 into equation (2) gives equation (4).
{a+b[H+]}[Fe(C2O4)33-][MnO4-] ……………………(4)
The first order dependence of [H+] suggests the existence of two parallel pathways for the reaction, involving a
protonated and unprotonated species. It has been noted that redox reactions of many oxoanions are strongly acid
dependent [11,12].
Ionic Strength The effect
-3
of ionic strength on the rate of the reaction was studied over the range 0.2-1.0, mol dm using NaCl, while
keeping all other conditions constant. The results presented in Table 1, show that the rate constants increase with
increase in ionic strength, suggesting a positive Bronsted-Debye salt effect [13,14]. This is an indication that the
rate determining steps, or one or more of such steps involve(s) charged ion(s) [15].
Dielectric constant
The dielectric constant was varied by varying acetone/water concentration. It was observed that that the
reaction rate was enhanced as a function of 1/D (D= dielectric constant). This is also an indication that the rate
determining step or one of the rate determining steps involves oppositely charged species [15].
Anion Dependence
-
The rate constant of the reaction was found to decrease with increase in concentration of CH3COO and
-
NO3 as tabulated in Table 2. This is suggestive of an outer sphere mechanism because outer sphere reactions are
expected to be affected by presence of anions, but they should have no effect on inner sphere reactions [16].
Test for free radicals
Addition of acrylamide to partially reacted mixture gave a gel in the presence of excess methanol. This
suggests that free radicals are important intermediate in this reaction.
Table 2: Effect of added anions on the second order rate constants for the oxidation of [Fe(C2O4)33-] by
- 3- -3 - -3 + -3 -1
MnO4 at [Fe(C2O4)3 ] =( 0.01 -0.045 ) moldm , [MnO4 ] = 0.001 moldm , [H ]= 0.2 moldm , I= 2.0× 10
-3 0
moldm (NaCl), T= 27.0±1.0 C and ë=420 nm.
X 102[x], moldm-3 103k1, s-1 10k2, dm3mol-1 s-1
189
CH3COO- 2.0 4.22 2.11
4.0 4.07 2.04
6.0 4.07 2.04
8.0 3.90 1.95
10.0 3.65 1.83
-
NO3 2.0 4.42 2.21
4.0 4.00 2.00
6.0 3.89 1.95
8.0 3.52 1.76
10.0 3.39 1.69
Michaelis-Menten plot
3-
Least squares analysis of 1/k1 versus 1/[Fe(C2O4)3 ] gave a straight line with a significant positive
intercept (figure 2), indicating the formation of intermediates prior to electron transfer. The presence of kinetic
evidence for precursor complex in this reaction indicates that the reaction most probably proceeded by an inner-
sphere mechanism.
Figure 1: Plot of 1/ k1 versus 1/ [Fe(C2O4)33-] for the oxidation of [Fe(C2O4)33-] by MnO4-
Reaction mechanism
On the basis of the kinetics and stoichiometric results, a plausible mechanism is proposed for the
reaction:
k1
- +
MnO4 + H HMnO4- ................................ (5).
k2
190
3- -
[Fe(C2O4)3 ] +HMnO4 products (slow) ……………………. (6)
k3
3- -
[Fe(C2O4)3 ] + MnO4 products (slow) …………………… (7)
From equations (6) and (7), the rate equation can be given as
3- - - 3-
Rate =k2 [Fe(C2O4)3 ] [HMnO4 ] + k3[MnO4 ][Fe(C2O4)3 ] ………………….. (8)
From equation (5),
[HMnO4-]= k1 [MnO42-][H+] ………………… (9)
Substituting equation (9) into (8), gives
3- - + 3- -
Rate = k2k1 [Fe(C2O4)3 ][MnO4 ][H ] + k3 [Fe(C2O4)3 ][MnO4 ] ………………… (10)
+ 3- -
Rate = (k3+k2k1 [H ]) [Fe(C2O4)3 ] [MnO4 ] …………………(11)
3 -1 -1
Equation (11) is analogous to equation (4) where k3 = a =0.12 dm mol s and
6 -2 -1
k1k2 = b = 1.35dm mol s .
A key point of interest is whether the reaction occurs via the inner or outer-sphere m e c h a n i s m o r a
combination of both. This issue was addressed as follows;
(a) Addition of acrylamide to the partially oxidized reaction mixture gave a gel in the presence of excess
methanol, suggesting the presence of free radicals in the reaction. This evidence is in support of the
inner sphere pathway.
(b) From table 2, it can be observed that the reaction rate decreased in the presence of acetate and nitrate
ions, which supports outer-sphere mechanism. 1
1
© A Lineweaver-Burk modification of Michaelis-Menten plot of versus [ Fe(C O ) 3-]
k1 2 4 3
was linear with a significant positive intercept on the
axis. This supports the inner-sphere mechanism.

(a) Also, acid-catalysis of the reaction was observed, which also is in favour of inner- sphere mechanism.
From the above reasoning (a)-(d), it can be inferred that the reaction occurred through
an inner-sphere mechanism.
REFERENCES
[1] Horacio, D.M, Nina, C. (2006). Kinetic studies of the oxidation of L- ascorbic acid by
tris(oxalate) cobaltate in the presence of CDTA metal ion complexes: Journal of Brazilian chemical society
vol 17(2)
[2] Fico, M and Treindl, L (1972). Kinetics of the oxidation of Tris(oxalato)chromate(III) i o n b y
manganese(III) ion. Chem. Zuesti, 26, 103-109
[3] Mamman, S and Iyun, J.F (2008). Kinetics of the Oxidation of 2-Methylpentane-2,4- diol by
3-
[Co(Ox)3] . African Journal of natural Sciences. 11, 9-12. Nigeria.
[4] Baran ,Y Lawrence, G. (2002). Kinetics of Cobalt(III) Siro-Octaamine . Turk Journal o f
Chemistry. 26,771-776.
[5] Shun,J.A (1994), Platinum-Palladium Photographic Printing: Journal of Photographic Science, 23
– 24, Footers Press. Glasgow.
[6] Ramanjaneyulu, J.V.S and Shukla, B.K (1956). Inorganic complexes in volumetric analysis- part 1: Use
of ferric oxalate complex in estimation of iodide. Springer. 151(1), 31-33.
191
[7] Teinkink, E.R.T; Whitehouse, M.N (1995): Handbook of Metal-ligand Interaction in Biological
Fluids. Marcel Dekker, New York.
[8] Sahul, K; Natrajan, L.V; Anwaruddin, Q (1997), Photopolymerization of Acrylamide in Aqueous
Solution by Potassium Trisoxalatoferrate(III): Journal of Polymer Science, polymer science letters, 15(10),
605-9
[9] Ogori, B.O and Lohdip, Y.N (2015). Kinetics and Mechanisms of the Redox Reaction o f
Potassium Trisoxalatoferrat(III) and Tetraoxosulphate(VI) Ion in Aqueous Hydrochloric Acid
th
Medium. Conference Paper at the 38 Annual International Conference of the Chemical Society of
Nigeria, Abuja 2015.
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Manual, Edward Arnold Limited. London.
[11] Iyun,J.F Ayoko, G.A and Lawal, H.M (1992). Kinetics and Mechanisms of the Oxidation of
iodide by Diaquotetrakis (2,2-bipyridine)-µ-oxodiruthenium(III) ion in acid medium. Transition Metal
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[12] Iyun, J.F, Ayoko, G.A and Lohdip, Y.N (1992). Kinetics and Mechanisms of the Oxidation of
Diaquotetrakis (2,2-bipyridine)-µ-oxodiruthenium(III) ion by Bromate in aqueous perchloric acid.
Polyhedron. 11(18): 2389-2394.
[13] Bronsted, J.N (1922). Physical Chemistry. 102, 160.
[14] Birck, J.P (1978). Inorganic Chemistry.17, 504
[15] Hassan, R.M. (1991). Kinetics and mechanism of oxidation of DL-alanine by acid perchlorate
ion media solution. Canadian Journal Chemistry. 69, 2018.
[16] Adegbite, A, Iyun, J.F and Ojo, J.F (1977). Kinetics and Mechanisms of Electron Transfer Reaction
between Uranium(III) and some Ruthenium(III) Amine Complexes. Journal of Chemical Society.
Dalton.115.
192
THE IMPORTANCE OF CHEMISTRY PRACTICE AND RESEARCH IN THE OIL AND GAS SECTOR
OGODO AEMURERHIMEN DICKSON (Snr.) (Chartered Chemist)(A2652)

(HND, PGD, M.Sc., Ph.D (In- View), AISLT, C.CHEM., MICCON, MCSN, MNES, MSPE)
School of Applied Science, Department of Science Laboratory Technology, Chemistry Laboratory, Delta State
Polytechnic, P.M.B. 1030, Ogwashi –Uku, Delta State, Nigeria: Tel: 08030738401, 08094921724, E-mail:
dicksonogodo@yahoo.com. Formerly, Pioneer Co-ordinating Chairman, Research Institute for Tropical
Agriculture, School of Science and Technology, Department of Science Laboratory Technology, Chemistry
Laboratory, Delta State Polytechnic, P.M.B.5,Ozoro, Delta State, Nigeria.
Abstract
This scientific research shows that the Oil and Gas industry was originally developed by Chemist and a clear
understanding of Fundamental Chemistry is essential for optimal operations of Upstream, Refining and
Downstream activities. It clearly reveals that Crude Oil first called “Rock Oil” in 1854 was first analyzed by a
Professor of Chemistry at Yale University - Professor Silliman (Jr.). It identifies the importance of Chemistry
practice and research and expresses the beliefs on the ability of the Science Laboratory Chemist to design the
correct drilling mud and chemical composition that would ensure the balance and control of formation
pressure to avoid blow-outs in Drilling fluids Engineering in the Exploitation/Exploration or Production of
Crude Oil while Production Chemist finds solutions to the myriads of chemical oriented problems
encountered in the course of drilling such as well simulation with evaluation and identification of chemicals
(Acids, Toluene and Additives) that would enhance Oil recovery from reservoir. The research dwells
extensively and specifically on the role of the Chemist in reaching out to other divisions of the petroleum
industry. Thus, the research emphasizes clearly that the Chemist is inextricably inter-twined in the Upstream,
Refining and Downstream Sectors and have equally proved their mettle in their various roles in
Polypropylene, Carbon Black and Linear Alkyl Benzene Plants. The research concludes that if not for
chemistry, the Oil Industry would certainly not exist as the extracted Crude Oil cannot be physically separated
into its several components. Secondly, the search for Oil and Gas and conversion to more useable products
such as Petroleum and Petrochemicals products cannot be optimally achieved without the in-depth
involvement of Chemists. The research recommends that though, a few Chemists are in position of Authority
in the Oil and Gas industry, Chemist must be sufficiently integrated at decisions making positions to ensure
comprehensive charity in technical policy prescriptions. Secondly, Corporate Organization structures must be
revisited for a carefully redesigned and articulated career growth policy.
Keywords: Chemistry Practice, Research, Oil and Gas Sector, Science Laboratory/Production Chemist,
Technical Policy Prescriptions.
Introduction
How the Chemists can be involved
1
Nigeria is one of the world top ten Oil Producers and Leading African and Natural Gas . Petroleum industries
activities are conveniently divided into two namely, Upstream and Downstream activities1,2 .Chemistry is
practiced and Chemists are needed in each of these areas. Downstream activities include refining,
petrochemical, production, transportation and other related activities. Upstream activities consist of
exploration (search for oil), drilling, completion, production, crude oil treatment and other related activities 3,4.
193
Chemistry and the Petroleum Industry
Another very important Company with wide application of Chemistry is the Oil industry. This industry involves
activities for the upstream sector. Both sectors contain numerous activities that encompasses sound
knowledge of Chemistry. Principal activities of the upstream includes: Exploration, Drilling, Construction of
infrastructures for drilling and Production facilities. At each of these stages, Quality Assessment and Control is
regularly carried out to ascertain the quality of what is being generated against plan. The role of Chemistry
permeates all these phases 5,4.
The downstream on the other hand comprises of refining and distribution. Here, the extracted crude is refined
to several constituent products such as domestic gas, petrol, Asphalt, bitumen, engine oil, etc. The principle
behind refining is solely separation based on differential distillation of the components. But for Chemistry, the
Oil industry would certainly not exist and crude Oil would have remained crude, as the extracted crude cannot
be physically separated into its several components 5,4.
Chemistry and the Petrochemical industry
The petrochemical industry is concerned with large scale production of petrochemicals and petrochemical
products: which includes Rubber, Solvents, Paints, Dye, Drugs, Agricultural input such as (Fertilizers,
Insecticide, Herbicides and Pesticides), Pharmaceuticals etc 4. The Products of Petrochemicals are used in
several Companies such as: Paper Industry: where the naturally occurring cellulose in plants gives only a low
grade news print papers, consequently, some appropriate products are incorporated into the cellulose to
produce the well desired high quality glossy paper
• Solvent Industry: Where indispensable solvents such as Ethanol, Methanol, Ether, Petrol, acetone as
well as Benzene (all Petroleum Products) are used as raw materials.
• Production of Plastic containers of various shapes, dimension and sizes.
• Several Products generated from the Petrochemical Industry are involved in the manufacture of
drugs .
5
• Production of Fertilizer is heavily dependent on refinery products and several other Industry .
Chemists in the Nigerian Oil Industry
The search for Oil and Gas and conversion to more useable products such as Petroleum and Petrochemicals
products cannot be optimally achieved without the in-depth involvement of Chemists in all facets of the Oil
industry. Consequently, the Chemists is inextricably inter- twined in the Oil industry and strategically dispersed
in the Upstream, Refining and Downstream sectors. Chemists are also to be found in the Transportation and
Marketing of crude Oil and Refined products. 2.
Fig. 1 Shows an Illustration of the Chemist Reaching out to the Various
Divisions of the Oil Industry.
EXPLORATION AND
DRILLING
MARKETING
AND TRANSPORTATION GAS ANALYSIS
LUBE PRODUCTION AND REFINING

LUBRICATION THE CHEMIST AND THE
OIL INDUSTRY PETROCHEMICALS
RESEARCH AND DEVELOPMENT
POWER GENERATION
ENVIRONMENT AND
POLLUTION CONTROL
SAFETY AND WASTES
MANAGEMENT
Figure 1
194
Upstream
Exploration and Drilling
In the exploitation or production of crude Oil, the optimum use and interplay of chemicals and additives is a
desideratum for success of production activities especially in the modern techniques of Enhance Oil Recovery
(EOR). The Chemist is thus well integrated in the art of Petroleum Engineering, usually called the Production
Chemist2 (e.g. Seplat Petroleum). He is expected to find solution to the myriads of chemicals oriented problems
encountered in the course of drilling. These include well stimulation which is concerned with evaluation and
identification of chemicals (acids, toluene and additives) that will enhance oil recovery from reservoirs. In
drilling fluid engineering, the Chemists has to design the correct drilling mud composition that will ensure the
balance and control of formation pressure to avoid blow-outs. Table 1 shows typical composition of drilling
mud's as used in the industry. The Chemists is also involved in the design of well casing and additives for use in
the cement mixture thus further reducing the risk of blow-outs6,2,4.
Table 1
Chemicals Functions
Barite, Calcium Carbonate Weighing agents
Bentonite, Xanthan, Polyacryl Viscosifiers
Amides
Lignosulphonate, Tanin Dispersants (Deflocculating Agent)
Carboxynethylcellulose, Starch Fluid Loss Additives
Polyacrylates
Emulsifiers, Sodium Dichromate Specialty Chemicals
Petrochemicals
Petrochemicals as the name implies are chemicals derived from petroleum and at the bench scale were
originally developed by Chemists2. The contribution of the petrochemicals industry to enhance living standards
worldwide and the multiplier effects so created are positive attributes, which can rightly be claimed by
Chemists. In the Nigerian petrochemical scene, the Chemists have equally proved their mettle in their various
roles in Poly-propylene, Carbon Black and Linear, Alkyl Benzene Plants 2,7,8.
Polypropylene
In Propylene Polymerization the Chemists first ensures that the Quality of the raw materials are in conformity
with the standard specification as stipulated by Plant licensors. These raw materials include Propylene (main
raw material) and other additives and catalyst materials. A typical Propylene specification is shown in the Table
2
2 below .
195
Typical Propylene Specification
Propane 6% “(max)
Hydrogen 10 -12 ppm wt.
Nitrogen 100 pp Max
Ethane 25 ppm max
Cyclopropane 10 ppm wt. Mx
Sulphur 1 ppm.
Raw propylene from the refinery is passed through the purification unit to meet the above specification which is
the polymerization grade quality. Chemists thus monitor the efficiency of the purification unit. consequently,
evaluating the extent of Hydrogen Sulphide, and Carbonyl Sulphide removal, among others2.
Carbon Black
Chemists in Carbon Black processing unit ensure that the main feedstock Decant Oil meets design specification
and also control the quality of the intermediate and final products in order to obtain marketable products with
desired and specified quality. A number of tests are thus performed at various stages of manufacture and
2
storage. Typical feedstock characteristics as evaluated by the Chemist is as shown .
Research and Development

Current activities for which the Chemists are involved include Chemicals and Process Development, Catalyst
Research, Core Analysis etc. Varied activities are performed but key points as reflected in their mission
2,9,10
statement has a focus on adaptive research utilizing local materials .
Chemists and Mechanical Maintenance

Most large complexes have heavy duty rotating equipment such as pumps, compressors, turbine, generators
etc. These are constantly lubricated. It is the task of the Chemist, in a new maintenance concept known as
Reliability Engineering, to monitor the State of these lubricants at periodic intervals and advise on Oil change.
Analysis of Oils also point the direction to possible trends and resolution of mechanical problems. Thus the
Chemist is involved in some aspects of preventive and predictive maintenance. Analysis covered in this
dimension include:
196
§ Viscosity at various temperature and viscosity index
§ Moisture: This encourages sludge formation and additive breakdown.
§ Total acid numbers: Oil additive breakdown due to oxidation is normally reflected in increase of acidity
flash point – Indicate lubricant dilution.
§ Infrared Spectroscopy: Antioxidants breakdown can be monitored at the molecular level.
§ Metals Analysis by Atomic Absorption Spectroscopy: Metals analysis could show trends of absorption
2,9,10
spectroscopy, gradual bearing failure .
The Role of the Chemist in Environmental Protection
The role of the Chemist in Environmental protection is deeply rooted in the Environmental Laws and
Regulations.
1. The Chemist must be the person to set Environmental Standard and Guidelines or put simply as the one
who must head the team of Scientists charged with responsibility of setting up the standard. The choice
of the Chemist is based on the fact that, he or she is the only trained professional who understands the
11
composition and properties of pollutants since pollutants are chemicals .
2. Environmental Impact Assessment (EIA) and Environmental Evaluation Report (EER) review and
approval is the statutory function of Federal Environmental Protection Agency (FEPA). (submerged
under Federal Ministry of Environment.) It must be carried out with Chemist as the head of the team,
since all other Environmental Scientists rely on the Chemist to interpret their result.
3. Chemist has a great role to play during spill, as in most cases they are required to carry out Qualitative
Analysis such as Fingerprinting.
4. Chemist play very vital role in the monitoring of waste emanating from industrial operations and
installation, as well as management for safe disposal.
11
5. The review of Standards and Guidelines cannot be executed without the involvement of the Chemist .
6. The Chemist is also better positioned to educate the public on the need to protect the environment.
Recommendation and Conclusion

1. The Oil and Gas Industry was originally developed by Chemist and a clear understanding of Fundamental
Chemistry is essential for optimal operations of Upstream, Refining and Downstream activities
2. If not for Chemistry, the Oil Industry would certainly not exist and CRUDE OIL would have remained
CRUDE, as the extracted crude oil cannot be physically separated into its several components.
3. The search for oil and gas and conversion to more useable products such as Petroleum and Petrochemicals
products cannot be optimally achieved without the in-depth involvement of Chemists in all facets of the
Oil Industries.
4. Chemists must be sufficiently integrated at decision making position to ensure comprehensive clarity in
technical policy prescriptions.
5. The Chemist plays a very crucial role in the protection of the Nigerian environment 11.
6. Chemists have been found in position of authority within the OIL industry.
2
7. It is also re-emphasized that the Oil industry is a multidisciplinary industry .
8. Corporate Organization structures must be revisited for a carefully redesigned and articulated career
growth policy.
References
1. O. Osibanjo (2006), Constraints in the Monitoring of the Environmental Issues of the Oil and Gas
Industries. Lead Paper Presented by Director, Basel Convention Regional Co-ordinating Centre for
197
Africa for Training and Technology Transfer in Hazardous Waste Management, University of Ibadan –
Federal Ministry of Environment Linkage Centre for Cleaner Production Technology and Hazardous
rd th
Waste Management. University of Ibadan, Ibadan, Oyo State, Nigeria. 23 – 25 November 2006.
Conference on: Chemical Engineering and Nigerian Content in the Industrialization of Nigeria.
Published in the Books of Proceedings of the 36th Annual Conference and Annual General Meeting of
the Nigerian Society of Chemical Engineers (NSChE), A Division of the Nigerian Society of Engineers
(NSE) 36:77-97.
2. E.E. Nkang (2001), Chemists in the Oil Industry. “The Delta Chemist” (A Biannual Magazine of the Delta
State Chapter of the Chemical Society of Nigeria (Delta CSN). Volume 3. September 2001. Pages 27 – 31.
3. S.E. Ofodile (2002), Oilfield Chmistry: Are You Interested? Chemistry in Nigeria (A National Magazine of
the Chemical Society of Nigeria. 2(1). September. Pages 16-17.
4. A.D. Ogodo (2008), Oil-field Chemistry, Petroleum and Petrochemical Industries: The Role and
Contribution of the Chemist. Conference on: General Studies: Goal to a Balanced Education. Ozoro
Journal of General Studies (OJOGS), Delta State Polytechnic, Ozoro, Nigeria, Volume 1 (2008) Published
by School of General Studies, Delta State Polytechnic, Ozoro. 1(353-362).
5. A. O. Igbuku (1999), Chemistry and Nigeria's Industrialization. “The Delta Chemist” (A Biannual
Magazine of the Delta State Chapter of the Chemical Society of Nigeria Delta CSN). 1 (1). October 1999.
Pages 35 – 38.
6. A. D. Ogodo (2011), Career Prospects and Opportunities for Scientists in Science Laboratory
Technology for the Service of Man through the Study of Science. Conference on: Prospects and
th th
Challenges of Science Laboratory Technology (SLT) Profession in Nigeria. 26 – 29 October, 2011).
th
Oduduwa Hall, Obafemi Awolowo University, Ile-Ife, Osun State, Nigeria. 27 Annual National
Conference/Scientific Workshop (IFE 2011) of the Nigerian Institute of Science Laboratory Technology
(NISLT). 34 Pages.
7 E. E. Nkang (1999), Chemical Industries: The Sure Way to Nigeria's Economic Independence, “The Delta
Chemist” Magazine. A Biannual Magazine of the Delta State Chapter of the Chemical Society of
Nigeria (Delta CSN). Publication NO.1 October 1999. Pages 26-27.
8. E. E. Nkang (1999), Refining Imperatives for the Next Millennium: The Nigerian Challenges. “The Delta
Chemist” Magazine. A Biannual Magazine of the Delta State Chapter of the Chemical Society of Nigeria
(Delta CSN) Publication No 1. October 1999. Pages 39–42.
9. A. D. Ogodo (2012), The Role of Science Laboratory Technologists on Local Content for Oil Field
Chemicals in Oil and Gas Industry. Conference on: Science Laboratory Technology (SLT) and
th th
Development of Oil and Gas Sector in Nigeria: Prosects and Challenges. 7 – 10 November 2012.
th
International Conference Centre, Univeersity of Calabar, Calabar, Cross Rivers State, Nigeria. 28
Annual National Conference/Scientific Workshop of the Nigerian Institute of Science Laboratory
Technology (SLT).
198
10. A. D. Ogodo and H. I. Esemuede (2013), The Relevance of Science Laboratory Technology in Oil and Gas
Exploration. Conference on: The Role of Science Laboratory Technology in Food and Security: Adding
Value to the Food Chain. 30th October – 2nd November 2013. The University Main Auditorium, Univeristy
of Illorin, Illorin, Kwara State, Nigeria. 29th Annual National Conference and Scientific Workshop (Illorin
2013) of the Nigerian Institute of Science Laboratory Technology (NISLT).
11. U. S. Siakpebru (1999), The Role of the Chemist in Environmental Protection. “The Delta Chemist” (A
Biannual Magazine of the Delta State Chapter of the Chemical Society of Nigeria (Delta CSN) 1 (1).
October 1999. Pages 48-49
199
THE EFFECT OF EXTRACTS OF CRINUM JUGAS ON THE ACUTE TOXICITY OF THE VERNONIA
AMYGDALINA ROOT POISONED GUINEA PIGS
*Ogbuanu Cyril C, Amujiogu Steve N, Nwagu Lauretta N, Chime Charlse C, and Arinze-Nwosu Uche L
1. Enugu State University of Science & TechnologyDepartment of Industrial Chemistry.

*Phone: +234(0)8067958630. Email: Cyril.ogbuanu@esut.edu.ng
2. Raw Materials Research and Development Council, Enugu

.
ABSTRACT
The toxicity of Vernonia amygdalina root and enzymatic activities of Crinum jagus extracts on guinea pigs were
investigated. The cold water, n-hexane, ethyl acetate and methanol extracts were used to evaluate the effect of
V A root poison on the the aspartate transaminante, alanine transaminante and alkaline phosphatise enzymes
activities in the serum/liver of guinea pigs. No death of guinea pigs were recorded neither in the control nor in
the extracts treated groups during the observation period. Visual observations of guinea pigs were made 1 and
2 hrs after poison and extracts administration respectively, including changes in respiration, motility,
behavioural pattern, food and water consumption and recorded systematically. These parameters changed in
groups 2, 3, 4, 5 and 6 guinea pigs compared to the control group 1, 2hrs after poison administration and
recovered to normal in groups 3, 4, 5 and 6 after extracts administration. The pathological examination of the
liver showed no significant abnormality in groups 1, 4, 5 and 6 while the enzymes in group 3 were inactive and in
group 2, the enzymes of the only surviving guinea pig was found to be dead.
Keywords: Crinum jagus, , enzyme activity, aspartate transaminante, alanine transaminate, alkaline
phosphate,
INTRODUCTION
The use of plants to treat ailments is as old as antiquity. Records of humans using plants to treat diseases
have been recorded as far back as 6000 – 4000 years ago when Agurvedic physicians started treating
tumors with extracts from Vinca roseus (Oguneyemi, 1979). The important of plants in medicine remains
even of greater relevance with the current global shift to obtain drugs from plant sources, as a result of
which attention has been given to the medicinal value of herbal remedies for safety, efficacy and
economy (Glombitaze et al. 1993).
Today natural products derived from plant are being tested for presence of new drugs with new methods
of pharmacological action. A special feature of higher plants is there capacity to produce a large number
of secondary metabolites (Castello et al, 2002). Recent studies are involved in the identification and
isolation of new therapeutic compounds of medicinal importance from higher plants for specific diseases
(Erturk et al, 2006).
Crinum jagus (Amaryllidaceae) is a bulbous plant with freshly, wide spreading rich green and glaucous
leaves reading 75cm long by 6cm wide. It grows well in damp site and is distinctly ornamental. The
plant attracts considerable attention due to various medical properties as anti-tumor, immuno-
stimulating, analgesic, anti-viral, anti-bacterial, anti-fungal and memory loss and other mental symptoms
associated with ageing using the in situ bioauthographic test for enzyme inhibition (Houghton et al.
2004, Burkill, 1985).
200
Augustine and Lycorine isolated from this plant showed moderate anti-malarial activity against
plasmodium falciparum but the selectivity was very low compared to anti-malaria control compounds
(Ghosal et al. 1985, Tran et al 2003). Some members of the family also contain pharmacologically active
chemical compounds, Crinamine from crinum jagus, possessed strong anti-bacterial activity, while
lycorine hamayne and 6-hydroxycrinamine were inactive (Adeanaya et al, 1992). Crisisine from bulb of
crinum asiaticum was also an effective insecticide (Tram et al 2002).
Ethnobotanical study (Ogunkunle and Olapade, 2011) of the plant revealed its potency in the treatment
of asthma and related coughs.
METHODS
Plant extract and Phytochemistry
The bulbs of Crinum jagus were collected from Oyofo-Oghe Ezeagu L.G.A., Enugu State Nigeria in
August 2011. The bulbs of the plant were shredded and sun dried for three weeks and crushed. The
identity of the plant was confirmed by Prof. J. C. Okafor of Department of Applied Biology and
Biotechnology, Enugu State University of Science & Technology, Enugu – Nigeria.
About 53.8g of the powdered plant bulb was successively extracted with n-hexane, ethyl acetate and
methanol. After extraction, the extract was evaporated to dryness under reduced pressure using a rotary
vacuum evaporator. Phytochemical screening of the crude extracts was carried out using standard
methods described elsewhere (Sofoware, 1993, Trease and Evans, 2002, Perinos and Quinby, 1967).
Each plant extract was screened for the presence of different classes of compounds including saponin,
saponin glycoside, steroid/triterpenoids, glycoside, anthracenes, digitals glycoside, tannins, pseudo
tannins, flavonoids, resins, alkaloids and volatile oils.
Plant extract preparation

Exactly 150cm3 of each extract was evaporated at reduced temperature and the result residue was
dissolved (macerated) in distilled water (75mg/ml) before oral administration to the guinea pigs.
Animal grouping
One hundred and twelve guinea pigs weighing 800-1053g were bought from Zoology Department,
University of Nigeria Nsukka, Nigeria. The guinea pigs were kept in a well ventilated house conditions
0
(temperature: 27 – 30 C) and fed for three days with sunrise feeds and tap water. They were housed in
plastic cages of dimension 165cm x 101cm x 96cm with cleaning done before each meal. They were
distributed into six groups of seven (7) guinea pigs. This include one control (group A), poisoned only
(group B), poisoned and dose with cold water extract (c), with n-hexane extract (D), with ethyl acetate
extract (E) and methanol extract (group F) respectively for Aspartate transminante, Alanine
transaminante and Alkaline phosphates.
Poisoning and Treatment of Animals

3
Guinea pigs in group A were administered orally with 1cm of distilled water respectively. While guinea
pigs in groups B,C,D, E and F were administered orally with 1cm3 of “food poison (prepared using the
tap roots of bitter leaf – vernonia amygdalina (1g/100cm3 in distilled water).
3
After 1h of administration of poison groups C, D, E, and F were given 3cm each of (anti-poison)
(75mg/ml) in distilled water of n-hexane, ethyl acetate, methanol and cold water extracts of crinum jagus
respectively for aspartate transminate, Alanine transminate and alkaline phosphates.
201
Enzyme Assays (Determination of enzyme activities)
Table 1:Results of phytochemical Screening of Crinum Jagus plant bulb

Saponin + - -
Saponin glycoside + - -
Steroid/triterpenoid - + +
Glycoside - - +
Digitalis glycoside - + +
Anthracene - - -
Tannins - - +
Hydrolysable tannins + - -
Pseudo tannins - - -
Flavonoids - - -
Resins - - -
Alkaloids - - -
Volatile oils - - -
202
The effects of the extracts of crinum jugas on the acute toxicity of the vernonia amygdalina poison on the guinea
pigs.
During the observation period, no death of guinea pigs was neither recorded in the control group A nor in
the treated groups of C,D,E, & F. visual observations of rats were made and recorded systematically 1 and 2h after
poisoning and extract administration, including respiratory, behavioral pattern, food and water consumption. Only
one out of the 7 guinea pigs in group B survived. The pathological examination of the liver showed no visual
abnormality in groups A, D, E and F while the enzymes in group C were inactive and in group B, the enzymes of
the only surviving guinea pig was found to be dead.
Table 2: Effect of Extracts of crinum jugas on Different Liver Biochemical Parameters of guinea pigs in International
unit per liter (1U/L)
Doses administered (cm3) Extracts AST ALT ALP
(1UL (1UL-1) (1UL-1)
Grou Poiso CW H EA M -1
)
ps n E E E E
A 0 0 0 0 0 18.12 7.01+ 32.37+
+
0.20 0.17
0.29
B 1 0 0 0 0 1.02 0.47 7.39
C 1 3 0 0 0 20.11 13.29+ 32.19+
+
0.23 0.22
0.20
D 1 0 3 0 0 9.14+ 7.18+ 28.23+
0.26 0.18 0.20
E 1 0 0 3 0 5.44+ 3.39+ 0.1 32.41+ 0.
0.24 1 20
F 1 0 0 0 3 15.33 5.29+ 59.28+
+
0.16 0.17
0.26
Key: CWE – Cold Water Extract; HE – n-hexane extract; EAE – ethyl acetate extract; ME – methanol
extract; AST – Asparate transaminante; ALT – alanine transanimante; ALP – alkaline phosphatase;
IUL-1 – International unit per liter.
The results of the various extracts of crinum jugas on different liver biochemicals of poisoned guinea
pigs were present in Table 2. An increase in the biochemicals assayed was noticed only in the group (c)
from the control group A. the only surviving guinea pig in group B showed a significant decrease in the
biochemcials assayed below the reference (standard) range (AST, 5-18, ALT, 3-12 and ALP, 15-92)
indicating that the biochemical are dead. Groups D, E and F showed results within the reference range
with only Alkaline phosphate biochemical having readings within that of the control. This shows that the
extracts of n-hexane, ethyl acetate and methanol of crinum jugas is effective in normalizing the effect of
the poison in guinea pigs or the liver injuries due to the poison is minimized may be due to the enzyme
regeneration capacity of the extracts.
Visual observation of the guinea pigs 1 and 2h after poison and extracts administration, including
changes in respiration, behavioral pattern, and food and water consumption. These parameters changed
in groups B, C, D, E and F in after the administration of the poison and reversed 2h after the
administration of the extracts in groups C, D, E and F just as in the control group A.
203
REFERENCE
Ogunyemi, A. O. (1979). The origin of herbal cure and its spread; proceeding of a conference on African
Medicinal plants. Sofowara. A (eds). University Press, Ile-Ife; pp 20-22.
Glombitza K. N.; Mahran G. H., Mirhom Y. W, Michael K. G. and Motawi T. K. G. and Motawi T. K.
(1993). Hypoglycemic and hyperglycemic effects of Zizyphus Spinachrist in rats. Planta Med., 60;
pp 244-247.
Erturk O., Kati Hi; Yali N; and Demirbag Z. (2006). Antimicrobial properties of Silene Muttifida (Adams)
Rohib. Plant Extracts Turk J. Biol; 30(1); pp 17-21.
Castello M. C; Phatak A, Chandra N; and Sharon M. (2002). Antimicrobial activity of crude extracts from
plants parts and corresponding Calli of Bixa Orelland L. Indian J. Exp. Biol; 40 (12); pp 1378 –
1381.
Houghton P. J., Agbedahunsi JM and Adegbulugbe A. (2004). Cholinesterase inhibition properties of
alkaloids from two Nigerian Crinum species. Phytochemistry; 65(21); pp 2893 – 2896.
Burkill H.M. (1985). The Useful Plants of West Tropical Africa (second edition). Royal Garhens KEW; 2; pp
641 – 643.
Ghosal S, Saini KS and Razadan S.(1985). Crinum Alkaloids: Their Chemistry and Biology. Review Article.
Phytochemistry; 24(10); pp 2141 – 2156.
Adesanya S.A., Olugbade TA, Odebiyi O.O, and Aladesanmi J.A. (1992). Antibacterial Alkaloids in Crunum
jagus. Inter. J. Phamacog; 4; pp 303 – 307.
Tram N. T. Titorenkova T.V.; Bankova V.S., HAndjieva N.V; and Popov S.S. (2002). Crinum L.
(Amaryllidaceae) Review. Fitoterapia 73; pp 183-208.
Ogunkule A.T.J. and Olopade O. R. (2011). Studies on the asthma coughs plant crinum jagus L.
(Amaryllidaceae) in Nigeria. African J. of plant Science; 5(2); pp 108-114.
Sofoware A. (1993). Medicinal Plants and Traditional Medicine in Africa. 2nd Ed. Sunshine House, Ibadan,
Nigeria: Spectrum Books Ltd; Screening Plants for Bioactive Agents; pp 134-156.
th
Trease G. E. and Evans W. C. (2002). Pharmacognosy. 15 Ed. London. Saunders Publishers; pp 2, 42-44,
221-229, 246-249, 304-306, 331-332, 391-393.
Perinos G. J. and Quimby M. N. (1967). Nigeria plants 111: Phytochemical Screening for alkaloids, tannins,
saponins, J. Pharm. Sci.; 56(2); p. 152.
Babson A. L., Greeley S. J, Cole man C. M, and Philip G.D, (1966). Phenolphthatein monophosphate as a
substrate for serum alkaline phosphatase. Clinical Chemistry; 12; pp 481-490.
Reitman, S, and Frankel S. (1957). A colorimetric method for the determination of serum glutamate –
oxaloacetic acid and glutamate pyruvic acid transminases. Amer. J. Clin Path. 28; pp 56-63.
204
Kinetics and Sorption models of Unmodified and Modified Okra Stem Derived Cellulose for the
Removal and Recovery of Hg and As Ions from Aqueous Environment
Obidiozor, C.J. and Okoro I.A

Department of Chemistry, Michael Okpara University of Agriculture, Umudike PMB 7267, Umuahia Abia
State, Nigeria
Abstract: The efficiency of modified and unmodified Okra (Abelmoschus Esculentus) stem was studied as
adsorbent for removal of heavy metals( Hg(II) and As(III) ions) from its aqueous solutions. Chemical
modification of the sorbent produced a larger surface area which enhanced the binding capacity of the biomass
though not significantly. The equilibrium adsorption data were analyzed by Langmuir, Freundlich, Dubnin-
Radushkevich and Temkin isotherm models. The equilibrium data indicates the following order to fit the
isotherm: Freundlich > Temkin > Dubnin-Radushkevich > Langmuir. Pseudo-first order and Pseudo- second
order were used to determine the sorption kinetics. The biosorption kinetics was found to follow Pseudo-second
order kinetic model, with correlation (R2 =1 and 0.99) for mercury and arsenic ions respectively. The results from
this study revealed that Okra stem both Unmodified and Modified could be employed as a low cost alternative for
removal of Hg(II) and As(III) ions from aqueous solutions.
Keywords: Hg ion, As ion, Okra derived cellulose biomass, Sorption isotherm, Sorption kinetics
INTRODUCTION
Water pollution by heavy metal ions has become a serious threat today and of great environmental concern due to
their toxicity and persistent nature (Upendra, 2006; Sari and Tuzen,2008).The heavy metal ions are not only toxic
to living organisms in water but also causes harmful effects to plants, animals and humans through food chain
transfers. The metals hazardous to humans includes Lead, Cadmium, Chromium, Arsenic, Mercury and Copper.
Arsenic and Cadmium can cause cancer. Mercury can cause mutation and genetic damage, while copper, lead
and chromium brain and bone damage. Therefore, it is necessary to eliminate such hazardous heavy metals in
wastewater before discharging it into the ecosystem. The removal of heavy metal ions from wastewater involve
many techniques such as ion-exchange, evaporation, precipitation, adsorption and membrane processes (Wang
and Chen,2009). However, each method has its merits and limitations in application and they are often limited by
technical and economical issues (Pacheco et al., 2006). In addition, these methods have some disadvantages such
as incomplete removal, production of toxic sludge or waste products that also require disposal and economically
unviable for the removal of heavy metals at lower concentrations (Aklil et al., 2004). This is the basis of the use of
biosorption, a process which have proven to be quite effective at removing metal ions from contaminated
solutions in a low cost and environment-friendly manner. Biosorption is the ability materials of biological
origin(dead or alive biomass) to accumulate heavy metals from wastewater. The use of dead biomass as
biosorbent has many advantages; they do not require a continous supply of nutrients, they can be regenerated and
re-used from many cycles. Moreover, dead cells have shown to accumulate pollutants to the same or greater
extent than growing cell (Fu and Viraraghavan, 2002). This is the basis fpr the use of agricultural wastes such as
yam peels, rice husks, coconut fibers, okra stem, maize husks among others as biosorbents for metal removal.
The two toxic metal ions: Hg ion and As ion such as the subject of this study are metal ions without biological
roles and are listed as priority pollutants (Abia and Igwe, 2005;Gang-Sun and Wexing Shi, 1998).
Modification and Recovery
The proper selection of additives for modification of the adsorbents play crucial role in ensuring better efficiency
and more cycles of usability. Chemical modification involves treatment of the adsorbent with an array of
chemicals for washing, enhancement of binding groups and elimination of inhibition (Park et al., 2010). The
205
chemicals used are minerals (hydrochloric acid, sulfuric acid, nitric acid), organic acids, bases, salts, alcohols,
aldehydes, ketones, oxidizing agent, aromatic compounds (benzene, toluene), chelating agents (Ngah and
Hanafiah, 2008;Park et al., 2010).
Technological application of microbial metal accumulation may depend on the ease of metal recovery either for
subsequent reclamation or for further containment of toxic waste. Heavy metals can be recovered by physical or
chemical treatment. Physical method involves microwaving and heating; whereas chemical method involves the
use of eluents as acids, alkali and organic solvents. NaOH, H2SO4, HCL, HNO3, NaNO3, EDTA etc are commonly
employed for desorption. Soleimani and Kaghazchi (2008) suggested that gold can be eluted from biosorbents
by improved striping method that involves alkali treatment followed by organic solvent.
Experimental
Okra stems were collected from a vegetable farm at Olokoro, Abia state and were identified by a Botanist Dr,
Omosun Garuba of plant science and biotechnology department of Michael Okpara University of Agriculture
Umudike. All reagents were analytical grades from BDH Chemicals, London.
Sample Preparation
Okra stems were air-dried, milled and ground into powder using a milling machine (binatone model). The
powdered samples were sieved using 0.05mm,0.5mm and 1mm sieve. The ground Okra stems were converted
into cellulose pulp by alkaline pulping method. Okra derived cellulose pulp was dried, sieved and termed
Unmodified. The pulped sample was divided into half and one part was further treated using toluene diisocyanate
(TDI). The TDI treated Okra derived cellulose was termed modified.
Metal ion solutions Preparation
1000mg/L solution of mercury and arsenic were prepared as stock from the salts HgCL2 and As2O3 respectively.
3
This was done by dissolving appropriate amount of each salt in 1000cm of de-ionised water, and made up to
mark of volumetric flask. Each of these solutions represents the metal ion in aqueous solution (wastewater) of
1000mg/L concentrations. From the stock solutions of 1000mg/L working concentrations of 10 to 50mg/L of
each of the metal ions were prepared by serial dilution. The initial concentration of metal ion solutions used for
adsorption study for investigating the effect of pH, contact time, particle size and temperature was 50mg/L. On
the other hand, metal ion concentrations of 50,40.30,20 and 10mg/L were used to investigate the effect of
variation in initial concentrations of metal ions on sorption.
Atomic Absorption Spectrophotometric (AAS) analysis
Each filterate collected was analysed for unsorbed Hg(II) using AAS instrument(Unicam model 969). The
difference between the initial concentration of Hg ion and the amount of Hg ion in the filterate gives the amount
of mercury ion sorbed. A similar procedure was used for As ion. A hollow cathode lamp containing Hg and
another hollow cathode lamp containing As were each used respectively during the AAS analysis (Okoro and
Ejike, 2007).
EFFECT OF MODIFICATION BY TOLUENEDIISOCYANATE ON THE OKRA STEM
The adsorption of metal ions by the adsorbent in aqueous solutions can be manipulated by pre-treating the
adsorbents with either alkalies, acids, aromatic compounds, heat etc. This may lead to increase adsorption. The
okra stem was first pre-treated with ethanolic sodium hydroxide and further modified with Toluenediisocyanate
(TDI) in order to create a large surface area for adsorption. Our results showed that chemical modification of okra
stem using TDI increased the sorption capacity of the biosorbent by enhancement of binding group, which
involves amination of hydroxyl groups thereby increasing the efficiency of the biosorption of mercury and
arsenic ions.
Sorption Kinetics
206
The kinetic studies of sorption of Hg ion and As ion using unmodified and modified okra derived cellulose
biomass carried out are reported in the following Tables.
Table 1 PSEUDO-FIRST ORDER CONSTANTS AND CORRELATION COEFFICIENTS(R2
VALUES) FOR SORPTION OF METAL IONS ONTO UNMODIFIED AND MODIFIED OKRA
STEM
Constants Unmodified Okra Stem(UOS) Modified Okra Stem(MOS) Hg(II)
As(III) Hg(II) As(III)
qeexp (mg/g) 9.998 9.991 9.999 9.998
qecalc (mg/g) -1.803 -1.640 -1.844 0.486
k1(min-1) 0.001 0.035 0.012 0.050
2
R 0.843 0.912 0.994 0.778
Table 2 PSEUDO-SECOND ORDER CONSTANT AND CORRELATION

COEFFECIENTS(R2 VALUES) FOR SORPTION OF METAL IONS ONTO UNMODIFIED
AND MODIFIED OKRA STEM
Constants Unmodified Okra Stem(UOS) Modified Okra Stem(MOS)
Hg(II) As(III) Hg(II) As(III)
qecalc(mg/g) 10 10.101 10 10.204
k2(g/mg/min) 10 0.064 10 0.034
ho(mg/g/min) 1000 6.489 0 3.488
2
R 1 0.999 1 0.999
Figure 1:Pseudo-first order plot of In(qe-qt) vs t for adsorption of the metal ions onto unmodified okra
stem
207
Figure 2: Pseudo-first order plot of In(qe-qt)vs t for adsorption of the metal ions onto modified okra stem
Figure 3: Pseudo-Second order plot of vs t for adsorption of the metal ions onto unmodified okra
stem
Figure 4: Pseudo-Second order plot of vs t for adsorption of the metal ions onto modified okra
stem
208
The study of sorption kinetics of heavy metal removal from wastewater is significant as it provides
valuable insights into the reaction pathways and into the mechanism of sorption reactions. To analyze
the mechanism of biosorption process, the experimental data were fitted to the pseudo-first and pseudo-
second order kinetic models as indicated by Equations 1 &2, respectively(Ho and McKay, 1998; Fu and
Viraraghavan, 2002).
In( - ) =In - k1t or Log( - )=Log - t 1
t/ =1/(k2qe2) +(t/qe) 2
2+ 3+
The rate constant (k1) using pseudo-first order model for Hg and As ions was obtained from the slope of the
linear plot of In(qe-qt) against time, t for each solute (Figure 1&2). The rate constants (k2) for these metals by
pseudo-second order was obtained from the slope and intercept of plots of t/qt against t (Figure 3&4). The rate
constants with the R2 values are represented in Table 1&2 for the two metal ions. It was observed that the
experimental data for the two metal ions for both unmodified and modified okra stem followed the pseudo-
second order model. Despite the fact that pseudo-first order plots showed good linearity, the calculated values
(qe,calc) showed a large difference with the experimental values (qe,exp) unlike in pseudo-second order model.
Therefore, the adsorption process did not follow pseudo- first order model. Also it is clear that the correlation
2
coefficients(R ) values for pseudo-second order model for both UOS and MOS respectively, are very high(in
most cases >0.99) when compared with pseudo-first order model. Therefore, it is likely that the sorption of
Hg(II) and As (III) by okra derived cellulose biomass are kinetically controlled as a second order reaction rather
than a first order process. The data obtained indicates that the adsorption of these metal ions onto unmodified and
2+ 3+
modified okra stem follows this trend Hg >As .
Sorption isotherm models
Langmuir isotherm model: This model is based on assumptions that adsorption occurs at specific homogenous
sites, energy of adsorption is constant and there is no transmigration of adsorbate in the plane of the surface, and
is written as:
Ce/qe =1/qmaxKL + Ce/qmax 3
Where is the adsorption capacity in mg of adsorbate per gram of adsorbent, Ce is the residual (equilibrium) metal
ion concentration in solution, qmax is the maximum adsorption capacity corresponding to monolayer coverage
and KL is the Langmuir isotherm constant, which expresses the intensity of the adsorption process.
Figure 5:Langmuir isotherm of plot of Ce/qe vs Ce for adsorption of metal ions onto Unmodified okra
stem
209
Figure6: Langmuir isotherm plot of Ce/qe vs Ce for adsorption of the metal ions onto Modified okra stem
Based on Figure 5&6 which showed low R2 values, Langmuir isotherm model was found not to agree well with
the experimental isotherm data.
Freundlich Equilibrium Isotherm

The Freundlich equilibrium isotherm assumes a heterogenous surface with a uniform distribution of heat of
adsorption over the surface. The linearised form of Freundlich adsorption isotherm is represented as:
In = In kf + In Ce 4
where K and n are Freundlich constants related to adsorption capacity and adsorption intensity respectively.
Figure 7: Plot of Inqe vs In Ce for adsorption of the metal ions onto Unmodified okra stem
210
Figure 8: Plot of Inqe vs In Ce for adsorption of the metal ions onto Modified okra stem
Dubnin-Radushkevich isotherm
The Dubnin-Radushkevich (D-R) model was applied to the equilibrium data to assess the nature of
adsorption process, i.e whether it is physical or chemical adsorption. The linearised D-R adsorption
isotherm is represented as (Bansal et al., 2009):
in =In –BD 5
) 6
-2
Where is the theoretical saturation capacity (mg/g), is a constant related to adsorption energy (K J ), R is
gas constant (KJ ) and T is the temperature (K).
Figure 9:Dubnin-Radushkevich plot of Inqe against [RTIn(1+1/Ce)]2 for adsorption of the metal ions
onto Unmodified okra stem
Hg(II) As(III)
2.5
1.5
y = -0.0030x+7.7481
1
y = -0.0033x+6.5513 R^2 = 0.9613
R^2 = 0.9922
0.5
0
0 500 1000 1500 2000 2500
2
Figure10: Dubnin-Radushkevich plot of Inqe vs [RTIn(1+1/Ce)] for adsorption of the metal ions onto
Modified okra stem
4.7.4 Temkin isotherm
The temkin isotherm assumes the effect of some indirect interactions amongst adsorbate particles and
suggests linear decrease in the heat of adsorption of all the molecules in the layer. Temkin isotherm
equation is given as (Mittal,2009);
qe = BT (InAT +In Ce) 7
where AT and BT are Temkin constants which can be obtained from intercept and slope of the linear plot of
versus In Ce, respectively.
211
Figure 11: Temkin isotherm plot of qe vs In Ce for adsorption of the metal ions onto unmodified okra
stem
Figure 12: Temkin isotherm plot of qe vs InCe for adsorption of the metal ions onto modified okra stem
Table 3: Comparison of correlation coefficients (R2) values for Langmuir, Freundlich, Temkin and
Dubnin-Radushkevich adsorption isotherm
Metal ions Okra Stem

Unmodified Modified
Langmuir Isotherm
2+
Hg -2.70 0.449
3+
As -3.22 0.143
Av. -1.332
Freundlich isotherm
2+ 3+
Hg 0.976 0.953 As
0.999 0.988 Av.
0.979
Dubnin-Radushkevich isotherm
2+ 3+
Hg -0.88 0.9613 As
-2.78 0.9922 Av.
-0.427
Temkin isotherm
2+
Hg 0.949 0.949
As3+ 0.952 0.966
Av. 0.954
212
2
A comparison of the correlation coefficient (R ) for the adsorption isotherm is shown in Table 3. However,
though it is obvious that three out of the four isotherms are good in describing the effectiveness of unmodified
and modified okra stem for removal of Hg2+ and As3+ ions from aqueous solution. It could be concluded from the
average values of R2 that the Freundlich isotherm is the best, followed by Temkin, Dubnin-Radushkevich and
the Langmuir isotherm for the adsorption process of this work.
Conclusions
The biosorption of Hg(II) and As(III) ions onto okra waste was proven to be an efficient process. The Langmuir,
Freundlich, Dubnin-Radushkevich and Temkin isotherm models were used for the mathematical description of
the biosorption equilibrium of heavy metal ions onto okra derived cellulose biomass. The Freundlich isotherm
model was found to provide the best fit of the experimental data, followed by Temkin and Dubnin-Radushkevich
which gave a fairly good fit and then the Langmuir which did not yield satisfactory description behaviour.
Adsorption kinetics was found to follow the pseudo second-order model, which relies on the assumption that
adsorption may be the rate limiting step involving valence forces through sharing or exchange of electrons
between adsorbent and adsorbate.
Results obtained from this study indicated that okra stem which is an agricultural waste and pollutant, could be
used as a potential sorbent for the removal of Hg(II) and As(III) ions from aqueous solutions. Conventional
methods of removal seems to be expensive, so the need for a sorbent material like okra stem as alternative should
be employed. This is because it is readily available, economically feasible, biodegradable and above all its in
tandem with the popularization of green chemistry, green environment and green technology for sustainable
development.
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Soleimini M, Kaghazi T (2008). Activ
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214
SYNTHESIS, CHARACTERIZATION AND CORROSION INHIBITORY OF Co (II), Ni (II), Cu (II), AND Zn (II)
COMPLEXES DERIVED FROM NICOTINIC ACID HYDRAZIDE.
*1
O.W.Salawu, 2R.A. Wuana, 3J.T.Ashimom
*1
Department of Chemistry, Federal University, Lokoja, Kogi State, Nigeria.
2&3
Department of Chemistry, University of Agriculture, Benue State, Nigeria.
*Correspondence author
*E-mail:
Abstract: Metal complexes formed by the reactions of Nicotinic acid Hydrazide (NAH) with copper (II),
Cobalt (II), Nickel (II), and Zinc (II) sulphate ions were prepared and characterized by their infrared,
electronic, magnetic moment, elemental analysis, solubility, molar conductivity, and melting point as
well as their corrosion inhibition properties using the Weight Loss Method (WLM).
INTRODUCTION
In a world where there is heavy dependence or reliance on the use of metals and alloys; protecting
metals from corrosion therefore, has been paramount on the hearts and minds of the chemists whose
concern has always been to help reduce or curb this threatening condition or phenomena over the past
decade. A review of literature shows that many hydrazides have been used widely in medicine, catalysis, and
[i-iii]
analytical chemistry . However, the use of hydrazide metal complexes to inhibit corrosion is scarce and so,
there is a need for a broad spectrum of hydrazide complexes to be synthesized and employed in the control
of corrosion.
Materials
Hydrazine hydrate (NH2NH2.H2O), Nicotinic acid, Absolute ethanol, Methanol, CuSO4.6H2O, NiSO4.6H2O,
ZnSO4.7H2O , CoSO4.7H2O, CaCl2, Concentrated HCl, Acetone, were of reagent grade and were obtained from
commercial sources (Sigma-Aldrich Chemical Limited) and used without further purification. All solvents
used were of analytical grade.
Preparation of the Ester and the Nicotinic acid hydrazide (NAH)
The ester was prepared using the Fisher-Speier's Method [iii]. Here, 37.0g (0.300mole) of nicotinic acid
was reacted with 36 mL (28.4 g; 0.616 mole) of absolute ethanol saturated with 19 mL (19.3 g, 0.539 mole)
o
of concentrated hydrochloric acid in a 250 mL round bottom flask and refluxed for 5hours at 80 C in a water
[iv]
bath and the NAH was prepared by the hydrazinolysis of ester with NH2NH2.H2O . The ester of exactly 35.0
g (0.232 mole) was reacted with 11 mL (11.3 g; 0.354 mole) of NH2NH2.H2O in a 250 mL round bottom flask
o
and heated for 15 minutes under reflux in a water bath at 80 C. After 15 minutes, 19 mL (15.0 g; 0.325 mole)
of absolute ethanol was added into the solution until a clear solution was observed. The mixtures were left
overnight to crystallise. The resulting crystals were recrystallized with a minimum quantity of ethanol. The
final crystals obtained were dried over CaCl2 in a dessicator and weighed. [Weight = 35.0g and Yield = 77.2%
for ester and Weight = 22.0g and Yield = 69.2% for NAH.
Preparation of the [Cu(H2O)3(NH)2SO4]; [Ni(H2O)3(NH)2SO4]; [Co(H2O)3(NH)2SO4];
and [Zn(H2O)(NH)2SO4] metal complexes
Xg (0.027 moles) of each of, CuSO4.6H2O, NiSO4.6H2O, CoSO4.7H2O, ZnSO4.7H2O respectively was
dissolved in 5 mL of distilled water then, 3.75g (0.027 moles) of NAH was also dissolved in 5 mL of absolute
ethanol. The metal salt solutions were added with constant stirring to the solution of the NAH and heated
for about 10 minutes. The mixture was left overnight to crystallise. The resulting crystals were recrystallized
with a minimum quantity of ethanol. The final crystals obtained were dried over CaCl2 in a dessicator and
weighed. The weights and yields were as follows: 6.5g and 62.3% for copper complex, 7.0g and 64.9% for
nickel complex, 9.3g and 82.3% for cobalt complexes, and 8.5g and 74.2% for zinc complex respectively.
215
Weight Loss Method (WLM) for Corrosion Testing.
[v]
The method as described in the literature was modified and adopted. Here, the mild
steel was perforated, cleaned and weighed. Two (250 mL) beakers were used to which 1.0M HCl
was added to each beaker and a coupon each and labelled I and II respectively. The first beaker
represented the control system and to the second beaker, 0.12g of the ([M(NAH)2SO4]n(H2O)) was
added also and this last process was carried out for all the ([M(NAH)2SO4]n(H2O)) metal
complexes. After 72 hours (3 days), the coupons were removed, washed and weighed on a
weighing balance in grams up to 0.000.
The melting point of the ligand and metal complexes are shown in Table 1. The results
o
show high melting points ranging from 155-205 C indicative of the high purity of the metal
complexes. The Conductance data on Table 1 shows that all the metal complexes are non-
electrolytic in nature and thus, exhibit weak ionic character. The hydrazide metal complexes
were soluble in ethanol but insoluble in other organic solvents as shown in Table 2. The
elemental analysis data in table 3 is given in mg/kg (mg/L). The concentration over the
absorption from table 7 on atomic absorption spectroscopic data gives the percentage
composition. The result shows that the metal to ligand ratio of the complexes is 1:2. The
corrosion inhibitions of the metal complexes in Table 6 shows that the cobalt hydrazide complex
showed higher inhibition abilities probably because, of the presence of more vacant, low-energy
electron orbitals for electron transfer or sharing with the metal [vi]. The Molar Magnetic
susceptibility data is given in table 7 and the magnetic moment was calculated using the
relationship as follows:
ì/ ìB = 797.5 √Xmol .T
where ì = Magnetic moment, ìB = Bohr Magneton, Xmol = Molar Magnetic susceptibility
and is given as the Magnetic susceptibility (Xgram) × Relative Molecular Mass, and T =
temperature.
The magnetic moment (ìeff) for the complexes is given in Table 5 and the value for the
Co hydrazide complex is 4.95 B.M observed for Octahedral Co2+ complexes [vii] and for the Ni2+
2+
2+ [viii] 2+
hydrazide complex is 3.11 B.M for octahedral Ni complexes . The (ìeff) for the Cu hydrazide
2+ [ix] 2+
complex is 1.93 B.M observed for an octahedral Cu complex . The Zn hydrazide complex has
the (ìeff) of 0.41 B.M observed for a tetrahedral Zn2+ complex. The hydrazide complexes were
synthesized using equi-molar quantities of the metal (II) sulphate salts of Co, Ni, Co and Zn
respectively as shown below:
II II
M SO4.nH2O + RCONHNH2 [M(H2O)n(R1CONHNH2)2SO4] +
nH2O Where M is Co, Ni, Cu or Zn and n = 1, 2, 3, ….7
Infrared Spectra
The relevant IR spectra assignment is shown on table 4. The relevant IR spectra for the
ligand and metal (II) complexes are explained as shown below.
216
Bands Frequency (cm -1) Assignment
v(O-H) 3431-3876 Water molecules
v(C=N) 1396-1453 Azomethine in bonding
v(N-H) 3231-3669 Azomethine confirmed
v(N-N) 1213-1242 Azomethine confirmed
v(SO42-) 1050-1153 Inner sphere coordination
v(C=O) 1600-1733 Conjugation with double bond and resonance effect
Electronic Spectra
The electronic spectra and the magnetic data are represented in Table 5. The electronic spectrum for
the zinc hydrazide complex [Zn(NAH)2SO4].H2O showed a band at 361.00 nm (27700 cm-1) assigned to 1A1 (ð-
[viii]
ð* due to the presence of C=C, C=O, or C=N groups) transition and suggestive of a tetrahedral geometry.
-1 4
The cobalt hydrazide complex [Co(NAH)2SO4].3H2O showed a band at 786 nm (12720 cm ) assigned to T1g -
2
T2g (n-ð* due to water molecules) transition and suggests an octahedral geometry [xi]. The copper hydrazide
complex [Cu(NAH)2SO4].3H2O showed a band at 419.00 nm (23900 cm-1) and assigned to 2Eg - 2T2g (ð-ð* due to
the presence of C=C, C=O, or C=N groups) transition which is suggestive of a distorted octahedral geometry
[xiii]
due to Jahn-teller effect . The nickel hydrazide complex [Ni(NAH)2SO4].3H2O showed a band at 783.50 nm
(12800 cm-1) due to 3A2g - 3T1g (n- ð* due to the presence of water molecules) transition and is suggestive of
an octahedral geometry [xiv].
Conclusion
On the basis of the physicochemical properties, the following structures are suggested for the 1:2
Octahedral and tetrahedral geometry for the Hydrazide Metal Complexes where M = Co (II), Ni (II), Cu (II)
and Zn (II) and R = C6H5N-
OH2
R O NH
R O NH C NH2
C NH2
M SO4. nH2O
. nH2O
M
H 2N C
H 2N C HN O R
HN O R
SO4
217
References
i. Nwabueze, J.N. and Salawu, O.W. (2012). Complexes of Ni(II) and Cu(II) Sulphates with Keto and Enol
Forms of Isobutryl Acetic Acid, 4-Amino Benzoic Acid and 4-Cyano Benzoic Acid Hydrazide. Advances
in Pure and Applied Chemistry. 1(2): 1-9.
ii. Salawu, O.W and Abdulslam, A.O (2011). Synthesis, and Spectroscopic Characterization of Schiff Base
Complexes of Cinnamaldehyde Hydrazone. Journal of Physical Science and Innovation. 3: 1-11.
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Edition, New York, John Wiley and Sons, Inc. Pp623-1257.
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20(6): 367-372.
v. Kavita, K. Givish, P. Rajewa, V. and Pingalkar, R.S. (2014). Corrosion Inhibition Property of Some SEDA-
Complexes on Mild Steel in Acidic Medium. Indian Journal of Applied Research. 4(4): 1-2.
vi. Papavinasam, S. (2000). Corrosion Inhibitors. CANMET Materials Technology Laboratory, Ottawa,
Ontario, Canada. Pp. 1-17.
vii. Walaa, H. M, Gehad, G. M., and Maher, M. I. (2014). Synthesis, Characterization and in vitro
Biological Activity of Mixed Transition Metal Complexes of Lornoxicam with 1,10-phenanthroline.
International Journal of Electrochemical Science. 9:1-20.
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Pp1049-1310.
ix. Salawu, O.W., Kolawole, S., and Akpah, A.F. (2011). Preparation and Characteristics of Some Metal
Ion Complexes. Pelagia Research Library. 2(2):1-8.
x. Ragab, R., Taj Al-ansi, Fathi, Al-Azab, El-Reedy, and Ahmed, A. (2015). Cooridnation Compounds of
Butane-1,1,4,4-Tetracarboxylic Acid Tetrakis-(Salicylaldehydene Hydrazide): Their Preparation,
Characterization and Structural Investigation. International Journal of Advanced Research. 3(3): 1-9.
xi. Vidya, V.G., and Mini, S. (2014). Synthesis, Spectroscopic Characterisation and Biological Studies of
Copper (II) Complexes Derived from Salycyloyl Hydrazide with Furfuraldehyde. Research Journal of
Recent Sciences. 3: 1-3.
xii. Ushma, J. and Sunil, I. (2014). Novel Hydrazide Compounds and their Metal Complexes. Archives of
Applied Science Research. 6(4): 267-270.
xiii. Rathakrishnan, S.A., Abdul Jameel, and M., Syed Ali Padusha (2014) Synthesis, Characterization and
antimicrobial studies of a chiral compound and its metal complexes. International journal of scientific
and Research publications. 4(7): 1-12.
xiv. Waeel Mohammed Hamud (2013). Synthesis, Characterization and Kinetic Study of Metal Complexes
with new acyclic Legend N2O2. Diyala Journal of Pure Science. 9(1): 1-19.
Table 1: Some Physical Constants, molar conductivity and the Melting point for the Ligand and Metal (II)
Complexes
218
Ligand/complexes Molecular Molecular Yield Colour Molar Temp.
(%)
formula weight Conduct.(ms/cm) (oC)
NAH C6H7N3O 137.14 69.2 Milky 0.80 155
[Cu(NAH)2SO4].3H2O C12H20N6O9SCu 487.83 62.3 Light Blue 0.14 205
[Ni(NAH)2SO4].3H2O C12H20N6O9SNi 482.99 64.9 Light 0.14 175
Green
[Zn(NAH)2SO4].H2O C12H16N6O7SZn 453.37 74.2 White 0.99 184
[Co(NAH)2SO4].3H2O C12H20N6O9SCo 483.21 82.3 Pink 0.08 195
Table 2: Solubility test for the ligand and Metal (II) Complexes
Ligands/Complexes Distilled Ethanol Acetone Methanol Ethyl DMSO Chloroform
water Acetate
NH NS S NS NS NS NS NS
[Co(NAH)2SO4].3H2O NS S NS NS NS NS NS
[Zn(NAH)2SO4].H2O NS S NS NS NS NS NS
[Cu(NAH)2SO4].3H2O NS S NS NS NS NS NS
[Ni(NAH) 2SO4].3H2O NS S NS NS NS NS NS
Note: S = Soluble, NS = Not Soluble, PS = Partially Soluble
Table 3: The Elemental Data for the Ligand/Metal (II) Complexes
Compounds %M %H2O % SO42-
NAH Found - - -
Calculated - - -
[Cu(NAH)2SO4].3H2O “ 12.95 10.97 19.58
“ (13.01) (11.08) (19.69)
[Ni(NAH) 2SO4].3H2O “ 12.21 10.89 19.90
“ (12.16) (11.18) (19.87)
[Zn(NAH)2SO4].H2O “ 14.38 4.07 21.22
“ (14.42) (3.97) (21.17)
[Co(NAH)2SO4].3H2O “ 12.15 11.25 19.90
“ (12.20) (11.18) (19.87)
Table 4: The Relevant Infrared Spectra Data for the Ligands/Complexes in KBr/NaCl Pellets in cm-1.
Ligands/Metal complexes N-H C=O C=N N-N M-N M-O SO42- H2O
NAH 3669 1733 1396 1242 - - - -
[Co(NAH)2SO4].3H2O 3282 1617 1507 1387 608 393 1079 3736
[Zn(NAH)2SO4].H2O 3265 1617 1511 1319 607 387 1070 3875
[Cu(NAH)2SO4].3H2O 3231 1610 1517 1302 616 466 1063 3996
[Ni(NAH)2SO4].3H2O 3277 1607 1510 1387 612 402 1072 3753
Table 5: Electronic Spectra and Magnetic susceptibility for the Ligand/Metal (II) Complexes.
Metal complexes µeff ëmax (nm) Assignment Assignment Geometry
(B.M)
219
NAH - 366.50 Charge - -
Transfer
4
[Co(NAH)2SO4].3H2O 4.95 786.00 T1g - 2T2g Paramagnetic Octahedral
1
[Zn(NAH)2SO4].H2O 0.41 361.00 A1 Diamagnetic Tetrahedral
2
[Cu(NAH)2SO4].3H2O 1.93 419.00 Eg - 2T2g Paramagnetic Octahedral
3
[Ni(NAH)2SO4].3H2O 3.11 783.50 A2g - 3T1g “ Octahedral
Table 6: % Inhibition Efficiency of the Hydrazide Metal (II) Complexes

s/no Compound Initial wt. Final Loss in wt. Corrosion % Inhibition
in g wt. in g in g Rate efficiency
1. Control (Blank) 2.967 2.748 0.219 0.00051 -

2. HCl + [Cu(NAH)2SO4].3H2O 2.910 2.811 0.099 0.00023 54.8%
3. HCl + [Ni(NAH)2SO4].3H2O 2.896 2.835 0.061 0.00014 72.2%
4. HCl + [Zn(NAH)2SO4].H2O 2.998 2.935 0.063 0.00015 71.2%
5. HCl + [Co(NAH)2SO4].3H2O 2.989 2.853 0.136 0.00032 37.9%
Table 7. The Atomic Absorption Spectroscopic Data for the Metal, Sulphate and Water constitutes in the
complexes as well as the Molar magnetic susceptibility Data (Mag. Suscep.)
M SO42- H2O Mag. Suscep.
Sample Conc. Abs. Conc. Abs. Conc. Abs. Xmol
[Cu(NAH)2SO4].3H2O 0.0065 0.0500 0.0050 0.0260 0.0021 0.0191 1.965 ×10-8
[Ni(NAH)2SO4].3H2O 0.0049 0.0400 0.0055 0.0280 0.0020 0.0184 5.103 ×10-8
[Zn(NAH)2SO4].H2O 0.0056 0.0450 0.0018 0.0085 0.0023 0.0565 8.869 ×10-10
[Co(NAH)2SO4].3H2O 0.0059 0.0490 0.0112 0.0560 0.0017 0.0151 1.293 ×10-7
220
PREPARATION AND CHARACTERIZATION OF ACTIVATED CARBON FROM Hura Crepitans
Linn SEED SHELLS
1
E. W. NSI1 , A. E. AKPAKPAN1 and U. D. AKPABIO
1
Department of Chemistry Akwa Ibom State University, Ikot Akpaden, Akwa Ibom State, Nigeria
Department of Chemistry University of Uyo, Uyo, Akwa Ibom State, Nigeria
ani4sucess@yahoo.com, ukanadakpabio@yahoo.com
ABSTRACT
Activated carbons were thermally prepared from Hura Crepitan L. seed shells. Zinc chloride (ZnCl2) and
Phosphoric acid (H3PO4) were separately used as the activating agents. The activated carbons obtained were
characterized by determining the percentage yield, moisture content, ash content and percentage fixed carbon.
The adsorption of methylene blue by the activated carbon was done using 0.1 to 0.5g of the activated carbon. The
results revealed that the percentage yield and ash content of H3PO4 impregnated activated carbon was higher than
ZnCl2 impregnated activated carbon. On the other hand ZnCl2 impregnated activated carbon had higher moisture
content and percentage fixed carbon. It was also revealed that ZnCl2 impregnated activated carbon had greater
adsorption capacity than H3PO4 impregnated activated carbon. However it was found that the higher the
adsorbent (activated carbon) dosage, the higher the adsorption capacity.
Keywords: Hura Crepitan L., activation, activated carbon, Zinc chloride and Phosphoric acid
INTRODUCTION
Activated Carbons are carbon of highly micro-porous form with both high internal surface area and porosity, and
commercially, the most common adsorbents used for the removal of organic pollutant from air, water and
industrial products (Tsai et al., 2001).
Food and pharmaceutical industries uses activated carbon in the removal of compounds that are responsible for
undesirable colour, odour and taste in their products. Activated carbon is also used in gas cleaning applications
where it is extensively used in air filters at industrial level as well as in air condition equipment (Vigayan et al.,
2012).
Commercial activated carbon is produced from bituminous or lignite coal. Because of the environmental
problems and increasing high cost associated with petroleum products, research have been reported on the
production of activated carbon from wood which is environmentally friendly and cost effective. However,
because of the environmental problem associated with deforestation, seed shells have been used to replace wood
for the production of activated carbon. These products are highly cost effective as most of the seed shells are
discarded as waste materials, and the activated carbon produced from them have similar potential in industrial
application.
The production of activated carbon from coconut shells, palm kernel shells, Cananium scheqinflurthi nut shell,
corn cob and other seed shells have been reported (Olawale and Ajayi, 2007; Diya et al., 2008; Chang et al, 2001
and Vijayan et al., 2012).
Hura crepitans L., the sand box tree also known as possum-wood, is an evergreen tree of the spurge family
Euphorbiacae. The wood is used for furniture under the name “Hura”. The oil extracted from the seed is used in
the production of alkyl resin (Ezeh et al., 2012) and metallic soap (Umoren et al., 2013). The seed pods (shells)
are not utilized for any industrial purposes; hence they are waste and thereby pollute the environment. These
shells are hard and therefore suitable for the production of activated carbon which can be used for the treatment of
221
waste water from textile industries and can also be used in food and pharmaceutical industries to remove
pollutants/contaminants.
Therefore, the aim and objectives of this work is to prepare activated carbon from Hura crepitans L. shells,
characterize and apply it in the removal of methylene blue from contaminated water.
2.0 MATERIAL AND METHODS

2.1 Sample Collection
The seed shells of Hura crepitans L. also known as sand box were collected from the University of Uyo Town
Campus. The samples were cut into chips of workable size.
2.2 Activation-Carbonization
Activation of the sample was carried out in a muffle furnace using the solution of zinc chloride (ZnCl2) and
phosphoric acid (H3PO4) separately as the activating agents. 400g of the sample chips were charred in the furnace
at 500oC for 2 hours. 30g of the charred sample was weighed into a beaker containing 300cm3 of 1M solution of
each activating chemical. The contents of each beaker was thoroughly mixed and heated until all the activating
liquor was absorbed by the sample.
o
The samples were then transferred into the crucible and dried in the oven at 100 C for 12 hours. The crucible was
then placed in the furnace and fired at 650oC for 2 hours.
The sample was then removed from the furnace and cooled in the desiccator, it was then washed with 0.5M
NaOH followed by distilled water to a neutral pH. The activated carbons obtained were dried in the oven at 100oC
to a constant weight.
2.3 Characterization of the Activated Carbon.
· Percentage Yield
The yield of the activated carbon was calculated using;
Percentage yield = M2 x 100%
M1
Where M1 = Weight of the charred sample
M2 = Weight of the activated carbon
· Moisture content
Moisture content of the activating carbon was calculated using :
Percentage moisture content = M1-M2 X 100%
M1
Where M1 = Weight of the activated carbon before drying in the oven
M2 = Weight of the activated carbon after drying in the oven
· Determination of Ash content and percentage fixed carbon

Ash content of the activated carbon was determined as follows: 2g of each activated carbon sample was put in a
o
crucible and fired at 900 C in a muffle furnace for 3 hour. The resultant ash was removed and allowed to cool in
the desiccator.
Ash content was calculated using:
% Ash content = W2 x 100%
W1
222
Where W2 = Weight of the ash
W1 = Weight of the activated carbon
Percentage fixed carbon was calculated from the ash content in accordance with the method reported by
Ogbonnaya (1992) and Duff and Ross, (1988).
2.4 Adsorption Study

The adsorption potentials of the activated carbon sample obtained from different activating agents were
determined according to the method reported by Odebunmi and Okelola (2001). In this method, batch sorption
experiments were carried out at room temperature. 0.1 to 0.5g of the activated carbon was added into a beaker
containing 100ml of 250mg/l methylene blue solution. The mixture was thoroughly mixed and shaken for 1 hour
using a mechanical shaker. After shaking, the suspensions were filtered and remaining concentration was
determined spectrophotometrically at ënm of 250nm using UV/V spectrophotometer. The amount of methylene
blue adsorbed from each solution was calculated as shown below.
Percentage methylene blue adsorbed = (Co – C) X 100

Co
Where:
Co (mg/L) = Initial concentration of methylene blue
C (mg/L) = Post-adsorption concentration of methylene blue3.0 RESULTS AND
DISCUSSION
The percentage yield, ash content, moisture content and percentage fixed carbon of the activated carbon
are presented in Table 1. These parameters are factors which could influence the adsorption capacity of the
activated carbon (Okeola et al., 2011).
Result revealed that the percentage yield of H3PO4 activated carbon was 44.90% and that of ZnCl2
activated carbon was 31.94%. The decrease in the yield of ZnCl2 activated carbon may be due to the reactions
between ZnCl2 and the sample.
The ash content and percentage fixed carbon compare favourable with those for the commercial
activated carbon. Also the ash content of H3PO4 activated carbon compare favourable with that of H3PO4, ZnCl2
and KOH activated carbon from maize cob (Odebunnin and Okeola, 2001) and NaCl activated carbon of
Jatropha curcas seed coats (Okeola et al., 2011). The ash content obtained for ZnCl2 activated carbon compare
favourably with the value obtained Coconut shell (Odebunmi and Okeola, 2001). However, ZnCl2 impregnated
activated carbon has greater percentage fixed carbon than H3PO4 impregnated activated. There is a correlation
between percentage fixed carbon and adsorption capacity. Sample with higher percentage fixed carbon are
expected to have high adsorption capacities (Ogbonnaya, 1992). The moisture content of ZnCl2 activated carbon
was higher than that of H3PO4 activated carbon, implying that ZnCl2 activated carbon has greater pore size for
absorption than H3PO4 activated carbon.
Table 1: Physical parameters of the activated carbon
223
Activating Agent H3PO4 ZnCl2
Yield (%) 44.90 31.94
Moisture content (%) 5 17
Ash content (%) 28 16.5
Percentage fixed carbon (%) 72 83.8
3.1Adsorption Studies
Methylene blue was chosen in this study because of its recognized usefulness in characterizing
adsorptive material (Itodo et al., 2010). The size of methylene blue molecule is much larger and its adsorption is
restricted mainly to the mesopores adsorbent. However a small portion is also found in larger micropores
(Cleiton et al., 2011).
The results of the methylene blue adsorption by the activated carbons are presented in Table 2 and 3. The
results revealed that the quantity of the adsorbent has greater effect on the adsorption of methylene blue from
aqueous solution. The result shows that the adsorption efficiency increased with increase in mass of the
adsorbent. The increase in adsorbent mass increases the contact surface area of the adsorbent particles which
means that it become more probable for solute molecules to be adsorbed on the adsorption sites and thereby
increasing adsorption efficiency (Okeola et al., 2012).
3.2 Effect of Activating Agents
With all the adsorbent dosages, ZnCl2 impregnated activated carbon sample has greater adsorption
efficiency than the one prepared with H3PO4. This is due to the fact that samples with high percentage fixed
carbon always have high adsorption capacities (Ogbonnaya, 1992). This result also suggests that ZnCl2
impregnated activated carbon sample have well developed mesoporosity favorably for adsorption of larger
molecules compared to H3PO4 impregnated activated carbon sample. However, the percentage adsorptions of
methylene blue by these activated carbons are low, this may be due to the fact that the size of the methylene blue
molecule is much larger and its adsorption is restricted mainly to the mesopores activated carbon (Cleiton et al.,
2011).
Table 2: Adsorption of Methylene Blue by H3PO4 activated Carbon

AC dosage (mg) Post Adsorption Conc. of MB Adsorption efficiency (%)
(mg/l)
100 201.67 19.33
200 197.54 20.98
300 180.64 27.74
400 167.36 33.05
500 140.58 43.77
224
Table 2: Adsorption of Methylene Blue by ZnCl2 activated Carbon
AC dosage (mg) Post Adsorption Conc. of MB Adsorption Efficiency (%)
(mg/l)
100 195.84 21.66
200 186.29 25.48
300 178.64 28.54
400 145.06 41.98
500 122.77 50.89
CONCLUSION
Hura Crepitan L. seed shells were successfully converted into activated carbon using H3PO4 and ZnCl2 as the
activating agents. The results of the physical properties of the activated carbon compared favourably with the
commercial activated carbon and with other activated carbons prepared from seed shells and cob. The adsorption
of methylene blue by ZnCl2 impregnated activated carbon was higher than H3PO4 impregnated activated carbon.
However H3PO4 impregnated activated carbon is recommended for used in food and pharmaceutical industrials
since it is non toxic activating chemical as it does not contain heavy metal while ZnCl2 impregnated activated
carbon can be apply for the treatment of waste water from textile and allied industries.
REFERENCES
Ajayi, O.A., and Olawale, A. S. (2009). A comparative study of thermal and chemical activation of Canarium
Schweinfurthii Nutshell. J. Appl. sci. Res., 5(12): 2148-2152.
Cleiton Nunes A., Guerreiru Mario C., (2011). Estimation of surface area and pore volume of activated carbons
by methylene blue and iodine numbers, Quim. Nova, 34(3) 472-476.
Diya udden, B. H., mohammed,I. A. Ahmed, A.S. and Jibril, B. Y. (2008). Production of activated carbon from
corncobs and its utilization in crude oil spillage clean up. Agricultural Engineering International: The
CIGR E-Journal. Vol. x: 1-9.
Duff, O.G. and Ross, S.M. (1988). Adsorption on activated charcoal. J of chem. Educ. 65(3) 815
Ezeh, I.E., Umoren, S.A., Essien , E.E. and Udoh. A.P. (2012). Studies on the utilization of Hura Crepitans
L. seed oil in the preparation of alkyl resins, industrial crops and products 36:94-99,
Itodo, A. U., Abdulrahman, F. W., Hassan. L. G., Maigandi, S. A. and Itodo, H. U. (2010). Application of
Methylene blue and iodine Adsorption in the measurement of specific area by four acid and salt treated
activated carbons. New York Science Journal, 3(5): 25-33.
Joshi, S., Adhikari, M., Pokharel, B. P. and Pradhanga, R. R. (2013). Effect of Activating agents on the Activated
carbons prepared from Lapsi seed stone. Research Journal of Chemical Sciences, 3(5): 19-24.
Odebunmi, E. O. and Okeola, O. F. (2001). Preparation and characterization of activated carbon from waste
material. J. Chem. Soc. Nigeria, 26(2): 149-155.
Ogbonnaya, O. (1992). Preparation and comparative characterization of activated carbon from Nigerian sub-
225
bituminous coal, palm kernel and cow bone. J. Chem. Soc. Nigerian, 17,11.
Okeola, O. F., Odebunmi, E. O. and Ameen, O. M. (2012). Comparison of Sorption capacity and surface area of
Activated carbon prepared from Jatropha curcas fruit pericarp and seed coat. Bull. Chem. Soc. Ethiop,
26(2) 171-180.
Olawale, A. S. and Ajayi, O. A. (2009). Thermal activation of Canarium Schweinfhurthi nutshell. Australian
Journal of Basic and Applied Sciences, 3(4): 3801-3807.
Tsai, W., Chang, C., Wang, Y., Chang C., Chien, F., Sun, H.(2001). Preparation of activated carbons from corn
cob catalyzed by potassium saltz and subsequent gasification with CO2. Bioresource technology 78(2) ;
203-208
Umoren. S.A., umoren, I. U. Johnson A.S.(2013). Metal soaps of Hura crepitans seed oil: potential stabilizer
for unplasticised polyvinyl chloride against thermal degradation. Journal of chemical Acta, 2:92-94
Vijayan, S. N., Makeshkumar, M. and Sridhar, K. (2012). Physical and chemical Analysis of Activated carbon
prepared from coconut shell charcoal and usage-A case study. International Journal of Advanced
Scientific Research and Technology, 2(3): 168-175.
226
PHYTOCHEMICAL AND ANTIMICROBIAL STUDIES OF Crossopteryx febrifuga LEAF EXTRACT AGAINST
METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS SKIN INFECTIONS
,1 2 2 3
Nma N. Y.* , Mann A. , Ndamitso M. M. and Muhammad B. M.
1
Departent of Science Laboratory Technology, Federal Polytechnic Bida
2
Department of Chemistry, Federal University of Technology Minna
3
Departent of Science Laboratory Technology, Niger State Polytechnic Zungeru
*Corresponding Author: ndakoyahayanma@gmail.com (08134196960)
Abstract
Crude extract of Crossopteryx febrifuga leaf, used traditionally for the treatment of wounds and skin infections
was phytochemically and antimicrobially screened .The phytoconstituents were qualitatively determined. The
crude methanol extract was partitioned into four soluble fractions namely n-hexane, dichloromethane,
ethylacetate and water fractions. These solvent soluble fractions were also phytochemically screened. Both
crude extract and its fractions were subjected to thin layer chromatography analysis as well as screened for
their Methicillin-resistant activity against Methicillin-Resistant Staphylococcus aureus(MRSA).The
phytochemical screening of the crude methanol extract and the solvent soluble fractions revealed the
presence of saponins, terpenes, flavonoids, steroids, tannins, carbohydrates, anthraquinones and
glycosides. The result of the antimicrobial screening of the crude methanol extract and solvent soluble
fractions showed potent activity against Methicillin-Resistant Staphylococcus aureus, however, the
ethylacetate fraction was the most active fraction against the studied microbe (160µg/cm3). GC-MS analysis
of the ethylacetate soluble fraction revealed fourteen constituents. Three major phytocompounds were
suggested to be 6-methylenebicyclo [3, 2, 0] hept-3-en-2-one (31.56%); 2, 6-dimethoxy- phenol (8.417%) and 2-
methoxy-4-vinylphenol (7.533%).
Key words: - Phytochemical, Antibacterial, Methicillin-Resistant Staphylococcus aureus, Crossopteryx

febrifuga, Thin-Layer Chromatography, GC-MS.
Introduction
The skin is the largest organ and the outer covering of the body which constitutes the first line of defense
[1].Skin diseases are mostly due to infections that have greater effects on most individuals especially in the
tropical areas [3]. Infectious skin diseases are caused by different microbes such as bacteria, fungi viruses and
parasites [4]. Methicillin is a narrow spectrum â-lactam antibiotic of the penicillin class that is used to treat
infections caused by gram positive Staphylococcus aureus, Staphylococcus pyogenes and Staphylococcus
epidermidis [6].Methicillin – resistant Staphylococcus aureus (MRSA) also known as super bug (a strain of
Staphylococcus bacteria) causes skin infections that is resistant to antibiotics commonly used to treat ordinary
Staphylococcus infections[8].Over the decades, strains of Staphylococcus have become resistant to some
antibiotics that once destroyed them.
Modern and novel approach to cure the increasing skin diseases is required and can be found in traditional
medicine. In this approach, plants (leaves, roots, barks, stem, fruits and seeds) or their combinations become
natural sources for tapping from the potentials of their phytochemical constituents [9]. These compounds
227
exhibited remarkable biological activities which include inhibitory, modulatory, protective effects against
allergies and as antioxidants [10].
Crossopteryx febrifuga (Rubiaceae) known as crystal bark in English is a deciduous savannah tree that is
traditionally used for treating many ailments in Nigeria and other West African countries such as Tanzania,
Zimbabwe, Ghana, Guinea and Malawi [11]. It is identified in different localities in Nigeria by different names, it
is called Kashinawaki in Hausa [12], Nambisunsun in Nupe, Ayeye in Yoruba [13] and Rima Jogohi in Fulani [13].It
is used as febrifuge for treating fever, dry coughs, gastrointestinal pains, skin diseases, swellings, diarrhea,
dysentery, gonorrhea, worms, cold, chicken pox and scabies in Nigeria, Ghana, Tanzania, Malawi, Sudan and
Zimbabwe [14]. It is widely used in Nupeland of Central Nigeria, for the treatment of wounds and skin infections
[15]. Previous pharmacological investigations of extracts of Crossopteryx febrifuga have been reported [16].A
number of phytoconstituents has been associated with its antimicrobial activity which includes alkaloids,
tannins and saponins [17]. This study aimed to investigate the effect of the crude extract and soluble fractions
of Crossopteryx febrifuga leaf on Methicillin-Resistant Staphylococcus aureus and identify the metabolites
associated with MRSA activity.

Sample Collection: - The leaves of Crossopteryx febrifuga (Afzel.ex.G.Don) Benth were collected along Bida –
Wuya, road in Niger State, Nigeria in May, 2015. The plant was properly identified and authenticated by Dr.
Abdullahi Mann of the Natural Product Unit, Chemistry Department Federal University of Technology, Minna.
Methods: - The phytochemical screenings of the crude and ethylacetate fraction of C. febrifuga were
carried out according to methods described by [18, 22]. The partitioning of the crude extract into four solvent
soluble fractions and column chromatography were determined by methods described by [19, 23] with slight
modifications and the TLC according to methods described by [20]. The antimicrobial activities were conducted
in Microbiology lab of the Federal University of Technology, Minna using methods described by [21] also with
slight modifications and the GC-MS analysis of the ethylacetate fraction was carried out in Chemical
Engineering Department of the University of Ilorin, Nigeria using a GC-MS machine (QP-2010; Shimadzu, India)
according to [24].
Table 1Phytochemical Screening of the Crude and the Ethylacetate Fraction
Secondary metabolites Tests Crude extract Ethylacetate fraction
1 Glycosides Fehling’s Test + +

2 Cardiac Glycosides (Keller-Killiani) _ +
3 Saponins Froth Test + +
4 Tannins Lead Acetate + +
Ferric Chloride + +
5 Flavonoids Shinoda + +
Sodium + +
hydroxide
6 Carbohydrates Molisch + -
Fehling’s + -
7 Steroids / Triterpenes Lieberman- + +
Burchard
Salkowski + +
8 Alkaloids Mayer’s - -
Wagner’s - -
Dragendorff’s - -
9. Anthraquinones Burntrager + +
228
Table 4.1Result of Antimicrobial Screening of the Crude Extract
Microorganism Crude Extract Concentration Concentration Of Standard
(mg/cm3) (mg/cm3)
MRSA
Crossopteryxfebrifuga Beecham Ampiclox
Zone of Growth 40 80 120 160 200 80
Inhibition
- - - 12 mm 18 mm 21 mm
Table 4.2Result of Antimicrobial Screening of the Solvent Soluble Fractions
Fractions (mg/cm3) 40 80 120 160 200
n-Hexane - - - 16 mm 19 mm
Dichloromethane - - - - 17 mm
Ethylacetate - - - 18 mm 21 mm
Aqueous - - 12 mm 16 mm 18 mm
Table 4.3Result of MIC and MBC of the Fractions

Fractions MIC ( mg/cm3 ) MBC ( mg/cm3 )
n-Hexane 160 200
Dichloromethane 200 200
Ethylacetate 160 160
Aqueous 120 160
Standard (Ampiclox) 80 80
Table 5: Result of GC-MS Showing Molecular Masses, Percentage Areas and Major Peaks of Some of the
Constituents of the Ethylacetate Soluble Fraction
Line Compound Mol. Mass Area (%) Peaks (m/z)
4 6 – methylenebicyclo 120 31.56 (120), 110, 91

[3, 2, 0] hept-3-en-2-one 64
5 Phenol 94 29.507 (94), 95, 96, 66,

65.
5 2-methoxy-4-vinylphenol 150 7.533 (150), 135, 107,
77, 51.
4 Benzylmethanol 108 4.559 (108), 97, 77
51.
3 Phenol, 1, 2-methoxy 124 6.308 (124), 109, 81
229
Discussion of Result: - The healing properties of medicinal plants were well known in the past and
nowadays scientifically proving due to their phytochemical constituents which form the basis of their
antimicrobial activities [26]. The result of phytochemical screening of the crude and ethylacetate fraction of
the Crossopteryx febrifuga leaves are presented in table 1. The result shows the presents of flavonoids,
tannins, saponins, anthraquinones and steroids similar to the report by [27]. These phytochemicals may be
responsible for the antimicrobial activity against Methicillin resistant Staphylococcus aureus (MRSA)
infections [28]. Reported that flavonoids, tannins and steroids found in the methanolic extracts of neem,
tulsi and aloe vera are the major active compounds against MRSA infections which also agreed closely with
the report by [29] on the antimicrobial activities of Rose and Oregano oils against MRSA and Methicillin
sensitive staphylococcus aureus (MSSA). Flavonoids are known for their anti-inflammatory and anti-oxidant
properties which inhibit the synthesis of nucleic acids in microbes, steroids are known for their anti-
inflammatory properties and tannins are known for their ability to precipitate amino acids and proteins
[30].
The result of the crude partitioning and their percentage yield in various solvents are presented in table 2. The
n-hexane, dichloromethane, ethylacetate and aqueous fractions yield 12.6g (2.52%); 10.2g (2.04%); 15.3g
(3.16%) and 15.8g (3.16%) respectively. The highest percentage yield of water may be attributed to it higher
polarity which enables it to dissolve most fractions in the crude while the least yield of dichloromethane
fraction may be due to its low polarity. The fractions shows different colors which may be attributed to their
different bioactive constituents.
The result of the Column Chromatography which was monitored by Thin-Layer Chromatography (TLC) analysis
is presented in table 3. The retention factors (Rf) and the number of spots present in each fraction with their
distinct colors indicate their active components. The n-hexane fraction presents two spots with Rfs of 0.89 and
0.31; the dichloromethane present three stops with Rfs 0.35, 0.52 and 0.95 while the ethylacetate has four spot
with Rfs 0.47, 0.53, 0.73 and 0.76 and the aqueous fraction presents two spots with Rfs 0.35 and 0.77
respectively. These are indications that the ethylacetate fraction has the highest of bioactive compounds.
The results of the antimicrobial activities of the crude and the solvents soluble fraction are presented in table
4.1 – 4.3. The MIC (160mg/cm3) and MBC (200mg/cm3) o the crude extract where close to 150mg/cm3 and
3
190mg/cm reported by [13] for methanolic crude extract of root bark of C. febrifuga while the MIC of the
3 3 3 3
fraction is in the range of 120mg/cm – 200mg/cm and MBC is in the range of 160mg/cm – 200mg/cm . Out of
the four fractions, the ethylacetate has the largest zone of growth inhibition (18mm - 21mm), these may be
attributed to the largest number of its TLC spots. These results are similar to those obtained from the ether
extract of Thyme vulgaris against multi-resistant Salmonella typhimurium and MRSA isolates reported by [31]
3 3 3
and by [32] for the ethanolic extract of neem (160mg/cm ); tulsi (180mg/cm ) and aloe vera (150mg/cm )
against MRSA respectively. However, these values are lower than that reported by [33] for the ethanolic extract
of lemon grass (512mg/cm3) againstthis resistant species of bacterial.
The result of GC-MS analysis of the ethylacetate fraction, major peaks, percentage areas and the suggested
fragments some of the major compound suggested by the GC-MS machine library are presented in table 5. The
result shows some of the major bioactive compounds, their percentage areas and major peaks present in the
fraction. Some of these bioactive compounds are similar to those reported by [30] to be responsible for the
antimicrobial activities of neem, tulsi and aloe vera and also for those reported by [34] to be present in Oregano
and Rose mary oils against MRSA.
Conclusion: - It is concluded that this research work has been able to scientifically prove the ethnobotanical
uses of Crossopteryx febrifuga leaf for the treatment of various Methicillin-resistant Staphylococcus aureus
skin infections.
230
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__
Key: + = Present = Absent
Table 2Weight and Percentage Yield of Various Fractions

Solvent Colour of Fraction Mass of Fraction (g) (%) Yield
n-Hexane Oily pale Green 2.6 2.52
Dichloromethane Dark Green 10.2 2.04
Ethylacetate Brownish Yellow 15.3 3.06
Aqueous Brown 15.8 3.16
Table 3 Result of Thin Layer Chromatography (TLC) of the Fractions
232
Fractions Number of Components Colour of Components Rf Values
n-Hexane fraction 2 Brown 0.89
Pale Yellow 0.31
Dichloromethane 3 Pale Yellow 0.35
Green 0.52
Brown 0.95
Ethyl acetate 4 Brown 0.47
Green 0.73
Yellow 0.53
Purple 0.76
Aqueous 2 Pale Yellow 0.35
Brown 0.77
233
Antibacterial And Antioxidant Activity of Justicia Spicigera Extracts: Activity Enhancement by
addition of Metal Salts.
Ohenhen O. N1., Njoku P. C2., Igara C. E3.
1
Department of Chemistry, Michael Okpara University of Agriculture, Umudike.
2
Department of Chemistry, Alvan Ikoku Federal College of Education, Owerri.
3
Department of Science Laboratory Tech., Akanu Ibiam Federal Polytechnic, Unwana, Afikpo.
E-mail: offinel2012@gmail.com
ABSTRACT
Justicia spicigera was screened for its phytochemicals. The results showed the presence of alkaloids, flavonoids,
tannins, phenols, saponins and steroids. Extracts of Justicia spicigera prepared with solvents of different
polarities (n-Hexane, Chloroform, Methanol, and Ethyl acetate) was used to analyze the antibacterial properties
in the absence and in the presence of metal salts. The antioxidant activity was analyzed using 2, 2-diphenyl-1-
picrylhydrazyl reagent (DPPH). Quantitative determination of total phenolics was carried out using
spectrophotometric method. The results indicate that the tested bacterial strains were most sensitive to the n-
Hexane extract but moderately sensitive to the chloroform and methanol extracts. The chloroform, methanol,
and ethyl acetate extracts showed potent radical scavenging activities. The ethyl acetate and methanol extracts
showed higher total phenolic content than the other extracts. Antioxidant activity was found in the extract-Fe3+
2+
ion combination. The extract-Zn ion combination showed enhanced antibacterial activity against tested
bacterial strains compared to the extract alone. The results scientifically establish the efficacy of using the plant
extracts and its metal salt combination as antibacterial and antioxidant agents.
Key words: Justicia spicigera, total phenolic content, antibacterial activity, antioxidant.
INTRODUCTION:
Justicia spicigera a shrub from the Acanthaceae family has been found to produce biologically active species
that are responsible for the plant therapeutic properties (Dominiguez et al., 1990; Euler et al., 1982). It is
commonly referred to as firecracker plant due to their narrow red flowers, mutile in Mexico and Iyip abasi by the
Efiks in Nigeria. J. spicigera has been used in traditional medicine for many years as a stimulant, for
gastrointestinal disorders, diabetes, inflammations, blood depurative treatment, uterine cancer, skin and kidney
infections (Marquez et al., 1999). The infusion of Justicia spicigera has characteristic red color.
Currently there is much interest in the protection of low density lipoprotein, important cells and organs as well as
food systems against oxidative damage caused by superoxide, hydroxyl and peroxyl radicals. This has led to an
increasing demand to evaluate the antioxidant properties of direct plant extracts or isolated products from plant
origin rather than looking for synthetic ones. Antioxidants may guard against reactive oxygen specie (ROS)
toxicities through the prevention of ROS construction by the prevention of ROS attack, scavenging reactive
metabolites and converting them to less reactive molecules or by enhancing the resistance of sensitive biological
targets to ROS attack (Sen, 1995). Free radicals, reactive oxygen species (ROS), and reactive nitrogen species
(RNS) are associated with many pathological conditions such as atherosclerosis, arthritis, ischemia, reperfusion,
injury of many tissues, central nervous system, gastritis, cancer and AIDS (Cook et al., 1996; Kumpluainen et al.,
1999). The anti-oxidative properties of phenolics are due to mechanisms such as scavenging of free radicals,
chelation of metal ions, and inhibition of enzymes responsible for free radical generation (Akinmoladun et al.,
2007).
Several studies have been made in order to determine the antioxidant properties of plants, especially those used
in traditional medicine (Jang et al., 2007; Surveswaran et al., 2007). This study is in support of other studies that
have been made to replace synthetic antioxidants that are carcinogenic with natural antioxidants from natural
sources.
234
MATERIALS AND METHODS.
The plant samples were collected and identified by comparing with the leaves inflorescence in standard text
(Akobondu and Agyakwa, 1998); it was further authenticated by Prof. Nmeregini of Forestry Dept., MOUAU.
Extraction: The extraction was performed according to standard method of (AOAC, 1995). The Methanol
extract was extracted using 80% methanol and labeled ME, the Ethyl acetate extract was prepared with Ethyl
acetate and labeled (EAE), Chloroform extract labeled CE and n-Hexane extract labeled (n-HE).
Phytochemical Determination: The Phytochemicals determination was done according to the standard method
by Harborne et al. (1998).
Determination of Total Phenolic content: The total phenolic content was measured by spectrophotometric
determination with a modified Folin-Ciocalteau method Tuberoso et al. (2007) using Gallic acid as a standard
phenolic compound. The total phenolic content was expressed as Gallic acid equivalent (GAE) in g/100g of dry
wt. of extract. Mixture of water and reagent was used as blank. Sample measurement was done in triplicates and
the mean and standard deviations calculated in each case.
DPPH Free radical scavenging activity: The free radical scavenging activity of the plant extracts and extracts-
metal ions was evaluated by the stable radical DPPH (2, 2-diphenyl-1picrylhydrazyl) reagent described by
Miliauskas et al. (2004). Radical scavenging potential was expressed as inhibiting conc. IC50 value. This
represents the sample conc. at 50% of the DPPH radicals scavenged. The assays were done in triplicate and the
results were expressed as mean values ± standard deviations.
Antibacterial Screening: Extracts of the leaves 1g was dissolved in 1ml of dimethyl sulphoxide (DMSO) to get
a concentration of 100mg/ml. This was used as stock solutions for the antimicrobial assay (Okigbo and
Omodamiro, 2006).
Four representative test microorganisms were used in testing the antimicrobial activity of the leaves extracts of
Justicia spicigera– n-Hexane (n-HE), Chloroform (CE), Methanol (ME) and Ethyl Acetate (EAE). The test
microorganisms used were Streptococcus pneumonia, Salmonella typhi,Escherichia coli and Aspergilus niger.
The test cultures were originally obtained from Federal Medical Centre (FMC) Umuahia, Abia State, Nigeria and
were maintained in nutrient agar.
The antimicrobial test used the filter paper disc diffusion method (Anderson, 1974). Ciprofloxacin drug 0.1g
dissolved in 1ml of distilled water was used on a separate disc. This served as a standard while the disc with
DMSO (100mg/ml) served as the control. The mean was taken as the diameter zone of inhibition. To increase the
synergistic effect the metal salts Fe(III) and Zn(II) ions was combined with the extracts for the antibacterial tests.
Aliquot containing solutions with a constant concentration of extract 10equivalent and freshly prepared metal
salts ZnSO4, FeCl3 solution with a stoichiometric equivalent (1-5eq) were added to give a final extract-metal ion
ratio of 10:01, 10:02, 10:03, 10:04, and 10:05. The obtained product was filtered and collected. The experiment
was repeated three times for each extract and the means of these values were recorded.
RESULTS:
Table 1: Total Phenolic content of Justicia spicigera
Extrac t Total Phenolic content (g GAE per 100g) dry wt. of extract
n - HE 3.28±0.01
EAE. 5.65±0.02
CE 3 99±0.02
ME 4.54±0.04
235
- Data are means± standard deviations from two extraction replicates
Table 2: Antioxidant activity of J. spicigera with and without metal ion
Sampl e DPPH (IC 50 )µg/m l
n - Hexane 34.18±0.50
3+
n-Hexane-Fe 34.01±0.15
2+ 36.73±0.80
n-Hexane-Zn
EAE 13.73±0.21
EAE-Fe3+ 11.64±0.18
EAE-Zn
2+ 12.68±0.39
ME 18.71±0.74
ME-Fe
3+ 18.01±0.12
ME- Zn2+ 18.50±0.19
14.94±0.16
CE 13.28±0.51
CE-Fe3+ 14.57±0.13
CE-Zn2+
14.94±0.30
AA (Ascorbic acid)
Data are means± standard deviations from two extraction replicates
Fig 1: Inhibition zone (mm) of extracts against four bacterial strains with standard drug (S=
Ciprofloxacin).
3+
Fig 2: Inhibition Zone (mm) of extract-Fe combination against four bacterial strains at concentrations
10:04 and 10:05.
236
Figure 3:Inhibition Zone (mm) of extract-Zn2+ combination against four bacterial strains at concentrations 10:04
and 10:05.
Discussion:
The phytochemical screening done on all the plant extracts revealed the presence of alkaloids, phenolics,
flavonoids in all the samples except n- Hexane. Steroids were present in all the extracts but more in the ethyl
acetate extract. Tannins and saponins were found in methanol and ethyl acetate extracts. The results of the total
phenolic content of Justicia spicigera is shown in table 1.There is a positive correlation between the total
phenolic content and antioxidant activity of Justicia spicigera with that reported for other plant species of
Acanthaceae (Sawadogo et al., 2006; Jang et al., 2007). The total phenolic content of Justicia spicigera ranged
from 3.28±0.01 to 5.65±0.02g GAE per 100g dry wt. of extractwith methanol extract richer in phenols than the
other extracts. The results of CE, ME, EAE were found to be significant at P<0.05 but not significant for n-
Hexane extract at P>0.05 when compared to the blank.
The results of the DPPH radical scavenging activity of the extracts and ascorbic acid are presented in Table 2. The
results show that the extracts exhibited a concentration dependent anti radical activity and the lower IC50 values
are indicative of a higher anti radical activity. At low concentrations (10:01,10:02) the extract-metal ion
combination showed results comparable to the extract alone but a discernable effect was observed of the free
radical scavenging activity at concentration 10:03. ME-Iron(III) exhibited highest percent inhibition of DPPH
radical as compared to other fractions. CE-Iron(III) ion and EAE-Iron(III) ion exhibited modest scavenging
activity than the extracts alone. The results of the radical scavenging activity for the chloroform extract,
methanol extract and ethyl acetate extract were found to be significant at P<0.05 but not significant for n-Hexane
at P>0.05 when compared to the reference standard ascorbic acid. The extracts-Zn(II) ions did not show
significant improvement in radical scavenging activity.
The antibacterial result of the extracts in the absence of metal salts is shown in figure 1. The results indicate that
the tested bacterial strains were sensitive to all the extracts. In comparison the extracts were most sensitive to the
n-Hexane extract, moderately sensitive to the chloroform and methanol extracts least sensitive to the ethyl
3+
acetate extract. The antibacterial result of extracts with metal salts at different concentrations with Fe is shown
in figure 2,the antibacterialresults for extracts with metal salts at different concentrations with Zn2+is shown
infigure 3. The results indicate that the extract-metal salt combination enhanced the antibacterial activity than the
crude extracts alone. The extract-metal salt combination possessed stronger antibacterial activity against Gram
237
+ve bacteria as compared to the Gram -ve bacteria. This may be because gram -ve bacteria have been reported to
be more resistant than gram +ve bacteria (Russel, 1991). At lower conc. of extract-metal salt combination (10:01;
10:02, 10:03 ratio) no recordable growth was observed. Further increase in metal ion salt conc. to (10:04, 10:05)
had significant effect but further increase above that resulted in almost same growth for all tested bacteria. This
confirms the study by Simon et al., (2009) which showed that antibacterial properties of natural products can be
enhanced by the addition of metal ions due to positive participation of extracts in coordination with the metal
ions.
Conclusion:
Many plant species are currently used as sources of nutritional additives because of their wide antibacterial and
antioxidant properties that increase immunity to some diseases. This study has looked at the biological
evaluation of the various extracts of Justicia spicigera and the enhanced synergistic activity by the addition of
metal salts. The results suggest that combination of Fe(III) ion with EAE, CE and ME could be considered as a
new potential source of natural phenolic antioxidant for food, pharmaceutical, and cosmetic industries. The
considerable antibacterial activity of n-HE-Zn(II) ion suggests that Justicia spicigeracan serve as selective agent
for the maintenance of human health.
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239
Comparison of Heavy Metals Pb, Cu, Ni, Zn and Al in Human Scalp Hair: A Case Study of Port-
Harcourt and Okigwe
*Ngwu, Comfort M and Onwu, Francis K.
Department of Chemistry, Michael Okpara University of Agriculture Umudike, P.M.B 7267 Umuahia, Abia
State, Nigeria.
ABSTRACT
The proliferation of industries has led to increased presence of heavy metals in our environment and, humans are
continually exposed to these heavy metals through their engagements in different industrial activities. The
current study is designed to make a comparative assessment of the level of heavy metals, Pb, Cu, Ni, Zn, Al in the
hair scalp of humans living in industrialized and unindustrialized cities of Port Harcourt and Okigwe
respectively. Male hair samples were randomly collected and analysed for their trace metals contents by atomic
absorption spectrophotometry (AAS). Higher geometric mean values for Zn, Ni and Pb were obtained in hair
samples of donors from Port Harcourt as compared to that of Okigwe which rather recorded high geometric mean
values for Cu and Al. It was observed that there was a significant difference in the mean concentrations of Pb
between Port Harcourt and Okigwe, with Port Harcourt having a very high concentration of 513.5 mg/kg while
no significant differences were observed in the mean concentrations of the other metals between these two cities
studied. For the general population studied, geometric mean values of 6.4 mg/kg, 118.9 mg/kg, 4.2 mg/kg, 35.9
mg/kg and 45.7 mg/kg were obtained for the metals Cu, Zn, Ni, Pb and Al respectively. Results showed that the
heavy metals found in human hair of the people living in the two cities were dependent mainly on environmental
factors, occupation, lifestyle and feeding habits.
Key Words: heavy metals, scalp hair, industrialized and unindustrialized area, environmental factors,
occupation, lifestyle.
Introduction:
Our bodies are gradually turning to dumping ground for thousands of toxic compounds that invade our air, water
and even the soil in which our food grows. It is estimated that our bodies cannot metabolize over 20000 of these
chemicals [1]. Some metals are naturally found in the body and are essential to human health, though they
normally occur at low concentrations and are known as trace metals [2]. Heavy metal poisoning is the toxic
accumulation of heavy metals in the soft tissues of the body. Hair analysis is one way of detecting heavy metal
poisoning in humans. Studies indicate that levels of potentially toxic elements in human hair correlates with
blood levels and body tissue deposition levels [3]. Hair is a site of excretion of essential, nonessential and
potentially toxic elements. Hair is also an excellent biopsy material. It is easy to sample, preserve and transport.
Hair has the advantage of long term memory. It can keep a “permanent record like tree rings” [4]. Therefore, hair
tissue mineral analysis (HTMA) is an analytical test which measures the presence of heavy metals, toxins, the
nutritional content of hair and the balance between the nutrient level and heavy metal toxin [4]. In this study, a
comparative assessment of heavy metals content in hair scalp of humans in two cities: PortHarcourt and Okigwe
was made. The two cities were chosen so as to make a comparison of the concentration of these metals among
two categories of people; those living in a densely populated industrialized city and those who live in an
unindustrialized city.
Sample Collection, Preservation and Treatment
The class of people selected for this study were the male population from two localities viz; PortHarcourt and
Okigwe and were of varying ages ranging from 10 – 40 years old. Male hair samples were chosen, since they
maintain a low haircut, and do not treat their hair like women. A primary data was used, since data was directly
gotten from donors through the assistance of barbers. A questionnaire was used as a research instrument, and
240
information on age, occupation and residence were obtained from donors. 10 samples were randomly collected
from youths and adults in PortHarcourt and the other 10 from youths and adults in Okigwe, so as to compare the
extent of exposure in these geographical areas.
A minimum of 2 g per sample of hair was collected from each individual. This was gotten from the occipital
region of the scalp. Sampling near the face was avoided due to increased likelihood of contamination from
sebaceous secretions and facial hygiene products. Hairs that have been bleached or dyed were not collected.
The hairs were washed with acetone and afterward washed with deionized water. Acetone is an effective fat
solvent and, partly a polar molecule, having the capacity to dissolve both fat soluble and water soluble elements
attached to the hair strands following external contamination.
The hair samples were further cut into lengths of approximately 0.5cm, soaked and washed in 2% nonionic
surfactant (persil) and rinsed several times. The hair samples were then soaked and washed in acetone to remove
oil and grease. Each washing lasted approximately 10 minutes with continuous stirring. The samples were dried
in electric oven at 60oC for 4 h. After drying, samples were left in a covered watch glass till all samples were ready
for treatment.
Sample digestion and Chemical Analysis
Exactly 0.5 g of the sample was weighed out for digestion. Wet digestion was used and the sample was digested in
a digestion beaker with 10 ml concentrated HNO3 (AnalaR grade) and 5 ml of 80 % (v/v) H2O2 at room
o
temperature to prevent foaming. The beaker was then heated up to 120 C till the content became amber as the hair
decomposed. Heating was continued for some time to allow the content of the beaker to evaporate to dryness.
The content was further treated with two portions of the HNO3/H2O2 and evaporated to near dryness. This was
necessary in order to oxidize the hair completely to give an almost clear to faint amber solution. This whole
process was done in a fume cupboard with the aid of a hot plate and a magnetic stirrer. The content of the beaker
was taken up in a prepared 1M HNO3 and the solution filtered through a pre-acid washed filter paper , collected in
a 10 ml volumetric flask, and the filtrate made up to volume with the 1M HNO3. The filtrate was then analyzed
using flame atomic absorption spectrophotometer. The same procedure was followed for the entire sample.
Chemical analysis was done for the heavy metals Cu, Al, Zn, Pb and Ni using flame atomic absorption
spectrophotometer (unicam 966 model).
Data Analysis Technique
The statistical parameter for comparison of the heavy metal concentrations between both cities was the student's
t-distribution test, at 0.05 confidence level (i.e., level of significance). T-test was used because it tries to confirm
if there is a significant difference in the level of heavy metal concentrations in both areas. Kari Pearson's product
moment of coefficient of correlation r was also used. The geometric mean values of the five metals were
determined. This is because it is more appropriate to report results of studies such as this using geometric mean
value, as the results are usually positively skewed and close to the normal log [5].
Results and Discussions
From the results, it was observed that a distinct pattern of distribution was observed for Pb and Al (Tables 4 and 5)
in Okigwe area. The order of increasing level for Pb and Al are the same i.e. 10 – 20 years > 21 – 30 Years > 31 –
40 Years while that for Cu follows the trend 10 – 20 years > 31 – 40 years > 21 – 30 Years as seen in Table 1. The
highest geometric mean values obtained for heavy metals in hair samples of those in age limit 10 – 20 years could
be attributed to feeding habits, lifestyle and environmental exposure. The highest geometric mean values were
gotten for Cu, Zn, Ni and Pb (Tables 6 – 9) in age limit 21 – 30 years while the highest geometric mean value was
obtained for Al in age limit 31 – 40 years (Table 10). The factors outlined above, could also be the reasons for the
high geometric mean values obtained in the different age limits.
A total hair concentration of Cu is seen to be higher in Okigwe than in Port Harcourt (Table 11) with a
241
total concentration of 70.8 mg/kg and a geometric mean of 6.9 mg/kg against 67.1 mg/kg and 6.0 mg/kg
respectively.
However, from the t-test conducted (Table 16), it is observed that there is no significant difference in the mean
concentrations of copper between both cities. But the high concentration of copper in Okigwe may be attributed
to the consumption of contaminated food or drinking water and also due to contamination of hair. In a similar
study in Sweden [6], a large proportion of young children obtained their daily requirements of copper solely from
drinking water due to copper in water pipes. According to age range, 21- 30 years of age had the highest copper
concentration, which could be due to accumulation of the metal from similar sources. And the order of increasing
level is 21- 30 years > 31- 40 years > 10 – 20 years.
Zinc concentration was the highest in all hair samples (Table 12). This could be due to the type of
treatment given to hair, or due to feeding habit. This is because zinc being an essential element tends to be high in
concentration in the human body. For the general population studied, the overall geometric mean value of 118.9
mg/kg obtained for zinc is similar to that obtained from a similar study in south-eastern Nigeria (146.2 mg/kg).
Very high total zinc concentration in Port Harcourt (1202.80 mg/kg) is slightly higher than that in Okigwe
(1189.9 mg/kg). The Zinc ratio between geometric mean values in both cities is 1.02. From the t-test conducted
(Table 17), no significant difference was seen in the mean concentrations of Zn between Port Harcourt and
Okigwe. Male population ranging from 21 – 30 years had the highest mean concentration of Zinc in their hair
compared to other age limits.
The hair concentration of nickel was observed to be higher in Port Harcourt than in Okigwe (Table 13), with a
total concentration of 105.0 mg/kg and a geometric mean of 3.6 mg/kg. Those within the range of ages 21- 30,
showed the highest geometric mean of 9.3 mg/kg compared to other age ranges for Ni. The high concentration of
nickel observed from Port Harcourt could be attributed to the consumption of food cooked with vegetable oil and
margarine. These cooked or fried food items are usually prepared with vegetable oil manufactured through the
hydrogenation of palm kernel oil (PKO) using Ni catalyst. A similar study in Pakistan made a similar observation
[7]. The ease of affordability of the hydrogenated oil in the industrialized area (PortHarcourt) could also be a
cause. Most families in Okigwe may prefer the readily available palm oil or palm kernel oil. Therefore, a
difference in cooking and eating habits could explain the Ni content in the hair samples from both cities studied.
Other reasons could be attributable to the consumption of chocolate, or food grown near industrialized areas, or
even those residing close to solid waste incinerators. However, the t-test in Table 18 showed no significant
difference in the mean concentrations of Ni between Port Harcourt and Okigwe.
From Table 14, a higher Pb concentration of 513.5 mg/kg was obtained for hair samples from Port
Harcourt compared to 296.9 mg/kg for samples from Okigwe. A distinct pattern of distribution was observed for
Pb for the age groups. The order of increasing level of Pb in the hair samples in Port Harcourt and Okigwe is 10 –
20 > 21 – 30 > 31 – 40. From Table 19, the t-test conducted showed a significant difference in the mean
concentration of Pb between both areas. High level of Pb observed in the hair samples from Port Harcourt being
an industrialized area may be primarily due to the deposition of Pb particulates from automobile exhaust fumes
on human hair. Occupational exposure and contaminated drinking water could also be other causes. This is
because acid and soft water may dissolve Pb from lead pipes or lead soldered copper tubes, thereby
contaminating the drinking water.
Aluminium hair concentration was observed to be higher in Okigwe (Table 15) with a total
concentration of 517.9 mg/kg and geometric mean of 49.2 mg/kg in Okigwe compared to 438.1 mg/kg and
geometric mean of 42.5 mg/kg in samples from Port Harcourt. A similar distinct pattern observed in the age range
for Pb was also observed in Al, where the order of increasing levels of Al in hair samples from Port Harcourt and
Okigwe is 10 – 20 > 21 – 30 >31 – 40 . Table 20 Shows from the t-test that there is no significant difference in the
242
mean concentration of Al between Port Harcourt and Okigwe. However, the high concentration of Al may be
attributed mainly to drinking water, intake of Al-containing drugs, and the application of Al-containing
deodorants, increased Al intake through juices and soft drinks has also been reported in Spain [8]. An overall
range of 21.8 – 81.1 mg/kg was obtained compared to 0.2 – 21.4 µg/g of that obtained from a similar study in
Sweden [6].
For the whole sampling material, strong positive correlations were observed for Cu – Zn
Age Limit No Range Mean (mg/kg) Geometric Mean Std. Dev
(mg/kg) (mg/kg)
10 – 20 3 6.6 – 7.7 7.2 7.2 7.1
21 – 30 4 4.8 – 8.7 7.0 6.8 1.5
31 – 40 3 5.2 – 9.8 7.1 6.8 2.3
Table 2Zn Content in hair samples from Okigwe
Age limit No Range(mg/kg) Mean(mg/kg) Geometric Mean Std. Dev

(mg/kg)
10 – 20 3 113.9 – 116.2 115.3 115.3 3.6
21 – 30 4 112.7 – 140.5 123.3 122.8 11.3
31 – 40 3 106.8 – 125.5 116.9 116.7 10.0
Table 3 Ni Content in hair samples from Okigwe

(mg/kg)
10 – 20 3 3.1 – 4.7 3.8 3.7 0.6
21 – 30 4 1.3 – 19.8 7.0 4.2 8.6
31 – 40 3 2.5 – 3.0 2.8 2.8 0.6
Table 4 Pb Content in hair samples from Okigwe

(mg/kg)
10 – 20 3 16.7 – 70.4 39.5 33.3 27.7
21 – 30 4 23.5 – 36.3 29.6 29.0 7.0
31 – 40 3 11.4 – 35.9 20.0 17.4 13.8
Table 5 Al Content in hair samples from Okigwe
243
(mg/kg)
10 – 20 3 53.5 – 81.1 63.0 61.8 15.9
21 – 30 4 35.1- 66.4 50.4 49.1 12.9
31 – 40 3 21.8 – 58.0 42.4 39.1 18.5
Table 6 Cu Content in hair samples from PortHarcourt

(mg/kg)
10 – 20 3 1.9 – 8.4 5.7 5.0 2.7
21 – 30 4 4.3 – 11.7 8.9 8.1 4.0
31 – 40 3 4.9 – 7.0 6.0 5.9 1.0
Table 7 Zn Content in hair samples from PortHarcourt
(mg/kg)
10 – 20 3 94.8 – 152.8 119.6 118.2 21.4
21 – 30 4 116.2 – 148.2 128.7 128.0 17.5
31 – 40 3 100.52 – 109.0 108.6 11.0
117.4
Table 8 Ni Content in hair samples from PortHarcourt
(mg/kg)
10 – 20 3 1.4 – 22.2 6.2 3.4 9.0
21 – 30 4 2.9 – 30.1 12.3 7.0 15.4
31 – 40 3 1.3 – 35.6 18.5 6.8 24.2
Table 9 Pb Content in hair samples from PortHarcourt

(mg/kg)
10 – 20 3 44.5 – 57.0 47.8 47.6 5.0
21 – 30 4 41.0 – 91.4 63.1 58.7 25.7
31 – 40 3 34.7 – 50.7 42.7 41.9 11.3
Table 10 Al Content in hair samples from PortHarcourt
244
(mg/kg)
10 – 20 3 29.7 – 60.4 42.1 41.0 11.2
21 – 30 4 39.1 – 57.5 43.3 41.9 13.4
31 – 40 3 37.7 – 60.0 48.9 47.6 15.5
Table 11 Total Copper Content in hair for the two cities
Sample No Range(mg/kg) Mean(mg/kg) Geometric Mean Std. Dev
(mg/kg)
Overall 20 1.9 – 11.7 6.9 6.4 2.3
result
PortHarcourt 10 1.9 – 11.7 6.7 6.0 3.1
Okigwe 10 4.8 - 9.8 7.1 6.9 1.4
Age 10 – 20 8 1.9 – 8.4 6.2 5.8 2.4
Age 21 – 30 7 4.3 – 11.7 7.8 7.4 2.8
Age 31 – 40 5 4.9 – 9.8 6.8 6.4 2.5
Table 12 Total zinc Content in hair for the two cities
(mg/kg)
Overall 20 94.8 – 152.8 119.6 118.9 14.0
result
PortHarcourt 10 94.8 - 152.8 120.2 119.0 18.4
Okigwe 10 106.8 – 140.5 119.0 118.7 8.8
Age 10 – 20 8 94.8 – 152.8 118.0 117.1 16.3
Age 21 – 30 7 112.7 – 148.2 125.6 125.0 13.3
Age 31 – 40 5 100.5 – 125.5 113.7 113.4 10.5
Table 13 Total Nickel Content in hair for the two cities

(mg/kg)
Overall 20 1.3 – 35.6 7.6 4.2 10.4
result
P.H 10 1.3 – 35.6 10.5 4.9 13.4
Okigwe 10 1.3 – 19.8 4.8 3.6 5.3
Age 10 – 20 8 1.4 – 22.2 5.3 3.5 6.9
Age 21 – 30 7 1.3 – 35.6 9.3 5.2 11.1
Age 31 – 40 5 1.3 – 35.6 9.1 4.0 14.8
Table 14 Total Lead Content in hair for the two cities
245
(mg/kg)
Overall 20 11.4 – 91.4 40.5 35.9 19.5
result
P.H 10 34.7 – 91.4 51.4 49.7 15.4
Okigwe 10 11.4 – 70.4 29.7 25.9 17.1
Age 10 – 20 8 16.7 – 70.4 44.6 41.6 16.1
Age 21 – 30 7 23.5 – 91.4 44.0 39.5 23.7
Age 31 – 40 5 11.4 – 50.7 19.4 24.7 29.5
Table 15 Total Aluminium Content in hair for the two cities
(mg/kg)
Overall 20 21.8 – 81.1 47.8 45.7 14.2
result
P.H 10 29.7 – 60.4 43.8 42.5 11.5
Okigwe 10 21.8 – 81.1 51.8 49.2 16.1
Age 10 – 20 8 29.7 – 81.1 49.9 47.8 16.3
Age 21 – 30 7 35.1 – 66.4 47.4 45.9 12.4
Age 31 – 40 5 21.8 – 6.0 45.0 42.3 15.6
Table 16 T- test Comparison of mean Concentrations of Cu between PortHarcourt and Okigwe.
No ∑x X S.D Cal. T-value Decision at PL.05

Zn Conc. In 10 67.1 6.7 3.1 -0.37 No significant
samples from difference
PH
Cu conc. In 10 70.8 7.1 1.4
samples from
Okigwe
Table 17 T- test Comparison of mean Concentrations of Zn between PortHarcourt and Okigwe.
246
Zn Conc. In 10 1202 120.2 18.4 0.185 No significant
PH
Cu conc. In 10 1190 119.0 8.8
samples from
Okigwe
Table 18 T- test Comparison of mean Concentrations of Ni between PortHarcourt and Okigwe.
Zn Conc. In 10 105.0 10.5 13.4 1.524 No significant
PH
Cu conc. In 10 47.8 4.8 5.3
samples from
Okigwe
Table 19 T- test Comparison of mean Concentrations of Pb between PortHarcourt and Okigwe.

Zn Conc. In 10 514 51.4 15.4 2.958 No significant
PH
Cu conc. In 10 297 29.7 17.1
samples from
Okigwe
Table 20 T- test Comparison of mean Concentrations of Al between PortHarcourt and Okigwe.
Zn Conc. In 10 438 43.8 11.5 -1.270 No significant
PH
Cu conc. In 10 518 51.8 16.1
samples from
Okigwe
247
CONCLUSION
This study was primarily intended to compare the level of heavy metal concentrations in human scalp hair
between Port Harcourt and Okigwe. Five metals were studied which include; Cu, Zn, Ni, Al, and Pb. Hair
samples were randomly collected from the male population from both cities under study. There was no
significant difference in the mean concentration of Cu, Zn, Ni, and Al between Port Harcourt and Okigwe, when
tested at 0.05 level of significance. A significant difference was observed in the mean concentration of Pb
between Port Harcourt and Okigwe. Higher concentrations and geometric mean values for Zn, Ni, and Pb were
observed in the hair samples from Port Harcourt, while higher concentrations and geometric mean values for Cu
and Al were obtained for hair samples from Okigwe.
It was also observed that the level of these heavy metals found in these two cities could result from feeding habits,
occupational exposure, environmental factors and lifestyle. For instance, the high concentration of Pb could be
probably due to high gasoline lead discharge from exhaust fumes. Results clearly show that there were some
heavy metals in the hair samples of donors. This indicates that one is constantly exposed to heavy metal
contamination throughout one's life. But the fact remains that heavy metals may be found in low concentrations
when adequate precautions are taken.
REFERENCES
1. D. Airey (1983). Nickel in Human Hair due to Environment and Diet: A review”, Environmental Health
Perspective, 52: 503 – 516.
2. W. Ashraf, M, Jaffer, K. Anwer and U. Ehsan (1995). Age-and-Sex-Based Comparative Distribution of
Selected Metals in Scalp Hair of an Urban Population from Two Cities in Pakistan, Environ. Pollut., 87:
61-64.
3. R.L. Carson, H.U. Eliss, and J.L. McCann (1987). Toxicology and Biological Monitoring of Metals in
Humans, Lewis Publishers, Chelsea MI. pp.27 – 33.
4. L. Daniel and A. Richard (1981). Sample Collection and Analysis, Environmental Research, New York,
Vol. 87: 61 – 66.
5. R. Peltersson, and F. Rasmassen (1999). Daily Intake of Copper from Drinking Water Among Young
Children in Sweden, Environ. Health Persp., 107: 441-446.
6. H. Petering, D. Yeager and S. Witherup (1973). Trace Metal Content of Hair. Arch. Environ Health, 27:
327 – 333.
7. R. Thatcher, and M. Lester (1982). Effects of Low Levels of Cd and Pb on Cognitive Function in
Children, Arch. Environ. Health, 37:159 – 166.
8. R. Tuthill (1996). Hair Pb Levels Related to Children's Classroom Attention Deficit Behavior, Arch.
Environ. Health, 51: 214 – 216.
248
GEOCHEMICAL ASSESSMENT OF THE PETROLEUM GENERATIVE POTENTIAL OF EKENKPON AND NKPORO
SHALE FORMATIONS IN THE CALABAR FLANK SOUTH EASTERN NIGERIA
by
P. A Neji1; H A Neji2; TNganje3;O.O Oli4;U J Ibok5

1.Department of chemical sciencesCross River University of technology Calabar Nigeria
2,3,4 5 University of Calabar, Calabar Nigeria.
ABSTRACT
A total of 43 outcrop shale samples were analyzed for the assessment of the petroleum generative potential of
both shale formations. The samples were subjected to various analytical techniques such as
soxhletextractionof thesolubleorganic matter(SOM),Walkley Black titration method for the determination of
percentage total organic carbon %TOC or (%Corg), chromatographic separation into saturates (SHC),
aromatics (AHC) and polar compounds(NSO), over Al2O3/Silica gel 1:2 column and gas chromatography-mass
spectrometry(GC-MS) analysis of the saturates for the assessment of the sources of organic matter
contributions into the shale sediments. The %TOC and the chromatograms obtained from the (GC-MS)
analysis portray that the sediments contain type 11 and type 111 organic matter with appreciable petroleum
generative potential and are mostly gas prone. The maturity status was also assessed to be immature for both
formations, this was supported by the low yields of SOM, SHC,AHC,and the TPH The Nkporo shale formation
portray a more positive petroleum generative potential than the Ekenkpon shale formation, though the
Ekenkpon shale formation appears to be organically richer than the Nkporo shale formation, as evident in the
%Corg .
KEY WORDS: Geochemical Assessment, Petroleum, Generative, potential, Ekennkpon, Nkporo,
Shale, Formations Calabar Flank.
INTRODUCTION:A lot of geological studies have been documented in theCalabar flank,

(Murat,1972 [1].Adeleye,1975 [2]Hoque,1977 [3].Adeleye and fayose ,1978 [4]Petters, 1980,1981,1982
[5,6,7].Nyong,1995.) [8]etc. From foraminifera and palynological data these authors arrived "at various
conclusions on the sedimenthology, mineralogy, lithostratigaphy and depositional history of the Calabar
flank in relation to Anambra sub basin and the Benue trough. The Ekenkpon shale formation is a product of
two trangressive cycles with black organic rich sediments suggesting deposition under anoxic
environment during the late Albian- Cenomenian and early turonian). Of interest is the fact that the
Cenoroenian marine sediments in Nigeria are restricted to the Calabar Flank. Rament(1964,1965)[9,10];
Murat,1972[1]. Kogbe, 1976[11]and Huque, 1977[4].
The Nkporo shale formation span from the late Campanian to Maaslrictian sediments, which is
characterized by dark grey shales, with thin band of mudstones and gypsum Rayment, (1965)[12] This
paper attemptsto establish the generative potential, maturity status and sources of organic matter
contributions or imput into the Ekenkponand Nkporo shale formations using Geochemical parameter
249
Experimental
Materials and methods.
Sample collection.
250
A total of 43 outcrops shale samples were collected in polyethylene bags which have never been used.
This was to prevent the samples from contamination from any other source. These shales were sampled at
kilometers 23,24.8,26,28,30,and 38.where the outcrops appear in pronounce sequence along Calabar -Ikom
and Calabar -Itu roads The samples were sun dried for about one month and then stored in polyethylene and
polypropylene bags until when needed for analysis.
Sample preparation.
The dried shale samples were crushed using a ball crusher model NO. retschs1000 in the Aluminum
Smelter Company of Nigeria limited (alscon limited)at IkotAbasi The crushing process was repeated 2 or 3 times
and then sieved to obtain particlessize fractions that pass through 200g mesh sieve using a Retch vibrator sieve
model NO.D-42781 Germany.
Soxhlet extraction.
Each sample (100g) was weighed and extracted for 48 hours using a mixture of acetone benzene,and
methanol in the ratio 8:1:1 by volume. Excess solvent was recovered from the extracts using a rotary evaporator
model-Buchrotavapo R-114. The extracts were collected in sample bottles previously washed with detergent
and rinsed with acetone and dried. The extracts were weighed to obtain the soluble organic matter (SOM) or
extractable organic matter (EOM) by difference
Precipitation of asphaltenes
The heavy fractions of the asphaltenes were precipitated following procedures describe by Schoel et
al(1983)[13]using petroleum benzene and dichloromethane (DCM) mixture of 3:1 ratio by volume .The light
fractions (the maltenes) were then collected using a dropping pipette and used for characterization into
saturates (SHC) aromatics(AHC) and the polar corripounds(NSO) by column chromatography.
Percentage total organic carbon (%TOC)determination
Two comparative methods were used,normally the Walkley –Black (1947)[14]titration methods and
the use of leco carbon analyzer model No. leco CHN -1000.
For the Walkley -Black titration method,the crushed samples (0.5g)was usedand oxidized with k2cr207,
and conc H2SO4 and allowed to stand for 30minutes before dilution with
distilledwater.ConcentratedH3PO4NaFand diphenylamine indicator were also addedto the mixture. Which was
then titrated against 0.5M ferrous ammonium sulphatesolution (Fe(NH4)(SO4)2.6H2O). to the diphenylamine
end point. Duplicate titrations were also carried out. The % TOC values were obtained by calculation from the
formula %TOC = 10(1-T/S)(0.3/W).
The crushed sample (0.2g) was used for the leco carbon determination. The samples were treated with
0.5M H2SO4 and allowed to stand for 20 minutes to remove all the inorganic carbon. The residue was allowed to
dry under the sun for 6 hours and then loaded into the leco carbon analyzer for the determination of organic
carbon content.
Characterization of extracts
The extracts (maltenes) were characterized by resolution over alumina/silica gel (1:2) column. The
following solvent polarities were designed for elution of the various fractions.
The saturated hydrocarbons (SHC) fraction was eluted with n-hexane, while the aromatic
hydrocarbons (ARC) was eluted with methyl-benzene. And the hetero components (NSO) were eluted from the
column using methanol/dichloromethane (2:1) mixture 50 ml of solvent was use during the elution of each
fraction, after the development of the column with 10ml of n-hexane. Excess solvent was allowed to evaporate
at room temperature under a fan. This was to avoid the denaturation of the fractions. All fractions were then
weighed and their yields obtained by difference. The fractions were all stored in a refrigerator after sealing the
sample bottlecorks with aluminum foil.
251
Gas chromatography-Mass spectrometry analysis of the aliphatic fractions of the shale extracts was
done in Germany.
RESULTS
The variations in the levels of various parameters with depth at various locations are presented in
tables 1.1 to 1.6. Figure 1.1 depicts the frequency distribution of %TOC from Ekenkpon and Nkporo shale
formations. The hydrocarbon generative potential base on the variation of TPH with % TOC is represented in
figure 1.2. The GC/ MS finger prints are presented in figures 1.3 and 1.4 The ternary and the pie chart plots
which show the maturity of organic matter are presented in figures 1.5 and 1.6 respectively.
16
14
12
Y 10
C
N
E
U 8
Q
E
R 6
F
0
0.5-1.3 1.4-2.2 2.3-31 3.2-4.0 4.1-4.9 5.0-5.8
Fig 1.1 Frequency distribution of total organic (% TOC) of Ekenkpon and Akporo shale formations in
the Calabar flank
252
Table I.I variations in the levels of the various parameters with depth at km 23 Ekenkpon
SAMPL E DEPTH EOM ASPH MALT SHC ARC NSO TPH % % % AHC % mg mg mg TPH
ppm ppm ppm ppm ppm NSO SHC
CODE IN ppm ppm TOC SHC AHC NSO in mg
NO ppm
01 M
TOP 107.0 223.0 52.0 53.6 114.4 105.0 3.95 23.3 24.0 51.3 5.20 HC/g
10.56
02 SAMP
13.5 330.0
766.0 128.0 638.0 24.4 35.4 .471.0 59.8 3.41 3.8 5.5 73.8. 2.40 5.36
3.54 11.44
47.10 5.94
03 22.5 689.0 112.0 577.0 33.2 66.1 433.0 99.3 33.03 57.5 11.5 75.0 3.32 6.61 43.30 9.93
04 32.5 488.0 113.0 375.0 52.0 41.9 268.0 93.9 3.51 13.9 14;2 71.5 5.20 4.19 26.80 9.39
05 41.5 429.0 104.0 325.0 39.6 48.4 225.0 88.0 1.73 12.2 14.9 69.2 3.96 4.84 22.50 8.80
06 50.5 285.0 44.0 241.0 17.0 42.0 179.4 59.0 1.73 7.1 17.4 74.4 1.70 4.20 17.94 5.90
07 59.5 350.0 40.0 310.0 18.0 39.6 247.4 57.6 3.92 5.8 12.8 79.8 1.80 3.96 24.74 5.76
08 68.5 670.0 53.0 617.0 26.1 52.6 469.0 78.7 3.57 4.2 8.5 76.0 2.61 5.26 46.90 7.87
09 77.5 950.0 174.0 776.0 102.0 854.0 300.0 456.0 3.18 13.0 45.6 38.7 10.20 35.4 30.00 45.6
Table 1.2 variations in the levels of the various parameters with depth at km 24.8 Ekenkpon.PH
253
SAMP DEPT EO ASP MAL SHC AH NSO TPH % % % % mg mg mg TPH
L
H M H T ppm C ppm ppm TOP SHC AH NSO SHC AH NSO inmg
E
in ppm ppm ppm ppm C C HC/g
M TOP
CODE
NO
10 TOP 350.0 46.0 304.0 18.8 51.2 146. 70.0 1.56 6.2 16.8 48.0 1.88 5.16 14.60 7.00
SAM 249 0
4.50 60.0 189.0 72.0 52.0 55.0 124. 0.66 38.1 25.5 29.1 7.20 5.20 5.50 12.40
11 0 0
12 9.0 205.0 28.0 177.0 15.7 23.6 128.0 39.3 4.83 8.9 13.3 72.3 1.57 2.36 12.80 3.93
13 18.0 482.0 97.0 385.0 94.7 112.8 171.5 207.5 207.5 24.6 29.3 44.5 9.47 11.28 17.15 20.75
14 22.5 282.0 54.0 228,0 13.5 46.9 129.0 60.4 60.4 5.9 20.6 56.6 1.35 4.69 12.90 6,04
15 31.6 163.4 49.1 114.3 26.0 29,5 53.2 55.5 55.5 22.8 25.8 46.5 2.60 2.95 5.32 5.55
16 40.5 740,0 83.0 657.0 12.5 51.6 508.0 64.1 64.1 1.9 7.9 77.3 1.25 5.16 50.80 6.41
17 41.0 910.0 168.0 724.0 18.7 55.7 537.0 74.4 74.4 2.5 7.5 72.4 1.87 5.57 53.70 7.44
18 46.5 1196 301.0 695.0 335.0 215.7 310.0 550.0 550. 37.4 24.0 34.6 3.35 21.5 31.0 24.65
19 56.5 514.0 90.0 424.0 39.0 98.0 196.0 128.0 1.5. 9.2 21.0 46.2 3.90 8.90 19.60 12.80
20. 61.0 445.0 125.0 321.0 14.9 68.7 101.0 83.6 1.42 4.6 21.4 31.5 1.49 6.87 10.10 8.36
Table 1.4 Variation in the levels of the various parameters with depth at km 26 Ekenkpon.
SAMPLE DEPTH in EOM ASPH MALT SHC NSO ppm TPH ppm % TOC % SHC % AHC % NSO mg SHC mg AHC mg TPH
CODE ppm NSO
M AHC ppm ppm inmg
254
21 TOP SAMP 601.0 180.0 421.0 18.3 302.0 89.6 1.48 4.3 16.9 71.7 1.83 7.13 30.2 8.96
22 4.0 301.0 66.0 235.0 18.8 155.0 69.7 2.20 5.9 23.6 66.0 1.38 5.59 15.5 6.97
23 8.0 484.0 63.0 421.0 22.4 226.0 112.4 3.35 5.3 23.7 53.7 2.24 9.97 22.6 12.21
24 12.0 616.0 150.0 466.0 85.0 315.0 142.0 3.33 18.2 12.2 67.6 8.50 5.90 31.5 14.20
25 16.0 430.0 113.0 317.0 19.5 174.0 89.0 2.24 6.2 22.0 54.9 1.95 6.97 17.4 8.90
SAMPLE DEPTH in EOM ASPH MALT SHC AHC NSO TPH ppm % % SHC % AHC % NSO mg SHC mg mg TPH
CODE ppm ppm AHC NSO
26 4.0 405.0 45.0 360.0 77.0 54.1 223.0 131.1 1.94 21.4 15.0 61.9 7.50 5.41 22.3 13.11
27 6.0 361.0 83.0 278.0 39.1 76.4 142.0 115.5 3.45 14.1 27.5 51.1 3.39 7.64 14.2 11.55
28 7.0 651.0 90.0 561.0 21.5 50.6 468.0 468.0 5.74 3.8 9.0 83.4 2.15 5.06 46.8 7.21
SAMPLE DEPTH in EOM ASPH MALT SHC AHCNSO TPH ppm % % SHC % AHC % NSO mg SHC mg mg TPH
CODE ppm ppm AHC NSO
29 M
TOP AHC 226.0
276.0 50.0 ppm 13.9
ppm ppm 129.0 60.8
46.9 TOC
1.48 6.2 20.8 57.1 1.39 inmg
4.69 12.9 6.08
30 6.0 488.8 73.0 415.0 40.9 64.4 187.0 105.3 2.46 9.9 15.5 45.1 4.09 6.44 18.7 10.53
31 7.0 249.0 39.0 210.0 13.6 69.8 118.0 83.4 3.69 6.5 33.2 56.5 1.36 6.98 11.8 8.34
Table 1.6 Variation in the levels of the various parameters with depth at km 38 Nkporo.
SAMPLE DEPTH EOM ppm ASPH MALT SHC AHC NSO TPH % % SHC % AHC % mg mg mg TPH
CODE ppm ppm NSO SHC AHC NSO
32 TOP 4567. 2965. 1602. 201. 172. 889. 373. 2.39 12.5 10.7 55.5 20.1 17.2 88.9 37.3
33 4.0 370.0 130.0 240.0 18.3 57.9 105.0 76.2 0.75 7.6 24.1 43.8 1.83 5.79 10.5 7.62
34 8.0 4948.0 4112.0 836.0 216.0 295.0 307.0 511.0 1.18 25.8 35.3 36.7 21.6 29.5 30.7 57.1
35 12.0 4534.0 3931.0 603.0 194.0 179.0 215.0 373.0 3,12 32.2 29.7 35.7 19.4 '17.9 21.5 37.3
36 14.0 2730.0 1362.0 1368.0 317.0 656.0 385.0 973.0 1.36 23.2 48.0 28.1 31.7 65.6 38.5 97.3
37 16.0 1231.0 278.0 952.0 309.0 391.0 220.0 700.0 2.09 32.5 41.1 23.1 30.9 39.1 22.0 70.0
38 20.0 1971.0 1208.0 770.0 147.0 516.0 101.0 663.0 1.39 19.0 67.0 13.1 14.7 51.6 10.1 66.3
39 24.0 5251.0 4483.0 768.0 35.7 72.6 606.0 108.3 1.39 4.6 9.5 79.0 3.57 7.26 60.6 10.83
255
DISCUSSION
Organic richness
The organic richness expressed as the total organic carbon contents (%TOC) is shown in tables 1.1 to 1.6
.The TOC of both the Ekenkpon and Nkporo formations varies significantly with depth. The Ekenkponshales have
TOC values ranging from 0.66-5.7% with an average TOC value of 2.78%. While the Nkporoshales have TOC
ranging from 0.75-3.12%.with an average TOC values of 1.64%.The frequency distribution of TOC shown in fig.
1.1 shows that most of the samples have TOC values higher than 0.5 /0.5g shale, which is usually taken as the
l o we r l i m i to fo rga n i c m atte r r i c h n e s s re q u i re d fo r p et ro l e u m s o u rc e b e d . ( T i s s o t a n d
welte,/1984a,1984b)[15,16].Tissot 1984 [17] The samples with the highest and least TOC content for the
Ekenkpon formation were obtained at kilometer 28 and kilometer 24.8 with values of 5.74 and 0.66%
respectively While for the Nkporo formation the samples with the highest and least TOC content were obtain at
kilometer 38 and had values of 3.12 and 0.75 % TOC respectively. Such high organic carbon contents arc
attributed to highly reducing environment of deposition and a high productivity of organic matter leading to
high preservation of organic matter.
Organic matter type

The organic matter type is assessed from the plot of % SHC + AHC VSTOC shown in fig 1.2 this is a plot of the
total petroleum hydrocarbon (TPH)againstthepercentage organic carbon. (%Corg) content.From this
plot,theEkenkpon and Nkporo samples could be classified as type 11
and type 111
256
organicmatter Consequently, samples 01, 02, 03, 04, 05, 06, 07, 08, 10, 14, 15, 17, 19, 20,21, 23, 24 25,
28, 30, 33, and 35, plot within the gas zone and are associated withtype 111 organic matter. Such organic matter
is of terrestrial or terrigenous plantsorigins, deposited under less reducing environment, with virtually gas
generativepotential .while samples 09, 11, 13, 18, 34 36 37 and 38 plot within the oil with gasand gas with some
oil zones. Such organic matter is often associate with type 11organic matter of mixed marine and continental
origin deposited under a strongreducing environment. Sample 12, and 16 plotted at the zone of little
hydrocarbonsource potential.
Sources of organic matter

Gas chromatography -mass spectrometry analysis of the aliphatic fractions of the shale extracts also
showed some correlation with the geochemical log plots in fig 1.2.The shale extracts were analyzed at Germany
using Acgmethod PAK8SCN. The chromatograms are shown infig.1.5 and fig 1.6 The chromatograms of the
Ekenkpon and Nkporo shale extracts were characterized by thedominanceof n-alkane members (above n-
C16compounds).The chromatograms showed the conspicuous presence of n-C23 members which is often
associated with marine derived autochthonous organic matter, also prominent in the chromatograms were the n-
C27n-C29 and n-C31 members. These members indicate the contributions of allochthonous terrestrial organic
matter from higher plant waxes into the sediments. The chromatogram of the Ekenkpon extract exhibited a
bimodal distribution pattern typical of mixed marine and continental organic matter input with maxima at n-C21
typical of planktonic origin and n-C27members (Moldowan et al,1985)[18]:The chromatogram .of Nkporo extract
did not exhibit a bimodal distribution but showed prominent peaks of n-C21n-C25 and n-C27-n-C31 typical of marine
and continental organic matter respectively, but with a more prominent representative of more marine organic
matter abundance.
According to Goth et al (1988)[19], pyrolyzates of the npnmarine algae tetraedron and of laminated
messel shale (Germany) containing densely packed tetraedron algae remains exhibited similar distribution of
n-alkanes and n-alkenes. Simoneit (1986)[20], used similar hydrocarbon distribution patterns to distinguish
marine from continental or terrestrial organic matter contribution to the sedimentary organic matter.The
chromatograms also exhibited very sharp n-alkanes distribution peaks without odd/even carbon numbers
preference.
257
Fig 1.5 Ternary diagram showing maturity of organic
Thermal maturity status

For the Ekenpon extracts, the asphaltene content was low with values' between 28.9-3-10pp and
samples 12 and 18 obtained at kilometer24.8 having lowest and highest asphaltene contents respectively the
Nkporo extracts yielded significantly high asphaltene content with values between 130-4483 .0 ppm also
corresponding to samples 33 - 39 respectively
The SHC contents were moderately low for the Ekenkpon shale Formation with values ranging between
12.5 - 335.0 ppm while the Nkporo shale Formation yielded SHC values ranging between 18.3 - 317.0ppm.
The AHC yields were also low for both formations but higher values were obtained compared to the
yields of the SHC and samples 09 and 12 obtained at kilometers .23 and 24.8 having least and highest values of
23.6 and 354.0 ppm for the Ekenkpon extracts. The Nkporo extracts yielded AHC values between 57,9-65.0
ppm corresponding to samples 33 and 36 having the least and highest values. The extracts from both
formations generally showed a strong predominance of the NSO sand AHC over the SHC with values between
55.0 -537.0 ppm for the Ekenkpon Formation and values between 101.0 - 8890 ppm for the Nkporo
Formation! This trend generally portrays immaturity of the samples. This is also supported by the ternary plots
in Fig 1.3 and the pie chart plots Fig 1.4 where, in the case of the ternary plots most of the samples plotted
towards the NSO.and non toward the SHC, while pie chart showed a clear dominance of NSO and UCM
(unresolved complex mixture) in the Nkporo samples The Ekenkpon samples showed a higher yield of the SHC
258
and little UCM Thus the Ekenkpon shale appears to be more mature than the Nkporo samples. This is expected
since the Ekenkpon shale Formation is of the Cenomenian - Turonian age and the Nkporo shale Formation is of
Maastrichtian age.
259
Generative Potential
The generative potential often assessed from the quantities of EOM, SHC, AHC and TPH obtained which
invariably depends on the organic matter type. The EOM of Ekenkpon extracts was moderately low yielding
values ranging between 163.4 -1196.oppm, with samples 15 and 18 obtained at kilometer 24.8.being the
poorest and richest sample respectively The Nkporo extracts yielded higher EOM values ranging between 370 -
5251ppm with samples 33 and 39 obtained at kilometer 38 being the poorest and richest samples respectively
The SHC yields were generally low for both Formations. The Ekenkpon Formation had SHC contents ranging
between 12 .5 -335ppm with samples 16 and 18 also obtained at kilometer 24.8 being the poorest and richest
samples respectively. The Nkporo Formation on the other hand had SHC content ranging betweenl8.3 -
317ppm with 33 and 36 being the poorest and richest samples.
The total petroleum hydrocarbon were particularly low for the Ekenkpon extracts which yielded TPH
.contents between 55.5 - 550.0ppm with samples 15 and 18 obtained at kilometer 24".8 having theJeast and
highest yields .while the Nkporo extracts yielded concentrations of TPH between 76.2 - 973.0 ppm also
corresponding to samples 33 and 36 respectively. These yields show no correlation with the % TOC values
obtained for both Formations. The values however portray a more positive generative potential for the Nkporo
than the Ekenkpon Formation.
CONCLUSION.
Both the Ekenkpon and the Nkporo shale Formations has been successfully studied from all
parameters analyzed, for the determination of the organic richness, organic matter .type, sources 'of organic
matter, thermal maturity status and the generative potential, for both Formations, we conclude that both
Formations are organically rich. With the Ekenkpon shale Formation being organically richer than the Nkporo
shale Formation. Organic matter type is of type 11 and type 111. Sources of organic matter contributions into
the shale sediments is from terrestrial or continental and marine origin. Both Formations have appreciable
petroleum generative potential with the Nkporo Formation portraying a more positive generative potential.
And mostly gas prone as most of the sample plotted under gas and gas with some oil zones.The thermal
maturity is of low status, or immature for both Formations. This is supported by the fact that all the samples
analyzed were outcrop samples and may not have attained a reasonable thermal stress.
REFERENCES
R. C., Murat. (1972) Stratigraphy and paleogeography of Cretaceous and lowerTertiary in Southern Nigeria in
Dessauavagie, T. F. J. and Whiteman, A. J.(eds) African geology, Ibadan, university press.
D. R., Adeleye., (1975).Nigerian Late cretaceous stratigraphy and paleogeographyam.assoc. pet. geol. Bull. V 59
No.l2}p2302 - 2313.
M., Hoque.(1977). Petrogaphic differentiation of tectonically controlled Cretaceous sedimentary Cycles. South
Eastern Nigeriain sediment Geology,17, 235 - 245
D. R.,Adeleye,and E. A., Fayose, (1978) stratigraphy of the type section of AwiFormation, Odukpani are, South
Eestern Nigeria Nig jour Min. Geol.15,35-37.
S. W., Petters. (1980),New cave sites in Cross River State of Nigeria W.AfrJr Arch. 10, 171 -176.
S. W., Petters. (1981) New cave lower karst near Calabar, Nigeria Fid 46, 49 - 20.
S. W., Petters. (1982) Central West African Cretaceous - Tertiary benthic foraminifera and stratigraphy.
PaJeontographic, Abt. A 179, 1 - 104
E.E., (Nyong, 1995).Cretaceous sediments in the Calabar flank in Geological Excursion Guide book to Oban
massif, calabar Flank and Mamfe Embayment South Eastern Nigeria.published in the 31st annual
260
conference of the Nigerian Mining and Geosciences society, Calabar.
R. A., Rayment. (1964), Review of Nigeria Cretaceous Cewojoicstratigraphy.Jour. Mining GeoLMetalSocvol.l,
No. 2 pp 61 - 80.
C. A., Kogbe. 1976The Cretaceous andpaleogene sediments of Southern Nigeria: in .Geology of Nigeria (Ed.C. A.
Kogbe )Lagos.Elizabethan publishing Company, Nigeria.
R. A., Rayment. (,1965) Aspects of the geology of Nigeria Ibadan Univ. press ,Ibadan,
M., Schoel.B., Durand, J. L.Oudin, M., Teschner , and H. Wehner, (19830) Maturity related biomarker and stable
isotope variations and their applications to Oil/source rock correlation in the mahakam Delta
Kalimantan. In Advances in geochemistry 1981 (Eds.M.Bjoroy et al )Wiley and sons, New York, 156 -
163.
A. E.,Walkley (1947) Organic matter content of the kimmeridgeshales Am. Assoc. of . Petro Geologists
bull.60.5001 -5002-
B. P., Tissot, and D. H., Welte, (1984a),Petroleum formation: A approach to oil and gas exploration.Berlin,
Spinger -Verlag 538.
Tissot and welte, D. H. (1984b) Petroleum formation and occurrence.2nd edititionSpingerVerlag.
B. P., (Tissort, 1984).Recent advances in petroleum geochemistry applied to hydrocarbon exploration. Bull Am.
Assocpetro.Geol. 68, 533 - 563.
J.M., Moldowan, E.J., Gallegos, and W.K., Seifert, (1985) Relationship between petroleum composition and
depositional environment of petroleum source rocks A. A. P. G. Bulletin vol 69,1255 - 1368.
K., Goth, J. W. De Leauw, W., Putmann, and E. W., Tegelaar. (1988).Origin of Messel Oil Shale Kerogen.Nature,
336,759-761
B. R. T.,Simoneit, (1986).Cyclic terpenoids of the geosphere. In Biological markers in the sedimentary Records
(R. B. Johns. Ed.) New York, Elsevier.
261
SYNTHESIS, CHARACTERIZATION AND AND BIOLOGICAL ACTIVITY OF NEW MANNICH
BASE 1-((4-CHLOROPHENYL)(2,5-DIOXOPYRROLIDIN-1-YL)METHYL)UREA
M.R. Abdullahi *1,S. Rajeswari2
1
Department of Pure and Industrial Chemistry, Bayero University Kano, P. M. B. 3011, Nigeria
2
Department of Chemistry, SRM University, Kattankulathur, Tamil Nadu, India.
E.Mail: mrabdullahi86@gmail.com
Phone Number: +2348039347945
Abstract:
A new mannich base 1-((4-chlorophenyl)(2,5-dioxopyrrolidin-1-yl)methyl)urea has been synthesized from the
condensation of succinimide, 4-chlorobenzaldehyde and urea. The compound has been characterized on the
basis of elemental analysis, FTIR, 1HNMR 13C NMR, and Mass spectroscopy. The compound was tested for its
antimicrobial activity against Pseudomonas aeruginosa, Staphylococcus aureus, Aspergillus niger, and
Candida albicans using agar well diffusion method.
Key words: Mannich base, succinimide, antimicrobial
1.0Introduction
Mannich reaction is one of the most fundamental and important C–C bond forming reactions in organic
synthesis. It is a three-component condensation between compounds containing an active hydrogen i.e.
compound capable of supplying hydrogen atom, with an aldehyde and primary or secondary amine (Tamil et
al.,2011). The reaction involves the elimination of water molecule and leads to the formation of compound
calledMannich base.
The formation of both C–C and C–N bonds in this amino methylation process makes the Mannich reaction an
extremely useful synthetic transformation. Mannich bases have wide application in the area of pharmaceuticals
and macromolecular chemistry (Tamil et al., 2011).In Mannich reaction, ammonia or primary or secondary
amines are employed for the activation of formaldehyde, tertiary amines lack an N-H proton to form the
intermediate imine (Muthumani et al., 2010).The active hydrogen is replaced by amino-methyl group and form
carbon–carbon and carbon–nitrogen bonds. This amino methylation process makes the Mannich reaction an
extremely useful synthetic transformation and the Mannich bases are highly reactive because of the basic
function which renders the compound soluble in many aqueous solvents (Babu and Pitchai, 2014)
The ligands containing N and O donor atoms possess a number of biological activities such as antibacterial,
antifungal, anticancer,anticonvulsant, andanti HIV (Venkatesa et al., 2010, and Sivakami et al., 2014).
Mannich bases have gained importance due to their applications in pharmaceutical chemistry as they are used as
local anesthetic, antimalarial, anti-inflammatory, analgesic, antioxidant and antitubercular activities (Sriram et
al., 2009).
Mannich bases have also found numerous applications in the treatment of natural macromolecular materials such
as leather, paper and textiles, and as additives used by the petroleum industry, water treatment, analytical
reagents, cosmetics, dyes, used in the removal heavy metals from effluents.They are also used as powerful
pesticides and insecticides apart from other biological applications and these properties are enhanced when
ligands form complexes with metals (Tharmaraj et al., 2013).
Numerous mannich bases were synthesized, characterized and analyzed for different applications.
Mannich base derived from the condensation of succinimide, benzaldehyde and urea together with their metal
complexes showed great antimicrobial activities (Venkatesa et al., 2010). Mannich base 1-(morpholino(phenyl)
methyl)pyrrolidine-2,5-dioneformed by the direct condensation of morpholine, succinimide, and
benzaldehydehas been synthesized and characterized (Rajeswari et al., 2010). Mannich base 1,3-bis-
(morpholin-4-yl-phenyl-methyl)-thiourea derived from the direct condensation of morpholine , benzaldehyde
and thiourea have been synthesized and the compound was used to inhibit mild steel corrosion in hydrochloric
262
acid medium to the great extent (Venkatesa et al., 2011). The synthesis of novel mannich base and it is metal
complexes derived from succinimide 4- methoxy benzaldehyde and thiourea have also been reported and the
compounds were screened for anti-microbial activity against E.coli and B. subtilis bacteria. The compounds
were also found to have a significant DNA binding ability
(Sivakami et al.,2014). In this Paper, we report the synthesis, characterization and biological activities of new
mannich base derived from succinimide, urea and 4-Chloro benzaldehyde.
2.0 Experimental
2.1 Materials and methods

All the reactants and solvents used for the synthesis of this compound were of analytical grade and commercially
available and used without further purification.
The Melting point of compounds was determined in an open capillary tube using SMP40 melting point
apparatus. Elemental micro analysis was carried out with a Carlo Erba 1108 Elemental Analyzer. Mass spectrum
was recorded on a shimadzu 2010s Mass Spectrometer. The IR spectra were recorded on Shimadzu FT- IR
instrument using KBr pellets. 1H NMR spectrum was recorded using DMSO-d6 as a solvent and TMS as internal
standard. 13C NMR spectrum was recorded in DMSO using a Bruker 500 MHz instrument.
2.2 Synthesis of Mannich base 1-((4-chlorophenyl)(2,5-dioxopyrrolidin-1-yl)methyl)urea (SCBU)

Succinimide, 4-chlorobenzaldehyde and urea were taken in 1:1:1 equimolar ratio. Succinimide and urea were
taken in a 250 mL beaker. Sufficient amount of ethanol was added to make the contents a homogeneous solution,
and then 4-chlorobenzaldehyde was added slowly in drops with continuous stirring of the solution. A white
powdery substance was formed immediately. After 2 weeks, the product was washed several times with distilled
water and dried in an air oven at 60 0C for 1hour and recrystallized from ethanol by slow evaporation. The scheme
for reaction is given below.
O O
O
NH
O
N NH 2
O
-H2O
NH
H2 N NH 2 O
O
Cl
Cl
Succinimide 4-chlorobenzaldehyde Urea 1-((4-chlorophenyl)(2,5-dioxopyrrolidin-1-yl)methyl)urea
3.0 Results and discussions

Table 1: Physical Characterization of the ligand
Melting Percentage Yield Colour
Point (0C) (%)
Ligand 157-1600C 83 White
Table 2: Solubility Test of the Ligand
263
Solvent Water Ethanol Methanol Chloroform Hexane DMSO DMF
Ligand IS IS SS IS IS S S
The analytical and spectral data obtained for the ligand are given below.
Analysis: Calculated for C12H12ClN3O3: C 51.16, H 4.29, N 14.92 % Found: C 50.97, H 4.28, N 14.87 %; FT-
-1
IR: (KBr, cm ): 3360 (N-H), 3198-3058 (Aromatic C-H), 2964 (Aliphatic CH2), 1699 (succinimide C=O), 1658
(urea C=O), 1175 (C-N-C); 1H NMR: (DMSO-d6, äppm): Multiplet 7.32-7.93 (Aromatic protons), 2.69-2.70
(succinimide CH2 protons), 5.68-6.0 (NH2 protons), 2.57 (Methine C-H proton), 10.01(N-H protons); 13C NMR:
(DMSO-d6, ppm): 29.5 (CH2 carbon of the succinimide), 40.0 (Methine carbon), 128.2-137.9 (Aromatic
carbon), 156.7 and 179.4 (Urea and succinimide carbon); Mass: Calculated for C12H12ClN3O3 m/z = 281.69,
found 282.
3.1 In vitro antimicrobial activity
The in vitro antimicrobial activity of the synthesized compound was tested against one gram negative
Pseudomonas aeruginosa and one gram positive Staphylococcus aureus bacterial pathogens and one mycelial
fungal pathogen Aspergillus niger and one yeast pathogen Candida albicans. These selected pathogenic strains
were obtained from Microbiological Division (Jayagen Biologics Analytical Laboratory, Chennai).
3.2 In vitro antibacterial activity
The antibacterial activity was determined by well diffusion methods About 25 mL of molten nutrient agar was
poured into a sterile Petri plate. The plates were allowed to solidify, after which 18 hrs grown (OD adjusted to
0.6) 100 µl of above said pathogenic bacteria cultures were transferred onto plate and made culture lawn by using
sterile L-rod spreader. After five min setting of the pathogenic bacteria, a sterile cork borer was used to make 5
mm well on the agar. The test samples were dissolved in 5% DMSO and loaded in to wells with various
concentrations such as 50 g/well, 100 g/well, 150 g/well and 200 g/well. The plates were incubated at 37°C in a
40 W florescent light source (~ 400 nm) for 24 hrs. The antibacterial activity was determined by measuring the
diameter of the zone of inhibition around the well using antibiotic zone scale (Holder and Boyce, 1994).The
results for the antibacterial activities are given in table 3.
3.3 In vitro antifungal activity
The agar plug of mycelia fungal inoculum was placed onto the potato dextrose agar individually. Whereas,
5
yeast pathogen at the inoculum concentration of 5×10 cfu was swabbed using sterile cotton swabs. The wells
were made using sterile cork borer (5 mm). The given sample was loaded as 50 g/well, 100 g/well, 150 g/well and
200 g/well concentration. The plates were incubated at 35°C and observed after 48 and 72 hrs. The inhibition
zones were read at the point of complete inhibition of growth (ZOI).The results for the antifungal activities are
given in table 4.
Table 3: Antibacterial activities of the ligand
264
Compound Zone of Inhibition (mm) Zone of Inhibition (mm)
Pseudomonas aeruginosa Staphylococcus aureus
Concentrations (µg/mL) Concentrations (ìg/mL)
50 100 150 200 50 100 150 200
SCBU 6 8 9 10 - 7 8 10
C12H12ClN3O3
Standard bacterial agent used: Chloramphenicol at 30 µg/ml, ZOI=20mm

From the results of antibacterial activity, theligand was found to be active against the tested microbes. The
activity of the compound increases with increase in concentrations, but the ligand showedno activity at lower
concentration (50 ìg/mL) against Staphylococcus aureus. The overall activities of both the ligand was found to
be lower than that of the Chloramphenicol at 30 µg/ml which was used as standard, with inhibition zone of
20mm.
Table 4: Antifungal activities of the ligand (SCBU)
Compound Zone of Inhibition (mm) Zone of Inhibition (mm)
Aspergillus niger Candida albicans
Concentrations (ìg/mL) Concentrations (ìg/mL)
50 100 150 200 50 100 150 200
SCBU - 6 7 8 6 7 7 9
C12H12ClN3O3
265
Standard antifungal agent used: Clotrimazole at 30 µg/ml. ZOI= 24mm
The results for the antifungal activities reveals that both the ligand shows no activity at lower concentration
against Aspergillus niger, but was found to show some activity against Candida albicans. The overall activities of
both the ligand was also found to be lower than that of the standard, Clotrimazole at 30 µg/ml with inhibition
zone of 24mm.
3.4 Conclusion
A new mannich base have been synthesized and characterized by spectral techniques and the compound have
been evaluated for its antibacterial and antifungal activities and the compound showed lower activity against the
tested microorganisms and also lower activity when compared with selected drugs used for the study.
REFERENCES
1. Babu. K and Pitchai. P(2014) An innovative synthesis of new mannich base, its metal complexes
and their antibacterial studies, international journal of pharmaceutical, chemical and biological
sciences, IJPCBS 2014, 4(2), 256-261 ISSN: 2249-9504
2. DevarajKarthik , D. Tamilvendan , G. VenkatesaPrabhu (2011) Study on the inhibition of mild steel
corrosion by 1,3-bis-(morpholin-4-yl-phenyl-methyl)-thiourea in hydrochloric acid medium. Journal
of Saudi Chemical Society (2014) 18, 835–844
3. D. Sriram, D. Banerjee, and P. Yogeeswari (2009) “Efavirenz Mannich bases: synthesis, anti-HIV and
antitubercular activities,” Journalof Enzyme Inhibition and Medicinal Chemistry, vol. 24, no. 1, pp. 1–5,
2009.
4. D. Tamil vendan S. Rajeswari S. Ilavenil K. Chakkaravarthy G. VenkatesaPrabhu (2011) Syntheses,
spectral, crystallographic, antimicrobial, and antioxidant studies of few Mannich bases, Medicinal
chemistry research, springer ISSN 1054-2523 19(7)617-716 Med Chem Res DOI 10.1007/s00044-011-
9944-2,springer vol.19 no.7 an international journal promoting bioactive compounds
5. Holder IA and Boyce ST. (1994) Agar well diffusion assay testing of bacterial susceptibility to
various antimicrobials in concentrations non-toxic for human cells in culture. Burns 20:426-429.
6. M.Sivakami, B.Natarajan, M.Vijayachandrasakar, S. Rajeswari(2014) Evaluation of DNA
Binding and Cytotoxic activity of a new Mannich base and its Metal Complexes. (IJIRSE)
International Journal of Innovative Research in Science & EngineeringISSN (Online) 2347-3207
7. P. Muthumani, Neckmohammed, R. Meera, S.Venkataraman, N. Chidambaranathan, P. Devi, C.A.
Suresh Kumar (2010) Synthesis and evaluation of anticonvulsant and antimicrobial activities of some
Mannich bases of substituted aminophenol and acetophenone.Int J Pharm Biomed Res 2010, 1(3), 78-
86. ISSN No: 0976-0350
8. P.SaravanaBhava, P. Tharmaraj, V.Muthuraj , M.Umadevi (2013) Synthesis, characterization and
biological activities of transition metal chelates of 2-aminothiazole-derived Mannich bases, IOSR
Journal of Engineering (IOSRJEN)ISSN: 2278-8719 Vol. 3, Issue 3 (April. 2013), PP 18-27
9. S.Rajeswari, D.Tamilvendan, G.VenkatesaPrabhu(2010) 1-[(2,5-dioxopyrrolidin-1-
yl)(phenyl)methyl] urea and 1-[(2,5- dioxopyrrolidin-1 yl)(phenyl)methyl] thiourea and its
transition metal (II) complexes: Synthesis, structural characterization and antimicrobial study.
10. S. Rajeswari G. VenkatesaPrabhu D. Tamilvendan V. Ramkumar (2010) Spectroscopic Studies and
Crystal Structure of 1-(Morpholino (Phenyl) Methyl) Pyrrolidine 2,5-Dione springer J
ChemCrystallogr (2010) 40:437–442 DOI 10.1007/s10870-009-9674-1
266
NANOTECHNOLOGY AND IMPLEMENTATION IN NATURAL PRODUCTS
GREEN CHEMISTRY PROCESSES
1 1 1 1 2
M.M. Adeyemi* , P.E. Omale ; O.A. Olawuyi A. Ngokat and A. Bhattacharya
1. Nigerian Defence Academy, Department of Chemistry, P.M.B. 2109, Kaduna, Kaduna State,
Nigeria.
2. Nanotechnology Section, Department of Biomedical Engineering, Rajiv Gandhi Institute of
Technology and Research Centre Cholanagar, R.T. Nagar Post, Hebbal, Bangalore-560 032,
India.
*Corresponding Author Email: esdupe@yahoo.com; Cell phone: +234-803 453 2419
ABSTRACT
Nanotechnology, already showing great potential for application in environmental protection
has been suggested to be able to provide green and effective alternatives for the management of
insect pests in agriculture and post harvest processes. This is in an effort to reduce the hazardous
effects of previously used non-biodegradable substances on humans and the environment. The
paper will discuss emerging nanotechnologies in antifeedant and pesticide formulations from
natural products including novel photodegradable insecticide involving nanoparticles. This will
include studies on synthesized nanoparticles used as larvicides, antifeedants, insecticides and
pesticides. Nanoscience and nanotechnologies have offered the opportunities to combine
organic constituents from natural products with elemental components in order to create
complex, innovative and specific material in chemical synthesis for future and immediate use in
green technologies.
1.0 INTRODUCTION
Nanotechnology has been defined as the manipulation of atom-by-atom interactions in nature
which can take part in chemical and biological functions and processes. Nanotechnology is
becoming a revolutionary field in its integration with the green chemistry approach. The current
approach involves the use of material that are effective at nanoscale such as gold and silver
because of their natural and chemical characteristics. The inorganic nanomaterials exhibit well-
adopted physical, chemical, and biological properties, though properties may vary in size based
on their character as well as on specific pH and temperature. Nanoscale metal oxides such as
TiO2, ZnO, and Al2O3 dendrimers (nanosized polymers built from branched units) are performed
with atom–atom interaction (1).
Nanotechnology has found a large field of application including agricultural food production
and post harvest storage, improvements in nanoporous zeolites for efficiency in the dosage and
release of required quantities of water and fertilizers in plants, nanocapsules for herbicide
delivery and vector and pest management and nanosensors for pest detection (2,34). Nanoparticles
have also been involved in the synthesis of insect repellents and deterrents, antifeedants,
267
insecticides and pesticides (5). Nanoencapsulation has been defined as a process through which
a chemical compound such as an insecticide is slowly but efficiently released to a particular
host plant for insect pest control. This process in conjunction with nanoparticles in form of
pesticides allows for proper absorption of the chemical into the plants unlike the case of larger
particles2. This process can also deliver DNA and other desired chemicals into plant tissues for
protection of host plants against insect pests (6). Release mechanisms of nanoencapsulation
include diffusion, dissolution, biodegradation and osmotic pressure with specific pH (7).
1.1 The Role of Nanoparticles in Agriculture
Agricultural production faces the challenge of enhancing crop production and providing
nutritionally adequate diets for the increasing population, under uncertain climatic extremes,
water scarcity, in limited and sometimes degraded land area, and mostly with poor quality
water and air, associated with rapid changes in natural biodiversity as well as trying to resolve
pre- and post- harvest losses associated with pests.
The use of nanotechnology in agriculture and green chemistry practices has been mostly
theoretical, but the impact of Nanotechnology in the control of insect pests in the agricultural
systems is intended to enhance food production (8). The effective use of nanotechnology in
agriculture sector could be multi-directional. It involves the delivery of encapsulated
pesticides, insecticides, antifeedants, etc in nanomaterials for controlled release, stabilization
of bio-pesticides, slow release of nanomaterial assisted fertilizers, bio-fertilizers and
micronutrients for efficient use in addition to other agrochemcial applications (9).
Stored nanoparticles in plants influence some of the insect-plant interaction roles and the fact
that plants have been observed to have the ability to produce secondary metabolites tend to
help facilitate the process. Plants which have ability to produce nano synthesizing materials
are solely responsible for producing metallic nano materials. The mechanism involved in
production of this eco-friendly nanoparticle have been found as effective methods for
preparation of nanoformulations as the plant producing nano particles are highly acceptable, as
because, it maintains uniform size and shape without aggregation or agglomeration. The
utilization of this key component from plants is a crucial factor for nanopesticides production
via biological processes, a pathway for future eco-friendly pesticides. There are several
methods that have been reported in literature for the synthesis of plant-nanoparticle-pesticide
core-shell conjugate which can be used to control agricultural insect pests. These conjugant
nano particles may be in different shape, size (2-150 nm) and various forms (10). Medicago
sativa (alfalfa) was the first reported plant material used in the synthesis of nanoparticles and it
was credited to been capable of synthesizing gold and silver nanoparticles. This has led to
several other dried plant material being screened and evaluated for nanoparticle synthesis (11).
Some of these are listed in Table I below:
Table I: Plants Utilized in Synthesizing Nanoparticles
268
S/No Synthesized Nano Particles Plants Utilized for the Synthesis
1. Gold Nanoparticles Emblica officinalis
Ocimum basilicum
Psidium guajava
Syzygium aromaticum
Tamarindus indica
Terminalia catappa
2. Silver Nanoparticles [AgNPs] Brassica juncea
Camellia sinensis
Coriandrum sativum
Cymbopogon exuosus
Ipomoea aquatic
Nelumbo nucifera
Parthenium hysterophorus
Tanacetum vulgare
3. Gold and Silver Nanoparticles Apiin (henna leaves)
Cinnamomum camphora
Emblica officinalis
Murraya koenigii
Some of these plants have found applications as larvicides, antifeedants, insecticides and pesticides, and it has
been established that plant-based green synthesis approach is one of the effective method for silver nanoparticles
(AgNPs) production. Other plants that have been used in the synthesis of AgNPs include Acalypha indica leaves,
Murraya koenigii (curry) leaves, Nicotiana tobaccum leaves, Eucalyptus, Jatropha curcas seeds, Rosarugosa,
Trianthema decandra roots, Ocimum sanctum stems and roots, Sesuvium portulacastrum leaves, Macrotyloma
uniflorum seeds, Ocimum sanctum (Tulsi) leaves, Stevia rebaudiana leaves (12); Others include some Acanthe
phylum, Citrus sinensis leaves, Ocimum tenuiflorum, Solanum trilobatum, Syzygium cumini, Centella asiatica,
Arbutus unedo leaves, Ficus benghalensis leaves, Mulberry leaves, Olea europaea leaves and Chenopodium
album leaves (13). The roots and leaves of Acorus calamus have the ability to synthesis silver nanoparticle in
addition to having antioxidant, antimicrobial and insecticidal activities (14).
Plant mediated metal nanoparticles are presently being introduced as insect pest control in agricultural sector
along with other several plants responsible for production of several secondary metabolites which helps
agricultural insect pest control loaded with nanomaterials. Synthesis involving the use of Nanoparticle has also
been considered as a means of irradiating the tobacco caterpillar, Spodoptera litura (Fabricius) (Lepidoptera:
Noctuidae) which is a major pest of grains and pulses with high mobility and reproductive capacity (15, 16, 17).
Inorganic nanoparticles such as CdS, Nano-Ag and Nano-TiO2 have also been used to control Spodoptera litura
which feeds on more than 120 host plants including tobacco, cotton, groundnut, jute, maize, rice, soybeans,
cauliflower, cabbage, capsicum, potato, castor etc (18). Latex of Alstonia scholaris and Hevea brasiliensis,
Calotropis gigantean, Musa paradisiacal and Achras sapota, also posses the potential to produce green
nanoparticles (19). This was also observed when nanoparticles loaded with garlic essential oil was effective against
the storage pest of maize and milled flour Tribolium castaneum Herbst (20).
Insecticides of synthesized natural products for vector control have been a priority in recent times. Larvicidal
activity of synthesized silver nanoparticles (AgNPs) utilizing aqueous extracts from Eclipta prostrata, a member
of the Asteraceae was investigated against fourth instar larvae of filariasis vector, Culex quinquefasciatus say
and malaria vector, Anopheles subpictus Grassi (Diptera: Culicidae). The stability of the silver nanoparticles can
be attributed to the formation of silver electride that may form a thin layer on the aqueous surface of the reaction
mixture. It seems quite probable that the phenols play an important part in the reduction of ions to AgNPs as the
269
concept of antioxidant action of phenol compounds is not new. Therefore, in combination with mosquito nets or
other vector control measures, such plants synthesized AgNPs may have significant impact on malaria and
filariasis incidence and can be potential candidates to be considered in integrated vector control programs. Plants
synthesized AgNPs, being readily available and their application methods being simple and affordable may be
useful in protection against malaria and filariasis. The pediculocidal and larvicidal activity of synthesized silver
nanoparticles using an aqueous leaf extract of Tinospora cordifolia was also reported to show maximum
mortality against the head louse Pediculus humanus and fourth instar larvae of Anopheles subpictus and Culex
quinquefasciatus (21, 22).
1.3 Controlled Release Formulations [CRFs]
The developed Controlled Release Formulations [CRFs] have been found useful for efficient pest management
in different crops and post harvest processes. It has been suggested that when a commercial formulation for
practical field applications are sought after, it must be ensured that compatible material with the proposed
applications: environment friendly, readily biodegradable, not generating toxic degradation by products and
low-cost are used. The use of several biopolymers, i.e., polymers that are produced by natural sources, which at
the same time have good physical and chemical properties and still present mild biodegradation conditions, are
an interesting approach to avoid the use of petrochemical derivatives that might be another source of
environmental contamination. Some of the recognized common polymers (synthetic and natural ones) used in
CRFs for insecticide and antifeedant applications are listed in Table II.
Table II: Some Polymers used in Nanoparticles for Biopesticides Controlled Release Formulations
[CRFs] production (23)
S/N Polymer Active Compound Nanomaterial
1. Polyethylene glycol B-Cyfluthrin Capsule
2. Polyethylene Piperonyl Capsule
Butoxide and
Deltamethrin
3. Polyethylene glycol Garlic Essential Capsule
Oil
4. Carboxymethylcellulose Carbaryl Capsule
5. Alginate-glutaraldehyde Neen Seed Oil Capsule
6. Alginate-bentonite Imidacloprid or Clay
Cyromazine
7. Polyamide Pheromones Fiber
8. Starch-based polyethylene Endosulfan Film
9. Carboxymethyl chitosanricinoleic Azadirachtin Particlea
acid
10. Vinylethylene and Pheromones Resin
vinylacetate
11. Glyceryl ester of fatty acids Carbaryl Spheres
1.4 Developing New Nanopesticides
Microencapsulation has been used as a versatile tool for hydrophobic pesticides, enhancing their
dispersion in aqueous media and allowing a controlled release of the active compound. Serving as
delivery systems, they convene more selectivity, without hindrance in the accessibility of the bioactive
compounds towards the target pathogen (24).
It has been observed that some nanopesticides have already been on the market for several years too.
Attempt to develop essential oil for pesticides and insecticides have been made in a variety of water-
270
soluble formulations such as nanoemulsion incorporated with B-cypermethrin (25) and essential oil-loaded
microcapsules for pest control (26).
Many attempts have been made to manage pests and insects, an example involves the use of biological control
which has a high timing rate factor. This led to the development of the controlled release systems which has been
termed a viable option. The Controlled Release Formulations (CRFs) associate the active compound with inert
materials. The latter are responsible for protecting and managing the rate of chemical compound release into the
target site in a defined period of time. The main purpose of controlled release systems is in controlling the
availability of the active compound after the application. Most of those controlled release biopesticides
applications were and are still been successfully made due to the advances in nanotechnology area.
Other advantages of the use of nanoparticle insecticides are the possibility of preparing formulations which
contain soluble compounds that can be more readily dispersed in solution. It reduces the problems associated
with drifting and leaching, due to its solid nature, and leads to a more effective interaction with the target insect.
These features enable the use of smaller amount of active compound per area, as long as the formulation may
provide an optimal concentration delivery for the target insecticide for longer times. Since there may be no need
for re-applications, they also decrease the costs), reduce the irritation of the human mucous-membrane, the
phytotoxicity, and the environmental damage to other untargeted organisms and even the crops themselves (27).
In summary, nanotechnology can be applied in several ways in order to enhance efficacy of insecticides in crops
and for post harvest storage processes. Nanotechnology would provide green and efficient alternatives for the
management of insect pests in agriculture without harming the nature. An objection to the use of previously
known synthetic pesticides is still ongoing as there is a great concern regarding the fact that such chemical
compounds have potential to exert hazardous effects on human and the environment (28) and when we have a
nano-pesticide, it becomes a double edged weapon. However, the question that comes to mind is how safe are
these Nanopesticides?
1.5 Safety of Nanopesticides
Due to growing concerns, presentations were made at the EPA Pesticide Program Dialogue Committee (PPDC)
meeting on the safety of the usage of Nanopesticides. The supposed potential Human Health Concerns were
categorized into:
(a) Dermal absorption (so small they may pass through cell membranes)
(b) Inhalation (go to the deep lung and may translocate to the brain i.e., could cross the blood brain barrier)
(c) Potential Environmental Concerns: High durability or reactivity of some nanomaterials raises issues on
their fate in the environment, i.e. biodegradability.
(d) Lack of information to assess environmental exposure to already engineered nanomaterials.
Through considerations for nanotechnology-based pesticides (NBPs) exposures, researchers have found that
effects of dermal exposure to pesticides may be localized at the immediate area of contact or the pesticide may be
absorbed into the bloodstream, depending on a variety of factors, most notably, whether or not the active
ingredient or carrier is lipid soluble.
In recommendations to assess exposure to Nanotechnology- based pesticides, reports according to the
Government Accountability Office, since 2007 stated that the EPA has received a few applications for
registration of various nanosilver pesticide preparations. Some of the applications have disclosed the fact the
pesticide included nanomaterials, while in other cases the agency was able to determine the pesticide contained
nanomaterials from the manufacturing processes.
It was also found that numerous studies and reviews will be required to address the exposure related issues.
There are several lines of existing nanotechnology and pesticide research that are directly relevant to NBP
exposure. While no single study will address all of the exposure-related concerns outlined above, it is evident that
much of the prior research conducted on pesticides and nanomaterials would be applicable to characterize
271
exposure to NBPs. This had led to the need to determine:
· If completely new active ingredients that are synthesized as NBPs are amenable to existing tests and
assessment
· If data from conventional active ingredients that may be reformulated into NBPs are suitable for
registration purposes, and
· If NBPs should be regulated under a completely different framework. (27)
Conclusion
The evolution of Nanoparticles is a resourceful and effective means for control of pests, but the deficiency of
required information on how their detailed mechanisms and actions and how they can be controlled are giving
regulators pause before allowing their release into the environment. Nanopesticides hold promise for reducing
the environmental footprint left by conventional pesticides (28).
Research on nanoparticles and insect control should be geared toward introduction of faster and ecofriendly
pesticides in future. It is time therefore that leading chemical companies try to focus on formulation of nano scale
pesticides for delivery into the target host tissue through nanoencapsulation. Nanoencapsulation is currently the
most promising technology for protection of host plants against insect pests.
At present, the toxicological and ecotoxicological risks linked to this expanding technology ("emerging
technology") cannot be assessed yet. Further developments in nanotechnology in this sector can be expected to
become the main economic driving forces in the long run and benefit consumers, producers, farmers,
ecosystems, and the general society at large. Over the next two decades, the Green Revolution would be
accelerated by means of nanotechnology. While nanotechnology is increasingly moving into the centre of public
attention, it is currently not yet linked to any great degree to concerns about health and the environment (29, 30).
The current level of knowledge does not appear to allow a fair assessment of the advantages and disadvantages
that will result from the use of some nanopesticides. It is clear that a great deal of work will be required to
successfully combine analytical techniques that can detect, characterize (e.g., through size, size range, shape or
nature, surface properties), and quantify the active ingredient and adjuvants emanating from nanoformulations,
and also to understand how their characteristics evolve with time, under realistic conditions.
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SORPTION KINETICS OF METHYLENE BLUE ON ADSORBENTS DERIVED FROM DELONIX
REGIA SEED PODS AND VIGNA SUBTERRANEA FRUIT HULLS
1* 2 2
L.K. Akinola ; Ali Ibrahim and Mohammed Mohammed
1
Department of Chemistry, Bauchi State University, Gadau, Bauchi State, Nigeria
2
Department of Chemistry, Federal University Dutse, Jigawa State, Nigeria
*
[ Corresponding Author, e-mail: lukman.akinola@yahoo.com & Telephone: +2348039690371]
ABSTRACT
Kinetics of methylene blue adsorption on adsorbents prepared from Delonix regia seed pods (DRSP) and Vigna
subterranea fruit hulls (VSFH) were investigated. Characterization of the adsorbents indicated that the points of
zero charge (PZC) of DRSP and VSFH are 5.30 and 4.80 respectively. Optimum removal efficiencies of
methylene blue by DRSP and VSFH occurred at pH of 8.00 and 11.00 respectively. For both DRSP and VSFH,
equilibrium was attained after 4 hours of agitation time, leading to over 80% removal of methylene blue at ionic
strength of 0.4 M and beyond. Among the kinetic models tested, the experimental data fitted well to pseudo-
second order model ( = 0.996 for DRSP and = 0.998 for VSFH), suggesting that the adsorption of
methylene blue on DRSP and VSFH involves chemisorption. The study demonstrates that DRSP and
VSFH can serve as alternative low-cost adsorbents for removing methylene blue from wastewater.
KEYWORDS: Adsorption, Delonix regia, Kinetics, Methylene blue, Vigna subterranea
ABBREVIATIONS AND SYMBOLS
= adsorbate initial concentration (mg/L)

= residual concentration of adsorbate in solution at equilibrium (mg/L)
= residual concentration of adsorbate in solution at time t (mg/L)
= intraparticle diffusion rate constant
= pseudo-first order rate constant
= pseudo-second order rate constant
m = mass of the adsorbent (g)
= final pH of solution
= initial pH of solution
? pH = difference between final and initial pH
PZC = point of zero charge
= amount of adsorbate sorbed on the adsorbent at any time t (mg/g)
= amount of adsorbate sorbed on the adsorbent at equilibrium (mg/g)
t = time of adsorption (minutes)
V = volume of the adsorbate solution (L)
á = initial sorption rate of adsorbate
? = desorption constant
275
INTRODUCTION
In recent time, there is a growing interest and concern among scientists and the general public over
increasing contamination of aquatic environment by chemical substances. This is partly due to the realization
that the presence of certain chemical contaminants in aquatic environment is potentially damaging to both
human and animal health. Methylene blue is a heterocyclic aromatic compound, known systematically as 3,7-
bis(dimethylamino)-phenothiazin-5-ium chloride (Fig. 1). In analytical chemistry, methylene blue is used for
spectrophotometric determination of hydrogen sulfide [1] and as redox indicator [2]. It is also used in medicine
for the treatment of refractory hypotension [3], methemoglobinemia [4–7], sepsis in immunosuppressed patients
[8] and as antidote for cyanide poisoning [9]. However, in spite of its vast medical and scientific applications,
consumption of excess amount of methylene blue has been shown to have neurotoxic effect on the central
nervous system [10]. Due to its widespread use, substantial quantity of methylene blue is released into aquatic
environment during production, usage, and disposal and this may result in excess amount of methylene blue
entering the food chain through bioaccumulation by aquatic biota. Besides its harmful impact on human health,
contamination of aquatic environment by excess methylene blue (or any other dye) causes reduction in the
growth of algae due to obstruction of light required for photosynthesis, which subsequently leads to ecological
imbalance in the aquatic ecosystem [11].
In view of the adverse effects associated with contamination of aquatic environment by excess
methylene blue, remediation techniques such as biodegradation [12-13], phytoremediation [14], adsorption [15-
16] and advanced oxidation process [17-20] have been studied for their viability in removing, decolorizing or
degrading methylene blue in wastewater. Among these remediation techniques, removal of methylene blue (and
other contaminants) by adsorption process is considered to be the safest because no toxic residue is left behind in
the treated water. Although adsorbents such as clay minerals [21-24] and commercial activated carbon [25] have
been employed for the removal of methylene blue from wastewater, research into the use of alternative low-cost
adsorbents derived from agricultural wastes is steadily growing. Advantages of using adsorbents obtained from
agricultural waste materials for the removal of toxic contaminants from wastewater include a partial reduction in
the volume of solid waste materials in the environment and a cutback in the cost of wastewater treatment [26].
Trees of Delonix regia (picture on the left of Fig. 2) are planted mainly for ornamental purpose while Vigna
subterranea (picture on the right of Fig. 2) is cultivated mainly because of its high nutritional value. In
northern Nigeria, large quantities of Delonix regia seed pods (DRSP) and Vigna subterranea fruit hulls
(VSFH) are usually disposed of as waste materials. Hence, the main focus of this paper is to study the
kinetics of methylene blue adsorption on adsorbents derived from DRSP and VSFH with a view to finding
out the suitability of removing methylene blue from wastewater by these adsorbents.
Fig. 1: Structural formula of methylene blue
276
Fig. 2: Pictures of Delonix regia seed pods (left) and Vigna subterranea fruit hulls (right)
Collection and Preparation of Adsorbents
Delonix regia seed pods were obtained from the main campus of Bauchi State University, Gadau, Bauchi
State, Nigeria and Vigna subterranea fruit hulls were obtained from a farm in Darazo Local Government Area of
Bauchi State, Nigeria. The seed pods and the fruit hulls were washed with water, air-dried, pulverized and sieved
into fine particles as previously described [27-28]. The fine particles derived from Delonix regia seed pods and
Vigna subterranea fruit hulls were stored in air-tight containers and labeled DRSP and VSFH respectively.
Preparation of Adsorbate and Reagent Solutions
Analytical grade reagents were used to prepare stock solutions containing 1000
methylene blue, 2 KNO3, 0.1 HNO3 and 0.1 NaOH solutions using
standard procedure [29]. This involves dissolution of appropriate amount of each reagent in doubly
distilled water and diluting the resulting solutions in 1000 volumetric flasks. These stock
solutions were later used in subsequent experiments described in this paper.

Determination of Points of Zero Charge
The points of zero charge (PZC) of DRSP and VSFH were determined using solid addition method
[30]. In this method, 1.0 g of DRSP or VSFH were added to ten 250 Erlenmeyer flasks
containing 50 of aqueous solution each. Prior to transferring these aqueous solutions into the
250 Erlenmeyer flasks, the ionic strengths were adjusted to 0.1 by adding
appropriate amount of 1.0 while the pH values were adjusted to values ranging
from 2.00 to 10.00 by adding appropriate amounts of 0.1 solution or 0.1
NaOH solution. The flasks were then agitated at 200 rpm for 1 hour using WSZ series Orbital
Shaker. Thereafter, the pH values of the supernatants were measured using JENWAY 3505 pH
meter. The difference between the initial and final pH was calculated using equation (1). Plots of
against were constructed and the points of interception on the axes gave values of PZC for
DRSP and VSFH.
= -
Effects of Operating Variables on Removal Efficiency
Effects of initial solution pH (in the range of 2.00 to 12.00), ionic strength (in the range of 0.2 to 1.6 M)
and contact time (in the range of 20 to 480 minutes) on the removal efficiencies of methylene blue by DRSP and
VSFH were investigated using a one-factor-at-a-time (OFAT) approach [31]. This approach consists of selecting
277
a baseline set of levels for each factor (pH = 2.00; ionic strength = 0.2 M; contact time = 300 minutes were
selected as baseline set in this study), and then successively varying each factor over its range with the other
factors held constant at the baseline level. For each treatment combination, 1.0 g of adsorbent was added to 50
3 3
cm of dye solution containing 50 mg/L methylene blue in 250 cm Erlenmeyer flask. The flasks were then
agitated at 200 rpm for appropriate periods of time using WSZ series Orbital Shaker. The content of each flask
was filtered and the residual concentrations of methylene blue in the flasks were determined at 664 nm using
UNICO UV-2100 Spectrophotometer (UNICO Instrument Co., Ltd, Shanghai, China). The removal efficiencies
of methylene blue (expressed as % removal) were then calculated using equation (2). Optimum condition was
selected for each operating factor by constructing a graph that indicates how the removal efficiencies are affected
by varying the factor with all other factors held constant.
% Removal = x 100% (2)
Investigation of Sorption Kinetics

3
Experiment for sorption kinetic study was carried out by adding 1.0 g of DRSP or VSFH to nine 250 cm
Erlenmeyer flasks containing 50 cm3 of dye solutions. The concentration of methylene blue in each flask was
maintained at a value of 50 mg/L while the pH and ionic strength of each solution were adjusted to the requisite
optimal values. The flasks were then agitated at ambient temperature (28 °C) for specific periods of time (20, 40
60, 80, 100, 120, 140, 160 and 180 minutes) using WSZ series Orbital Shaker at 200 rpm. The content of each
flask was filtered and the residual concentrations of methylene blue in these filtrates were determined at 664 nm
using UNICO UV-2100 Spectrophotometer (UNICO Instrument Co., Ltd, Shanghai, China). The amount of
methylene blue adsorbed onto the adsorbent at various time intervals was calculated using equation (3). This
experiment was carried out twice and the fitness of the average data obtained was tested using intraparticle
diffusion model (equation 4), pseudo-first order model (equation 5), pseudo-second order model (equation 6)
and Elovich equation (equation 7)
= ( ) (3)
= (4)
= log - t (5)
= + t (6)
= ln (á ) + ln t (7)
The point of zero charge (PZC) for DRSF and VSFH obtained from the plots of ? pH against pHi
displayed in Fig. 3 are 5.30 and 4.80 respectively. PZC is the pH at which the surface charge of a material is equal
to zero at some ambient temperature, applied pressure, and aqueous solution composition [32-33]. At pH values
below PZC, the surface charge on an adsorbent is negative but the surface charge becomes positive at pH values
above PZC [34-35]. PZC reported in literature for adsorbents prepared from other agricultural wastes are 7.10 for
oil palm fruit fibre [36], 5.00 for sugarcane bagasse [37], 6.00 for watermelon shell [38], 4.18 for orange peel [39]
and 3.50 for groundnut shell [40].
Optimum Operating Variables
Results showing the variation of percents removal of methylene blue by DRSP and VSFH as functions of
initial solution pH, total ionic strength of the solution and contact time are presented in Figs. 4, 5 and 6
respectively. As shown in Fig. 4, higher removal of methylene blue occurred at higher pH values, reaching
278
maximum values of 94.72 % at pH of 8.00 for DRSF and 95.70 % at pH of 11.00 for VSFH. The increased
removal efficiencies of methylene blue by the DRSF and VSFH at pH values above 5.30 (value of PZC for
DRSP) and 4.80 (value of PZC for VSFH) is consistent with the fact that methylene blue molecules exist as
positively charged ions in aqueous solution, and therefore exhibit enhanced electrostatic attractions for the
adsorbents since the surface charges on the adsorbents are negative at pH values above their PZC. The plots
presented in Fig. 5 show that at ionic strength of 0.2 M, 71.88 % and 59.38 % of methylene blue were removed
from aqueous solution by DRSF and VSFH respectively. However, beyond the ionic strength value of 0.2 M,
adsorptions of methylene blue on both DRSF and VSFH increased slightly above 80.00 % but remained
essentially constant in solutions with ionic strength values ranging from 0.4 to 1.6 M. In Fig. 6, the percent
removal of methylene blue by DRSF increased gradually from 38.85 % at 20 minutes to 89.22 % at 240 minutes.
Beyond 240 minutes, the removal efficiency of methylene blue by DRSF remained more or less constant.
Similarly, Fig. 6 also shows that percent removal of methylene blue by VSFH increased gradually from 29.09 %
at 20 minutes to 84.42 % at 240 minutes, remaining roughly constant beyond 240 minutes. In view of these
findings, the kinetic experiments involving DRSF and VSFH were carried out at pH values of 8.00 and 11.00
respectively. The ionic strengths of all solutions were adjusted to 1.0 M and all the kinetic data were collected
within the first 3 hours.
Sorption Kinetics
The charts displayed in Figs. 7, 8, 9 and 10 are plots of intraparticle diffusion model, pseudo-first order
model, pseudo-second order model and Elovich model for testing the fitness of sorption kinetic data of
methylene blue on DRSP and VSFH. The coefficients of determination (
values) and the rate constants obtained from these plots are presented in Table 1. Among the four
kinetic models tested, the experimental data fitted well to pseudo-second order model for both
DRSP and VSFH as indicated by the values of coefficients of determination ( = 0.996 for DRSP
and = 0.998 for VSFH in Table 1). Pseudo-second order model is based on the assumption that
the adsorption process involves chemisorption, which requires sharing or exchange of valence
electrons between the adsorbent and the adsorbate [41]. The pseudo-second order rate constants
obtained in the present study (2.155 for DRSP and 2.873 for VSFH in
Table 1) are several order of magnitude higher than 0.0059 reported for the
adsorption of methylene blue on NaOH-modified dead leaves of plane trees [42], 0.0012
reported for the adsorption methylene blue on NaOH-modified durian leaf powder [43]. The
implication of these findings is that, the rates of removal of methylene blue from aqueous solution
by DRSP and VSFH are faster than the rates of removal of methylene blue from aqueous solution
by adsorbents derived from other agricultural waste materials in the literature cited.
Fig. 3: Plots of ? pH against pHi showing points of zero charge of DRSF and VSFH
279
Fig. 4: Plots of removal efficiencies against initial solution pH
Fig. 5: Plots of removal efficiencies against ionic strength
Fig. 6: Plots of removal efficiencies against contact time
Fig. 7: Intraparticle diffusion plots for methylene blue adsorption on DRSP and VSFH
280
Fig 8: Pseudo-first order plots for methylene blue adsorption on DRSP and VSFH
Fig. 9: Pseudo-second order plots for methylene blue adsorption on DRSP and VSFH
Fig. 10: Elovich plots for methylene blue adsorption on DRSP and VSFH
Table 1: Coefficients of determination and rate constants of methylene blue adsorption on DRSP and VSFH
Intraparticle diffusion model Pseudo-first order model Pseudo-second order model Elovich model
Adsorbent
á ?
DRSP 0.386 0.001 0.303 0.000 0.996 2.155 0.488 125.0
VSFH 0.610 0.000 0.674 0.000 0.998 2.873 0.519 -333.3
CONCLUSION
The search for alternative low-cost adsorbents from locally-available agricultural waste materials
provided the impetus for this research work. Adsorbents prepared from the seed pods of Delonix regia and the
fruit hulls of Vigna subterranea were characterized and the suitability of these adsorbents, vis-à-vis their ability
to remove methylene blue from aqueous solution, were evaluated by studying the kinetic behaviors of the
adsorption systems. Findings suggest that adsorptions of methylene blue on both adsorbents were adequately
described by pseudo-second order model, suggesting that the adsorptions involve sharing or exchange of valence
electrons between the adsorbents and the adsorbate. This study demonstrates that the adsorbents prepared from
281
the seed pods of Delonix regia and the fruit hulls of Vigna subterranea may be suitable for removing methylene
blue from contaminated wastewater.
ACKNOWLEDGEMENTS
The authors wish to acknowledge the technical support rendered by Mal. Isah Suleiman and Mal. Salihu Yunusa
of Chemistry laboratory, Bauchi State University, Gadau, Nigeria.
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284
QUANTUM CHEMICAL STUDIES ON THE EFFICIENCY OF
2-HYDROXYETHYLETHYLENDIAMINE AND DIETHYLENETRIAMINE AS CORROSION
INHIBITORS
1 ?,
K. J. Uwakwe , P. C. Okafor ,
1
Corrosion and Electrochemistry Research Group, Department of Pure & Applied
Chemistry, University of Calabar, Calabar, Nigeria
ABSTRACT
The corrosion inhibition potential of 2-hydroxyethylethylendiamine (HEED) and diethylenetriamine (DETA)
was studied by means of the density functional theory (DFT) approach BLYP/DNP calculations using the
software Material studio from Accelyrs Incorporation San Diego California U.S.A. Quantum chemical
calculations and other electronic properties such as highest occupied molecular orbital energy (EHOMO), lowest
unoccupied molecular orbital energy (ELUMO), energy gap (ÄE), dipole moment (µ), electron affinity (A),
ionization potential (I), global hardness (? ), global softness (S), Fukui functions f(r), natural atomic charge and
bond length analysis were performed on the molecule at geometry optimization. The inhibition efficiency is in the
order of HEED > DETA, with the active sites for adsorption on the molecules being the nitrogen and oxygen
heteroatoms in HEED and the nitrogen heteroatoms in DETA based on the natural atomic charge, distribution of
frontier molecular orbital, Fukui functions or indices and the bond length analysis. The molecules can be said to
act as corrosion inhibitor.
Keywords: Corrosion inhibitor, DFT, 2-hydroxyethylethylendiamine, diethylenetriamine, Adsorption.
1. INTRODUCTION
Iron and its alloys are widely used in many applications, which have resulted in research into their
resistance to corrosion in various environments. In efforts to mitigate metal corrosion, the primary strategy is to
isolate the metal from the corrosive agents [1, 2]. Among the different methods available, to mitigate corrosion, is
the use of corrosion inhibitors [2]. As a protection technique, the addition of corrosion inhibitors are widely used
in many industries, such as petroleum, chemical industry, construction, etc. And it is also one of the most
economical ways to protect the metal [3]. Theoretical studies at molecular level have been reported to aim at
gaining insight on the molecules chemical activity [4] and in particular to study corrosion inhibiting processes
[5] taking into account model molecules and their structural and electronic properties. Studies on chemical
reactivity [4] showed that the heteroatom from the molecules plays an important role within the inhibiting
mechanism leading to a better understanding of this issue. The objective of this work is to determine the
inhibitive/adsorption ability and the active sites of 2-hydroxyethylethylendiamine and diethylenetriamine by
applying the Density functional theory (DFT) through the knowledge of the structural and electronic properties
of the molecules involved. The reason for choosing these compounds is because Amines and their salts are said to
be the most economic and effective corrosion inhibitors for oil and gas wells [6]. The names, abbreviations,
chemical formula, structures, molecular weights and melting points of the amine used in this work are shown in
Table 1.
Table 1. Names, abbreviation, chemical formula, structure and molecular weight of the studied compound
Name Abbreviation Chemical Structure Molecular
formula Weight (g/mol)
2-hydroxyethylethylendiamine HEED C4H12N2O NH 104.15

HO NH2
Diethylenetriamine DETA C4H13N3 NH 103.17

H2N NH2
285
2. COMPUTATIONAL DETAIL
The density functional theory (DFT) calculations was performed using BLYP (from the name Becke for the
exchange part and Lee, Yang and Parr for the correlation part) functional method and the double numeric with
polarization (DNP) basis set. This basis set is used because it is the best set available in Dmol3 [7].Among the molecular
properties that are well reproduced by DFT/BLYP include the energy of the highest occupied molecular orbital (EHOMO),
energy of the lowest unoccupied molecular orbital (ELUMO), global hardness (? ) and softness (S), electron affinity (A),
ionisation potential (I) etc. These quantities are often defined following Koopmans' theorem [8]. Where EHOMO and ELUMO
is related to the Ionization potential and the electron affinity according to and Thus the
value of the global hardness (? ) according to Pearson, operational and approximate definitions is given as [9]
The global softness (S) is the inverse of the global hardness (? ) [10] . The local reactivity was analyzed by
determining the Fukui indices which indicate the reactive regions, in the form of the nucleophilic and electrophilic
behavior of each atom in the molecule. The Fukui function f(r) is defined as the first derivative of the electronic density
ñ(r) with respect to the number of electrons N at a constant external potential v(r). Thus, using a scheme of finite
differences [11], we have (electrophilic attack) and
(nucleophilic attack). Where f- -, f +, ñN+1，ñN, and ñN-1 are the Fukui negative, Fukui positive, electronic densities of
anionic, electronic densities of neutral species and electronic densities of cationic species, respectively. The N is the
number of electrons in the molecule. N+1 is the anion, with an electron added to the LUMO of the neutral molecule. N-
1 is the cation with an electron removed from the HOMO of the neutral molecule. Figure 1 gives the optimized
geometry structures for the studied compounds with its atoms labeled. The colour codes for the atoms in the molecules
studied are gray for carbon, blue for nitrogen, red for oxygen and white for hydrogen.
(a) (b)
Figure 1. The optimized geometries for (a) HEED and (b) DETA with labeled atoms
3. RESULT AND DISCUSSION
Table 2 gives the calculated quantum chemical parameters using the BLYP/DNP density functional
theory approach.
Table 2.Calculated quantum chemical parameters for the two molecules studied
Molecule EHOMO (eV) ELUMO (eV) ? E (eV) µ(debye) I (eV) A (eV) ? S
HEED -5.041 0.484 5.530 3.251 5.041 -0.484 2.763 0.362
DETA -4.906 0.849 5.760 1.371 4.906 -0.849 2.878 0.347
According to the frontier molecular orbital theory (FMO) of chemical reactivity, transition of electron is due to
interaction between highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital
(LUMO) of reacting species [12]. The electron donating ability of a molecule is usually attributed to EHOMO, which
is how well a molecule can give out electrons to the vacant d-orbital of a metal. ELUMO is usually attributed to the
ability of a molecule to collect electrons present in a metal's d-orbital, resulting in the creation of another bond
[13]. From Table 2, DETA has the highest value of EHOMO, while HEED has the lowest value of ELUMO. The negative
286
signs observed on the values of EHOMO shows that the adsorption is physisorption [14]. The HOMO and LUMO
orbital plot on the molecules studied is shown in Figure 2. Note that the blue and yellow isosurfaces depict the
electron density difference; the blue regions show electron accumulation, while the yellow regions show
electron loss. The value of ÄE (ELUMO – EHOMO) is a vital stability sign. Higher ÄE values, gives good stability and
low reactivity and hence bad inhibition efficiency. Low value of ÄE gives better inhibition efficiency, for the
reason that lower excitation energy will be required to take away an electron from the last occupied orbital [15].
HEED has a lower ÄE value compared to DETA. The dipole moment is another important electronic parameter
that results from non-uniform distribution of charges on the various atoms in the molecule [16]. The high value of
dipole moment probably increases the adsorption between the chemical compound and the metal surface [17].
HEED has a higher dipole moment value than DETA this also confirms the higher inhibition effect of HEED.
Another important property to measure the molecular stability as well as reactivity of a molecule is the absolute
hardness and softness. A hard molecule has a large energy gap and a soft molecule has a small energy gap [18].
Normally, the inhibitor with the least value of global hardness (hence the highest value of global softness) is
expected to have the highest inhibition efficiency [19]. From Table 2 HEED has the lowest hardness and highest
softness. This also confirms the higher inhibition efficiency of HEED compare to DETA.
To determine the active site of a molecule, three influencing and controlling factors: natural atomic
charge, distribution of frontier molecular orbital and Fukui indices have to be considered [20]. Local reactivity is
analyzed by means of the condensed Fukui function. Condensed Fukui functions allow us to distinguish each
part of the molecule on the basis of its distinct chemical behavior due to the different substituent functional
groups. The nucleophilic and electrophilic attack is controlled by the maximum values of f + and f- -. The
calculated Fukui indices for nucleophilic and electrophilic attack for the three selected inhibitors are tabulated in
Table 3 (only C, N, and O are quoted) and their active sites (Fukui indices plots) are shown on the molecules in
–
Fig 2. From Table 3, O1, N4 and N7 atoms in HEED has high f values, while N1, N4 and N7 atoms in DETA also
show high f – values. With the main site for f – being the atom with the highest Fukui indices value which is N4 for
HEED and DETA. All atoms in the molecules studied posses negative Fukui indices values for f +, but the ones
+
with the highest values of f in the molecules studied is the N4 atom in HEED and the N7 atom in DETA and they
are considered to be the main sites for nucleophilic attack on the molecules.
Molecules HOMO LUMO f- f+
HEED
DETA
287
– +
Figure 2. HOMO, LUMO, f and f orbital plots for (a) HEED and (b) DETA
Table 3. f – and f + indices values for HEED and DETA

Molecules f (r) Atoms
O1 C2 C3 N4 C5 C6 N7
HEED f- 0.031 -0.014 -0.063 0.195 -0.055 -0.044 0.125
f+ -0.079 -0.123 -0.070 -0.002 -0.045 -0.023 -0.008
N1 C2 C3 N7 C5 C6 N7
DETA f- 0.076 -0.028 -0.044 0.134 -0.052 -0.038 0.140
f+ -0.091 -0.054 -0.063 -0.086 -0.061 -0.040 -0.025
Bond length or bond distance is the average distance between two bonded atoms in a molecule. The bond lengths
in Armstrong (Å) of the molecules are shown in Fig 3 (a) and (b). It is observed that the heteroatom's (nitrogen
and oxygen) bonded to a carbon atom show a shorter bond length than the carbon to carbon bond length. Shorter
bond length requires high dissociation energies to break the bond. This means that the closer the nuclei of the
bonding atoms are a greater supply of energy is needed to separate the atoms due to large force of attraction
between the atoms and hence the higher is the reactivity of the bond which can also lead to higher inhibition
efficiency. From classical chemical theory, all chemical interactions can be by orbital interactions or by
electrostatic interaction. The presence of electric charge in the molecule is the propelling force of an electrostatic
interaction. Local electric charges have been proven to be vital in several chemical reactions as well as
physiochemical characteristics of compounds [20]. Fig 4 (a) and (b) shows the natural atomic charges in
coulombs (C) for the two molecules studied. The heteroatoms show a lower atomic charge than the carbon atoms
thereby making them possible sites for adsorption on the molecules. It is observed that the oxygen atom is more
negative than any other atom in HEED and the carbon atom close to it (C2) is very much electron deficient, this
may be due to the highly electronegative nature of oxygen. The hydrogen atoms in both molecules studied are all
positively charged.
Figure 3. Bond length analysis for (a) HEED and (b) DETA
288
Figure 4. Natural atomic charge for (a) HEED and (b) DETA
4. CONCLUSION
Using the DFT/BLYP method, the inhibition efficiency of HEED and DETA is investigated leading to the
following conclusions
· Quantum chemical calculations such as ? E, µ, and S gives the inhibition efficiency to be in the order of
HEED > DETA.
· From the distribution of frontier molecular orbital plots, natural atomic charge values, Fukui indices
plots and values and also from the bond length analysis, the active sites for adsorption on these
molecules are the O1, N4 and N7 heteroatoms in HEED and the N1, N4 and N7 heteroatoms in DETA.
· HEED and DETA can be used as corrosion inhibitor, with HEED being more preferred.
ACKNOWLEDGMENT: K.J. Uwakwe is grateful to Dr P.C. Okafor for providing the software which was
used to perform this research work.
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[16] O. Kikuchi (1987), Systematic QSAR procedures with quantum chemical descriptors, Quant. Struct,-Act.
Relat, 6:179.
[17] X. Li, S. Deng, H. Fu and T. Li (2009), Adsorption and inhibition effect of 6- benzylaminopurin on cold
rolled steel in 1.0 M HCl, Electrochim. Acta. 54, 4089
[18] N.O. Obi-Egbedi, I.B. Obot, M.I. El-Khaiary, S.A. Umoren, E.E. Ebenso (2011) Computational simulation
and Statistical analysis on the relationship between corrosion inhibition efficiency and molecular structure of
some Phenathroline derivatives on mild steel surface, Int. J. Electro. Chem. Sci, 6:5649-5675.
[19] I. Lukovits, E. Kalman, F. Zucchi (2001), Corrosion inhibitors correlation between electronic structure and
efficiency, Corrosion 57: 3-8.
[20] S. Xia, M. Qui, L. Yu, F. Lui, H. Zhao (2008), Molecular dynamics and density function theory study on
relationship between structure of imidazoline derivatives and inhibition performance, Corros. Sci, 50:2012-
2029.
290
COMPARATIVE STUDIES OF THE INHIBITIVE POTENTIALS OF RED PEANUT SKIN
EXTRACT AND LASIANTHERA AFRICANA P. BEAUV. (NKANKA LEAF) MUCILAGE ON MILD
STEEL CORROSION IN SULPHURIC ACID SOLUTION.
a b c
A.O. James, Kalu Nnennaand Akaranta O.
a, b & C
Department of Pure and Industrial Chemistry, University of Port Harcourt,
P.M.B. 5323, Port Harcourt
a
Email address:abosede.james@uniport.edu.ng
b
Email address: nellyebony@gmail.com
c
Email address: akaranta.onyewuchi@uniport.edu.ng
a
Phone Number: 08057314332
b
Phone Number:08063485608
c
Abstract:
The inhibition efficiency of red peanut skin extract and lasianthera africana P. beauv. mucilage on mild steel
deterioration (corrosion) in 2M sulphuric acid was studied by the method of weight loss. The extracts were
prepared by immersing the red peanut skin and nkanka leaves in hot water (1000g/l and 100g/0.5l of water
0 0 0
respectively). The inhibition efficiencies were studied at various temperatures: 30 C, 40 C and 50 C. Results
obtained indicated that mild steel corrosion in sulphuric acid was mitigated significantly by the extracts. As the
inhibitor concentration in the acid increased, the inhibition efficiency also increased while temperature increase
resulted to a decline in the inhibition efficiency. The difference in the inhibition potency of the two extracts is
explained in terms of the difference in the molecular structures of their active component. The active component
in red peanut skin extract is catechin while that of lasianthera africana p. beauv. (nkanka leaf) mucilage is pectin.
Keywords:Corrosion; Red peanut skin; Mild steel; Mucillage; Weight loss.
INTRODUCTION
Corrosion is the weakening in the structure of a material due to interactiob with its surroundings. It can also be
described as the destructive attack of materials mostly metals, by electrochemical reactions (Natty, 2010).
Corrosion processes are redox reactions involving oxidation and reduction. (Lebo, et al., 2001).
Mild steel is exposed to the action of acid in industrial processes in which acids have important functions, for
example in oil well acidification, acid pickling, acid cleaning, and acid descaling. Corrosion inhibition of mild
steel in acid solutions has become one of the most urgent and severe challenges in acid pickling process (Shukla
and Qurarishi, 2010; Aljourani et al., 2009).
Corrosion inhibitors are substances added in small quantities to an environment to prevent corrosion of metals
(Quarishi, 1999).The use of inhibitors is one of the best options of protecting metals and alloys against corrosion.
The selection of inhibitor is controlled by its economic availability, its efficiency to inhibit the substrate material
and its environmental side effects.
Most of the efficient acid inhibitors are organic compounds containing nitrogen, sulphur and/or oxygen atoms in
their molecule (Sudhish and Quraishi, 2010; Eddy and Ebenso, 2008; Sharma et al., 2008). The toxicity of
organic and inorganic corrosion inhibitors to the environment has prompted the search for safer corrosion
inhibitors such as green corrosion inhibitors as they are biodegradable and do not contain heavy metals or other
toxic compounds. In addition to being environmentally friendly and ecologically acceptable, plant products are
inexpensive, readily available and renewable (Eddy and Ebenso, 2008).
291
In this study, the inhibition efficiency of mild steel corrosion in red peanut skin extract and lasianthera africana
mucilage in sulphuric acid at different concentrations and temperatures wasinvestigated by the method of weight
loss.
EXPERIMENTAL PROCEDURE
Preparation of the Metal Coupons
The metal coupons were purchased at steel market, Mile 1, Port Harcourt. They were mechanically press-cut into
4 x 3cmcoupons with a hole, 2mm in diameter at the tip for suspending the metal into the acid solution. The mild
steel coupons were not polished any further but was degreased in absolute ethanol and dried in acetone; then
stored in a dessicator to prevent any form of contamination before the experiment.
Preparation of Various Concentrations of Red Peanut Skin Extract

The red skin of roasted peanut was obtained locally from a groundnut seller at Choba junction in Port Harcourt,
Rivers State. 1000g of the red peanut skin was boiled in 1litre of water for about 3 hours. Filtration was done after
cooling of the mixture. The acid solution (2M H2SO4) was added to varied amounts of the extract solution in %v/v
using the following procedure:
10%v/v concentration was prepared by adding 90ml of H2SO4 to 10ml of the extract in a 250ml beaker; same
procedure was followed to prepare other concentrations: 20%v/v, 30%v/v, 40%v/v and50%v/v. Another beaker
containing 100ml H2SO4 was used as blank for the weight loss experiment.
Preparation of Mucilage From Nkanka Leaves (Lasianthera Africana)
Nkanka leaves (Lasianthera africana) were obtained locally from a farmland in Etchie Local Government of
Rivers State. The leaves were washed, dried, chopped and weighed. 100g of the leaves were completely
o
immersed in 0.5litres of hot water (100 C) for 3 hours till the water turned cold. The mucilage (slimy liquid) was
squeezed out of the leaf and filtered with a sieve to remove tiny leaf particles. 2M H2SO4 solution was added to
varied amounts of the extract solution in %v/v.
Weight Loss Determination

The various concentrations (blank, 10%v/v, 20%v/v, 30%v/v, 40%v/v, and 50%v/v) of both extracts of red
peanut skin and mucilage in 2M H2SO4 were maintained at 300C. The mild steel coupons which had been weighed
were placed in the corrodent-inhibitor solutions. Each coupon was recovered from these solutions every 24hours
consistently for 168hours (7 days). Each retrieved coupon was immediately dropped into distilled water to stop
the corrosion reaction, washed, dried and reweighed. The coupon weight difference was recorded as the weight
o o
loss. The temperature was raised to 40 C and 50 C and the experiment was performed again in a thermostatic
water bath.
Efficiency of the inhibition (%E) was computed from the outlined equation below:
%E = WB – Wi X 100 ------------------- (1)

WB 1
Where ? WBand ? Wi are the weight loss values of the mild steel coupons in the absence
(blank) and presence of an additive respectively.
292
The results obtained from the weight loss experiment with inferences drawn from them are
discussed below:
Inhibitory Effect of Red Peanut Skin Extract on Mild Steel Corrosion
Fig. 1: Variation of Weight Loss with Time for Mild Steel Coupons in 2.0M H2S04 and various concentrations of
0
red peanut skin extract solution at 30 C.
From the variation of weight loss with time of exposure of mild steel in 2M sulphuric acid (blank) at 300C (Fig. 1)
compared with those containing the additives, there is a remarkable decrease in weight loss signifying corrosion
inhibition.
The same observations were made at 40oC and 50oC. This shows that the red peanut skin extract is very
economical since it could inhibit corrosion greatly even at small amounts.
Effect of Temperature on the Inhibition Efficiency of Red Peanut Skin Extract
293
Fig. 2: Variation of inhibition efficiency with inhibitor concentration of red peanut skin extract for mild steel
corrosion in 2M H2SO4 solution at different temperatures.
0 0
From fig. 2, it can be seen that as the temperature of the system was increased from 30 C to 50 C, the inhibition
0
efficiency decreased. But even at this temperature (50 C), the red peanut skin extract still inhibited corrosion
significantly.
Effect of lasianthera africana mucilage on mild steel corrosion in 2M H2SO4 solution:

The figures below show the effect of lasianthera africana mucilage on the weight loss of mild steel in
sulphuric acid solution at the temperatures studied.
Fig. 3: Variation of Weight Loss with Time for Mild Steel Coupons in 2.0M H2S04 solution containing
0
different concentrations of the mucilage extract from lasianthera africana (nkanka leaves) at 30 C.
Fig. 3 shows the effect of the addition of the mucilage the acid, it was observed that as the additive concentration
increased, the metal weight loss decreased significantly, with 10%v/v showing the highest weight loss while the
highest additive concentration 50%v/v showed the least weight loss at the different temperatures studied. This
o
signifies inhibition of the metal dissolution in sulphuric acid.Same behaviour was recorded at 40 and 50 C.
Effect of Temperature on the Inhibition Efficiency of lasianthera africana mucilage
294
Fig. 4: Variation of inhibition efficiency with inhibitor concentration of lasianthera africana mucilage for mild
steel corrosion in 2M H2SO4 solution at different temperatures.
From fig. 4, it can be seen that as the temperature of the system was increased from 30 to 500C, the inhibition
efficiency decreased. But even at this temperature (500C), the mucilage still inhibited corrosion significantly.
The active components in the extracts and mucilage.

The active component in red peanut skin extract is catechin while that of lasianthera africana p. beauv. (nkanka
leaf) mucilage is pectin. Both possess electron-rich sites for their adsorption unto the metal surface leading
corrosion inhibition.
OH
OH
HO O
OH
OH
Figure 5: The structure of Catechin (Red Peanut skin).
295
Figure 6: The structure of Pectin (lasianthera africana mucilage)
Comparison of the corrosion inhibition potentials of the extract and the mucilage.
Table 1: The Inhibition Efficiency (%)for mild steel corrosion in 2M H2SO4 solutionby lasianthera africana
mucilage and red peanut skin extract at the temperatures studied
Inhibitor Inhibition Efficiency (%)

Concentration
(% v/v) Lasianthera africana mucilage Red peanut skin extract
30oC 40oC 50oC 30 Co
40oC 50oC
10 79.71 75.11 65.62 74.66 70.04 61.54
20 86.07 81.45 69.95 80.73 72.73 68.85
30 90.39 86.30 80.65 85.86 80.62 79.01
40 93.55 90.28 85.91 89.45 84.11 83.63
50 96.49 94.13 91.68 93.73 90.07 88.98
The inhibitive effect of the extract and the mucilage may be explained by considering the adsorption of their
active compounds through the electron rich oxygen atoms in the molecules on the corroding metal surface. The
active ingredient in lasianthera africana mucilage is Pectin while that of red peanut skin extract is catechin.It is
evident that the inhibition efficiency of the two compoundsdepends mainly on the molecular size of their
structures, the charge density on the adsorption sites,electron clouds, mode of interaction with the metal surface
and formation of metallic complexes. Adsorption is expected to take place primarily through the oxygen atoms
on the molecules. Considering the structures of the compounds (figs.5-6), there are numerous electron-rich
oxygen atoms in them, hence the high percentage inhibition efficiencies recorded (Table 1).
A comparative study of the two extracts shows that lasianthera africana mucilage recorded higher inhibition
efficiency than red peanut skinextracts (Table 1) at all the temperature studied. This may be due to the fact that
the oxygen atoms in pectin are more than those in catechin. Thus creating more adsorption sites and increased
inhibitive potency.
296
CONCLUSION
The following conclusions were made from this study:
§ Red peanut skin extract and lasianthera africana mucilageare good corrosion inhibitors for mild steel in
2M H2SO4 solution.
§ Inhibition efficiency decreased with increased temperature.
§ In both extracts, increase in inhibitor concentration led to an increase in the inhibition efficiency.
§ The mucilage extract exhibited greater inhibition efficiencies (max. = 96.49%) than the red peanut skin
extract (max. = 93.73) under the same conditions. This may be attributed to the size and nature of their
active components; larger structures with larger number of heteroatoms always give a higher inhibition.
Red peanut skin extract and lasianthera africana mucilage should be employed as eco-friendly corrosion
inhibitors and be used to replace toxic organic corrosion-inhibitors.
REFERENCES
1. Aljourani, J., Raeissi, K., Golozar, M.A. (2009). Benzimidazole and its derivatives as corrosion
inhibitors for mild steel in 1M HCl solution. Corros. Sci. Journal, 51:1836-1843.
2. Eddy, N.O., and Ebenso, E.E. (2008). Inhibitive and adsorption properties of ethanol extract of Musa
sapientum peels as a green corrosion inhibitor for mild steel. Afr. J. Pure Appl. Chem. 2(6):46-54.
3. James, A.O., Akaranta, O. and Awatefe, K. J. (2011). Red peanut skin: an excellent green inhibitor for
mild steel dissolution in hydrochloric acid solution. Alfa Universal, An International Journal of
Chemistry, 2(2): 72 – 78.
4. Lebo, S. E., Gargulak, J. D. and McNally, T. J. (2001). Encyclopedia of Chemical Technology; 7th ed.
John Wiley & Sons Inc. NewYork. pp. 420 – 424.
5. Natty, K. (2010). Corrosion and its Mitigation in the Oil and Gas Industry; Leonardo Journal of
Science, 23(6): 12, 19.
6. Quraishi, M. A. (1999). “Investigation of some green compounds as corrosion and scale inhibitors for
cooling systems,” Corrosion Journal,55(5) 493–497.
7. Shukla, S.K., and Quraishi, M.A. (2010). The effects of pharmaceutically active compound
doxycycline on the corrosion of mild steel in hydrochloric acid solution. Corros. Sci. 52(1): 314-321.
8. Sudhish, K.S., and Quraishi, M.A. (2010). Cefalexin drug: A new and efficient corrosion inhibitor for
mild steel in hydrochloric acid solution. Mater. Chem. Phys.120:142-147.
297
In vitro antioxidant activity of Heinsia crinata root extracts.
*Ita, Basil N and Willie, Akaninyene H.
Department of Chemistry, University of Uyo, Uyo, Akwa Ibom State, Nigeria.
* Corresponding author; E-mail: basil_ita@yahoo.com. Tel: +234 80 2334 8014.
Abstract
Heinsia crinata is consumed as a vegetable and its organs are used traditionally in the treatment of various
ailments. In this study, we evaluated the total phenolics, flavonoid and in vitro antioxidant activity of the hexane
and methanol extracts of Heinsia crinata roots. The antioxidant activity was determined by measuring the metal
chelating, ferric reducing power and DPPH scavenging activity of the extracts. Total phenolics ranged between
78.34 - 123.26 mgGAE/g, with a higher content in the methanol extract (123.26mgGAE/g), while flavonoids was
also higher in the methanol extract (89.96 mgQE/g) than the hexane extract. In addition, the methanolic extract
was a better free radical scavenger (EC50 = 0.4mg/ml), had good reducing power (EC50 = 1.01mg/ml) and could
chelate ferrous ions effectively (EC50 = 0.21mg/ml) than the hexane extract. These results imply that the methanol
root extract of Heinsia crinata is a good antioxidant and suggests support for its use in traditional medicine.
Keywords: Total phenolics, flavonoids, antioxidant activity, Heinsia crinata.

Introduction
Heinsia crinata (Rubiaceae) grows abundantly in the South – South region of Nigeria. The leaf is
consumed as a vegetable and has a distinctive aroma. The plant is also used traditionally to treat various ailments
[1-3]. The nutrient composition of the plant has been evaluated [1] and its leaf extracts have been reported to
possess numerous bioactivities including antimicrobial, antioxidative, antiplasmodial, antidiabetic,
hypoglycaemic and antihypertensive effects [4-13]. In addition, the in vitro neuroprotective potentials of the leaf
extracts have also been reported [14]. However, there are less research outcomes on the root of this medicinal
plant. We report in this work, the content of total phenolics and flavonoids as well as the in vitro antioxidant
potentials of the hexane and methanolic root extracts of Heinsia crinata.
Plant material
Roots (876g) of Hiensia crinata were harvested from a local farm in Nsit Ibom, Akwa Ibom State,
Nigeria, in December 2015 and identified by a taxanomist in the Department of Botany and Ecological Studies,
University of Uyo. The roots were air dried, powdered, and extracted at room temperature consecutively with
petroleum ether and methanol (2.1L x 3) respectively. Filtration and concentration (in vacuo) gave the petroleum
ether and methanolic extracts respectively.
Evaluation of Antioxidant activity
Antioxidant activity of the hexane and methanol root extracts of H. crinata was evaluated by measuring its
DPPH radical scavenging ability, ferric reducing power and metal chelating activity.
(a) DPPH Assay: 1ml of varying concentrations (0.1 – 5.0 mg/ml) of the extracts were mixed with 1ml of
0.004% methanol solution of DPPH. The mixture was shaken vigorously and allowed to stand for 30 min at
room temperature in the dark. The reduction of the DPPH radical was determined by measuring the absorption at
517nm. The procedure was repeated for the blank and control. The radical scavenging activity was calculated
using the equation:
DPPH scavenging effect (%) = [(Ablank – Asample)/Ablank] x 100.
Concentration providing 50% activity (EC50) was calculated[15]. BHA and Vitamin E were used as positive
controls.
298
(b) Evaluation of metal chelating activity: Metal chelating ability was determined according to the method of
Decker and Welch [16], with some modifications. Briefly, 0.5ml of each extract (0.1 -5.0mg/ml) was mixed with
0.05ml of 2mMFeCl2 and 0.1ml of 5mM ferrozine. The total volume was diluted with 2ml methanol. Then, the
mixture was shaken vigorously and left standing at room temperature for 10mins. After the mixture had reached
equilibrium, the absorbance of the solution was measured spectrophotometrically at 562nm. The percentage of
inhibition of ferrozine – Fe2+ complex formation was calculated using the formula:
Scavenging activity (%) = [(Acontrol – Asample)/Acontrol] x 100,
2+
where Acontrol = absorbance of ferrrozine – Fe complex,
and Asample = absorbance of sample.
EDTA was used as a positive control.
(c) Evaluation of Ferric reducing powery: The reducing power was determined according to the method of
Oyiazu [17]. Varying concentrations of each extract (0.1 – 5.0mg/ml) in ethanol (2.5ml) was mixed with 2.5ml of
200mM sodium phosphate buffer (pH 6.60) and 2.5ml of 1% potassium ferricyanide, and the mixture was
o
incubated at 50 C for 20 minutes. Later, 2.5 ml of 10% trichloroacetic acid (w/v) was added, and the mixture
centrifuged at 200g for 19 minutes. The upper layer(5ml) was mixed with 5ml of deionised water and 1ml of
0.1% ferric chloride and the absorbance was measured at 700nm against a blank. A higher absorbance indicated a
higher reducing power. EC50 value (mg/ml), the effective concentration at which the absorbance was 0.5 for
reducing power was obtained by interpolation. Ascorbic acid and BHA were used as positive controls.
Determination of Total Phenolics

Concentration of total phenolics in the extracts was determined using the Folin- Ciocalteu reagent
[18]. 0.1ml of each extract (1mg/ml) was dissolved in 0.5ml(1/10 dilution) of the Folin-Ciocalteu reagent and
1ml of water/methanol (1:2) was added. The solution was mixed and incubated at room temperature for 1 min.
After 1 min, 1.5ml of 20% Na2CO3 solution was added. The final mixture was shaken and incubated for 2 hrs in
the dark at room temperature. The absorbance was measured at 760nm using a UV-Vis spectrophotometer. Gallic
acid was employed as the standard and the results expressed in mg gallic acid per gram (mgGAE/g) .
Determination of Flavonoids
A slightly modified version of the spectrophotometric method was used to determine the flavonoid
content. Each extract (0.1g) was mixed with 20ml of 80% aqueous methanol and then filtered with Whatman
filter paper No. 42 to obtain a clear filtrate. A 0.5ml aliquot of this filtrate was taken in a test tube and 3ml of
distilled water and 0.3ml of 0.5% sodium nitrite were added. The solution was mixed and allowed to stand at
room temperature for 5 min. To this solution, 0.6ml of 10% aluminium chloride was added. After 6 min, 2ml of
1M sodium hydroxide was added. The solution was then diluted with distilled water to make the final volume up
to 10ml. The absorbance was read at 510nm. Flavonoid content was calculated using a standard calibration curve
299
Concentration (mg/ml)
Hexane extract Methanol extract BHA Vitamin C Vitamin E EDTA
Total phenolics (mgGAE/g) 78.34 + 0.19 123.26 + 0.27 - - - -
Flavonoids (mgQE/g) 41.13 + 0.15 89.96 + 0.21 - - - -
a
DPPH activity 1.30 + 0.03 0.40 + 0.02 0.06 + 0.01 - 0.06 + 0.01 -
a
Ferric reducing activity 2.06 + 0.02 1.01 + 0.02 0.05 + 0.01 0.06 + 0.01 - -
a
Metal chelating activity 1.51 + 0.02 0.21 + 0.01 - - - 0.05 + 0.01
a
EC50 value, the effective concentration at which 1,1-diphenyl, 2, picrylhydrazyl (DPPH) activity was 50 %; the absorbance was 0.5 for
reducing power and the DPPH radicals were scavenged by 50%. EC50 value was obtained by interpolation from linear regression
analysis. BHA, Vitamins C, E, and EDTA were used as positive controls.
Antioxidant activity.
Antioxidant activity of the extracts was determined by measuring its DPPH radical scavenging activity,
ferric reducing power and metal chelating activity.
Scavenging ability on DPPH radicals.
The extracts exhibited variable DPPH activities with concentration (Fig. 1). At 0.1mg/ml, the
scavenging abilities on DPPH radicals were 37.4% and 41.2 % for hexane and methanol extracts respectively.
This increased with increasing concentration of the extracts. At 5.0mg/ml, the scavenging abilities of the extracts
were 69.1% and 77.5% respectively. Generally, the methanol extract was a more potent radical scavenger (EC50 =
0.4mg/ml) than the hexane extract (EC50 = 1.3 mg/ml). This result suggest that the methanolic extract is richer in
primary antioxidants which act to scavenge free radicals by donating hydrogen atoms or electrons that convert
them into more stable products [20]. Our results were higher than reports for roots of Turnera subulata [19].
Ferric reducing power

The ferric reducing power of the extracts varied significantly with concentration. (Fig. 2). This assay treats
the antioxidants contained in samples as reductants in a redox-linked colorimetric reaction and the value
reflects the reducing power of the antioxidants [21-22]. At 0.1mg/ml, the reducing power of the hexane
extract was 0.27 while that of the methanol extract was 0.39. At 5 mg/ml, the methanol extract had a reducing
power of 0.78, while the hexane extract had a reducing power 0.58. Generally, the higher the reducing power
of a substance, the higher is its absorbance. EC50 values (Table 1) indicated a significant reducing power for
the methanol extract (EC50 = 1.01mg/ml), suggesting a high content of compounds capable of converting
3+ 2+
Fe to Fe . It may be implied that the methanol root extract of H. crinata had notable ferric ions reducing
ability and electron donor properties for neutralizing free radicals. However, the reducing power of BHA and
vitamin C (EC50 = 0.05mg/ml and 0.06mg/ml) were more pronounced than that of the extracts.
300
Metal chelating activity
The presence of transition metal ions in biological systems could catalyse the Haber- Weiss and Fenton-
type reactions, resulting in generation of hydroxyl radicals (OH*). However, these transition metal ions could
form chelate with the antioxidants, which result in the suppression of OH * generation, and inhibit the
peroxidation process of biological molecules. At 0.1mg/ml, the chelating abilities of the hexane and methanol
root extracts of H. crinata were 36.7% and 47.2% respectively (Fig. 3) and at 5mg/ml, that of the methanol extract
was 71.3%. However, EDTA exhibited excellent metal chelating activity for ferrous ions and its chelating ability
was > 85% at 0.1mg/ml. Our result indicate that the methanol extract had notable chelating abilities ( EC50 =
0.21mg/ml), suggesting that its action as an antioxidant may be related to its iron binding capacity, and may act as
a secondary antioxidants (Nurliyana et al., 2010). Nevertheless, our values were higher than reports for Coleus
aromaticus [23].
301
Conclusion
It is concluded from this study that the methanol root extract of H. crinata had the highest content of total
phenolic compounds and flavonoids. It also exhibited promising free radical scavenging activity, had good
reducing powers and could chelate ferrous ions effectively. These properties bestows on the extract good
antioxidant potentials. To fully understand the mechanisms of some of these phenolic compounds, fractionation
of the extract and further identification are in progress. However, based on our results, the methanol root extract
of H. crinata may be found useful in enhancing human health through protection against oxidative damage.
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303
EVALUATION OF MEDICINAL COMPOSITION OF CYATHEA LATEBROSA LEAVE EXTRACT
Ikpa, C.B.C1*., Ikezu, U.J. M2 and Ugochukwu, M.I3
Department of Chemistry Imo State University Owerri, Nigeria
Email- ikpacbc@gmail.com
Abstract
Phytochemical analysis of chloroform extract of leaves of Cyathea latebrosa gave alkaloid
2 2 2 2
(0.08×10 ±0.02), saponine (0.16×10 ±0.02), tannin (0.11×10 ±0.01), flavonoid (0.16×10 ±0.01) and
phenol (0.18×102±0.01). The FT-IR (KBr) cm-1 analysis showed -NH-(3433.41weak), -OH- (3851.01sharp ),
-C-F (1408.8strong) –SO-(1101.39) and Ar (1646.3weak) GC/MS characterization shows that the extract
contained [2-Aziridinylethyl]amine (70.7%), Benzeneethamine, 3-fluoro-â,5-dihydroxy-N-methyl
(14.0%), N-Methyltaurine (10.7%) and (R)-(-)-2-Amino-1-propanol (3.15%). The presence of these
compounds showed that the leaves of Cyathea latebrosa can be used for the treatment of alleviated cold,
cancer, malaria and to regulate immune functions.
Key Words; Phytochemical, Characterization, GC/MS analysis, Treatment,
Introduction
Medicinal plants have been used by all mankind as source of medicines since ancient times. In the recent times,
there have been growing interest in exploring the biological activities of different medicinal herbs due to their
1
natural origin, cost effectiveness and lesser side effects . Interest in medicinal plants as a re-emerging health aid
in the maintenance of personal health and well-being has been fuelled by rising costs of prescription drugs and
the bio prospecting of new plant-derived drugs2. Generally, plants which produce constituents having medicinal
values are called medicinal plants. These substances differ from plant to plant, thus the plant kingdom provides a
large store of various chemical substances with potential therapeutic properties which have been utilized in
treatment and cure of human and other animal diseases including relieving of pains, convulsion and
cardiovascular diseases3. Drugs of natural origin are considered to be less toxic and free from adverse effects than
synthetic ones. Even though active compounds of many herbal drugs are unknown, they have been widely
4
prescribed by the practitioners of the traditional medicines due to their minimal adverse effects and low cost .
Cyathea latebrosa is a common and widespread species of tree fern which grows in a wide range of habitats,
including forest, secondary forest, and plantations, from sea level up to an elevation of about 1500m5. Cyathea
latebrosa is a popular plant of gardens and parks6. Cyathea latebrosa which is commonly called 'Paku' in Malay
7
simply means tree fern .The uses of fern ranges from decorative to health purpose though little about Cyathea
latebrosa has been recorded. C. Latebrosa is used to relieve pains, alleviate cold and flu symptoms8. It is also
used for the treatment and prevention of heart failure9. Indians use fern leaves powder to cure centipede bites,
10 11
wounds, skin disease and cough . Maiden hair fern act as contraceptives and anti-hyperglycaemic agent . Ferns
12
are one of the most common wild plant food collected by people around the world especially in China and
Hawaii13. The non toxicity of ferns like Cyathea latebrosa14 was proved by the use of it to feed herbivores like
goat, sheep and cow15.
Methodology
Identification and preparation of plant leaves; Matured leaves of Cyathea latebrosa, identified by Prof. Onu,
M.O. of IMSU Owerri Nigeria was harvested from a palm plantation at Ehime Mbano L.G.A. Imo state, Nigeria.
The leaves sample was dried at room temperature and grinded with new (unused) mortar and pestle. The
powdered leaves 300g was soaked with 1.5 litre of ethanol and filtered. The crude ethanolic fraction was
partitioned with water and chloroform and the organic fraction (chloroform) was further analysed using GC/MS
and FT-IR to obtain the chemical composition.
Phytochemical Analysis:
304
Determination of alkaloids: This was done by gravimetric method. Powdered leaves of C.latebrosa 5g was
soaked in 50ml ethanol containing 10% acetic acid. The mixture was filtered after 4h and concentrated by
evaporating 75% of the solvent. Conc. NH4OH was added drop-wise to precipitate the alkaloids. The alkaloid
was filtered, washed, weighed and the percentage determined.
Determination of tannins; Powdered leave sample 0.5g was extracted with 10ml of H2O and added to 35ml of
H2O in 50ml conical flask. 5ml of tannic acid was treated in the same manner. The mixture was allowed to stand
for 1.5h and the absorbance was taken at 760nm to obtain concentration of tannin.
Determination of Phenols; 0.2g of sample was soaked with 10ml methanol and filtered. 1ml of the filtrate was
mixed 1ml of Folin Ciecalteau reagent and 2ml of 20% Na2CO3 solution was added. A standard phenol was
treated in the same way and the colour absorbance was recorded at 560nm.
Determination of saponins; The powdered leave sample 10g was soaked in 100ml of 20% ethanol. The
suspension was heated in a hot water bat for 4h with continuous mechanical stirring at 550C. The combined
0
extract was reduced to 20ml over water bath at 90 C. The concentrate was transferred into a separating funnel and
10ml of diethyl ether was added and shaken vigorously. The aqueous layer was recovered while the ether layer
was washed twice with 5ml of 5% aqueous NaCl. The remaining solution was heated in water bath to evaporate
it. After evaporation, the residue was weighed dried in an oven to a constant weight and the percentage saponin
determined.
Determination of flavonoids; The powdered sample 20mg was extracted with 50ml of 80% aqueous methanol at
room temperature. The solution was transferred into a crucible and evaporated to dryness over a water bath and
weighed.
-1
IR, GC/MS Analysis; IR spectra were recorded on KBr (cm ) with SHIMAZUFT-IR 8400s FOURIER
TRANSFORMER INFRARED SPECTROPHOTOMETER NATRIC Zaria. GC/MS analysis was recorded
with Agillant 7890AGCMS of Halden Company Port Harcourt, Rivers State.
Results
Table1; Results of the Phytochemical analysis of C.letabrosa
Phytochemicals % concentration
2
Alkaloid 0.08×10 ±0.02
Flavonoid 0.16×102±0.01
Saponin 0.02×102±0.02
2
Phenol 0.18×10 ±0.01
2
Tannin 0.11×10 ±0.01
The phytochemical results showed the presence of alkaloids, flavonoids, saponin, phenol and tannins at different
concentrations.
Table 2; Results FT-IR Analysis of C.latebrosa leaves
S/N Functional groups Peaks Comments
1 -SO- 1101.39
2 R-F 1408.08 strong
3 Ar 1646.3 weak
4 -NH- 3433.41 weak
5 -OH- 3851.01
305
FT-IR analysis of the chloroform fraction of C.latebrosa leaves indicated the presence of the following
functional groups; -SO-, R-F, Ar, and –OH. The peaks that are stretched and pointed are said to be strong while
the dwarf peaks are referred to as weak peaks.
Table 5. Result of GC/NS Analysis

Compds Name M.weight M.formula Base peak R. index % content R. Time/min
1. (2-Aziridnylethyl)- 86 C4H10N2 45 14533 70.7 30.75min
Amine
2. Benzeneethanamine- 185 C9H12FNO 42 14308 14.0 30.755min

3-fluro-â-5-dehydroxy
-N-methyl
3. N-methyl-taurine 139 C3H9NO3S 64 14722 10.7 26.936min
4. 2-amino-I-propanol 75 C3H9NO 42 3386 3.15 30.755min

M.weight=molecular weight, M.formula=molecular formula, R.index=Retention index,
R.Time/min=Retention Time/min
GC/MS characterization shows that the extract contained [2-Aziridinylethyl]amine (70.7%), Benzeneethamine,
3-fluoro-â,5-dihydroxy-N-methyl (14.0%), N-Methyltaurine (10.7%) and (R)-(-)-2-Amino-1-propanol
(3.15%).
Discussion
The result of the phytochemical screening (table 1) showed the presence of flavonoid which are potent water
soluble, antioxidants and free radical scavengers,which prevents cell damage and have anti cancer properties16.
Tannin properties contribute to the use of the plant leaves in the treatment of ulcer and wounds. Alkaloids and
17
phenols support the traditional use of plant to treat bacteria and fungi infections .
The indications of –OH-, -NH-, R-F, -SO- functional groups on FT-IR result (table 2) support the medicinal
properties of the plant leaves.
GC/MS analysis (Table 3) gave the presences of the following four compounds which have some pharmaceutical
properties.
Compound 1:
H2N
N
[2-Aziridinylethyl]amine
[2-Aziridinylethyl]amine is an amine compound that has been used to synthesis many pharmaceutical
compounds. [2-Aziridinylethyl]amine derivatives has been applied in the treatment and prevention of many
18
diseases .
Compound 2: OH
HO NH
306
Benzeneethamine, 3-fluoro-â,5-dihydroxy-N-methyl
Benzeneethamine, 3-fluoro-â,5-dihydroxy-N-methyl is applied medicinally in the treatment of cold, pains and
weight loss.
Compound 3:
O
O OH
S
HN
N-Methyltuarine
N-Methyltaurine showed chemo preventives against colon and hepatic cancer, used as anti bacteria, anti malaria
19
and regulates immune functions .
Compound 4: NH2
OH
(R)-(-)-2 -Amino -1 -propanol
(R)-(-)-2-Amino-1-propanol is used to boost immunity and also for water treatment20.
Conclusion
This study supports the usefulness of Cyathea latebrosa leaf and it's medicinal properties. This partly explains the
use of this plant material in herbal medicine for treatment and prevention of the symptoms of some diseases.
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Energy Available from Undergrowth in oil Palm Plantation for Rumunant Production” J. Agro forestry system
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gangetic plains of west Beengal India”97; 19-29.
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Oxford; Blackwell Scientific.
17. R.N.Bennett and R.N.Wallsgrove. (2006). “Secondry Metabolites in Plants Deffence Mechanism” New
Phytol 127. 617-633.
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compounds in Buckwheat sprouts by GCMS. Asian J. Of chem.. vol.26(3). 77-782.
19. R.Atla-Ur., M.I.Choudhary and A.B.Reitz (2009). Frontiers J. Of medicinal chem. 4.
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Relationship of -2-substituted-2-amino-1,3-Propanediol” Biomed J 5:(5) 853-856.
308
DETERMINATION OF HEAVY METALS IN TELFEIRIA OCIDENTALIS
(PUMPKIN LEAVES) GROWN AT ORIMEKPANG – OCHOR VILLAGE, BOKI
LOCAL GOVERNMENT AREA,
CROSS RIVER STATE, NIGERIA.
1 2 2
IKEZU, U. J., UDEOZO, I.P. and ODOK, J.M.
1
Department of Chemistry, Imo State University,
2
Department of Chemistry, Tansian University, oba
Corresponding author: Joymag16@gmail.com
ABSTRACT
The concentration of heavy metals was investigated in the leaves of Telfeiria ocidentalis. The heavy
metals (Cu, Fe, Cd, Pb and Zn) concentration were analyzed using Atomic Absorption Spectrometer (AAS) in two
different farms at both late dry season and raining season. The result showed a remarkable increase in the
concentration of Fe in the two farms A and B which ranges from 0.771 – 3.693 ppm and 0.00 – 27.379 ppm
respectively from late dry season to raining season. The level of Pd in both season for farm B were 0.053 ppm and
0.001 ppm, but wasn't detected in farm A, Cu concentration values for both dry and raining season in farm A was
(0.577 ppm and 0.532ppm) and farm B was(0.994 and 0.259 ppm). These values are within the FAO/WHO
permissible standard. The concentrations of Fe in farm A for the two seasons, farm B during raining season, Zn at
late dry season in both farms including Cd in farm B for the same period were higher than the maximum
permissible limit. The consumption of pumpkin leaves as food may constitute possible health hazards to human at
the time of the study. These may be due to environmental pollution and there need to be caution in fertilizer, waste
water and other agro-chemical usage as they could be attributed to the elevated levels of some metals in pumpkin
in these farms at different seasons.
Keyword: Telferia ocidebtalis. heavy metals, concentration.,toxic element.
INTRODUCTION
Pumpkin leaf Telferia ocidebtalis is one of the vegetable that are usually grown on low land of
tropics, the plant has lobed leaves and long twisting tendrils. Usually, the plant thrives well in soils rich in
organic matter. The vegetable is planted in the forest farmlands in the rural areas and most gardens around the
residential compounds in both rural and urban settlements. Pumpkin is known to be the most frequently
consumed vegetables because it is a rich source of vitamins, mineral, fiber and also have beneficial
antioxidative effects [1]. The leaves of this plant are normally consumed as delicacies particularly in the
southern region of Nigeria, specifically the popular Ediikanyikong soup in Akwa-ibom and Cross River states
in Nigeria. Heavy metal contamination of vegetables cannot be underestimated as these foodstuff are
important component of human diet however intake of heavy metal contaminated vegetable may pose a risk to
human health [2]. Ingestion of vegetable containing heavy metal is one of main ways in which these element
enter the human body, once entered, they are deposited in bones and fat tissue, overlapping noble minerals
slowly released into the body which can cause array of diseases .However Knowing the concentrations of
heavy metals absorb by this plant is important in this research as heavy metals are known to be non-
biodegradable and persistent environmental contaminants which may be deposited on the surface and then
absorbed into the tissues of the vegetables. These metals are require in trace amounts by living organisms,
some are essential for certain metabolic activities, component of enzymes and pigments in living system while
others have adverse effect on human health [3]. Plants growing within the heavy metals contaminated area
usually take-up heavy metals through absorbing them. Potentially harmful metal contents in soil may come
not only from the bedrock itself but also from anthropogenic sources like solid or liquid rock deposit,
agricultural inputs and fallout of industrial and urban emission [4]. Several reports showed that heavy metals
309
can accumulate in various plant tissue and cause membrane depolarization and acidification of the cytoplasm
[5]. The accumulation of heavy metal in soil is of major concern in agricultural production due to the adverse
effects on food quality and health [6]. Vegetables especially the leafy ones accumulate higher amount of heavy
metals, while roots and leaves of herbaceous plants retain higher concentrations of heavy metal than stem and
fruits[7,8]. Excessive accumulation in agricultural soils may result not only in soil contamination but has also
consequences from food quality and safety so it is important to monitor food quality given that plant uptake is
one of the main pathway through which heavy metals enter the food chain[9]. Vegetables take up these metals
and accumulate them in their edible and non edible parts at quantities high enough to cause clinical problems
to both animals and human consumption of contaminated vegetables can seriously deplete some essential
nutrients in the body causing s decrease of immunological defense, disability associated with malnutrition and
a high prevalence of upper gastrointestinal cancer [10,11]. These heavy metals include active metals like
Thorium, Uranium, Chromium, Nickel, Iron, Copper, Zinc, Cadium, Lead, Plutonium etc. Some industrial
actives also introduce heavy metals into the environment. These activities include smoke from chimneys of
some industrial manufacturing companies and other waste water effluents. They are also discharge into the
environment from mining activities [12]. With increasing population and high industrial activities, the
accumulation of heavy metals in the environment arise from waste disposal, waste incineration, rural, urban
and industrial effluents is of increasing concern. This is because of the potential health problems they pose to
humans and all living organism if they are present in the environment at toxic levels. Heavy metal pollution of
soil, water and atmosphere represents a growing environmental problem affecting food quality and human
health in cities [13]. Plant uptake of these metals from the soil depends on varied factors such as solubility,
soil pH, plant species and soil type [14].Therefore this work aimed to investigate the concentration of some of
these heavy metals in the most frequently consumed vegetable of these region in different months and
compare them with the permissible limits established by FAO, WHO and NAFDAC.
Hazardous Effects of Heavy Metal
Heavy metal contamination of vegetables cannot be underestimated as these foodstuff are important
component of human diet, vegetables are rich sources of vitamins, minerals and fiber and also have beneficial
antioxidant effect, however intake of heavy metal contaminated vegetable may pose a risk to human health[2].
Soil and vegetables contaminated with Pb and Cd significantly decreases human life expectancy with 9-10
years. They are the most toxic element for man in terms of environmental concentration. Pb is the heavy metal
closest to level in which toxic signs manifest than any other substances [15]. Other elements like Cr , Cu and
Ni although essential for men but at concentration higher than recommended may cause metabolic disorder.
Cd is a dangerous element because it can be absorbed via alimentary tract, penetrate through placenta during
pregnancy and damage membrane and DNA once in the body, it may remain in the metabolism from 16 to
33year and is connected to several health problems such as renal damage and abnormal urinary excretion of
proteins, decrease in bone calcium concentration and increase in urinary excretion of calcium have also been
attributed to exposure to Cd, eventually causing death. It also affect reproduction and endocrine systems of
women [16]. Vegetables may contribute to about 70% of Cd intake by human, varying according to the
consumption level [17] .Toxic effect of Pb focus on several organs like liver, kidney, spleen and lung causing a
variety of biochemical defects. Nervous system of infants and children is particularly affected by toxicity of
these heavy metals, adult exposed occupationally or accidentially to excessive levels of Pb exhibit
neuropathology. There is association between Pb in human body and the increase of blood pressure in adult
[15]. High Pb concentrations observed in many vegetables, although they do not pose a risk to human health
may be attributed to crops located near roads of heavy traffics. The main source of Pb to human is inhalation
of airborne Pb from vehicle emission and from direct atmospheric deposition or soil, water and crops
constituting the gateway into the food chain [9]. Phosphate fertilizers are the major source of soil
310
contamination by trace metal especially Cd as it is naturally found as an impurity in phosphate rock[18].
Knowedge of Zn toxicity is minimal, the most important information reported is its interference with Cu
metabolism. Symptom that an acute oral zinc dose may provoke include, tachycardia, vascular shock,
dyspeotic nausea, vomiting, diarrhea, pancreatic and damage of hepatic parenchyma. Cu and Zn act as
micronutrient for the growth of animals and human being when present in trace quantities. Air pollution may
pose a threat to post-harvest vegetable during transportation and marketing causing elevated levels of heavy
metal in vegetables [19].

Sample Collection
The samples used (leaves of the pumpkin) were collected from Orimekpang – Ochor in Boki local
government area of Cross River state. The samples where respectively harvested in the second week of March
and fourth week of May 2015 from two different farms at 100m distance from each other. Only fresh
vegetables in good condition were collected.
Sample Preparation
The leaves were arranged into four groups and dried at room temperature for 48 hours to remove
moisture, four non-disposable petri-dishes were weighed empty using an electronic analytical balance as 2g of
each sample was weighed into the petri dishes, dried to constant weight in an oven maintained at 105oc and
pulverized to fine powder using an electric blender. The finely grinded powdered sample were collected in
labiates plates and placed in desiccators. 2g of each sample was carefully weighed into clean platinum
crucible and heated in a furnace for 2 hours, at 500oc to ash then cooled to room temperature in a desiccators.
The ash, was digested with 20ml, 20% H2SO4 and filtered with filter paper as filtrate was collected in a 50 ml
volumetric flask as deionized water was added to mark up to the mark of the flask [20].The metal
concentration were determine using atomic absorption spectrophotometer(AAS) model Varian 240 AA.
RESULTS
SAMPLING SEASONS MONTH HEAVY ME TAL (mg/l)

SITES
Cadmium Iron Copper Zinc Lead
FARM A LATE DRY MARCH 0.044 0.771 0.577 6.762 0.00
RAINING MAY 0.052 3.693 0.532 4.894 0.00
FARM B LATE DRY MARCH 0.071 0.00 0.994 8.175 0.053
RAINING MAY 0.058 27.379 0.259 4.371 0.001
FAO/WHO Minimum Permissible Limit 0.06mg/l 0.02mg/l 5.0mg/l 5.0mg/l 0.3mg/l
NAFDAC Minimum Permissible Limit 0.003mg/l0.3mg/l 0.5mg/l 3mg/l 0.01mg/l
311
Table 1: Analysis result on the concentration of some heavy metals in Telferia occidentalis from two
farms at different seasons. DISCUSSIONS
From the result, the analysis indicated a slight fluctuation in concentration of cadmium in the two farms
and at different months which ranges from 0.044 – 0.052 mg/l and 0.071 – 0.058 mg/l respectively. Although, the
values recorded for cadmium are within the permissible levels given by the FAO/WHO in three samples but one
exceeded the limit which may pose a health hazard to the consumers. A remarkable increase in iron concentration
from 0.771 – 3.693mg/l and 0.00 – 27.379 mg/l in both farms respectively from late dry season to raining season.
The values except one were significantly higher than the permissible levels which possess big risk in the
consumption of these pumpkin vegetables. However, the elevated levels of iron in the farm could be attributed to
excessive usage of fertilizers and other agro-chemical as well as environmental factors in the area. The
concentration level of copper found in the samples collected from the farm A and B ranged from 0.577 – 0.532mg/l
and 0.994 – 0.259mg/l are within the world health organization MPL and therefore safe. The concentration of lead
in farm B was within the minimum permissible limit as it was not detected in farm A.
The result indicated a decrease in zinc concentration in both farms during the period of research as may
be attributed to the change in seasons. This proved that the samples collected from both farms in the month of May,
during the raining season were safer for consumption than samples collected by March which is the late dry
season. And possibility of plant absorption of metal based pesticides, industrial pollutants may be higher during
dry season.
CONCLUSION
Food safety is known to be one of the major public health concerns all around the world. The vegetable
Telfeiria occidentalis grown at Orimekpang – Ochor contained variable levels of heavy metals (Fe, Cu and Cd)
and indicated Pb in trace amounts. There was remarkable increase in the concentration of iron from late dray
season than during the raining season. Whereas, decrease in concentration of zinc and copper were observed
during period of research at both farm A and B samples collected. The high levels of some metals in pumpkin
samples suggest that the vegetable plant were capable of absorbing heavy toxic metals in their tissues from the soil
or from the natural environment during the dry season than raining season.
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[11]Antonious, G., and Kochhar,T.,(2009) Mobilty of Heavy Metal from soil into Hot Pepper Fruits, a
Field Study, Bulletin of Environmental Contaminants and Toxicology 82:59-63.
[12]. Lenntech (2010), Heavy Metals. www.lenntech.com pp 1-3.
[13].Nriagu, J.O., (1990) Global Metal pollution. Environment, 32 (7),7-33
[14]. Yargoli, B., and Andazimi, A.A.,(2008) Investigation of Cadmium Absorption and
Accumulationin Different parts of Some Vegetables. Americian Eurosian Journal of Agricultural and
Environmental Sciences,3(3),357-365.
[15] Fernando, G, Anderson, R.T., Takashi, M, and Nerideness, C.M., (2012) Heavy Metal in
Vegetables and Potential Risks for Human Health, Sci agric ,Vol 69 no 1.
[16] World Health Organization (WHO) (2004) Evaluation of Certain Food Additives and
Contaminant in sixty first Report of the Joint FAO/WHO Expert Committee on Food Additives, WHO Geneva
Switzerland (WHO Technical series, 922)
[17] Wagner, G.J., (1993) Accumulation of Cadium in Crop Plant and its Consequences to Human
Health, Advance in Agronomy 51:173-212
[18] Mohhamed, H.H, Khairia, M.A.,(2002) Assesment of Some Heavy Metals in Vegetables, Cereals
and Fruits in Saudi Arabia Market, Egyptian Journal of Aquatic Research, Vol 38 (1):31-37
[19] Salgueiro, M.J. , Zubillaga, M., Lysionek, A., Sarabia, M.I.., Caro, R., Paoli, T.D., Hager, A., Neill,
R.,and Boccio, J.(2000) Zinc as Essential Micronutrient: a review. Nut Res 20(5)737-755.
313
Proximate Analysis and Production of Bioethanolfrom Post-Harvest Pineapple and Watermelon
Waste Fruit
G.U. Igelige, P. Ekwumemgbo, G.I, Ndukwe and J.I Achika
Department of Chemistry Ahmadu Bello University Zaria
*Corresponding Author: igeligeuzezi@gmail.com
ABSTRACT
The proximate composition was determined using standard method. The mean value obtained for moisture, ash,
crude protein, crude fiber, fat and carbohydrate was 91.52 ± 0.01 %, 0.22 ± 0.01 %, 0.45 ± 0.05 %, 0.19 ± 0.01 %,
0.18 ± 0.01 %, 7.18 ± 0.05 % respectively for watermelon and also the mean obtained for moisture, ash, crude
protein, crude fiber, fat and carbohydrate was 82.20 ± 0.07 %, 1.23 ± 0.15 %, 0.45 ± 0.07 %, 0.70 ± 0.01 %, 0.47 ±
0.03 %, 4.06 ± 0.20 % respectively for pineapple waste. The reducing sugar assay of the pineapple waste gave 7.8
g/100 g and watermelon gave 4.5 g/100 g. Effect of yeast concentration, duration of fermentation, pH and
temperature as they relate to the optimization of the ethanol production were also investigated. The result showed
that the fermented fruits waste produced ethanol: 5.88 ± 0.03 % and 1.91 ± 0.01 % (v/v) for pineapple and
watermelon respectively using direct fermentation and 14.18 ± 0.03 %, 3.07 ± 0.01 % (v/v) for pineapple and
watermelon respectively using simultaneous saccharification and fermentation, the result shows that the rate of
ethanol production by fermentation of both waste fruits by baker's yeast (Sacchromyces cerevisiae) increases
with fermentation period and maximum ethanol yield was at 72 h.
INTRODUCTION
One of the oldest biological processes known to mankind is the fermentation of biomass into bioethanol. The
process has been known since prehistoric time for the production of alcoholic drinks but has lately gained
growing importance due to the interest in producing fuel ethanol. Industrial production of fuel ethanol started in
Brazil during the oil crisis in the 1970s, and has ever since been widely important there. The know-how and
achievements from Brazil have been used and further developed around the world. Today ethanol is produced
from all sorts of biomass rich in sucrose, starch and cellulose. These are easily hydrolyzed by acids or enzymes to
C6 sugars (mainly glucose), which are fermented into ethanol and CO2. The largest producers of bioethanol are
Brazil and the USA using sugarcane and corn respectively as feedstock. Mainly wheat crops and sugar beet have
so far been used in Europe. The world production of fuel ethanol in 2007 was around 50 billion liters [1]. The
energy in this ethanol corresponds to less than 1.2 % of the total fuel consumption of 88.2 quadrillion Btu in 2005.
In the USA (the largest producer of bioethanol) the production of bioethanol has more than doubled from 2002 to
2007 [1].
Sample collection
Waste fruits 2.00 kg each of pineapple and watermelon were collected from Sabo Gari fruit market in Zaria,
Kaduna State. The experiment of the production of ethanol from waste fruit was carried out in the laboratory of
Chemistry Department, Ahmadu Bello University, Zaria.
Proximate analysis
This was carried out using the method of Association of Official Analytical Chemist AOAC 2005. The
314
parameters determined for proximate analysis include ash, moisture, protein, fat, fiber and carbohydrate
Direct Fermentation of the Fruits (Whole)
Sample preparation
Whole ripped watermelon and pineapple fruits were obtained from the market in Sabo Gari, Zaria, Kaduna
State, Nigeria. They were washed, cut in small pieces with their outer coats in place using a knife and then
blended with distilled water in ratio of 1:10 respectively using electric blender and stored in a refrigerator prior
to use.
Fermentation
Fermentation was carried out on the samples (fruit paste) by using baker's yeast (Saccharomyces Cerevisiae).
3 3
100.00 g of the sample was weighed into 250.00 cm conical flask and 100.00 cm of distilled water was added.
The pH was adjusted with 0.3moldm-3 NaOH to 4.5 and then pasteurized. Fermentation was monitored for 7 days
at 30 oC. At the end of the fermentation, the alcohol was recovered by simple batch distillation using laboratory
distillation unit. The alcohol content was determined by the determination of the specific gravity using a
standard alcohol density table [2].
Simultaneous Saccharification and Fermentation
Sample preparation
The fruits were washed, and then peeled, the pulp was cut in small pieces using a knife and then blended with
distilled water in ratio of 1:10 respectively using electric blender and stored in refrigerator prior to use. The
fresh ripened watermelon and pineapple fruits peels were chopped into small pieces (2.00 – 3.00 cm) and dried in
a hot air oven at 65 ºC for 48 h. The dried materials (peel) were ground in a dry compartment blender to pass
through a 1.00 mm screen and the powdered materials were stored at room temperature for further analysis [3]
Acid hydrolysis of fruits
10.00 cm3 of 43 %, 50 %, 60 %, 70 % and 77 % (v/v) diluted sulfuric acid was added to the non-soluble peel
component, the fruit peels was then hydrolyzed in a thermostatic water bath varying the temperature between 33
0 0
C to 70 C and also varying the time from 1hr 30 min to 7 h. After hydrolysis, it was neutralized with 10.00 M
NaOH until the pH show 7, the solid particles from the liquid in the hydrolyzate were separated using
centrifugation (to remove the non-fermentable lignin portion). After separating the solid part, it was washed with
distilled water twice. The washing was performed in order to extract all soluble sugars from the waste fruit peel,
the soluble component with the previously blended pulp solution was mixed together for the next procedure
(fermentation) for both fruits [3]
pH Adjustment
Before addition of yeast to the above prepared samples, the pH of the samples was 3.50 ± 0.40 and 5.53 ± 0.20 for
-3
watermelon and pineapple respectively which was adjusted using 0.3 moldm of NaOH. Otherwise the yeast will
be denatured in hyper acidic or basic state. A pH of around 5.0 - 5.5 was maintained. The mixed pretreated,
hydrolyzed solution was filtered and shaken. The substrate was primarily checked for pH using a digital pH
meter. The pH then was adjusted to 5.0 - 5.5, the mixed samples (pretreated and hydrolyzed) were acid
hydrolyzed. Then sodium hydroxide solution was added drop wise to the flask containing the samples with
constant stirring until the pH reached a range of 5.0 - 5.5 [3]
Fermentation
3 3
100.00 g portion of the slurry (pulverized) was weighed into 250.00 cm conical flask and 100.00 cm distilled
water was added. This was pasteurized by boiling in a water bath for 15 min. It was allowed to cool and the
yeast added. The fermentation was then monitored for 7 days. Different concentrations of the yeast
were used and the pH of sample was also adjusted with 0.5 moldm-3 NaOH to5.0, at the end of the
fermentation, the fermented sample was poured into a cheese cloth to drain out the fermented broth. This
315
procedure was carried out in triplicate [4]
Distillation
In this experiment separation were used by rotary evaporator at a temperature of 78.5 oC for 3 h. Some of the
components of the distillation set up include; distillation vessel, special top-fit of distillation vessel, condenser,
diverting glass that fits at the end of condenser and the top of harvesting vessel, condenser tubing, harvesting
vessel, stands and fixing screws, beaker, thermostat, the thermostat supporting flat metal bar. All distillation
experiments were carried out at a temperature of 78.5oC and a distillation time of 3 h.
Alcoholic Content
With the aid of the specific gravity values of the distillate the percentage alcohol was determined using a
standard alcohol density table [5]. The overall distillate obtained was then pooled together and rectified. The
percentage purity of the rectified (distillate collected at 78-80 0C) was determined [5].
RESULTS
Proximate Composition
Table 1: Proximate composition of Watermelon and Pineapple
Fruits Moisture Ash Crude Crude Fat CHO

(g/100 g) (g/100 g) Protein fibre (g/100 g) (g/100 g)
(g/100 g) (g/100 g)
Watermelon 91.52 0.22 0.45 0.19 0.18 7.18
± 0.01 ± 0.01 ± 0.05 ± 0.01 ± 0.01 ± 0.05
Pineapple 82.2 1.23 0.44 0.70 0.47 4.06

±0.07 ± 0.15 ± 0.07 ± 0.01 ± 0.03 ± 0.20
Optimisation of Ethanol Production with Direct Fermentation

Table 2: Ethanol yield (%) at different duration of fermentation and different yeastconcentration of
pineapple waste fruit
Yeast
Concentration
(%) 1 day 2 days 3 days 7 days
1 2.58 ± 0.04 3.30 ± 0.01 4.75 ± 0.08 5.23 ± 0.04
2 2.59 ± 0.05 3.33 ± 0.01 4.90 ± 0.03 5.50 ± 0.10
5 2.52 ± 0.04 4.39 ± 0.03 5.73 ± 0.01 5.45 ± 0.04
10 2.70 ± 0.02 4.55 ± 0.02 5.88 ± 0.03 5.30 ± 0.05
Results are means of duplicate experiments. Temp = Ambient temperature (27 ± 3 0C). Initial pH:5.53
Table 3: Ethanol yield (%) at different duration of fermentation and different yeast concentration of
watermelon waste fruit
316
Yeast
Concentration (%)
1 day 2 days 3 days 7days
1 0.38 ± 0.04 0.60 ± 0.01 1.58 ± 0.08 1.77 ± 0.04
2 0.25 ± 0.05 0.70 ± 0.01 1.88 ± 0.03 1.29 ± 0.10
5 0.42 ± 0.04 0.77 ± 0.03 1.91 ± 0.01 1.10 ± 0.04
10 0.80 ± 0.02 0.96 ± 0.02 1.76 ± 0.03 1.27 ± 0.05
0
Results are means of duplicate experiments. Temp = Ambient temperature (27 ± 3 C). Initial pH: 3.5
Table 4: Percent Ethanol formed with varying Concentration of Yeast on the Pineapple and
Watermelon for 72 h.
Concentration of yeast (g) % ethanol from % ethanol from water melon
pineapple
1.0 4.69 ± 0.03 0.68 ± 0.04
2.0 4.76 ± 0.02 0.77 ± 0.04
3.0 4.80 ± 0.04 1.84 ± 0.02
4.0 5.77 ± 0.01 1.88 ± 0.03
5.0 5.85 ± 0.02 1.65 ± 0.31
6.0 5.70 ± 0.01 1.91 ± 0.03
7.0 5.68 ± 0.04 1.84 ± 0.03
8.0 5.88 ± 0.02 1.90 ± 0.03
5.1. Proximate Composition
The result of the proximate composition is show in table 1. This study shows that watermelon contains more of
moisture compared pineapple. This is due to an increase in water content of the pulp, derived from carbohydrates
utilized during breathing and osmotic transfer from the peel to pulp due to increase in the sugar content in the
pulp[6].The pineapple fruit showed highest ash content of 1.23 % and the watermelon had the lowest ash content
of 0.15 %. This observed result is in agreement with the literature findings of Hammond et al., (1996) [8] in the
desert banana with ash content between 1.10 – 1.52 %.The carbohydrate content of 7.13 to 7.23 % in watermelon
and 4.04 to 4.08 % in pineapplewas observed in this present study and similar to the literature findings [6, 8]. This
is as a result of the degradation of starch to free sugars during the ripening process by the action of several
enzymes. The high protein content and carbohydrate content observed in pineapple and water melon is in
agreement with literature findings of [9]. This result explains the main source for fermentative ethanogenic
microbial growth. The highest lipid content of 0.47 % was observed in pineapple fruit and the lowest was
obtained in watermelon (0.18 %).The results obtained in the present study on the proximate composition are in
agreement with the literature findings on pineapple varieties with fat content between of 0.38 – 0.50 % [6].
Optimisation
The initial pH of the substrates was 3.5 for watermelon, this is lower than the recommended pH range of 4.5 – 5.0
for optimum ethanol yield and 5.53 for pineapple [10]. The reducing sugar assay of the pineapple waste gave 7.8
g/100 g and water melon gave 4.5 g/100 g of the sample which shows that not all carbohydrate in the samples are
available for fermentable sugar. The ethanol yield obtained from the substrate when fermented at its initial pH
and room temperature (27 ± 3 0C) at different duration of fermentation and different yeast concentrations are
shown in Table 2 and 3 for and pineapple and watermelon respectively. From these results, it is observed that
ethanol can be produced from waste pineapple even at initial pH of 4.3 and also that of watermelon which was
buffered to 4.3 to allow for optimum fermentation. Also, the yield was increased when the concentration of the
yeast was increased, it was observed from Table (2 and 3) that fermentation was optimum on day 3 (72 h) for
direct fermentation.
317
Table 4, shows that there is a significant difference from the increasing yield of ethanol in pineapple with added
yeast than when compared to the result obtained from water melon.
It was found that when 8.0 g of yeast was added to the substrate of both fruits a significant yield was optimised,
giving 5.88 % and 1.90 % for both pineapple and water melon respectively [4].
Stepwise Simultaneous Saccharification and Fermentation of Fruits

In this section, the stored and blended pulp solution in addition with the hydrozylate (which becomes slurry) was
fermented at room temperature and a hydrometer was used to monitor the difference in their specific gravity.
Saccharification of the dried, pulverized peels and hemicellulose was done at varying temperature (approximate
optimum temperature for yeasts), the simultaneous saccharification and fermentation (SSF) process yielded
higher results for pineapple and watermelon than when direct fermentation (DF) process was used.
The test for the percentage of ethanol formed from the slurry of pineapple and water melon was done by
3
subjecting the slurry to acid hydrolysis, 10.00 cm of the prepared sulphuric acid was used in each case. An
experimental design was setup using the design expert software 7.08 in order to make its analyses easier in
finding the most efficient condition or profound method for the successful production of bioethanol from the
these fruits.
Conclusively, the present work has clearly shown that stepwise simultaneous saccharification and fermentation
of waste materials from pineapple and watermelon to ethanol by a digesting sugar fermenter (S. cerevisiae) is
feasible. Simultaneous saccharification and fermentation has been found to effectively remove glucose, which is
an inhibitor to cellulase activity, thus increasing the yield and rate of cellulose hydrolysis.
References
1. RFA (Renewable Fuel Association). (2008). World ethanol fuel production. Available at
http://www.ethanolrfa.org/industry/statistics/#E.
2. AOAC (1970). Official Methods of Analysis, Harwits, W. (Ed.) 13th Edn. Association of Official
Analytical Chemist, Washington DC.
3. N.A.Amenaghawon, C.O. Okieimen and S.E. Ogbeide, (2012). Kinetic Modelling of Ethanol Inhibition
during Alcohol fermentation of Corn Stover using Saccharomyces Cerevisiae. International Journal of
Engineering Research. Vol. 2, Issue 4, pp. 798-803.
4. B.C.Akin-Osanaiye, , H.C. Nzelibe, and A.S. Agbaji, (2008). Ethanol production from Carica papaya
(Pawpaw) fruit waste. Asian Journal of Biochemistry 3(3):188-193.
5. AOAC (1974). Official Methods of Analysis, Harwits, W. (Ed.) 13th Edn. Association of Official
Analytical Chemist, Washington DC.
6. T. H.Emaga, , Andrianaivo, , R. H.B.Wathelet, , &M. Paquot, (2007). Effects of the stage of

maturation and varieties on the chemical composition of banana and plantain peels.
7. J. B., Hammond, R.Egg, , D. Diggins, andC.G Coble.1996). Alcohol from bananas,

Bioresource Tech., 56:125-130.
318
8. C.A., Regina, and M.B.AGloria,. (2005). Bioactive amines and carbohydrate changes during ripening
of 'Prata' banana (Musa acuminate x Musa balbisiana). Food Chem.,90:705 – 710. Food Chemistry,
103, 590–600.
9. AIEssien, RUBEbana, Udo HB (1992). Chemical evaluation of pod and pulp of the fluted pumpkin
(Telfairia occidentalis) fruit. Food Chem45: 175-178.
10. W.Morris, and R.PSarad,. (1990). Biotechnol. of Biomass Conversion: Fuels and Chemicals from
Renewable Resources. pp. 235.
319
EVALUATION OF CHEMICAL COMPOSITION OF Murraya koenigii (Linn) SPRENG LEAF
1 2 3 1 1
C. E. IGARA, D. A. OMOBOYOWA, A. A. AHUCHAOGU, N. U. ORJI AND M. K. NDUKWE
1
Chemistry Research Unit, 2Biochemistry Research Unit, Department of Science Laboratory
Technology, Akanu Ibiam Federal Polytechnic Unwana, Ebonyi State, Nigeria
2
Michael Okpara University of Agriculture, Umudike, Umuahia, Abia State, Nigeria.
ABSTRACT
MurrayaKoenigii (Linn) Spreng has been used for flavouring and spicing of food since ancient time. Its
medicinal value has also been identified. This work investigated the phytochemical screening, proximate
composition, vitamin and mineral contents of the plant leaves using standard procedures. The result of the
phytochemical screening revealed the presence of the bioactive constituents comprising flavonoid 7.43 ± 0.03%,
phenols 4. 25 ± 0.04%, saponins 2.50 ± 0.01%, alkaloids 1.90 ± 0.01%, tannins 0.86 ± 0.02% and glycosides 0.11
± 0.01%. The proximate composition was found to be as follows; carbohydrate 39.44 ± 0.04%, moisture content
23.42 ± 0.10%, crude fibre 6.30 ± 0.05%, ash content15.60 ± 0.21%, fats 6.48 ± 0.22% and protein 8.38 ± 0.02%.
The analysis of the vitamin content showed the presence of vitamin A (â-carotene) 6.04 ± 0.02 mg/100g, vitamin
C (ascorbic acid) 0.04 ± 0.002 mg/100g, thiamin 0.89 ± 0.01 mg/100g, riboflavin 0.09 ± 0.001mg/100g, niacin
2.73 ± 0.02 mg/100g and vitamin E 0.03mg/100g. The plant leaf was found to contain some important minerals;
Calcium 19.75 mg/100g, magnesium 49.06 ± 0.02 mg/100g, sodium 16.50 ± 0.21 mg/100g, potassium and zinc
0.04 ± 0.001 mg/100g each. Curry plant leaves contain some substantial amount of important phytochemicals
which possess anti-oxidant properties and some nutritive vitamins and minerals thus supporting its use as
medicinal plant and as food flavoring and spicing condiment.
Keywords: Phytochemicals, Minerals, Vitamins, Curry leaf, Anti-oxidant

INTRODUCTION
Humans have relied mostly on plants for nutritional and medicinal needs, Herbal plants provide most of the
medicinal needs. Important herbal products include spices, herbal teas, functional foods, medicinal raw
materials, essential oils,flavouring and dietary supplements1. The medicinal use of plant is as a result of the
phyto-constituents present in them.Some of these chemicals are bioactive and produce definite physiological
and biochemical actions in humans and animals. They are known as secondary metabolites or
phytochemicals and comprises alkaloids,flavonoids, tannins, phenolics,saponin, steroids,
glycoside,terpenesetc2. The use of plants in traditional medicine is of global interest.Traditional and herbal
medicines have been promoted as a source of less expensive and comprehensive medicare especially in
3
developing countries . Some of these medicines are easily available, cheaper and safer than these modern
synthetic drugs4. This led to the resurgence in the use and demand for medicinal plants as they play roles in
prevention and management of some health ailments such as diabetes , cancer, arthritis , degenerative
3
disorders like Parkinson and Alzheimer's diseases . The medicinal value of many plants remain unexploited.
Investigations are ongoing to discover novel drugs or templates for the development of new therapeutic
agents.
320
Phytochemicals Values (%)
Alkaloids 1.90±0.01
Saponins 2,50±0.01
Flavonoids 7.43±0.03
Tanins 0.86±0.02
Phenols 4.25±0.04
Glycosides 0.11±0.01\
Values are triplicate determinations ±std deviation
Table 2: Result of Proximate composition of Murraya koenigii plant leaf
Proximate composition Values (%)
Moisture content 23.4±0.10
Protein 8.38±0.02
Carbohydrate 39.44±0.04
Fats 6.48±0.22
Ash content 15.60±0.21
Crude fibre 6.30±0.05
Table 3: Result of vitamin content Murraya koenigii plant leaf
Vitamins Values (mg/100g)
Vitamin A (B-carotene) 6.04±0.02
Vitamin C (ascorbic acid) 0.04±0.002
Vitamin E (tocopherol) 0,03±001
Vitamin Bi (thiamin) 0.89±0.01
Vitamin B2 (riboflavin) 0.09±0.002
Vitamin B3 (niacin ) 2,73±0.02
Values are triplicate determination ±std deviation
Table 4: Result of Mineral elements of Murrayakoenigii plant leaf

Minerals Values (mg/100g
Calcium 19.73±0.02
Iron 0.16±0.01
Magnesium 49.06±0.02
Sodium 16.50±0.21
Zinc 0.04±0.001
Potassium 0.04±0.001
The phytochemical composition of the Murraya koenigii leaf is shown in Table 1.Flavonoid content was found to
be the highest7.43±0.03% whileglycoside was detected to be the least. Flavonoids have been shown to possess
anti-fungal and anti-bacterial activity9 . Flavonoids have strong anti-oxidant and anti-inflammatory properties.
10
They have ability to scavenge hydroxyl radicals, superoxide anions and lipid peroxy radicals . Thesefree
radicalshave been implicated in causing some age-long diseases such as diabetes, cardiovascular, Parkinson and
Alzheimer's diseases.The presence of flavonoids in the curry leaf may account for its use in treatment of these
diseases. Saponin was detected in an appreciable amount 2.50±0.01%. Saponins are known as immune booster.
Plants rich in saponins have been shown to demonstrate anti-inflammatory,cholesterol lowering and ant-cancer
properties11. Alkaloid was found to be 1.90±0.01% in the curry leaf. Alkaloids have anti-microbial properties
owing to their ability to intercalate with DNA of the micro-organisms12. Phenol was found to be 4.25±0.04%.
13
Urquiaga and Leighton reported that phenols and phenolics have anti-tumour and antioxidant effects . They
also have anti-inflammatory and anti-carcinogenic properties and play roles in scavenging H2O2 by donating
electrons to the
321
14
peroxide thus neutralizing them to water .
Table 2 reveals the proximate composition of Murraya koenigii leaf. The plant leaves contain a substantial
amount of carbohydrate 39.44±0.04% and moisture 23.42±0.10%. This shows that the leaf is a good source of
carbohydrate while the moisture content is high indicating a low shelf –life,so the plant material can be preserved
by drying it to retain other useful components. The low amount of fat indicates that the vegetable is not a good
source of lipid accumulation which can cause artherosclerosis and aging15 . The proteinsin the leaf can make fair
contributions to protein diet as proteins are involved in formationof hormones, enzymes and structural
membranes.M. koenigii contains a good amount of dietary fibre. Dietary fibre lowers cholesterol level, risk of
16
coronary heart disease, diabetes and cancer . Vitamin content of the M.koenigii is shown in Table 3. Vitamins
are found in trace amounts. Niacin vitamin B3 was found to be high 2.73±0.02mg/100g compared to other
vitamins.Vitamins A and C were found to be low 6.04±0.02mg/100gand 0.04±0.02 respectively. Niacin helps to
17
lower and regulate cholesterol level and helps in maintaining good blood circulation . Vitamin A helps to
provide good vision and healthy immune system. It also fights cancer by inhibiting the production of DNA in
cancerous cells18. Vitamins C and E are strong anti-oxidants. They are good in fighting cancer through their
scavenging of free radicals. Other B-groups vitamins B1 thiamine, B2riboflavine were also detected. Riboflavine
helps in production of red blood cells and is important for growth and healthy body. The presence of these
vitamins in the plant leaf has shown that the leaf possesses strong medicinal and nutritive values.
Table 3 gives the mineral content of the curry plant leaf. Magnesium was found to be the highest
49.06±0.02mg/100g followed by calcium 19.75±0.02mg/100g while zinc and potassium were found to be low
0.04±0.001mg/100g each. Mineral and trace elements play important roles in immune function and health.The
high content of magnesium proves that the curry leaf can be a good source of magnesium which is an activator of
many enzyme systems and maintain electrical potential in nerves19. Calcium was detected in an appreciable
amount. The M. koenigiileaf can serve as food supplement for calcium. Calcium combines with phosphate to
20
enhance strong bone and teeth formation . Sodium content was also found to be high 16.50±0.21mg/100g.
Sodium and potassium are needed in body fluids and nerves for transport and balance. The plant leaf contains
zinc ,though in small amount, zinc is important in the body as it enhances immunity, cell growth and insulin
regulation. It is used in restoring dehydration and stoppage of diarrhea. The presence of the mineral elements in
the plant leaf shows that consumption of M.koenigii can be used as a supplement for these essential nutrients.
CONCLUSION: This study has shown that Murraya Koenigii (curry) leaf used as spice and flavouringagent in
food contains substantial amount of phytochemicals and phytonutrients. The proximate analysis showed high
content of carbohydrate, protein ,fibre and fat. The presence of the vitamins and mineral elements in the plant
leaves showed that it could be consumed to supplement these scarce nutrients. With its content of these anti-
oxidants flavonoids, phenols,vitamins E and C, curry leaf possess anti-cancer and cardio-protective agents
supporting its use as medicinal plant.
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medicinal plant. Asian Journal of Pharmaceutical and Clinical Research 5(4): 5-14.
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4) Summan, S., More, P.K. &Sandhya, M.M. (2014).Curry leaves (Murrayakeonigii LinnSpreng) a
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322
5) Kirupa, S.L.S.&Kariitha, R. (2015).Anti-oxidant enhancing property of curry leaf powder M.
keonigii in type ll diabetes mellitus. Intl Journal of Phamacy&Biosciencs 6 (1)
507-514.
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ed
7) A.O.AC.(1994). Official method of analysis 15 , Association of Official Analytical Chemist,
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Caryophyllus and their anti-fungal activity, Phytochem. Lett1: 44
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323
Studies on the Energy Properties and Fuel Potentials of Selected Indigenously Processed Plant-Based
Bio-Resources Used as Sources of Sustainable
Flame in North Central Nigerian Communities
1 1 1
J. S. Gushit* , J. T. Shimuan , M. O. Ajana
1
Chemical/Petroleum Option, Department of Science Laboratory Technology (SLT),
University of Jos, Nigeria.
*Correspondence Author's e-mail: john.g445@gmail.com
In this work, the stems of two matured shrubby plants identified as wild bitter leaf (Vernonia colorata) & kenaf
(Hibiscus canabinus) were studied for their energy properties and fuel potentials. The indigenously processed
and dried stems of these plants have historically been used as sources of sustainable bright flame in
communities within north centrals Nigeria. The two samples were collected, dried, pulverized and extracted
using appropriate solvents to isolate the volatile combustible compounds believed to responsible for the
combustion to produce the bright flame. The isolates are being purified to be detected using appropriate
instrumentations such as GCMS and LCMS (ongoing). Prior to isolation, the physical energy properties which
include; total volatile organic compounds (TVOC), fixed carbon, ash content, calorific value, moisture contents
(MC) were determined. The gases emitted by these plants materials on combustion were also detected using
multigas detector (Altair 5x Multigas). The results obtained indicate the concentrations of life threatening gases
such as CO, SOx and, NOx from the processed substances on combustion are within acceptable standard limits
set for these gases, which suggest it is environmentally friendly. The technology employed in making plants
provide more sustainable flame for the light will eventually be studied by collecting samples of substances used
in the process (water, mud, algae) for characterization.
Keyboard: Energy, fuel, bio-resources, sustainable flameINTRODUCTION
The looming global energy crisis has prompted the search for alternatively energy sources globally in recent
times. This is attributed to daily energy demand from various places like schools, homes, business areas and
industries. It is forecasted that global energy demand could rise to 50% by the year 2025, with major part of
Harvesting & Drying Matured
Dipping in muddy Stream-
De-barking to remove Fibre [used
Drying Stem - Produces BRIGHT FLAME
324
The indigenous process technology used by these communities involves harvesting the matured plants. This is
eventually dried and dipped in muddy water for 4-5weeks, during which it is believed to have undergone some
catalytic and microbial transformations (Figure 1). The processed stem has a very good combustion property
as it is observed to burn continuously for some time with sustainable bright flame which acts as source of light
in the dark with little smoke to pollute the environment. Like every other flammable material, it is expected
that there should be some volatile components in the processed plant stem products which is responsible for
the sustainable burning quality, unlike the unprocessed plants stem which does not produce such a
sustainable bright flame. This is in line with Chaula et al.'s (2014) work, who confirmed that, those biomasses
that contain a high content of volatile components, burns with a thick and bright flame. The aim of the
research is to investigate the energy potential and the fuel properties of plants widely used as sources of
sustainable flame by indigenous communities in North Central Nigeria.
METHODOLOGY
Hibiscus canabinus and Vernonia colorata were harvested around November and December when they were
fully matured and sun-dried. After drying, it was soaked in muddy water for at least 4-6 weeks, it was debarked
and sun dried. The dried stems were crushed with mortar and pestle and ground. The pulverized sample was
sieved to obtain a particle size of 0.50 -1.00mm. The following analysis were done on the samples based on
methods adapted (Efomah, et al. 2015, Saheed et al., 2015 and Mitchual et al. 2014); density/bulk density,
moisture Contents, gross calorific value, ultimate analyses, volatile matter, ash content, fixed carbon, dry
matter. Similarly, method used by Wang, et al (2014) was slightly modified and used for characterization of
the volatile organic compounds (VOCs).

The proximate analysis of H. canabinus and V. colorata processed stems are presented in Figure 2.
The volatile matter content which is one of the parameter of interest in this research is 83.35% and 77.18% for
the H. canabinus and V. colorata respectively. The relatively high volatile matter in both plants justify why
they are widely used as sources of sustainable flame. Loo et al (2008) opined that when the volatile matters
detected is high; it signifies easy ignition and a proportionate increase in flame length. It also indicates that
during combustion, most plants will volatilize and burn in the gaseous phase. However, the H. canabinus with
volatile components of 83.35% will produce more flame. The relatively high dry matter contents of 85.00% for
H. canabinus and 90.92% for V. colorata further justify the high combustibility potentials of the plants.
325
Percentage ash content for both H. canabinus (8.60%) and V. colorata (4.65%) gives the
amount of the inorganic matter left out after complete The ash content is a lot higher than European
normal standard of <1.5% for biomasses reported (Mitchual et al, 2014). While the moisture contents
(MC) lies between 7.53% and 9.48 % which is well below the limit of 10-12% recommended by Austria
and German standards for fuel pellets and briquettes (ONORM M7135, 2003 and DIN 51731, 1996).
The low MC enhances the ignition quality which further heighten the combustion quality and helps
reduce objectionable smoke and toxic gases released (Chaney, 2010 and Werther et al, 2000) as cited
by Godbout et al.(2012).
Fixed carbon contents was 8.69% for V colorata and 0.23% for H. canabinus, which is quite low
in both samples. This is low when compared to a work reported for rice husk ash of 15.7% (Efomah and
Gbabo, 2015) and other related biomasses (kenaf grass -17.0%, neem wood -12.9%, mango wood-
11.36%, and corncob-11.36%) (Saidur, et al, 2011). Fixed carbon is not necessarily equal to the total
amount of carbon (ultimate carbon) in the plants because a significant amount is released as volatile
hydrocarbons components. This is confirmed by the value of elemental composition reported as
ultimate carbon to be 57.34% and 45.54% in V. colorata and H. canabinus respectively. This parameter
provides a rough estimate of the heating value of a fuel and acts as the main heat generator during
combustion. Thus, the higher the carbon content of a biomass the more likely that the species would
have higher heating value (Mitchual et al, 2014).
Figure 3 presents the ultimate analysis of the V colorata and H. canabinus stem obtained from proximate
analysis. Ultimate analysis involves the estimation of important chemical elements that makes up the
biomass, namely carbon, hydrogen, oxygen nitrogen and sulphur and are responsible for the heat
generating potential of the biomasses.
326
The results for V colorata is 57.34%, 6.56%, 35.50%, 0.56% and 0.04% for C, H, O, N and S
respective, while that of H. canabinus is C : 45.54%, H : 4.02%, O :31.32%, N : 0.20% and S : 0.01%. The
results obtained is within the limits of a similar work by Chaney (2010 showing the composition of the
principal constituents as carbon between 30 to 60% , oxygen 30-40%, hydrogen between 5-6%, and
nitrogen and sulphur normally makes up less than 1% of dry matter biomass. The sizable quantity of
carbon and hydrogen contents of these samples is very satisfactory as they contribute immensely to
the combustibility of the biomass (Musa, 2007) cited by Efomah and Gbabo, (2015). The slightly higher
hydrogen content as indicated from the H/C ratio of 0.11 for V. colorata when compared to H.
Canabinus with H/C ratio of 0.09 confirmed it has higher heating value. The nitrogen contents in both
plants is within the limit of ≤ 0.6% set by the Austria National Standard for Pellet and Briquettes
(Austria ÖNORM M7135 , 2003), and further conforms to limit of ≤ 0.3% set by the German National
Standard for Fuel Pellet, (Germany DIN 51731/DINplus, 1996). The sulphur contents was lower than
the limits set by the Austria and German national standards for fuel pellet and briquettes (Sulphur
content ≤ 0.08%). The low sulphur and nitrogen contents in both samples are low (Figure 3) which is
good as there will be minimal release of sulphur (SOX) and nitrogen oxides (NOx) into the atmosphere
on combustion, this is confirmed from the low concentration of SO2 averaging 3.13 ppm (Figure 6) for
both plants detected from the smoke emitted. Similarly H2S gases detected from the burning the H.
Canabinus and V. colorata stems were 6.50 and 23.00ppm which are still low compared to EPA
Maximum Standards 76.00ppm (REF). This further point to the fact that the use of the plants as sources
of light owing to the bright flame they produced to be environmentally friendly.
Calorific or heat value of both samples is 15 mJ/kg for H. canabinus and 31.52 mJ/kg for V. colorata. This is
within the range of a value reported in coconut shell-20.00mJ/kg, groundnut shell-19.20mJ/kg, eucalyptus-
19.35mJ/kg, softwoods-19.80mJ/kg, hardwoods-19.00mJ/kg, switch grass-19.90mJ/kg (Saidur et al, 2011).
But slightly below Gmelina arborea = 32.79mJ/kg, Terminaliasuperba = 32.69mJ/kg, Triplochitonscleroxylon
= 32.79mJ/kg reported Adegoke et al, (2014). The reason for high calorific value could be traced to high
carbon (ultimate carbon) content suggested by Kricka et al, (2012), who recounted that the calorific value of
fuel increases with similar increase in carbon level. Significance of the O: C and H: C ratios on the calorific
value of solid fuels was reported by Mckendry, (2002) and Dutta et al, (2013) using a Van Krevelen diagram
from which higher proportion of hydrogen and oxygen reduces the energy value of biomass fuel than fossil
fuel (coal). The average O:C and H:C ratios of 0.68 and 0.10 in both samples gives a fairly low level of
carbon which give a low calorific value in plants which has a remarkable influence on the heat values of the
plant under study. The C: N ratio helps in determining the appropriate energy conversion process in the
biomass the value obtained (Figure 4) which help in proposing the conversion of the energy of this plants
into more useful energy source.
327
From Figure 5, the loosed, loosed oven-dry and tapped oven-dry bulk densities of V. colorata are higher
than that of H. canabinus. This suggests that V. colorata is more likely to have higher energy per unit
volume. If all other factors remain constant, it will burns for a long period of time then the H. canabinus and
other biomasses used for that purpose.
328
CONCLUSION
Amongst the energy parameters detected in both the processed stems of H. canabinus and V. colorata,
the volatile matter contents of 83.35% and 77.18% respectively stands out as the major parameter that
justify the use of these plants by the indigenous communities as sources of light owing to the bright
flame the produce. The further characterization of the volatile matters which is ongoing will reveal the
chemical components that support the combustion and enhance sustainability of the bright flame.
ACKNOWLEDGEMENT
The Royal Society of Chemistry (RSC) is acknowledges for support and assistance for this work through
the Research fund grant which came timely.
REFERENCES
E.B. Agbro and N.A. Ogie (2010). A Comprehensive Review on Biomass Resources and Biofuel
production. Research Journal in Engineering and Applied Sciences. 1(3). pp.149-155.
Z. Chaula, M. Said, G. John (2014). Thermal Characterization of Pine Sawdust as Energy Source
Feedstock. Journal of Energy Technology and Policy. Vol. 4. No. 4. 57-65.
A. M. Saheed, S. W. Ahmed, M. Kazmi, M Mohsin and N. Feroze (2015). Impact of torrefaction
technique on moisture contents, bulk density and calorific value of briquetted biomass. Polish Journal
of Chemical Technology. 17 (2). 23-28
A. N. Efomah and A. Gbabo (2015). The Physical, Proximate and Ultimate Analysis of Rice Husk
Briquettes Produced from a Vibratory Block Mould Briquetting Machine. International Journal of
Innovative Science, Engineering & Technology, Vol. 2 Issue 5, May 2015. 814-822
T. Wang, L. Ding, Y Jia, H. Jin, R. Shi, L. Zhu and Y Ma (2014). Simultaneous
determination of six volatile components in Longhu Rendan pills using gas chromatography
coupled with triple quadrupole mass spectrometry. Anal. Methods, 6, 3713-3719
https://www.epa.gov/criteria-air-pollutants/naaqs-table
Hydrogen sulfide. Geneva, World Health Organization, 1981 (Environmental Health Criteria, No. 19).
08/06/2016
Similarly, other energy properties of the plant is determined in line with the work of Mitchual et al, (2014), who
evaluated the fuel properties of six tropical hardwood timbers species (Triplochitonscleroxylon,
Ceibapentandra, Aningeriarobusta, Terminaliasuperba, Celtismildbreadii and Piptadeniaafricana) for
briquettes, which includes; density, moisture content, volatile matter, ash content, organic carbon, elemental
composition and gross calorific value.
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329
Pollution index of a crude oil producing community in Niger Delta: A case study of Ibeno local government
area of Akwa Ibom State, Nigeria
1 1 2
*Godwin Asukwo Ebong , Helen Solomon Etuk and Imeh Edet Essien
1 2
Chemistry Department, University of Uyo, Physics Department University of Uyo
* Corresponding Author: Email: g_ebong@yahoo.com; goddyebong2010@gmail.com
Abstract
Surface soil samples were obtained from five (5) communities namely: Mkpanak; Iwou-Achange, Ukpenekang,
Okoroutip and Inua-Eyet Ikot within Ibeno, Akwa Ibom State, Nigeria. Control samples were also obtained in
Etinan local government area of the same State where impact of oil activities is not common. These samples and
Control were digested and analyzed for their lead, iron, chromium, cadmium, copper, vanadium and nickel
contents. Results obtained revealed that mean concentrations of Pb; Cr; Cu; V and Ni were within their
permissible limits in Nigerian soil while concentrations of Fe and Cd were above their critical limit. The mean
concentrations of these metals in studied soils were higher than their corresponding levels in Control indicating
anthropogenic inputs in the studied areas. Degree of contamination and pollution load index confirmed Mkpanak
as the location with highest level of contamination while Okoroutip was the least contaminated area studied. This
study also identified that concentrations of Pb; Cr; Cd; Cu; V and Ni were mainly from human activities within
areas studied while iron concentrations were mostly from natural source. Range of vanadium-nickel ratio
obtained signified that of a typical crude oil forming soil. Significant positive correlations were also observed
among these trace metals in the studied soils signifying a common anthropogenic source for them.
Key words: Trace metals, Ibeno, Enrichment factor, pollution load index, degree of contamination and
vanadium-nickel ratio.
Introduction
The origin of soil contamination and subsequent pollution by trace metals can be traced to the existence of human
on earth. Man in his quest to improve upon his living conditions has elevated the levels of metals (both the
essential and non-essential) significantly to obnoxious limits. Thus, with the inherent toxic and persistent nature
of metals in soil, their concentrations should be evaluated especially in developing nations of the world such as
Nigeria where environmental regulations are not strictly enforced. Ibeno as an area where intensive oil activities
is going on has attracted many oil related industries, commercial activities and thereby elevating the population.
Consequently, the level of toxic metals in the area is expected to be higher than normal and periodic evaluation of
the environment should be encouraged to forestall metal bioaccumulation with its associated health implications
in the area. Studies have shown that environmental problems associated with oil activities in an area are
enormous and alarming [1- 4]. Previous studies have also indicated that soils within Ibeno have elevated levels of
toxic metals [2, 5-7]. These high levels of toxic metals will eventually affect human beings through food chain
[8]. Hence, these reports coupled with the incessant oil spill within the area have necessitated the evaluation of
Ibeno environment to ascertain the status and avert any impending environmental disaster. This research aimed
at determining the pollution status of trace metals and locations in Ibeno studied. It also aimed at establishing the
source of these metals, the relationship among these metals and associated environmental implications. The oil
vanadium-nickel ratio will be employed to ascertain the extent of oil pollution within Ibeno.
Study area
Surface soil samples were obtained from Mkpanak, Iwou-Achange, Ukpenekang, Okoroutip and Inua-Eyet Ikot
? ! ? !
communities within Ibeno local government. Ibeno is a coastal area and lies between latitude 4 .30 N and 4 .36 N
and longitude 7?.48! E and 8?.18! E. It is a major oil producing community in Niger Delta region of the country.
Mobil producing Nigeria's operational base and the crude oil loading terminal are located in the area.
330
Sample collection and treatment
Top soil (0-15cm) samples were obtained from the five communities (Mkpanak, Iwou-Achange, Ukpenekang,
Okoroutip and Inua-Eyet Ikot) within Ibeno using soil auger. Control samples were obtained from a virgin forest
in Etinan local government of Akwa Ibom State. These samples and Control were obtained between the months
of November 2015 and January, 2016 following the procedures of Anake et al. [9]. A total of forty five (45) sub-
samples and five (5) composite samples were obtained for this study. These samples and Control were sun dried
for three days, ground and sieved through 0.5mm sieve. The sieved samples were stored in well-labeled
polyethylene containers for digestion and analysis. The sieved samples and Control were digested on a hot plate
using a mixture of concentrated HNO3 and concentrated HClO4 in the ration of 2: 1 [10]. The extract were
eventually analyzed for their Pb, Fe, Cr, Cd, Cu, V and Ni contents using Unicam Solar AAS 969 model.
Contamination factor (CF) was determined using equation 1: CF = C0-1/Cn----------------------------------------------
------------------------------------ (1) where CF is the contamination factor, C 0 -1 is the mean
content of metals from studied soils and Cn is the concentration of individual metal in control site. The degree of
contamination of the different locations by trace metals was assessed using the equation: Cdeg = ∑Cif -------------
-- (2) where ∑ Cif is the sum of the contamination factors for all the elements at a particular location.
Enrichment factor (EF) of trace metals was calculated using equation 3 indicated below:
EF = (M/Fe) sample / (M/Fe) Control ------------------------------------------------------------------------- (3) where (M/Fe)
sample is the ratio of metals and iron concentration in studied soil and (M/Fe) Control is the ratio of metals and iron
concentration in Control site. Pollution load index (PLI) of each site studied: was determined using the equation
4 below:
1/7
PLI = (CFPb × CFFe × CFCr × CFCd × CFCu × CFV × CFNi) --------------------------------- (4) where CF
indicates contamination factor of each trace metal studied.
Vanadium-nickel ratio was calculated using the equation: V/Ni = [V]/[Ni] ----------------------- (5) where [V] and
[Ni] denote mean concentrations of vanadium and nickel respectively at each location studied. The mean,
standard deviation; median and correlation coefficients of trace metals were analyzed using SPSS for windows
(version 20.0).
Results for the concentrations of trace metals in soil samples and Control are shown in Table 1. The
Contamination factor (CF), Degree of contamination (Cdeg) and pollution load index (PLI) of trace metals in soil
are indicated in Table 2. Table 3 shows the results for enrichment factor (EF) of trace metals in soil. The
correlation coefficients of studied trace metals in soils are shown in Table 4.
*Table 1: Distribution of trace metal concentrations (mg/kg) in surface soil Ibeno communities within Akwa
Ibom State.
Location Pb Fe Cr Cd Cu V Ni V/Ni
Mkpanak 14.15 1631.72 1.75 4.53 7.73 0.08 3.96 0.02
Iwou Achange 12.76 1509.84 1.86 4.21 6.14 0.05 3.84 0.01
Ukpenekang 12.68 1473.29 1.64 4.48 6.29 0.09 3.25 0.03
Okoroutip 11.21 1336.61 1.68 3.96 5.31 0.04 2.77 0.01
Inua-Eyet Ikot 11.36 1371.21 1.57 4.13 5.17 0.05 2.28 0.02
Mean 12.43 1464.53 1.70 4.26 6.13 0.06 3.22 0.02
Minimum 11.21 1336.61 1.57 3.96 5.17 0.04 2.28 0.01
331
Maximum 14.15 1631.72 1.86 4.53 7.73 0.09 3.96 0.03
Range 2.94 295.11 0.29 0.57 2.56 0.05 1.68 0.02
SD 1.20 117.43 0.11 0.24 1.02 0.02 0.71 0.01
Median 12.68 1473.29 1.68 4.21 6.14 0.05 3.25 0.02
Control 5.15 1213.48 1.25 2.76 3.86 0.01 0.84 0.01
* Mean of three determinations.
Results in Table 1 indicate that the distribution of total lead in surface soil of studied Ibeno communities
varied between 11.21mg/kg recorded in Okoroutip and 14.15mg/kg in samples from Mkpanak. This range is
higher than 0.01 – 0.07 mg/kg reported by Nwadinigwe et al. [7] but lower than 1.60 – 54.40 mg/kg obtained by
Etim et al. [6] in the studied area. However, the mean concentration of total Pb obtained (11.97±1.72 mg/kg) is
higher than 5.15 mg/kg recorded at the Control site. Thus, the activities of oil and oil related industries in the
studied area may have resulted in the accumulation of lead in the host communities. Nevertheless, the mean
concentration of lead reported in this study is lower than the critical value of 100 mg/kg by Lacatusu [11] and
DPR [12] recommended limit of 85 mg/kg in soil. Consequently, concentrations of total lead in the area may not
pose serious health risk though periodic assessment should be done to avoid bioaccumulation. Concentrations of
lead in the studied soils as indicated in Table 1 exhibit higher median than mean (negatively skewed).
Total Iron concentrations in surface soils varied between 1336.61 mg/kg at Okoroutip and 1631.72
mg/kg in samples from Mkpanak (Table 1). This range is higher than 135.17 – 245.20mg/kg reported in the same
area by Etim et al. [6] but lower than 1746.40 – 2839.40 mg/kg by Iwegbue et al. [2]. The mean concentration of
Fe (1464.53±117.43 mg/kg) is higher than 400 mg/kg recommended limit by FEPA [13] for Nigerian soil. The
mean concentration of Fe is also higher than 1213.48 mg/kg obtained in the Control sample indicating
anthropogenic inputs of the metal in the studied area. Thus, iron even though is known to be essential for plants
and animal growth could be considered a nuisance at the reported level. This is in agreement with the report by
Adefemi et al. [14] that Nigerian soils have high concentrations of iron. Studies have shown that iron
concentration is higher in acidic soils thus; the acidic nature of the studied area may have resulted in the high
concentrations of iron reported [4, 15]. Generally, concentrations of iron in soils within Ibeno showed a higher
median than mean (negatively skewed) (Table 1).
Concentrations of total chromium in the studied area ranged between 1.57 mg/kg obtained at Inua-Eyet
Ikot and 1.86 mg/kg at Iwou-Achange. This range is lower than 0.80 – 2.40 mg/kg reported by Ndeh et al. [16] in
soil within the area but higher than 0.005 – 0.200 mg/kg reported by Amos-Tautua et al. [17]. The mean
concentration of chromium (1.70±0.11 mg/kg) is lower than 1.25 mg/kg reported in Control which is an
indicative of anthropogenic addition of Cr to the studied soils. However, the obtained mean is lower than the
critical level of 50.00 mg/kg by Lacatusu [11] and DPR [12] recommended limit of 100 mg/kg in soil. Thus,
concentrations of Cr recorded at the different locations studied may be considered safe and useful for the proper
functioning of plants [18]. Chemistry of chromium in soil is mostly dependent upon the forms of chromium
which is a function of redox potential and pH of the soil. In soils of most places, chromium is present
predominantly in the chromium (III) oxidation state which is very low in mobility and low toxicity in living
organisms [19, 20]. However, under oxidizing conditions of soil chromium (VI) may be present and this form of
Cr is relatively soluble, mobile and toxic to living organisms [21]. Nevertheless, the acidic nature of the study
area could facilitate the reduction of chromium (VI) to chromium (III) making the element less toxic in the area
[22, 23]. The distribution of total chromium in studied areas skewed to the right with a higher mean than median.
Recorded total cadmium concentrations varied between 3.96 mg/kg at Okoroutip and 4.53 reported at
Mkpanak. This range is higher than 0.01 – 0.27 mg/kg and 0.00 – 0.35 mg/kg reported in soils within Niger Delta
by Etim et al. [6] and Nwadinigwe et al. [7] respectively. The mean total concentration of Cd (4.26±0.24 mg/kg)
332
is higher than 2.76 mg/kg reported in the Control indicating additional source of the metal other than natural to
the area. This mean is also higher than 0.80 mg/kg recommended limit for Nigerian soil by FEPA [13]. The high
concentrations of cadmium in studied locations may be attributed to the low pH range of soils within Ibeno [5,
24]. These high concentrations of Cd in Ibeno may be attributed to anthropogenic sources of the metal including
oil activities in the area as confirmed by enrichment factor value above 1 [25]. The elevated concentrations of Cd
recorded in this work may also be attributed to the low concentrations of zinc in studied area as reported in
literature [3, 7, 16, 24, 26]. This could be very risky as cadmium is known to be very toxic and highly mobile [27].
Concentrations of total cadmium in studied locations showed a positively skewed distribution as indicated in
Table 1 with a lower median than mean.
Concentrations of total copper in the studied areas ranged between 5.17 mg/kg in samples from Inua-
Eyet Ikot and 7.73 mg/kg obtained at Mkpanak (Table 1). This range is higher than 2.95 – 3.45 mg/kg but lower
than 23.20 – 1661.30 mg/kg reported for contaminated soils by Etim et al. [6] and Al-Momani [28] respectively.
Though, the mean concentration of copper (6.13±1.02 mg/kg) obtained is lower than the critical limit in soil
(50.00 mg/kg) by Lacatusu [11] and 10.1 mg/kg stipulated by FEPA [13] for Nigerian soils. Thus, copper at the
level reported in this study may be useful for optimum plant growth. However, the mean concentration of copper
obtained in higher 3.86 mg/kg recorded in the Control thus revealing some anthropogenic source of the metal in
the studied area. This should be monitored and controlled to forestall copper toxicity and its attendant's effects.
The low concentrations of copper in the studied areas may be attributed to the high sand and iron contents in soil
of the area as studies have shown that high levels of these properties in soil could reduce copper concentrations
significantly [29]. The distribution of copper in studied soils as shown in Table 1 has almost the same mean and
median (normal distribution).
Levels of vanadium obtained in this study ranged between 0.04 mg/kg at Okoroutip and 0.09 mg/kg in
samples from Ukpenekang. This range is lower than 0.00 – 1.13 mg/kg reported by Nwadinigwe et al. [7]. The
mean vanadium concentration (0.06±0.02 mg/kg) obtained is lower than a normal range in soil of 30 - 150 mg/kg
by kabata-Pendias [30] and 10.83 mg/kg by WHO [31]. Consequently, vanadium may not be considered as a
pollutant in the studied area. However, the mean is higher than 0.01 mg/kg recorded at the Control site which is
an indicative of anthropogenic addition of the metal to environment investigated. Generally, concentrations of
vanadium obtained were low and this may be attributed to low clay content, acidic nature of the studied soils and
high iron content [32 -34]. The distribution of total vanadium in soils evaluated showed a normal distribution as
the mean value is almost the same with median.
Nickel concentrations varied between 2.28 mg/kg at Inua-Eyet Ikot and 3.96 mg/kg reported in samples
from Mkpanak. This range is lower than 0.02 – 22.80mg/kg obtained in soil by Edet et al. [5] but higher than 0.00
– 0.99 mg/kg reported by Nwadinigwe et al. [7]. The mean concentration of total Ni obtained in this study
(3.22±0.71 mg/kg) is lower than 0.84 mg/kg reported in Control, the critical limit (50.00 mg/kg) by Lacausta [11]
and 35.00 mg/kg recommended limit by FEPA [13] for soils in Nigeria. Thus, nickel may not be at the nuisance
level in studied soils but due to persistence nature of metals in soil its availability should be monitored to avoid
bioaccumulation and associated health implications in the studied soils reported by Raymond [35]. Relatively,
the general levels of nickel in soils assessed were low and this could be as result of low soil pH and high sand
content which may accelerate mobility and leaching [5, 36]. Variations in the concentration of nickel within the
study area indicated a negatively skewed distribution with higher median than mean (Table 1).
Table 2: Contamination factor (CF), Degree of contamination (Cdeg) and pollution load index (PLI) of trace
metals in soil.
333
Location Pb Fe Cr Cd Cu V Ni Cdeg PLI
Mkpanak 2.75 1.35 1.40 1.64 2.00 4.00 4.71 17.85 46.00
Iwou Achange 2.48 1.24 1.49 1.53 1.59 2.50 4.57 15.40 18.00
Ukpenekang 2.46 1.21 1.31 1.62 1.63 4.50 3.87 16.49 26.00
Okoroutip 2.18 1.10 1.34 1.44 1.38 2.00 3.30 12.41 6.00
Inua Ikot Eyet 2.21 1.13 1.26 1.50 1.34 2.50 2.71 12.65 6.00
Mean 2.42 1.21 1.36 1.55 1.59 3.10 3.83 14.96 20.00
The pollution status of trace metals in studied soil environment was assessed using contamination factor
(CF) as reported by Mmolawa et al. [37]. Results obtained in Table 2 indicate that, Pb, Fe, Cr, Cd and Cu were in
the moderate contamination class. Hence, these trace metals could have contaminated studied soil environment
moderately. However, vanadium and nickel contaminated the studied soil environment considerable since their
mean CF values were above 3 as shown in Table 2. Thus, V and Ni could have impacted negatively and
considerably in soils within the studied communities in Ibeno.
Degree of contamination (Cdeg) was employed in this research to examine the extent of site to site
contamination by trace metals. Results obtained for Cdeg in Table 2 show a range of 12.41 - 17.85 with Mkpanak
having the highest Cdeg value while Okoroutip was the location lowest Cdeg value. Results obtained also revealed
that Iwou Achange, Okoroutip and Inua Ikot Eyet were moderately contaminated while Mkpanak and
Ukpenekang were considerably contaminated by the studied trace metals [38].
The severity of metal contamination and its variations from one site to the other was evaluated with
pollution load index. Table 2 indicates that pollution load index (PLI) of trace metals assessed ranged between
6.00 and 46.00. As reported by Ra et al. [39] when PLI is higher than 1.00, there is contamination in the study area
however, if PLI is less than 1.00 then there is no contamination by trace metals assessed. Results obtained
revealed that soils within Mkpanak, Iwou Achange and Iwou Achange were severely polluted with the studied
trace metals while Okoroutip and Inua Ikot Eyet were moderately contaminated by these metals. Results of PLI
analysis confirmed the observations of Cdeg on the pollution status of different locations studied. This study has
also shown that trace metals which may have resulted from oil exploration and exploitation activities in the
studied area may have impacted negatively and seriously on the area, thus remediation works are recommended.
The vanadium-nickel ratio according to Mirza et al. [40] used as indicator of oil pollution in soil ranged
between 0.01 and 0.03. The highest V/Ni ratio was reported in Ukpenekang where oil facilities are located in the
area while the lowest ratio was recorded in Iwou-Achange and Okoroutip. This study therefore revealed that oil
activities in the studied region may have contaminated the soil environment with crude oil and its chemical
compositions to a certain degree. The range of V/Ni ratio obtained in this study is lower than 0.09 – 0.41 reported
by Olarjire and Oderinde [41] in Nigerian crude oil but higher than zero values obtained by Adebiyi et al. [42].
The range of V/Ni ratios obtained in this research signifies an oil-forming continent environment.
Table 3: Enrichment factor (EF) of trace metals in soil
Location Pb Fe Cr Cd Cu V Ni
Mkpanak 2.07 1.00 1.07 1.21 1.48 3.06 3.47
Iwou Achange 2.01 1.00 1.23 1.21 1.27 2.07 3.63
Ukpenekang 2.05 1.00 1.11 1.32 1.33 3.82 3.15
Okoroutip 2.00 1.00 1.26 1.29 1.24 1.87 2.96
Inua Ikot Eyet 1.97 1.00 1.15 1.31 1.18 2.28 2.38
Mean 2.02 1.00 1.16 1.27 1.30 2.62 3.12
334
Enrichment factor (EF) of studied trace metals in Table 3 indicates that Pb had a mean EF value of 2.02.
Consequently, lead enrichment may have been mainly from anthropogenic inputs as reported by Hernandez et al.
[43]. Chromium, cadmium and copper indicated mean EF values of 1.16, 1.27 and 1.30 respectively. Thus, their
inputs may have been from anthropogenic source though minimal since their mean EF values are above 1.00
[25]. However, Iron used a normaliser element in this study recorded a mean EF value of 1.00 indicating that the
main source of the element was natural. The mean EF values for vanadium and nickel are 2.62 and 3.12
respectively indicating that, V and Ni were mostly from the anthropogenic source. The high mean EF values of
these metals may be closely related to oil activities in the studied environment since these metals are related to oil
[40].
Table 4: Pearson correlation coefficient among trace metals in soil.
Pb Fe Cr Cd Cu V Ni
Pb 1.000 0.899 0.915 0.986 0.904 0.803 0.953
Fe 1.000 0.809 0.859 0.983 0.831 0.934
Cr 1.000 0.853 0.783 0.627 0.956
Cd 1.000 0.870 0.909 0.905
Cu 1.000 0.861 0.922
V 1.000 0.785
Ni 1.000
Red is significant at P ≤ 0.01; green at P ≤ 0.02 and blue at P ≤ 0.10
Results of Pearson Correlation analysis for trace metals in Table 4 revealed significant positive
correlation between the following pair of metals at P ≤ 0.01: Pb/Fe; Pb/Cr; Pb/Cd; Pb/Cu; Pb/Ni; Fe/Cr; Fe/Cd;
Fe/Cu; Fe/V; Fe/Ni; Cr/Cd; Cr/Ni; Cd/Cu; Cd/V; Cd/Ni; Cu/V and Cu/Ni (r values are indicated in Table 4
above). Strong positive correlations also existed in Pb/V; Cr/Cu and V/Ni but at P ≤ 0.02 with r value of 0.803;
0.783 and 0.785 respectively. However, the significant positive correlation between chromium and vanadium
existed at P ≤ 0.10 with r value of 0.627. Generally, significant positive correlation existed among the entire
trace metals studied. Thus, the presence of the studied trace metals may have originated mainly from a common
anthropogenic source which could be oil related activities [44, 45]. This indicates that the activities of oil
companies and other downstream industries may have contributed some substantial amounts of these trace
metals into the studied soil environments.
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338
SYNTHESIS AND BIOLOGICAL ACTIVITIES OF METAL COMPLEXES OF SOME ORGANIC
BASES
Emwanta D. O.* and Ngochindo R. I.
Department of Chemistry, University of Port Harcourt, Rivers State, Nigeria.
ABSTRACT
Metal complexes of organic ligands such as imidazole and Schiff bases have been found to be biologically active.
Cr(III), Co(II), Fe(II), Cu(II) and Ni(II) complexes of theses ligands have been studied and they show
bioactivities as antimicrobial, antidiabetic, antioxidant and antifungal agents.
INTRODUCTION
Azoles are 5-membered monocyclic heterocycles with multiple heteroatoms with at least one of such atoms
being Nitrogen. Imidazole, a typical diazole is by far the most studied azole, an aromatic heterocyclic
organic compound with the formula C3H4N2. It is amphoteric in nature as it acts as a base via the azomethne
nitrogen with lone pair of electrons, and as an acid via the imino nitrogen with a proton attached.
N
N
H
Fig. 1 Imidazole
The compound is aromatic due to the presence of a sextet of ð-electrons, consisting of a pair of electrons from the
imino nitrogen and one each from the remaining four atoms of the ring. Imidazole derivatives have occupied a
unique place in the field of medicinal chemistry and have broadened scope in remedying various kinds of
ailments in clinical medicines. Typical examples include; 2-nitroimidazole (azomycin) and metronidazole
which are antitricomonal agents, uroconic acid – an antibiotic, Methimazole - an antithyroid agent, to mention a
few. Imidazole is present in important biological building blocks, and many drugs contain the imidazole ring
Schiff bases are condensation products of primary amines and aldehydes or ketones. They are chiefly
synthesized from other organic precursors as they are not found in naturally occurring compounds. Schiff bases
have a wide range of applications as polymer linkers, corrosion inhibitors, potentiometric sensors, extraction
solvents and other synthetic applications. Their use as biologically active agents has gained popularity in recent
times.
Synthesis of Halogenated Imidazoles

Imidazole undergoes both electrophylic and nucleophilic substitution with more of electrophilic reactions at
both the carbon and nitrogen atoms. Electrophilic substitutions with halogens on carbon atoms are quite
complex, giving rise to various halogenated imidazoles. They are synthesized with the use of appropriate
halogen containing compounds such as sodium hypochlorite, bromine and iodine. Typical examples are the
reactions of imidazole with NaOCl to give 4,5-dichloroimidazole, reaction of iodine with imidazole to give
2,4,5-triiodoimidazole and reaction of bromine with imidazole to give 4,5-dibromoimidazole.
339
Cl Br I
N N N
Cl N Br N I N I
H H H
4,5-dichloroimidazole 4,5-dibromoimidazole 2,4,5-triiodoimidazole
Fig.2 Halogenated Imidazoles
Pharmacological Properties of Halogenated Imidazoles and Their Metal Complexes
Common halogenated imidazoles are the chloro-, bromo- and iodoimidazoles. Chlorine, Bromine and Iodine
substitute directly into the ring. The fluorine substituent in the ring is, however, uncommon. Common
fluorinated imidazoles are compounds with fluorine on a substituent attached to the ring or highly substituted
imidazoles (1). These halogenated imidazoles have been synthesized and found to be bioactive. However few
researches have been reported.
Antimicrobial agents. 4,5-dibromoimidazole and 2,4,5-triiodoimidazole were found to be bioactive against
microbial strains Stahphilococcus aureus and Stahphilococcus pyogenes. Their antimicrobial potencies were
significantly enhanced by complexation with Co(II) and Cu(II) (2, 3)
Antifungal agents. 4,5- dibromoimidazole and 4,5-dichloroimidazole were found to show biological activities
against fungal strains - Aspergilla niger and Candida albicans. Their bioactive potencies were also enhanced by
complexation with Co(II), Cr(III) (2, 4).
Synthesis of Schiff Bases

Schiff bases are generally synthesized from the condensation reactions of a primary amine with an aldehyde
or ketone. The aromatic derivatives are more stable than the aliphatic derivatives
N N
3
aliphatic Schif f Base aromatic Schif f Base
Fig. 3. Structures of Schiff Bases

Pharmacological Properties of Schiff Bases and Their Metal Complexes
Schiff bases have been found to show biological activities as antimalarial, antibacterial, antifungal, and
anticancer agents. Fe(II), Cu(II) and Ni(II) complexes of 1-phenyl-N[(1E,2E)-3-phenylprop-2-en-
1ylidene]methanamine show significant bioactivity against some fungal and bacterial strains (5). Mn(II), Co(II),
Ni(II) and Cu(II) metal complexes of 4-acetyl-3-methyl-1-phenyl-2-pyrazolin-5-one and 4-Benzoyl-3-methyl-
1-phenyl-2-pyrazolin-5-one show biological activities as antimicrobial agents (6, 7). Mn(II), Co(II), Ni(II) and
Cu(II) metal complexes of the ligand (E)-N'-((7-hydroxyl-4-methyl-2-oxo-2H-chromen-8--
yl)methylene)benzohydrazide show antidiabetic and antioxidant activities (8)
Transition Metals and Associated Bioactive Properties
Metals are known to greatly enhance bioactivity of bioactive compounds. Some organic compounds were found
to posses bioactivities in various ways and showed enhanced activities when complexed with transition metals.
Organic complexes with metals such as cobalt, copper, nickel, iron, chromium and manganese show bioactivities
against gram positive microbial strains – Stahphilococcus aureus, Stahphilococcus pyogenes, Bacillus subtilis
340
and gram negative microbial strains - Escherichia coli, Proteus spp, Salmonela typhii (9, 10, 11, 12) Organic
ligands complexed with zinc, cobalt and nickel show antibacterial activities (12, 13, 14) Organic complexes with
chromium and cobalt show bioactivities against fungi strains – Aspergilla niger and Candida albicans.(15).
Organic and inorganic complexes with vanadium show bioactivities in diabetic screening, reducing the
concentration of glucose in streptozotocin induced diabetic rats. (15, 16, 17, 18) Vanadium also shows
bioactivity in antitumor screening by the inhibition of hepatocellular preneoplasia in experimental rats with
hepatocarcinogenesis. (19, 20, 21)
CONCLUSION
Quite a few researches have been reported on halogenated imidazoles metal complexes with their bioactivities.
Metal complexes of imidazoles and Schiff bases have been found to show various levels of bioactivities as
antimicrobial, antidiabetic, antioxidant and antifungal agents at low concentrations and also at higher
concentrations. Vanadium and Zinc complexes of organic compounds have shown antidiabetic, antitumor and
antimicrobial activities, respectively. Complexation of these halogenated imidazoles with these metals is
expected to further diversify the applications of organic bases as bioactive substances.
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19. Bishayee A., Oinam S., Basu M., Chatterjee M. (2000); Vanadium chemoprevention of 7,12-
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Molecular basis of vanadium-mediated inhibition of hepatocellular preneoplasia during experimental
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Assessment of Rainwater Quality of Bichi Local Government Area of Kano State of Nigeria
Emmanuel Bernard
Department of Chemistry, Nigeria Police Academy, Wudil. P.M.B 3474 Kano – Nigeria. E-mail:
royalhoodem@yahoo.com.
Abstract
The physiochemical and bacteriological quality of rain water in Bichi Local Government Area of Kano State
was studied using standard method of analysis. The result obtained indicated the mean and standard deviation of
o
Temperature, pH, Turbidity, Total Alkalinity, Calcium, Magnesium were found to be 25.03 ± 0.82 C, 6.08 ±
0.34; 3.13 ± 0.64 NTU; 22.85 ± 6.45 mg/l; 5.76 ± 3.41 mg /l; 1.70 ± 0.65 mg /l; 0.61± 0.00 mg /l respectively.
The value of others parameters such as chloride, free Carbon dioxide and conductivity were found to be 5.41±
1.98 mg/l; 4.53 ± 2.27 mg/l; 27.71± 11.81 s/cm respectively, while coli form and E.coli were absent. It was
observed that the rainwater samples as at collection were within WHO's standard. The study is significant in
order to establish campaign and creating awareness for possible cautions to minimize environmental pollution
that could contaminate rainwater and to reduce the incidence of water borne diseases in and around environment
and Kano State in general.
Keywords: Assessment, Bacteriological, Bichi, Drinking, Properties, Rainwater, Water
1.0 Introduction
Rainwater is an important source of fresh water especially for those living in rural areas, where portable water is
scarce or where surface and underground water quality is poor. (Chukwuma, et al 2012) As a natural occurrence,
it is seasonal. Although it is always available in season, it can be a source of water for domestic, agricultural and
industrial purpose when harvested and stored when in season. (Dinrifo, et al 2010).
Developments in science and technology have brought improved standard of living, but have also
unsuspectingly introduced some pollution into our environment. In a limited number of areas, specific
industries or very heavy traffic emissions may affect the chemical quality of rainwater, (Abdul, et al, 2009).
The prevailing activities occurring from local industries, uncontrolled refuse disposal, drying up of rivers,
ponds at times regulated by negative ecological distribution could contribute immensely to the occurrence of
the diverse chemical and microbial contaminations observed in water, (Bernard and Nurudeen A , 2012;
RAIN, 2008)
Many researchers have considered the effect of acid rain on human health. These acid pollutants can be deposited
in a dry form through dust. Pollutant that contributes to acid rain may be carried hundreds of miles before being
deposited on the earth. Because of this, it is sometimes difficult to determine the specific sources of these acid
rain pollutants. However, excessive amounts of acid gases like SO2, NO2 and CO2 that can dissolve in rainwater
as it is falling from atmospheric cloud are responsible for acid rain. (Dinrifo, et al 2010; Olobaniyi and Efe,
2007)
Rainwater harvesting for domestic use is becoming increasingly popular as the availability of good quality water
is declining. This is further aggravated by the adverse impacts of climate change on water supply sources.
Consequently, water authorities around the world are keen to explore alternative water sources to meet ever-
increasing demands for potable (i.e., drinking) water. (Abdul,, et al, 2009; Gardner, et al, 2011).
Previous studies on the quality of water resources in the tropical African environment have largely been
restricted to surface and groundwater to the negligence of rainwater. This is predicated on the assumption that
rainwater is usually very pure and therefore needs very little investigation. However, studies have shown that
rainwater quality can be significantly impaired in industrialised districts. Akharayi, et al, (2007) from Ondo
State, Ubuoh, (2012) from Akwa Ibom State and Olowoyo, (2011) from Delta State.
Kano State remains Nigeria's second commercial city and the commercial hub of the sub Sahara Africa.
343
Many of the commercial outfits and factories freely release their effluents and emissions into the
environment. This study is aimed therefore at evaluating the properties of rainwater being collected for use in
Bichi Local Government Area of Kano State for possible database and creating awareness of possible
cautions to minimize environmental pollution that could contaminate rainwater and reduce the incidence of
water bone diseases.
2. Materials and Methods

2.1. Description of Study Area
Bichi Local Government Area covers an area of 612 km² with a population of over 277,099 at the 2006 census
and lies between latitude 12°35' and 36.5°N and longitude 7°48' and 06°E. It is about 45Km from the commercial
centre and hosted by Kano state which is ranked second in population with about 9.0million people and lies
between latitude 11°30' and 11.5°N and longitude 8°30' and 8.5°E. Nigeria is located approximately between
o o
latitude 4 and 14 North of the equator, and between longitude 2° 2' and 15° east of the Greenwich meridian.
2.2. Collection/Treatment of Water Samples
The rainwater samples were collected between May and September in an open space on an elevated platform in
order to ensure that the water have no contact with any object before getting into the plastic container. This was
transferred into 200 ml capacity poly propylene containers, which had been previously washed with detergents
and soaked for 24 hours in 1 % HNO3. Few drops of concentrated nitric acid (analytical grade) were then added to
the samples for metallic ions determination. This is to maintain the stability of the oxidation state of the various
elements in solution and prevent precipitation to ensure metals remained in solution. Measurement of parameters
H
were made using various corresponding meters (e.g. Luvibond 1000, P Comparator), DR 3 Spectrophotometer
model HACH Drel / 5 and titrimetric method as described in standard analytical procedures as earlier reported by
APHA, 1998. The media used for microbiological analysis included multiple tube technique and violet red bile
agar-media for E-coli determination and the Coliform counts were carried out by means of the standard plate
count technique using MacConkey agar.( APHA, 1998)
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Figure 1. Map of Bichi Local Government Area of Kano State of Nigeria.
3. Results and Discussion

The results obtained from the physicochemical and microbiological analysis of rainwater samples collected
from Bichi Local Government Area is presented in Tables 1.
Table 1. Mean and standard deviation values of physicochemical/microbiological parameters of rainwater in

Bichi Local Government Area of Kano State
Parameters Mean Std WHO
Deviation Limit
Temperature ( oC) 25.03 0.82 20.0 – 32.0
PH 6.08 0.34 6.5 – 9.5

Turbidity (NTU) 3.13 0.64 5
Alkalinity-Methyl Orange 22.85 30.0 – 50.0
6.45
(mg/l)
Calcium – Ca 2+ (mg/l) 1.70 0.65 100 – 300
Magnesium-Mg 2+ (mg/l) 0.61 0.00 100
Chloride ion-Cl- (mg/l) 5.41 1.98 250
Free Carbon Dioxide-CO2 4.53 6.0 – 60.0
2.27
(mg/l)
Conductivity (ìs/cm ) 27.71 11.81 1000
Coliform (cfu/ml) 0.00
E. coli (cfu/ml) 0.00
From the result presented, it can be seen that the temperature of the rain water was found to be mean and
standard deviation of 25.03 ± 0.82. This value was within WHO's permissible limit. The values obtained are
as expected as the area under study is not an industrial area. According to Bhatia (2009) areas prone to
discharge of industrial wastes usually have temperature ranges above those of their surrounding
o
environments this was played out in oil producing areas were higher temperature values of 30.4 C at Delta
o
State (Dami et, al., 2012) and 31 C at Bayelsa State (Ezenwaji, et al., 2013) were reported.
The pH values of rainwater samples collected varied between 5.7 and 6.5 with a mean and standard
deviation of 6.08 ± 0.34, it was within the limit prescribed by WHO. Similar results were reported by Ubuoh,
(2012); Olowoyo, (2011) and Akharayi, (2007) for Akwa Ibom state Delta state and Ondo state of Nigeria
respectively. However, lower values were recorded (5.46 – 5.98) for Anambra State a commercial city with
high vehicular and industrial pollutants. (Chukwuma, et al, 2012) and 5.10-6.35 in oil producing area of
Nigeria (Olobaniyi and Efe, 2007). However more acidic value range of 4.20 – 4.94 was report in
highly industrial areas of Lagos state by Igwo-Ezikpe and Awodele (2010). PH is an important operational
water quality parameter (WHO 2011). In the study area the lowest pH values of 5.7 was below the acceptable
limit, this value was observed from the early rain collections.
The values for Turbidity obtained for the study area had a mean and standard deviation of 3.13 ± 0.64 NTU,
which was within WHO prescribed limits. Lower range of 2.42 - 1.19 NTU was reported by Dami, et, al.,
(2012) and Olobaniyi and Efe, (2007) from their study obtained higher range of 3.45 – 7.00 NTU from the
oil producing area. High turbidity levels are often associated with higher levels of disease-causing
microorganisms such as viruses, parasites, bacteria and dissolved chemicals in most waters; it may also be
due to colloidal and extremely fine dispersions and is a measure of the extent to which light is either
absorbed or scattered by suspended materials in water. (Gardner, et al., 2011).
Total alkalinity values for samples was found to be of mean and standard deviation of 22.85 ± 6.45 mg/l
345
which is within permissible thresholds. Dami et, al.,(2012) from Delta state reported a range value of 6.70 -
8.90 mg/l which was less than the reported value. Alkalinity of water is its capacity to neutralize a strong
acid and it is normally due to the presence of bicarbonate, carbonate and hydroxide compound of calcium,
sodium and potassium. Hydroxide ions are always present in water, even if the concentration is extremely
small. (Dami et, al., 2012)
2+ 2+
Calcium (Ca ) and Magnesium (Mg ) are directly related to hardness. From Table 1, calcium concentration
ranged was with mean and standard deviation of 1.70 ± 0.65mg/l and magnesium content in the investigated
samples had a mean standard deviation of 0.61 ± 0.00mg/l. Both were found to be below WHO limit. Dami et, al.,
(2012) reported similar ranges of 1.22 - 1.93 mg/l for Ca2+ and 0.65 - 1.08 mg/l for Mg2+ respectively. Hardness is
the property of water which prevents lather formation with soap and increases the boiling points of water. It
mainly depends upon the amount of calcium or magnesium salts or both, their absence may indicate that nutrient
supplement is of great need if the water is harvested for the purpose of human consumption (Chukwuma, et al,
2012). However, there is insufficient scientific information on the benefits or hazards of long-term consumption
of very low mineral waters to allow any recommendations to be made (WHO, 2011). The values obtained were
far less than the WHO prescribed limit for drinking water. However such water is best and excellent where
hardness is not needed as in the case of soap production.
In the present analysis, chloride concentration was found with mean and standard deviation values of 5.41± 1.98
mg/l and range of 2.94mg/l – 7.89mg/l. Dami et, al., (2012) reported lower range of 0.10 – 0.20 mg/l but
Chukwuma, et al, (2012) and Olobaniyi and Efe, (2007) reported higher value of 8.00 - 16.00 mg/l, and 19.50 –
26.95 mg/l respectively. The values were all within the limit accepted and this can be attributed to the mode of
sample collection.
The level of Carbon (IV) oxide (CO2) in the rain water samples had a mean and standard deviation of 4.53 ±
2.27mg/l and range 2.00 – 7.00mg/l, the value was in agreement with Chukwuma, et al, (2012) of 3 – 4mg/l.
Although this is still within the WHO upper limit, it however showed that there were appreciable emissions of
CO2 gas from vehicles and domestic activities.
The conductivity concentrations range was 10 – 40 ìs/cm with a mean and standard deviation of 27.71 ±
11.81 ìs/cm which were below standard for drinking water. Electrical conductivity is a measure of water
capacity to convey electric current. Olobaniyi and Efe, (2007) reported similar value range of 14.05-42.10
ìs/cm from Delta State and 76 ìs/cm was reported by (Ezenwaji, et al., 2013)
346
Figure 2: Variation of the physicochemical properties with time under the studied period
The micro-biological parameters assessed showed negative confirmation test to coliform and E-coli, suggesting
that it was devoid of pathogens as earlier highlighted by Olobaniyi and Efe, (2007), Chukwuma, et al, 2012 and
Dami et al., (2012) in their research.
The absence of coliforms, E-coli and low concentration of parameters can partly be explained by the mode of
collection. Rainwater samples were collected directly into containers as it fell through the atmosphere. Usually
serious microbial contamination only begins after contact with collection surfaces. (Olobaniyi and Efe, 2007.
Gardner, et. al, 2011),
Rainwater need to be monitored since safe rainwater can be easily contaminated after collection. Hygiene
education and monitoring of the operation and maintenance of the system, along with sanitary practices, are
essential if rainwater supplies are to provide clean water. Creating awareness on personal and environmental
hygiene and issues related to rainwater is crucial. (Olowoyo, 2011, UNESCO, 2002)
From Figure 2, it was observed that the first few rainfalls at the beginning of raining season in the month of
May contained the highest proportion of the pollutant load. This may be attributed to the deposition and
accumulation of pollutants during dry season periods. The longer the dry period, the greater the probabilities
of higher pollutant load in the first rain. (Chukwuma, et al, 2012, WHO, 2011.). The month of September
equally had values higher than those of the month of July when there were more frequent rains, and this can
be attributed to the break during the month of August leading to accumulation of pollutants
4. Conclusion
Rainwater is relatively clean and the quality is usually acceptable for many purposes with little or even no
treatment. In this study the quantities of parameters recorded were within WHO's standard. It was not high
enough to cause health hazard but as industrial and vehicular traffic grows in number and age, the quantity of gas
pollutants that will be released in the near future will probably be high enough to make a difference.
To solve the problem of little or no portable drinking water in urban and rural communities, the chemicals,
physical and biological study of available water sources is of paramount importance, for establishing campaign
and creating the awareness of possible cautions to minimize environmental pollution that could contaminate
sources of water to reduce the incidence of water borne diseases.
5. References
[1] Abdul, H. A., Ahmed, A. A., and Shaqlof, H. Assessment the quality and quantity of
harvested rainfall from different catchments systems study case: North West of Libya. Thirteenth
International Water Technology Conference, IWTC 13, 2009, Hurghada, Egypt.
[2] Akharayi, F.C, Adebolu, I. T and Abiagom. M. C, A Comparison of Rain Water in Ondo
State, Nigeria t0 FME Approved Drinking Water Quality Standard. Journal of Microbiology
2007, Vol 2 No 11 pp.807 – 815.
[3] American Public Health Association (APHA) Standard Methods for the Examination of
Water and Wastewater 19th edition. American Water Works Association. 1998, (AWWA), Washington
DC.
[4] Bhatia, E. (2009) Water Pollution in Chemical Industries. John Welsh Publications, India.Pp. 658
[5] Chukwuma, E, C., Nzediegwu, C, Umeghalu, E.C and Ogbu, K.N, Quality Assessment of Direct
Harvested Rainwater in parts of Anambra State, Nigeria. Nigerian Association of Hydrological
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Sciences, Publication. 2012.
[6] Coombes P J. "Energy and economic impacts of rainwater tanks on the operation of
regional water systems". Australian Journal of Water Resources. 2007. vol. 11, No2, pp 177–191.
[7] Dami, A., Ayuba, H. K, and Amukali, O..(2012) Effects of Gas Flaring and Oil Spillage on
Rainwater Collected for Drinking in Okpai and Beneku, Delta State, Nigeria. Global Journal of
Human Social Science (B) Volume 12 Issue 13 Version I Year 2012
[8] Dinrifo, R. R, Babatunde, S.O.E, Bankole, Y. O, and Demu, Q. A. Physico- Chemical
Properties of Rain Water Collected from Some Industrial Areas of Lagos State Nigeria. European
Journal of Scientific Research, EuroJournals Publishing, Inc, 2010, Pp.383-390.
[9] Egereonu, E. E, Physicochemical Assessment of Rainwater from Two Rain Gauge Stations in the
Rain Forest Region, Anambra State, Nigeria,. J. Chem Soc. Nigeria, 2006, Vol. 30 No. 1 & 2, Pp. 43
– 48.
[10] Bernard E, and Nurudeen A., (2012) Physicochemical Analysis of Groundwater Samples of Bichi
Local Government Area of Kano State of Nigeria: World Environment, Scientific and Academic
Publishing 2(6): 116-119
[11] Ezenwaji, E., Okoye, A. C and Otti, V. I.,(2013) Effects of gas flaring on rainwater quality in
Bayelsa State, Eastern Niger-Delta region, Nigeria. Journal of Toxicology and Environmental
Health Sciences. Vol. 5(6), pp. 97-105, June 2013
[12] Gardner, T., Ahmed, W and Toze, S. Microbiological Quality of Roof-Harvested
Rainwater and Health Risks: A Review. J. Environ. Qual. 2011, 40:1–9
[13] Igwo-Ezikpe, M.N and Awodele, O. (2010) Investigation of Some Physico-chemical and
Microbiological Parameters in Rainwater Collected from Industrial Areas of Lagos
State, Nigeria. AJPSP 1: 26-38.
[14] Olobaniyi, S. B and Efe, S. I. Comparative Assessment of Rainwater and Groundwater
Quality in Oil Producing Area of Nigeria: Environmental and Health Implications. Chatered
Institute of Environment Health. 2007, Vol 6 No. 2.
[15] Olowoyo, D. N. Physicochemical characteristics of rainwater quality of Warri axis of
Delta state in western Niger Delta region of Nigeria Journal of Environmental Chemistry and
Ecotoxicology 2011, Vol. 3 No.12, pp. 320-322.
[16] Rainwater Harvesting Implementation Network (RAIN) Guidelines and practical tools on
rainwater qualityNetherlands. 2008. www.rainfoundation.org.
[17] Ubuoh E. A., Akhionbare, W.N., Akhionbare, S.M.O. and Ikhile, C.I. Evaluation of the
Spatial Variation of Rainwater Quality in Parts of Akwa Ibom State of Nigeria Using
Chloropleth Map. American Based Research Journal. 2012, Vol 1:2
[18] United Nation Committee on Economic, Cultural and Social Rights. General Comment
No. 15. The right to water (articles 11 and 12 of the International Covenant no Economic, Social and
Cultural Rights). Twenty-ninth Session Geneva E C. 2002.
[19] World Health Organization: Drinking Water Standards and Health Advisories Office of
Water U.S. Environmental Protection Agency Washington, DC. 2011.
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EFFECT OF ADVANCE ORGANIZERS IN THE TEACHING OF CHEMISTRY IN SECONDARY SCHOOLS: ANAMBRA
STATE AS A CASE STUDY
Enekwechi E.E.
08135339971 E-mail: emeldakwechis@gmail.com
Department of Natural Sciences,
Federal Polytechnic, Oko.
Abstract
This study examined the effect of advance organizers on secondary school students' academic achievement
in chemistry and their scientific attitude. It used the pre-test post-test control group quasi-experimental
design with eighty-four senior secondary 2 (SS2) chemistry students as subjects selected from two sampled
schools. Chemistry Achievement Test (CAT) and Scientific Attitude Questionnaire (SAQ) were the
instruments used for collection of data. The reliability of the CAT was finally determined using Spearman-
Brown Prophesy Formula which yielded a coefficient of 0.82 while the Cronbach Alpha reliability coefficient
for SAQ was 0.91. Analysis of covariance (ANCOVA) was used as the statistical technique for the data
analysis. The hypotheses were tested at 0.05 level of significance. The findings indicated that: students
taught chemistry with advance organizers achieved better and had a higher level of scientific attitude than
their counterparts taught with the conventional method. The study therefore recommended that chemistry
teachers should adopt the use of advance organizers in order to improve students' achievement and
scientific attitudes.
Key Words: Advance Organizers, Chemistry, Instructional Strategy, Achievement, Scientific Attitudes.
Introduction
It is very necessary that developing countries should grape every opportunity provided by science and
technology for alround development. United States of America is one of the developed nations that boast
of so many scientific inventions hence it is one of the nations rated as the world power. A nation without
science and technology (chemistry inclusive) is regarded as undeveloped and backward.
Knowledge in chemistry plays a vital role in most aspects of human life. It's applications in pharmaceutical
industries has resulted in high level of good health. This has made a fundamental contribution towards
control of a wide range of diseases (Ezekannagha, 2013). Knowledge in chemistry has also been applied in
agriculture which made a tremendous contribution towards meeting the demand of food requirements for
the ever growing human population. Other areas where skills and knowledge in chemistry has been applied
include transportation, cosmetics and beverages.
Federal government of Nigeria has realized these facts and some efforts are been made which aim at
boosting the quality of science and technology education. For instance, secondary school chemistry
curriculum has gone through series of reforms in order to meet up with some of the country's challenges.
The chemistry curriculum ensures continuity, and flow of theme, reflects depth, appropriateness and
interrelatedness of the curricula contents. Emerging issues like value orientation, peace and dialogue, HIV
and AIDS education, entrepreneurial skills etc were also incorporated into the relevant contents of the new
curriculum. Activities for both teachers and learners, teaching and learning materials, evaluation guide are
also provided in the curriculum (FME 2007).
In a nutshell, as Nigeria desires to be identified with contemporary development worldwide, it then
becomes imperative to organize the contents of the chemistry curriculum around four themes of
349
·The chemical world
·Chemistry and environment
·Chemistry and industry
·Chemistry and life (FME 2007)
The chemistry curriculum is packaged with contents that lead to self-actualization by students, focuses on
practical activities with emphasis on locally available materials. This will enable learners to develop the
spirit of enquiry as well as achieve maximum potential in the subject of chemistry and its various
applications. This implies that whether students proceed to tertiary institutions or not, the learning of
chemistry ought to inculcate in them the scientific attitude which will be beneficial to them throughout life.
Some attitudes like open-mindedness, logical thinking and objectivity will enable the students react more
intelligently to the challenges they may come across in their life time. Moreover, scientific attitude is not
limited to those in chemistry but is needed by everybody for decision-making in everyday endeavour. Other
scientific attitudes include patience, curiosity, suspended-judgment, rationality, skepticism, honesty.
However, inspite of all these reforms, there seen not to be much improvement in students' performance in
chemistry. Several researchers including (Okeke, 2008; Orasi, 2007 & Okoli, 2011) have reported low
academic achievement and low enrollment of senior secondary school students in chemistry. The root of
this problem may be attributed to poor instructional approach. While examining the instructional strategy
factor, Oka (2008) pointed out that the type of instructional plan used by teachers determine how effective
learning can be, hence good teaching makes learning more meaningful. He is of the view that poor teaching
would lead to poor learning and poor performance. This demands a re-examination of instructional
strategies used in teaching senior secondary school students chemistry. There is need for chemistry
teachers to shift from standardized teacher-directed instruction to a more active learning environment
where students can participate actively and make use of their creative minds rather than remain passive.
Teaching approaches in which learners are actively involved would likely lead to meaningful learning and
not rote learning. Some researchers have observed that most chemistry teaching in Nigeria senior
secondary schools is done by the talk-and-chalk method (Oku, 2008; and Okeke, 2008). This method of
teaching encourages rote learning and regurgitation of facts (Okoli, 2011). Therefore, there is a need to take
a closely look at the ways chemistry could be taught in secondary schools in order to improve performance.
Advance organizer is one of such instructional strategies that encourages activity learning (Shihusa, 2009).
An advance organizer is information presented prior to learning and that can be used by the learner to
organize and interpret new incoming information. It is a kind of cognitive bridge which teachers can use to
help learners make a link between what they know and what is to be learnt. It can equally be referred to as
a short arrangement of material introduced to the learner before the lesson.
Ausubel supports the use of advance organizer as a mechanism to help link new learning material with
existing related ideas. As advance organizer is not a summary of a previous lesson rather it provides a
structure for students' thinking. Advance organizers are therefore framework that enable students learn
new ideas/information and meaningfully link these ideas to the existing cognitive structure. They can be in
form of short stories, questioning, reading material, demonstrations etc.
Ausubel's theory of advance organizers fall into two major categories namely comparative and expository
organizers. Comparative organizers are used when the material resembles and differs from that which is
350
already known (Curzon, 1990). This enable the students recognize that the topic they are beginning to learn
is not completely new, but rather can be related to something they are already comfortable with. Expository
organizers are used when the new learning material is unfamiliar to the learner. They emphasize context
and link the essence of the new material with some relevant previously acquired concepts.
Advance organizers also include metaphor, analogy, model as well as concept maps. Graphic organizers
provide a visual holistic representation of facts and concepts and their relationship within an organized
frame. They can exist in many forms like flow charts, story map, venn diagrams, sequence chain, tables,
matrix. Graphic organizers may be effectively used before instructional activities like reading or viewing a
film to activate prior knowledge in order to create/provide a conceptual framework for integrating new
material and encourage student prediction (Shihusa, 2009). During instruction, advance organizers enable
students to actively process and re-organize information. Novak (1980) asserts that creative teaching, when
done well, includes the selection and use of good advance organizers.
Curzon, (1990) points out that Ausubel's own research suggests that the use of advance organizers can
enhance the relationship between cognitive structure and new material, thereby facilitate teaching and
learning. Nwogbo, (2006) in his study demonstrated the effectiveness of using advance organizers in the
teaching of physics in secondary schools. The use of advance organizer is a highly effective instructional
strategy for all subject areas where the objective is to achieve meaningful assimilation of concepts.
Mayer, (1979) suggests that the most effective advance organizers are those that:
· Allow the students to generate all or most of the logical relationships in the material to be learnt
· Point out relationship between familiar and less familiar material
· Are relatively simple to learn, and
· Are used in situations in which the learner would not spontaneously use them
A study by Willerman, (1992) investigated the effect of advance organizers in students' conceptualization of
pollution in biology. The study used concept mapping as an advance organizer and revealed that students
are not passive listeners in the learning process. In the present study, teachers of the treatment groups
were trained by the researcher on how to use the advance organizer teaching strategy to ensure that they
successfully incorporated it in their instructional process.
Statement of the Problem

The implementations of every reform in secondary school chemistry curriculum starts in classroom where
teaching and learning takes place. Effective teaching and learning of science in secondary schools in Nigeria
has become a problem and needs urgent attention. This issue has been observed to be affecting students'
performance adversely especially in external examinations. It was equally found out that the poor academic
performance of students in chemistry is associated with the use of conventional lecture method in teaching
the students. This study therefore investigated the effect of advance organizer on senior secondary school
students' academic achievement in chemistry and scientific attitudes.
Purpose of the Study

The purpose of this study was to ascertain the effect of advance organizer on students' academic
achievement in chemistry and scientific attitude. Specifically, the study attempted to find out the
351
effectiveness of using advance organizers on students' achievement in chemistry as compared to the
conventional lecture method.
Research Questions
The following research questions guided the study:
1. What is the mean achievement scores of chemistry students taught with advance organizers and
those taught with conventional approach?
2. What is the effect of advance organizers and conventional teaching approach on students' scientific
attitude?
Hypotheses
The following null hypotheses were tested at 0.05 level of significance
1. There is no significant difference in the means achievement scores
2. There is no significant effect of treatment on students' scientific attitude scores
Methodology
Research Design
This study adopted a pre-test post-test quasi-experimental design. The experimental group had instructions
through advance organizers and the control group was taught through the conventional teaching method.
The choice of this design lies on its ability to identify cause and effect relationship to treatment. The cause
and effect which this study intends to establish is between the two teaching strategies and
achievement/scientific attitude.
The design was equally chosen because the unit of sampling a class was already constituted and therefore,
it was unethical to re-constitute one randomly. The design involved a random assignment of intact classes.
Sample and Sampling Technique

The sample for the study consisted of eight-four (84) senior secondary school chemistry students drawn
from two schools in Awka South Local Government Area of Anambra State, Nigeria.
Instrumentation
The instruments used for data collection are:
(a) Chemistry Achievement Test (CAT)
(b) Scientific Attitude Questionnaire (SAQ)
Chemistry Achievement Test (CAT)

The researcher designed this instrument to measure the SS2 students' achievement in chemistry. The CAT is
made up of twenty multiple-choice items based on periodic table and chemical reactions. These items
measured students' ability to identify, recall facts, interpret and apply such facts in real life situations.
The instrument was validated by three experts and their comments and suggestions were incorporated into
the final draft of the instrument. The test was administered to twenty SS2 chemistry students in a
secondary school not involved in the study sample. Test-retest of the instrument was done within an
interval of two weeks to establish the reliability using Pearson correlation which yielded a reliability
coefficient value of 0.82.
352
Scientific Attitude Questionnaire (SAQ)
The researcher designed this instrument (SAQ) in order to measure the chemistry students' scientific
attitude. The instrument consist of sixteen (16) items based on the following types of scientific attitude –
honesty, humility, objectivity, rationality, persistence, curiosity, open-mindedness and suspended judgment.
Cronbach's Alpha was used to determine the reliability of the instrument and a reliability coefficient of 0.91
was obtained.
The Construction and Use of Instructional Materials

The researcher conducted a training programme for the classroom teachers that were involved in the study.
The programme was an interactive session that lasted for three days. It aimed at acquainting the teachers
with the advance organizers teaching strategy before the experimental period. A pre-test was administered
to the groups then followed by the experimental period of four weeks. A post-test was administered to the
groups at the end of the treatment period.
The advance organizers adopted for the study were charts, concept mapping and handout. The students in
the experimental group were provided with all these advance organizers prior to classroom instruction.
They were expected to study these materials and give an explanation on what is involved in the periodic
table and chemical reactions. Groups and Periods, blocks elements, different families, s-block, p=bock, d-
block, f-block, atomic radii and ionic radii shown in the chart and concept mapping.
In the classroom instruction, the learners were give time to say what they have understood concerning the
materials given to them. Learners were expected to explain the message presented by the chart and the
concept mapping. During the instructional process the learners were engaged actively in a discussion in an
effort to interpret the chart and the concept mapping. The handout contained information on collision
theory and types of chemical reactions. In this case also, students were required to read the text and come
up with explanations concerning them.
Data Analysis
The research questions were answered using mean and standard deviations. The hypotheses were analyzed
using Analysis of Covariance (ANCOVA) to determine the experimental effects with pre-test scores as
covariates.
Analysis of Covariance (ANCOVA) was used to cater for the initial differences among the groups.
The results are presented in line with the two research questions and two hypotheses that guided the
study.
Research Question One

What is the means achievement scores of chemistry students taught with advance organizers and those
taught with conventional method? The research question one was answered using achievement mean
scores and standard deviation scores. The results are presented in Table 1.
Table 1: Students' achievement mean scores and standard deviation scores
353
GROUP PRE-TEST SCORES POST-TEST SCORES
N X SD X SD Means gain score
Experimental 43 8.28 4.21 13.09 4.30 4.81
Control 41 6.41 2.60 8.39 3.36 1.98
The above table revealed that the achievement means scores of both groups improved in their performance
after the treatment, however, the means gain score for the experimental group was 4.81 while that of the
control group had mean gain of 1.98 which is considerably low if compared with the experimental group.
The result therefore showed that the experimental group achieved higher than the control group.
Research Hypotheses One

There is no significant difference in the mean achievement scores in periodic table and chemical reaction of
students taught using advance organizers and those taught with conventional teaching approach. The
results for testing hypotheses one are displayed in Table 2.
Table 2: Summary of analysis of covariance results for post chemistry achievement test (achievement of
mean scores with reference to method by gender). Test of Between-Subject Effects
Dependent Variable: Post-Test

Source Type III Sum of Df Mean F Sig. Partial Eta
Squares Square Squared
Corrected model 78.955 4 18.381 2.052 .073 .068
Intercept 20.714 1 20.714 2.000 .103 .020
METHOD 20.201 1 20.201 2.012 .103 .020
Pre-test 15.453 1 15.453 1.721 .153 .015
Treatment 43.301 1 43.301 3.628 .031 .043
Error 878.450 101 8.563
Total 18820.000 106
Corrected Total 1011.634 105
a. R Squared = .068 (Adjusted R Squared = .030)
Table 2 above showed F-cal value of 3.63 for treatment which is significant at 0.031. This 0.031 value is less
than 0.05 significant level set for this study. Thus, the null hypotheses is rejected. This means that there is a
significant difference between the achievement means scores of students taught Period Table and chemical
reaction using advance organizers and those taught using conventional teaching method.
354
Research Question Two
What is the effect of advance organizers and conventional methods on student's scientific attitude? The
research question two is answered using mean scores and standard deviation. The result is presented in
Table 3.
Table 3: Extent of scientific attitude of students taught chemistry using advance organizers and conventional
teaching strategy
GROUP PRE-TEST POST-TEST MEAN GAIN

SCORE
N Mean SD Mean SD
Experimental 43 10.32 2.43 14.67 3.18 4.35
Control 41 8.88 1.59 12.13 2.66 3.25
Table 3 showed mean scores of 10.32 and 8.88 for experimental group and control group respectively for
pre-test in scientific attitude, and 14.67 and 12.13 for the post-test. The mean gain score for experimental
group is 4.35 while that of the control group is 3.25. This shows that experimental group had a higher gain
score than the control group.
Research Hypotheses Two

H2: There is no significant effect of treatment on students' scientific attitude scores.
Pre attitude
20
Post attitude
n
Me a
15
10
0
Advance organizer Conventional
Teaching Strategy
Fig 1: Comparison of pre-tet and post-test attitude scores according to treatment groups.
The above Fig 1 shows that the advance organizer teaching strategy improved students' scientific attitude
more than the conventional teaching strategy. Therefore, chemistry students taught with advance organizers
had a better scientific attitude than those taught with conventional method.
Table 4: Summary of analysis of covariance results for post-test attitude score
Dependent Variable: New post attitude
355
Source Type III Sum of Df Mean F Sig. Partial Eta
Squares Square Squared
Corrected model 80.501 3 20.132 20.42 .80 .072
Intercept 22.304 1 22.304 2.192 .108 .022
Pre-test 17.510 1 17.510 2.201 .121 .022
Treatment 22.342 1 22.342 3.640 .038 .015
Error 983.518 103 8.983
Total 18635.000 106
Corrected Total 1063.766 107
a. R Squared = .072 (Adjusted R Squared = .040)
From Table 4 above, the F-cal value of 3.64 for treatment is significant at .038 which is less than 0.05 level
set for this study. This means that the null hypotheses is rejected which shows that, there is significant
effect of treatment on students' scientific attitude scores in chemistry.
Discussion
Table 1 showed a significant main effect of treatment on students' achievement in chemistry. Table 2 also
revealed that the advance organizers facilitated learning more than the conventional teaching approach.
This agrees with the assertion that the conventional teaching strategy is unproductive (Okeke, 2008) and
ineffective in developing conceptual understanding of subject matter. The use of advance organizers in this
study engaged learners' interest to be active cognitively. Students in the experimental group performed
significantly better in the chemistry achievement test than their counterpart in the control group. This
supports the argument of Orasi (2007) that learners' active participation during instruction aids
understanding and higher cognitive achievement. This implies that science activities tend to arouse and
sustain learners' interest, make learning real and facilitate the understanding of chemistry concepts.
In the treatment group, where there were adequate instructional chemistry activities, the students had
significantly higher post-scientific attitude mean score. This finding is applaud being that the students were
exposed to these activities for only a short period. The students in the control group were not exposed to
the advance organizers but to 'chalk-and-talk' method of teaching. This method is commonly used in
schools for science teaching, (Okeke, 2008), which does not expose students to scientific process like
manipulation of instruments, measurement, formulation of hypothesis, among others. These activities tend
to promote students' scientific attitude.
Recommendation and Conclusion

The findings indicated that advance organizers teaching strategy is better than the conventional teaching
method. The use of activity-based teaching strategy like advance organizer, make students to be more
active, hence meaningful learning is achieved. This creates an environment for the inculcation of scientific
356
attitude. Thus advance organizers enhance students' achievement in chemistry compared to conventional
teaching approach. It is hereby recommended that chemistry teachers should adopt advance organizer
teaching strategy to enhance achievement in chemistry and scientific attitude.
References
Ausuble, D.P. (1960). The Use of Advance Organizers in Learning and Retention of Meaningful Material.
Journal of Education Psychology 51, 267-272.
Curzon, L.B. (1990). Teaching in Further Education: An Outline of Principles and Practice, London. Cassel
Education Publishers Ltd.
Ezekannagha, G.N. (2013). Basic Chemistry Concepts and Applications for Polytechnics. Adson
Educational Publishers, Onitsha.
Federal Ministry of Education, (2007). National Curriculum for Senior Secondary School, Chemistry for
SS1-3, Abuja.
Mayer, R.E. (1979). Twenty years of Research on Advance Organizers: Assimilation Theory is still the best
predictor of results. Instructional Science 8(2), 133.
Nwogb, E. (2006): Using Advance Organizers in the Teaching of Physics in Secondary Schools in Oyo
State, Nigeria. Unpunished Matters Thesis, University of Ibadan, Ibadan.
Okeke, U.J. (2008). Effect of Cooperative Learning on Students' Interest and Achievement towards
Chemistry. Unpublished Ph.D Thesis, University of Nigeria, Nsukka.
Okoli, B.C. (2011). Preliminary Development and Validation of Academic Input and Output Instructions for
Senior Secondary Students in Chemistry. Unpublished Ph.D Thesis, Enugu State University of
Technology.
Orasi, P.O. (2007). Effects of Heuristic Problem Solving Strategy on Senior Secondary School Students'
Learning Outcomes in Mathematics in Enugu State, Nigeria. Unpublished Ph.D Dissertation, University of
Ado-Ekiti, Nigeria.
Oku, E.A. (2008). Science Teaching in Nigeria Secondary Schools. Nuel Centi Publisher and Academic
Press Ltd. Awka.
Shihusa, H. (2009). Using Advance Organizer to Enhance Students' Motivation in Learning Biology.
Retrieved 18, July 2015 from http://www.Eurasialearning.org.
Willierman, J. (2009). The Concept Map as an Advance Organizer. Journal of Research in

Science Teaching, 28, 705-712.
357
REMOVAL OF CONTAMINANTS FROM INDUSTRIAL WASTEWATER USING PHOTOCATALYST
EKWERE, IFIOK OKON
Chemistry Department, Akwa Ibom State University, Ikot Akpaden
ifiokekwere@hotmail.com
08061168633
ABSTRACT
Innovative processes for treating polluted industrial wastewater involve technologies for reduction of toxicity in
order to meet technology-based treatment standards. In mixed wastewater stream, the presence of metals ions
can retard the destruction of organic contaminants and also, the efficiency of recovery of the metal is reduced
by the presence of organic species. Thus, special considerations must be made for the effective treatment of
such waste water. This paper is a review article that focuses on waste water treatment using photocatalysts.
Photocatalysts have the advantage of simultaneous removal of organic and inorganic pollutants through
oxidation and reduction mechanisms respectively. The photocalysts of interest in this paper is titanium dioxide,
though it could be doped with other compounds like hydrogen peroxide, zinc oxide etc. This paper will also
review the two main photocatalytic reactor design namely the fixed bed or slurry bed reactor design; and also
their application to achieve the aim of the article.
Keywords: adsorption, photocatalysis, heavy metals, wastewater, pollutants, titanium dioxide,

photocatalytic reactors.
BACKGROUND
The treatment of water contaminated with traces heavy metals of toxic organic compounds is a common
problem throughout the world. In most cases, industrial waste effluents contain a variety of pollutants which
can be characterized as organic or inorganic in nature. These pollutants pose serious hazardous effect on human
health, thus the need for their removal before effluent are discharged into the environment. In addition to
inorganic pollutants, photocatalysts are also effective in the removal of organic pollutants in contaminated
water.
Photocatalytic process can be used to achieve rapid and efficient destruction of environmental pollutants. The
mechanism is as follows: upon illumination of semiconductor electrolyte interface with light energy greater
- +
than the semiconductor band gap, electron-hole pairs (e /h ) are formed in the conduction and valence band of
the semiconductor, respectively. These charges which migrate to the semiconductor surface are capable of
reducing or oxidizing species in solution having suitable redox potential. Various semiconductors have been
used: TiO2, ZnO, CeO2, CdS, ZnS, etc. According to Mills et al; 1993, of all the semiconductor photocatalyst tested,
TiO2 appears to be the most active; therefore TiO2 will be the focus in this work. The dual functions of
photocatalyst in elimination of pollutants in waste water are: mineralization of organic waste and reduction of
inorganic waste like heavy metals. The removal of organic waste through the mineralization process can be
summarized in the following equations:
TiO2 + hv eCB- + hVB+
hVB+ + R intermediates CO2 + H2O
. -
H2O + hVB OH + H
.
OH + R intermediates CO2 + H2O
- .-
eCB + O2 O2
358
Removal of inorganic pollutants, like metal ions, occurs through the adsorption process. Adsorption of metal ion
to the surface of TiO2 is possible because the surface of TiO2 is electron rich enabling the oxide to attract the
cationic metal ion
M+ + e- M(s)
This paper aims to highlight the use of TiO2 in the removal of heavy metals and other organic pollutants from
their solutions, and it is significant because it supports the cleanup of hazardous waste from the environment.
Photocatalytic reactors
Photocatalytic reactors can be classified based on the deployed state of the photocatalyst, i.e., suspended or
attached. Photocatalytic reactors can use either UV or solar radiation. Solar photocatalytic reactors have been
of great interest for the photoxidation of organic contaminants in water. Such kind of reactors can be divided
into concentrating or nonconcentrating reactors. Both the reactor types extend certain advantages and
disadvantages. For example, non-concentrating reactors have negligible optical losses and therefore can use
direct and diffuse sun irradiation but are larger in size compared with the concentrating reactors and have high
frictional pressure losses. However, the use of solar radiated photoreactors is limited due to the intrinsic nature
of the TiO2 particles. Details of the two main type of reactor considered in this paper are given below;
Slurry reactors
The slurry reactors show the largest photocatalytic activity, compared with the immobilized photocatalyst and
provide a high total surface area of photocatalyst per unit volume, which is one of the most important factors
configuring a photocatalytic reactor. However, these reactors require separation of the sub-micron TiO2
particles from the treated water which complicates the treatment process. Several techniques were proposed
to achieve post-treatment separation such as the use of settling tanks (overnight particle settling) or external
cross-flow filtration system. However the use of filtrations systems increases the cost of the treatment process.
· Immobilized TiO2 reactors
Photocatalytic reactors with immobilized TiO2 are those in which catalyst is fixed to support via physical surface
forces or chemical bonds. These reactors extend the benefit of not requiring catalyst recovery and permit the
continuous use of the photocatalyst.
However, these reactors possess drawbacks such as low surface area to volume ratios, catalyst fouling and
significant pressure drop.
LITERATURE REVIEW
Also, some work though limited have been done on the treatment of waste water with TiO2. Samarghandi, et al;
2006, worked on the removal of phenol, lead and cadmium by means of UV/TiO2/H2O2 processes. Results
indicated that the optimum efficiencies of phenol and Cd removal were 76% and 97.7% at ph =11, respectively,
and for lead, it was 98.8% in all ph. The photocatalytic reactor design used was the slurry bed reactor design.
Javad & Shahrezaei, 2012, recorded over 78% reduction of COD in petroleum refinery wastewater using
nanotitania photocatalyst and UV light. In this work, slurry bed reactor was used. In a presentation by Endang, et
al; 2011, the removal of toxic metals in wastewater by photocatalytic method was reported using the slurry bed
reactor.
Some other researches on the degradation of pollutants using immobilized TiO2 and a fixed bed reactor have
been carried out by different scientists. Examples include Abhang, et al; 2011; Shivaraju, (2011)
CONCLUSION
The need for the elimination of pollutants from wastewater cannot be over emphasized; thus the relevance of
this paper. Previous researches on environmental cleanup in Nigeria has focused mainly on the biosorption
technique in the removal of pollutants, however, this technique has the disadvantage of being ineffective on
359
organic pollutants. Photocatalytic degradation, on the other hand, is capable of eliminating both organic and
inorganic pollutants in the environment; thus it is more practical in environmental cleanup process. Thus, there
is need for adequate research to be carried out in this field to determine the best parameters and conditions
that will give optimum experimental results. Such parameters include ph, temperature, catalyst concentration,
contact time, exposure to sunlight, elution time, etc.
REFERENCES
i. I. Oboh, E. Aluyor & T. Audu (2009),Biosorption of Heavy metals ions from aqueous solution
using a biomaterial, Leonardo Journal of Sciences, (4), 58 – 65
ii. D.O. Olukanni, J.C. Agunwamba & E.I. Ugwu (2014), Biosorption of heavy metals in industrial
wastewater using mico-organisms (Pseudonomonas aeruginosa), American Journal Of
Science and Industrial Research, 5(2):81 – 87
iii. H.P. Shivaraju (2011), Removal of Organic Pollutants in Muncipal Sewage Water by TiO2 based
Heterogenous Photocatalyst. International Journal of Environmental Sciences, 1(5): 976 –
4402
iv. T.W. Endang, H.A. Nural, H. Hatimah, & M.W. Afifah (2011), Removal of Toxic Materials in
rd
Wastewater. Presentation on the 3 Conference on Environmental Pollution and Human
Health working Group (EPHHWG) in London, 21st – 25th Nov. 2011.
v. R.M. Abhang, D. Kumar & S.V. Taalkar (2011), Design of Photocatalytic Reactor for Degradation
of Phenol in Wastewater, International Journal of Chemical Eng. & Appl. 2(5)
vi. M.A. Barakat (2011), New Trends in Removing Heavy Metals from Industrial Wastewater,
Arabian Journal of Chemistry, 4: 361 - 377
vii. M. R. Samarghandi, J. Nouri, A. R. Mesdaghinia, A. H. Mahvi, S. Nasseri, F. Vaezi (2007)
Effective Removal of phenol, lead and cadmium by means of UV/TiO2/H2O2 processes,
International Journal of Environmental Science Tech., 4 (1): 19 - 25
viii. S. Javad & F. Shahrezaei (2012), Organic Pollutants Removal from Petroleum
Refinery Wastewater with Nano titania Photocatalyst and UV Light Emission,
International Journal of Photoenergy 703074
360
Levels, spatial distribution and ecological risk assessment of Cadmium and Lead in surface soil of Nsukka
Industrial Cluster Areas, South-East Nigeria
N.R.EKERE *, B.U. NGANG, J.N IHDIOHA, P.O.UKOHA

Department of Pure & Industrial Chemistry, University of Nigeria, Nsukka.
Corresponding author e-mail: nwachukwu.ekere@unn.edu.ng. nwachukwuekere64@gmail.com;
Abstract
Surface soil samples collected from four Industrial work sites in Nsukka Urban Industrial cluster areas, South-
East Nigeria were investigated for levels, spatial distribution and ecological risk of Cadmium (Cd) and Lead
(Pb). A total of 200 samples were analysed using Flame Atomic Absorption Spectrophotometer (AAS) following
wet digestion using 65% Nitric acid. The results revealed Cadmium and lead concentration levels in surface soil
from the area studied are unevenly distributed spatially showing a range of 1.855-2.798 mg/kg for Cd in
Automobile Work.sites (AM), 0.068 – 0.084 mg/kg in Motor Parts Fabrilation sites (MP), 0.000 – 0.020 mg/kg in
wood Work Sites (WW) and 0.218 – 0.259 mg/kg in Aluminium work sites (AW). The respective mean
concentrations for Pb were 1.913 ? 1.410, 1.384 ? 1.114, 0.086 ? 0.074 and 1.020 ? 0.099 mg/kg. Apart from
WW samples, Cd levels in all the samples were above the maximum allowable unit of 0.07mg/kg set by World
Health Organisation (WHO). Pb levels exceeded the maximum allowable unit of 0.05mg/kg in all the samples
suggestive of increased Pb content in soil due to anthropogenic interactions. The RI of automobile worksite
(AM) showed high risk while motor parts fabrication worksite (MP) had moderate ecological risk. The
contribution of Cd to the risk potential was much.
Key Words: Industry, surface soil, pollution, Cadmium, lead, ecological risk, worksite, Nsukka.
INTRODUCTION
There is several rising global concern on the impact of toxic of heavy metals in the environment [1 - 3]. In 2010,
an estimated 400 children were reported to have died in Zamfara State, Northern Nigeria due to lead poisoning
from mining activities and many surviving children tested high levels of lead in their blood [4]. Another report in
2015 mentioned at least 65 cases of lead poisoning identified in Niger State, Nigeria and 28 children died from
the menace [5].
From the various studies investigating the sources of toxic metals exposures and their effects on human health,
soil has been identified as a potential pathway of human exposure to these metals especially Cd and Pb [6, 7, 8].
Apart from direct dermal contact with high levels of toxic metals in soils, food incorporated metals via plant
uptake is also a major potential threat to human health.
Routine analyses of soils to determine Cd and Pb in populated areas is vital since elevated levels of these toxic
metals are primarily found in urban areas due to industrial and human activities. This study establishes base-line
data on the concentration of cadmium and lead from industrial areas of Nsukka. The associated ecological risks
of these metals were also investigated.
Experimental
Area of study
This study was carried out in Nsukka Industrial Areas of Enugu State, South-East Nigeria. The area is
characterized by tropical humid climate and distinct dry and wet(rainy) seasons; usually beginning from
November to March and April to October respectively. The geographical co-ordinates of Nsukka are 6052'N and
0
7 23'E.
361
Sampling Technique
A total of 200 surface soil samples at 0 - 5.0cm depth were collected from 20 sample stations. 10 samples were
collected from each station worksite based on hexagonal grids mapped out in the area of about 1km2. Based on
land use, the industrial site was divided into four zones. In each sampling site, four subsamples were collected
2
within an area of 50m and mixed thoroughly to obtain a representative/composite sample. The collected samples
were dried at 60 oC, sieved through a 2.0mm Nylon sieve to remove plant debris, stones, gravels, metal scraps and
stored in polythene bags prior to analysis.
Digestion of Samples
One gram of each sample was put into 150 mL conical flask, a mixture of concentrated HN03: HClO4: HF in the
ratio 3:1:3 was added according to literature [9]. Distilled water was added to make up the mark. Blank solutions
were prepared and used to zero the instrument used for sample determination and also carried through the
digestive process. The concentrations of Cd and Pb were determined using Atomic Absorption
Spectrophotometer (ARIAN model AA4). Each sample was determined in triplicates.
Quality Control
The precision and accuracy of the method was tested by spiking the soil samples with standard materials and
determining recoveries. The % recovery was determined using the equation.
S- Y
% R =´ 100
Z
Where S = concentration of metal in spiked sample
Y = concentration of metal in unspiked sample
Z = spiking concentration in mg/L
Potential Ecological Risk Index
The degree of ecological risk of heavy metals in soils is estimated using the Risk index (RI)
proposed by Hakanson [10]. RI is calculated by the equations:
C Di ------------------------ (1)
C if =
C Ri
E ri =
Tr ´ C if ------------------------ (2)
n
i
RI =
å
E
l=
i
r ------------------------ (3)
Where C if = contamination coefficient

i
CD = concentration of heavy metal in soils.
i
CR = background concentration of heavy metal in soil.
Tr = toxic-response factor for a given heavy metal.
E ri = potential risk of individual heavy metal
RI = sum of potential risk of individual heavy metal
i
Using Hakanson approach, the Tr for Cd and Pb are 30 and 5 respectively [9]. The C Ri used in this
calculation is the world average elemental concentration value of Cd = 0.30 mg/kg and Pb = 20mg/kg
because Nigeria has no reliable background values for these metals [10].
Hakanson defined five categories of C ri and four categories of RI.
A recovery range of 99.8 – 101.1% for Cd and 98.3 – 99.9% for Pb obtained in the analysis gave an indication that
the analytical technique would sufficiently estimate the metals in the matrix.
The results of the metallic concentrations in the surface soils expressed as range and mean plus standard
deviation (Mean S.D) are presented in Table 1.
362
Table 1: Result of Cd and Pb analysis in the surface soil samples (mg/kg)
Sample Cd Pb Mean ?
S.D
Range Mean ?
S.D Range
Automobile
Worksite (AM) 1.855-2.798 2.269 ?
0.933 0.00-3.361 1.913 ?
1.410
Motro parts
fabrication sites 0.068-0.084 0.077 ?
0.006 0.176-2.864 1.384 ?
1.114
(MP)
Wood work sites 0.00-0.020 0.006 ?
0.009 0.004-0.178 0.086 ?
0.074
(MP)
Aluminum work 0.218-0.259 0.231 ?
0.019 0.901-1.145 1.020 ?
0099
site (AW)
Blank < 0.001 < 0.001 < 0.001 <0.001
WHO
Acceptable Limit 0.07 0.05
LOD
LOQ 0.001 0.001
0.003 0.003
LOD = Limit of detection ; LOQ = Limit of quantification

Cadmium and lead concentration levels in surface soil from the area studied are unevenly distributed
spatially. In AM, the mean value of Cd is higher than the World Health Organisation (WHO) accepted limit
of 0.07 mg/kg Cd in soil [12]. The high level of Cd can be attributed to the wide use of Cd – Ni batteries to
power tools in spite of the ban in most countries . In MP, the concentration of Cd ranged from 0.068 to 0.084
mg/kg with a mean value of 0.077 0.006 mg/kg, slightly above WHO acceptable limit. WW showed a Cd
concentration range of 0.00 to 0.020 mg/kg with a mean value of 0.006 0.009 mg/kg. This was the least
mean concentration of Cd in all the industrial work sites studied. AW had a mean Cd content of 0.231
0.019 kg/mg and ranged from 0.218 to 0.259 mg/kg all higher than the recommended WHO limit of Cd
concentration in soils [11]. The sequence of mean Cd concentration with respect to the industrial work sites
is as follows: AM>AW>MP> WW. A Cd concentration of 0.5 mg/kg in soil is said to reflect human
influence [11]. Cadmium adsorption and distribution in soils is principally influenced by competition from
other metals, hydrous metal oxides content, presence of organic and inorganic ligands, soluble organic matter
content, clay content and pH [13].
As shown in Table 2 the mean concentrations of Pb in AM, MP, WW and AW were 1.913 1.410, 1.384
1.114, 0.086 0.074, 1.020 0.099 respectively. The values were above WHO acceptable limit of lead in
soil (0.05) mg/kg) [13]. The trend of mean concentrations of Pb in the industrial worksites is viz: AM > MP
> AW > WW.
The result is comparable to those from studies of road side soils in Jos metropolis [14] and soils of Nasarawa
metropolis [15]. The use of leaded fuels in automobile engines, lubricants, etc. may account for high value of Pb
observed in AM. Several studies have recorded high lead concentrations in soil dust due to vehicular emissions,
lubricating oil/fuel spillage or some other factors due to industrial activities [13,16, 17].
Both Cd and Pb are categorized as class – 1 elemental impurity and thus require high consideration during risk
assessment across all potential sources of elemental contaminants [18]. Ecological Risk Assessment
The calculated value of risk index of the metals contamination is shown in Table 2.
Table 2: Potential ecological risk of Heavy metals index of soil samples.
363
Sample Element C
i
f
C
i
n
RI Grade of
Station soil
Auto Cd 7.56 226.0 226.50 High risk
mobile Pb 0.10 (High Risk)
(AM) 0.50
(Low Risk)
Motoparts Cd 2.57 77.10

Fabrication (moderate risk) 77.45 Moderate
(MP) Pb 0.07 0.35 Risk
(low risk)
Wood Cd 0.20 6.00

work site (Low Risk)
(WW) Pb 0.004 0.02 6.02 Low risk
(low risk)
Cd 0.77 23.1 23.125 Low risk

Aluminum (low risk)
worksite Pb 0.05 0.25
(AW) (low risk)
The RI of automobile worksite (AM) showed high risk while motor parts fabrication worksite (MP) had moderate
ecological risk. The contribution of Cd to the risk potential was much and would be of concern contributing up to
99.8% to total RI. The other two sites had low potential ecological risk. Due to high toxic response factor of Cd,
the contamination of the surface soil by Cd ought to be given urgent remediation. The result of this work is
consistent with the finding of several earlier works which reported high ecological risks mainly due to high Cd
load in the soils [18 - 20].
CONCLUSION
The concentration values obtained for both Cd and Pb exceeded the maximum permissible limits recommended
by World Health Organization most of sites studied. This is of concern since inhalation of Cd and Pb results in the
accumulation of the toxic metals in body organs with chronic effects and probable carcinogenic effects as well as
suppression of reproduction.
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364
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365
LOW COST/WASTE CATALYST FOR FATTY ACID METHYL ESTER PRODUCTION
a,* b b
M. O. Ekeoma , P. A. C. Okoye , V.I.E., Ajiwe
a
Department of Chemistry, College of Physical and Applied Sciences, Michael Okpara
University of Agriculture, Umudike, P.M.B. 7267, Umuahia, Abia State, Nigeria
b
Faculty of Physical Sciences, Department of Pure and Industrial Chemistry, Nnamdi Azikiwe
University, Awka, Anambra State, Nigeria
*Corresponding author. Tel: 284 8030766890;

E-mail address: ekeoma.moses@mouau.edu.ng; mosesoekeoma@yahoo.com
(M.O.Ekeoma)
Abstract
Non-edible crude Karanj (Pongamia pinnata) oil (CKO) with high free fatty acid (FFA) content was used
as effective renewable feedstock for fatty acid methyl ester (FAME) production. Calcium feldspar clay, a rare
compositional variety of plagioclase clay, a low cost, abundant Earth resource, containing over 90 % CaO and
belonging to the class of anorthite clay was used as heterogeneous catalyst in direct conversion of high FFA crude
karanj oil to fatty acid methyl esters. The efficiency of the catalyst was made possible by the structural
rearrangement of the mixed metal oxides' content of the catalyst at prolonged high temperatures, a behaviour
characteristic of glass transitions and properties of amorphous phases of plagioclase feldspar, and thus were
transformed into solid acid particles such as acidic mesoporous aluminum silicate mixed oxides. Optimum
FAME yield of 98.97 % was obtained at 4 h reaction time, 6 wt% catalyst loading, 9:1 methanol to CKO molar
ratio and at methanol reflux temperature. More than 90 % catalyst was recovered after four runs which still
o
exhibited strong activity after further calcination at 500 C for 1 h after each run. From the result of
transesterification, non-edible, high FFA crude CKO is a potential renewable feedstock for the production of
FAME using thermally treated, low cost, anorthite clay-derived catalyst.
Keywords: Anorthite clay; Fatty acid methyl ester; Crude karanj oil; Transesterification.
1. Introduction
Heterogeneous acid catalysts such as zirconium oxide (ZrO2), zinc oxide (KF/ZnO) [1, 2],
titanium oxide (TiO2), tin oxide (SnO2), zeolite oxides [2, 3], sulfonic ion-exchange resin,
sulfonic modified mesostructure silica, sulfonated carbon-based catalyst, heteropolyacids
(HPAs); as well as heterogeneous alkaline catalysts such as calcium oxide (CaO), potassium
phosphate oxide (K3PO4) and mixed oxides (Ky(MgCa)2xO3 [4-7]; have been found invaluable
in biodiesel production as a result of many challenges presented by homogeneous acid and base
catalysts. For instance, heterogeneous catalysts are non-corrosive and environmentally benign,
presenting fewer disposal problems, with easier separation and recovery of products, catalysts
and the co-product. Thus, solid base catalysis are economically and ecologically important
fields in catalysis and the replacement of liquid bases with heterogeneous catalysts is becoming
more and more important in the chemical industry [8, 9].
The need for biodiesel production at competitive pump price with petro-diesel has prompted the recent
366
search for biodiesel production from cheap or low cost resources for both feedstock and catalyst materials. Clay
constitutes about 60 % of the Earth's crust and therefore a very abundant natural resource. Varieties of clay
minerals abound with varying chemical compositions and have been applied as catalysts for esterification and
transesterification. Few attempts in the application of clay-based catalysts for biodiesel production were made
by Manut Jaimasith and Satit Phiyanalimat who impregnated Suratthanee black clay (SB) and Ranong kaolin
(RK) with CaO and concluded that at short reaction time, calcium oxide impregnated on Renong kaolin had
better catalysis performance in the transesterification of palm oil over that of Suratthanee black clay [10]. Also,
Olutoye and Hameed (2013), obtained 96 % FAME yield from waste cooking oil using iron-rich local clay
catalyst characteristic of montmorillonite [11, 12]. There is no record so far, however, of the application of clay-
based catalyst for direct conversion of high FFA crude karanj oil to fatty acid methyl ester. This is the reason why
we have selected the Ca-rich natural clay-based catalyst, calcium feldspar clay, for direct conversion of high FFA
to produce FAME.
Moreover, since it is estimated that 70- 85 % of the cost of biodiesel is determined by the cost of
feedstock alone; and from literature, current commercial production of biodiesel depend mainly on edible oils
such as soybean oil [13] , rapeseed oil [14], sunflower oil [15], safflower oil [16], jatropha oil [14], canola oil,
palm oil , kernel oil [17], beef tallow [18] and fish oil [19]; the competition for edible oils for biodiesel production
has made oil prices incredibly high. The result is the high cost of both food and biodiesel. Hence, current research
is focusing mainly on non-edible sources of feedstock such as used cooking oils, yellow greases, brown greases
and fats, Oils and grease (FOG), recovered from sewer pipes and waste water treatment pipes [20].
A single-step esterification of High FFA crude karanj (Pongamia pinnata) oil (CKO) over the reusable
SBA-16 supported molybdophosphoric acid catalyst for the synthesis of fatty acid methyl ester (FAME) reported
that the success of the direct conversion was attributed to the Bronsted acidity, Keggin type heteropolyacids
(HPAs) [21].
It is therefore reported in this contribution, the use of non-edible, high free fatty acid crude karanj
(Pongamia pinnata) oil (CKO) as a potent renewable feedstock in direct conversion to fatty acid methyl ester
(FAME), using a novel, low cost, naturally abundant, Ca-rich clay-based heterogeneous catalyst (anorthite clay
mineral).
Materials and method

2.1 Materials
Calcium-rich clay (Calcium Feldspar clay) was obtained from Xuzhou Sushang Chemical Technology
in Shanghai, China. The clay was in powdered form and was used as supplied without further modifications.
CKO was sourced from Telaga Madu Resources Sdn. Bhd., Malaysia. Methanol (99.9%) and GC grade n-hexane
of A.R. grade were purchased from Merck, Germany. Methyl heptadecanoate used for GC internal standard,
99.5% came from Sigma-Aldrich, Malaysia. The CKO was merely filtered to separate suspended matter while
the rest of the chemicals were used without further treatment.
2.2. Catalyst preparation and characterization

Known quantities of clay (10-12 g) were charged into ceramic crucibles and placed into a muffler
furnace for calcination at temperatures ranging from 500- 900 oC for 4 h at 4 oC/min ramping temperature.
Calcined catalysts were stored in sample bottles, labeled according to their calcination temperatures such as
o o
CaF-600, CaF-700, etc, for calcium feldspar clay calcined at 600 C, 700 C, etc while CaF-Raw stands for the
raw catalyst and all were placed in a desiccator.
Preliminary runs of transesterification reactions showed the catalyst calcined at 800 oC as having the
367
best activity, all characterizations were based on this catalyst to save cost. The first step was to determine the
strength of acid and basic sites of CaF-800 after calcination using temperature programmed desorption (TPD)
technique in a Micromeritics AutoChem 2920 II equipment. A quatz reactor was loaded with 50 mg of catalyst
and dried over He gas flowing at 400 oC for 1 h which was followed by a dry He sweep for 1 h. On cooling to 100
o
C, the sample was exposed to NH3/He gas mixture (90 vol. % He) for 1 h to make room for NH3 to be adsorbed by
sample. The sample was degassed of NH3 by flushing with pure He to allow for accurate detection of desorbed
NH3. The measurement of NH3 in sample was done by simply heating the sample to 850 oC at a heating rate of 10
o
C/min over He gas while the desorbed NH3 molecules were detected accordingly. The same process was
repeated using CO2 gas to determine the Basic strength. In comparison, surface acidity and basicity tests were
carried out for CaF-800 using acid base titration technique with NaOH and HCl in accordance with procedures
recorded in literature, and results obtained were correlated with results from temperature programmed
desorption technique.
The developed catalyst (CaF-800) and the raw clay catalyst (CaF-Raw) were subjected to textural
characteristics to determine the surface area and pore volume using Brunnauer Emmett Teller (BET) method
over Micromeritics ASAP 2020 surface area and porosity analyzer (Micromeritics Instruments Corporation,
USA) at 77 K using liquid nitrogen. The sample was thoroughly degassed at 200 oC before measurement began.
Powder X-ray diffraction (XRD) patterns were acquired on Philips PW 1710 X-ray powder
diffractometer over 10 ≤ 2è ≤ 90 using Cu Ká radiation (ë=1.5406 ) at 40 kV and 40 mA at a scan rate of 0.02
o o o
o
/min.
Scanning electron microscopy (SEM) was performed on the catalysts to determine the morphology and
to analyze the porous structure of catalysts using a Zeiss Supra TM 35 VP scanning electron microscopy (Zeiss,
Jena, Germany) coupled with FEI as a source of electrons accelerated at 300 kV. Energy-dispersive X-ray (EDX)
spectroscopy was performed simultaneously with SEM to determine the catalyst surface composition.
2.3 Catalytic reaction

The catalytic activities of catalysts were tested by their application in the conversion of high free fatty
acid (FFA) crude karanj oil (CKO) with methanol to fatty acid methyl ester (FAME). A set of four glass reactors,
comprising 250 ml round-bottomed flasks equipped with spiral glass condensers placed on heating mantles with
magnetic stirrer and thermostat was used for the transesterification reaction. In each run using crude karanj oil
(CKO), 36.81 g of methanol and 3.4 g of calcined catalyst (CaF-800) were firstly mixed to homogeneity and
refluxed at 65 oC for 0.5 h as the catalyst activation step. Then, 113.19 g of crude karanj oil was added to
transesterify between 1-7 h reaction time. The weight ratio among methanol: catalyst: CKO was 11:1:33. At the
end of reaction time, the reaction was allowed to cool to room temperature to allow for complete separation of
glycerol and ester layers. The product was decanted to obtain the FAME layer, centrifuged at 3000 rpm for 10
mins and sent for Gas Chromatographic (GC) analysis. Over 90 % of the clay catalyst material could be
recovered after each run, as some of it dissolved in the reaction process with methanol. The recovered catalyst
o o o
was washed severally with warm water, dried at 80 C for 6 h and re-calcined at 500 C for 1 h at 4 C/min ramping
temperature in a muffle furnace. The calcined catalyst was re-used 4 reaction cycles with a minimum yield of
88.67 %.
3. Results and Discussion

3.1 Catalyst characterization
Fig. 1 shows the scanning electron micrographs of both raw (CaF-Raw) and calcined (CaF-800) calcium
feldspar catalysts, at 3.00 k x magnifications (bars at lower left equal to 2µm). Amorphous, prismatic, massively
disordered, wood-like structure was observed for raw calcium feldspar (CaF-Raw), while more spherical
368
Fig. 1: SEM images of (a) CaF-Raw and (b) CaF-800 at 3.00 k x magnification.
From elemental compositions (Fig.2), verified by using energy dispersive X-ray (EDX) detector, calcined
calcium feldspar catalyst (CaF-800) was observed to contain over 80 - 90% by mass of calcium atoms with over
90 % of the calcium in the form of calcium oxide.
Element Wt% At%
OK 11.60 24.74
CaK 88.40 75.26
Fig. 2: EDX result of calcined catalyst, CaF-800.

Further investigation by chemical composition analysis revealed the presence of other metal
oxides Table 1. Such metal oxides include Al2O3 (1.1070 wt %), MgO (2.3025 wt%) and SiO2 (3.5204
wt%), among others. The presence of the metal oxides in calcined catalyst suggested increased active
sites for catalyst activity. The metal oxides, under prolonged calcination temperature, transformed into
mixed metal oxides similar to mesoporous aluminum silicate mixed oxides, expected to produce the
overall acidic character needed for direct conversion of high free fatty acid- crude karanj oil to produce
fatty acid methyl ester [23].
369
Table 1: Results of SFP (BULK) Semi-quantitative Analysis of CaF-800.
Compoundds MgO Al2O3 SiO2 P2O5 SO3 Cl K2O CaO TiO2 Cr2O3 MnO
% by weight 2.30 1.11 3.52 0.008 0.028 0.011 0.35 91.911 0,035 0.029 0.015
The above attribute was corroborated by the work of Lin and co-workers. which opined that, whereas many
currently known base catalysts were destroyed by the presence of high free fatty acids, the kiln dust catalyst
system could conveniently convert, even animal fats and various restaurant oils directly to biodiesel, when in the
presence of an alcohol simply by adding an acid, such as a solid acidic particle , for example, an acidic mesoporous
aluminum silicate mixed oxide . Acidic mesoporous aluminum mixed oxides can be used as a solid acid to
sequester alkaline materials in a reaction mixture. Clay catalysts could also act as molecular sieve used as drying
agents to help facilitate the esterification or transesterification reactions, in addition to helping to preserve the
recyclability of the catalysts [22].
Table 2: BET results of CaF-Raw and CaF- 800.

CaF-Raw CaF-800
2
BET surface area, m /g 1.162 9.386
Pore volume, cm3/g 0.0023 0.0445
Average pore size, Å 78.00 189.66
BET results of raw (CaF-Raw) and calcined (CaF-800) catalysts are presented in Table 2. A primary requirement
of an ideal solid catalyst for biodiesel synthesis is large interconnected pores that would minimize diffusion
limitations of molecules with long alkyl chains [24, 25]. A typical triglyceride molecule has a diameter of
approximately 58 Å. It follows that the poor catalyst performance of CaF-Raw could be attributed to its low
2 3
textural characteristics with surface area of 1.1624 m /g and pore volume of 0.0023 cm /g as against the
improved textural characteristics observed in calcined clay catalyst (CaF-800) with surface area increasing by
over 13% on thermal treatment and the pore size increasing by over 42% thereby justifying improved catalyst
performance of CaF-800 over CaF-Raw.
Fig. 3 is the powder X-ray diffraction plots of CaF-800 and CaF-Raw catalysts showing the major phase
compositions. The major crystalline phases in the catalysts include Calcium oxide (CaO), calcite (CaCO3),
Hatrurite (Ca3SiO5), aragonite (CaCO3), Calcium Aluminium oxide (CaAl2O4) and Calcite Magnesium
(Mg0.03Ca0.94)(CO3), etc. Though clay is a complex material and the main peaks of XRD patterns are usually
overlaid to one another, CaF-800 and CaF-Raw existed as hexagonal crystals CaCO3 located around the region
2è = 29 o- 48.5o. However, a change in crystal phase was observed at elevated pretreatment temperatures as
o
catalyst was transformed to more active calcite, syn. CaCO3. The intense peak observed at 2è =29.5 was
attributed to CaCO3 present in both CaF-Raw and CaF-800. The presence of the CaCO3 peak in the catalyst after
calcination, though with a major drop in intensity, is attributed to incomplete evolving of molecules of CO2 and
organic matter by thermal treatment after initial calcination at 800 oC. This was in good agreement with the DTA
o
findings by Sharma and co-workers on the weight loss observed in eggshell re-calcined at 700 C, which was
o
attributed to remaining CaCO3 decomposition and loss of organic impurities after initial calcination at 900 C.
While adsorbed CO2 and water molecules were evolved at about 250 oC to 650 oC, further treatment at higher
temperature served to transform oxygen anions and metal cations into more active centers.
370
Fig. 3: XRD patterns of CaF-Raw (3a) and CaF-800 (3b).
Four transform-infra-red (FT-IR) spectroscopy, as an effective tool for a semi-quantitative estimation of
structural information in complex solids is used to investigate the functional groups in the catalysts. The result of
-1
FTIR of CaF-Raw and CaF-800 are presented in Fig. 4. The distinctive broad band at 3444.87 cm observed in
both CaF-Raw and CaF-800 were attributed to the O-H stretching, indicative of the high amounts of water
adsorbed on the catalyst surface. While that of CaF-Raw appeared at 34.45 %T, the peak in CaF-800 appeared at
17.74 %T suggesting that much of the physisorbed water molecules were evolved by calcination and the O-H
stretching in the calcined material may be attributed to chemisorbed water molecules. The bands at 1415 cm-1 and
1608 cm-1 were attributed to carbonate species while the bands at 1114 cm-1, 1053 cm-1 and 883 cm-1 are
-1 -1 -1
characteristic of TO4 tetrahedral groups (T= Ca, Al, Si) and are similar to 875 cm , 806 cm and 702 cm in the
-1
raw catalyst. Also the band at 1114 cm is associated with T-O bond asymmetric stretching vibrations (this bond
provides information on the degree of crystalinity of sample, while the one centered at 883 cm-1 was associated
with T-O bond internal deformation vibration.
371
Fig.4: FTIR spectra of CaF-Raw (a) and CaF-800 (b).
The FTIR result was in good agreement with XRD patterns presenting with crystal monoclinic Hatrurite
(Ca3SiO5), triclinic calcium silicate (Ca3SiO5) and monoclinic calcium aluminum oxide (CaAl2O4) containing the
elements, Ca, Al, Si and O characteristic of anorthite clay with molecular formula CaAl2Si2O8. The bands of T-O
-1 -1 -1 -1
bond at 702 cm , 808 cm and 875 cm present in the raw catalyst were weaker bonds and shifted to 883 cm , 943
cm-1 and 1067 cm-1 in the calcined catalyst respectively and are attributed to the bending vibration modes
indicating that new bonds were indeed formed by the catalyst pretreatment at higher temperature resulting in
structural rearrangements to give more active species. The stretching vibrations due to –OH groups of water
-1 -1
molecules adsorbed on the catalyst surface could be partly responsible for the bands at 1608 cm , 3444 cm and
3633 cm-1. Nevertheless, the band at 3633 cm-1 could be partly attributed to the out - of - plane stretching
vibrations and phase symmetric stretching of T-O-T which were considered to be the active sites of the basic
species.
3.2 Effect of calcination temperature

From the results of transesterification of CKO using calcium feldspar catalysts calcined at various
temperatures (Fig.5), it was observed that raw clay catalyst (CaF-Raw) gave very low conversion of triglycerides
to FAME while thermally treated clay catalysts showed improved performance with increase in calcination
temperature with optimum FAME yield of 98.97% recorded at 800 oC. The low catalyst activity observed for the
catalysts treated at temperatures below 800 oC were due to the saturation of the catalyst pores with CO2, water and
other volatile matters which occupied available active sites meant to be centers of reaction and only partially
removed at lower calcination temperatures.
Fig. 5: Effect of calcination temperature on catalyst activity (methanol/oil mole ratio 9:1; catalyst loading/oil
weight ratio 6 wt%; reaction time 4 h; reaction temperature 65 oC and stirring speed 700 rpm).
Catalytic activities at various temperatures of calcination are represented by the graphs above as
o o o o
follows: a = raw sample, b = calcination at 500 C, c = 600 C, d = 700 C, e = 800 C and f = 900
o
C.
372
To obtain high activity of alkaline earth oxide, it is essential to remove adsorbed molecules such as
carbon dioxide, adsorbed organic matter and water. The catalyst activities of metals depend mainly on the
o
pretreatment temperature and time. When pretreatment temperature was low (below 600 C), the catalyst showed
as low as 11% yield. The activity increased with increase in calcination temperature, optimizing at 800 oC and
declined with higher pretreatment temperatures.
This calcination effect, coupled with the rearrangement of the surface structure of the mixed oxides in
anorthite clay at prolonged pretreatment temperatures for 4 h, resulted in the formation of sufficient acid centers
needed for the transesterification of high FFA crude karanj oil (CKO) with methanol. From TGA-DTA result (not
shown), the depression at 250 oC indicate the evolving of adsorbed CO2 and H2O molecules, while the prolonged
o
thermal treatment of catalyst to 800 C for 4 h was necessary to generate stronger basic and acid sites. As
pretreatment temperature increased, desorption of adsorbed molecules such as carbon dioxide occurred and
oxygen anions became available for the reactants. Desorption of adsorbed molecules started from weaker basic
sites and more severe pretreatment was required to generate stronger basic sites. At the same time, pretreatment
at higher temperature caused the rearrangement of the surface structure of catalyst to more active phases. These
two factors induced complex dependence of the catalytic activities with pretreatment temperature.
3.3 Tests of activities of catalysts using CKO
The catalytic activities of both the raw and calcined calcium feldspar-derived catalysts, (CaF-Raw and
CaF-800), were tested in the transesterification of high FFA crude karanj oil (CKO) for the production of fatty
acid methyl ester (FAME). Since the main objective of this work was the production of fatty acid methyl ester at
o
reduced cost, the reaction temperature was kept constant at methanol reflux temperature of 65 C while no
chemical modification of the catalyst was considered. Varying the reaction time from 0.5 -7 h; but under similar
reaction conditions of 6 % (w/w) catalyst loading and methanol/ oil mole ratio of 9:1, it was generally observed
that the FAME yield using the raw catalyst was far below 10 % while those of thermally pretreated catalysts
o o
improved with increase in calcination temperatures from 500 - 800 C (Fig. 5). The catalyst calcined at 900 C
gave a lower FAME yield than that at 800 oC. This trend agreed with the work by Boro and co-workers [26] . The
poor catalyst performance below 700 oC was attributed to the inhibition of the catalyst active centers by adsorbed
molecules like CO2, H2O, SO2, PO4 and other volatile matter present in the raw catalyst (CaF-Raw). The
improved activity observed with increasing calcination temperatures was attributed to the gradual but steady
evolving of the adsorbed molecules with steady increase of the active centers.
3.3.1 Effect of methanol/oil mole ratio

Investigating the effect of methanol to oil mole ratio on FAME yield, 6 wt% of catalyst loading was used
at 4 h reaction time and at a stirring rate of 700 rpm while methanol /oil mole ratio was varied from 6:1, 9:1, 12:1,
15:1 and 18:1. Although stoichiometrically, transesterification of 1M of oil required 3M of alcohol, excess of
alcohol improved conversion.
From the result of transesterification in Fig.6, it could be observed that FAME yield increased initially
with increasing methanol to oil ratio to a maximum at 9:1 which gave a FAME yield of 98.97% after 4 h. Further
increase from 12:1 - 15:1 gave no significant increase in FAME yield. Hence, for a 6 wt% CaF-800 catalyst
loading, 9:1 oil to methanol mole ratio was sufficient for good yield. A higher ratio would therefore be wasteful.
Also, poor phase separation between the fatty acid methyl ester and glycerol by-product was observed when
higher methanol to oil mole ratio was used.
373
Fig. 6: Effect of methanol/oil mole ratio on catalyst activity (catalyst loading/oil weight ratio – 6 wt%; reaction
temperature 65 oC; reaction time 4 h and stirring speed 700 rpm).
3.3.2 Effect of catalyst loading

The effect of catalyst loading was tested for the transesterification of CKO using 9:1 methanol/oil mole
ratio and 4 h reaction time at 700 rpm stirring rate while catalyst loading was varied from 3 - 9 % by wt. kg-1 oil.
A close observation of Fig.7 revealed an increase in FAME yield to a maximum of 98.97% obtained for 6
wt% after 4 h reaction time. Further increase in catalyst loading beyond 6 % by wt. showed no appreciable
increase in FAME yield.
The observed low FAME yield below 6 % by wt. catalyst loading could be attributed to insufficient
availability of catalyst active surface needed to drive the reaction for efficient conversion. Above 6 % by wt., the
reactants formed a viscous emulsion which limited the mass transfer of components to the catalyst active surface
causing saturation and diffusion effects which hindered interaction. This effect caused the observed decline in
FAME yield beyond 6 % by wt.. Therefore, further increase of catalyst beyond 6 % by wt. would amount to waste
of valuable resource.
374
Fig. 7: Effect of catalyst loading on FAME yield of crude karanj oil (methanol/oil mole ratio 9:1; reaction time
o
4 h; reaction temperature 65 C and stirring speed 700 rpm).
3.3.3 Effect of reaction time

To evaluate the effect of reaction time on FAME yield, other operational parameters such as catalyst
loading (6 % by wt.), methanol to oil mole ratio (9:1), methanol reflux temperature were kept constant while
duration of reactions were varied from 0.5 - 7 h. The result of the investigation is graphically presented in Fig. 8.
o
Fig. 8: Effect of reaction time on catalyst activity (methanol/oil molar ratio 9:1; reaction temperature 65 C;
catalyst loading/oil weight ratio 6 wt% and stirring speed 700 rpm).
It could be observed from the graph that the highest FAME yield was recorded after 4 h reaction time for
CaF-800 with a yield of 98.97%. The FAME yield could scarcely hit 50 % after 2 h reaction time. However, a
steady increase in conversion of triglycerides was observed until an optimum of 98.97% after 4 h duration. No
further significant increase in FAME yield was observed with prolonged reaction time. Hence, under the present
reaction conditions, prolonged reaction beyond 4 h reaction time would mean a waste of energy.
4. Conclusions
Non-edible, crude karanj (Pongamia pinnata) oil (CKO) with high FFA has been identified as potential
renewable, low cost feedstock for the production of FAME. Whereas many currently known base heterogeneous
catalysts were destroyed by high free fatty acids content in feedstock, the calcium feldspar catalyst system has
been shown here to excellently convert high FFA CKO directly to fatty acid methyl ester (FAME). This was
attributed to the conversion of some metal oxide content of the clay catalyst to acidic mesoporous aluminium
silicate mixed oxide centers by prolonged calcination. The clay material also functions as mesoporous sieve to
help facilitate conversion. The best FAME yield (? 98.97 %) was obtained at methanol reflux reaction
temperature, 6 % by wt. catalyst loading, methanol to oil mole ratio of 9:1 after 4 h reaction time.
Acknowledgement
The authors acknowledge the research grant provided by the University of Science Malaysia, under
Research University (RU) grant (Project No: 814126).
375
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377
COORDINATION CHEMISTRY IN OIL INDUSTRY
BY
*EHIRIM, Appolinus I. and OGWUEGBU, Martin O.C.;
Department of Chemistry, Federal University of Technology, Owerri.
* Corresponding author: 07030561903
ABSTRACT
In oil industry, achieving an optimal production of valuable products is an extremely important goal. The
processes of mining, refining, cracking and reforming, polymerization and production of petrochemicals are
major methods employed in oil industry. Production chemistry is a discipline in the oil/gas industry that deals
with issues that occur as a result of the chemical and physical changes in the overall oil/gas production and
processing systems (including enhanced oil recovery), especially changes to the well fluids as they are
transported from the reservoir to the sale's point. This paper will discuss the role of coordination chemistry in
proffering solutions to petroleum production chemistry issues, with the aim of facilitating the realization of this
goal. The success of some of these processes is ascribed to inhibition involving the ability of the chemical
reagents to form stable coordination complexes with metal ions present in the crude oil; or those in the reagents
introduced in the production and processing system.
INTRODUCTION
Coordination Chemistry refers to the chemistry of compounds formed by metals (Lewis acids) bonded to
inorganic ions or molecules, or organic molecules, or their ions (Lewis bases) through donation of lone pair of
electrons (Ogwuegbu & Chileshe, 2000). These coordination compounds or complexes, as they are often called,
may be ionic or neutral, containing central metal ions/atoms closely surrounded by electron donor groups called
ligands or coordinating agents (Ogwuegbu & Maseka, 1998; Huheey, Keiter & Keiter, 1993). The ligands must
have an available free electron pair on one or more of the donor atoms, while the orbital system of the central
metal atom (M), must be of the right energy and capable of accepting the electron pair from the ligand to form a
n+
coordinate covalent bond between itself and the ligand (L). A complex metal ion of the type, [M)H2O)6] is an
example of a coordination complex because M is bonded to water molecules by coordinate covalent bond. The
ligand (H2O) provides the electron pairs that are shared between it and the central metal atom (M). The term,
3+ + 2- 2-
complex, is used not only for the common ions such as [Co(H2O)6] , [Ag(NH3)2] , [Cd(CN)4] , [PbI4] and
uncharged species like Pt(NH3)2Cl2, but also for other types of coordination compounds and species such as the
ions, BF4-, PO43-, ClO4-, etc.
Each metal atom (or ion) has a characteristic maximum number of coordinate bonds it can form directly
with the donor atoms under normal conditions. This number is called the Coordination Number (CN).
Coordination number therefore, is the maximum number of donor atoms that coordinate or bond directly to the
central metal ion (Mn+) at any point in time. Coordination numbers of 2, 4, 5, 6, 7 and 8 are the most common and
useful. Polydentate ligands are ligands that coordinate through two or more points or donor atoms at the same
time. They are mostly organic molecules or ions and could be bidentate, tridentate, quadridentate,
quinquidentate, or sexadentate, containing 2, 3, 4, 5, or 6 coordinating electron pairs (or donor atoms)
respectively. When a ligand possesses a second or more donor atoms appropriately situated with reference to the
first, it may form two or more bonds simultaneously with the metal atom/ion (Mn+). In this situation, coordination
results in the formation of a cyclic (ring) complex termed a 'chelate' (a Greek word, meaning crab's claw or crab-
like; referring to spencer-like action of the ligands on the metal ions); such a ligand is called a chelating agent and
they hold on to the metallic ion tenaciously (Anzlik, 1976). This type of action is illustrated by the reaction
2+
between Nickel (II) ion (Ni ) and ethylenediammine (en) molecules:
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NH2 NH2 2+
2+
H2C NH2 H2C CH2
Ni + 2 Ni
H2C NH2
H2C CH2
NH2 H2N
In this reaction, a coordination compound is formed in which nickel forms a 5–membered chelate ring system
with each of the ethylenediammine (en) molecules.
Other examples of polydentate ligands are; acetylacetone, 2-2-dipyridine (dipy),1-10-phenanthroline (o-
phen), ethylenediamminetetraacetic acid (EDTA), porphine, crown ethers (e.g., 18-crown-6),
diethylenetriammine (dien), etc.
Several factors affect the formation of a stable metal coordination complex. They include: (i) the chelate
and ring size effect, (ii) the nature of the ligand, (iii) the nature of metal ion, and (iv) the steric effect (Ogwuegbu
& Chileshe, 2000).
The areas of influence of production chemistry start from the reservoir, to wells, flowlines, processing
facilities, and sale's point. On a project life cycle perspective, it enters the project during early design phase and
stays until the field reaches the end of its life and the production system is decommissioned.
In general, production chemistry issues include the following types:
· Problems with solid precipitation and deposition – asphaltenes, scales, wax, gas hydrates,
naphthenates, bio-growths, etc.
· Problems with facility processing - foaming, emulsion, water quality (including injection water quality
in case of waterflood), hydrogen sulphide, etc.
· Problems with material integrity - material corrosion from production fluids, microbial activities,
artificially introduced fluids (completion fluids, chemicals, etc.)
· Problems with Health, Safety and Environment (HSE) - water discharge, toxicity, radioactive scales,
H2S, etc.
P r o d u c t i o n C h e m i s t r y S o l u t i o n s
The solutions to production chemistry issues can be mechanical, chemical, or even physical.
Mechanical Examples are
insulation, pigging, used corrosion resistant alloys, proper separator sizing, etc. The solutions to the
mechanical issues could be handled elsewhere, while this paper discussed the solutions to the chemical issues via
coordination chemistry.
Chemical Scale Inhibitors,
asphaltene inhibitors, paraffin inhibitors, demulsifiers, defoamers, water clarifiers, H2S
scavengers, corrosion inhibitors, hydrate inhibitors, biocides, etc are reagents applied in the oil
industry to enhance efficiency and optimum recovery.
SCALE INHIBITORS
The use of scale inhibitors is very important in oil industry. The term Scale inhibitor is used to refer to a
chemical that stops or interferes with inorganic scale nucleation, precipitation and adherence to production
conduit, completion system or processing facilities; the three key elements susceptible to scale problems. Scale
Inhibitor performance is impacted by the pH, temperature, calcium and magnesium ion levels, and the presence
of other chemicals (e.g., corrosion inhibitor) in the scaling brine mixtures. Its threshold level for a defined level
of inhibition is called minimum inhibitor concentration (MIC), which is typically 0.5 to 20 ppm (Sorbie & Laing,
2004). However, the MIC for high temperature high pressure (HTHP) fields that have severe scale problems can
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be on the order of a few hundred ppm (Fan et al., 2011).
In oilfield, scale inhibitors are typically continuously injected via capillary or subsea umbilical tubes to the tree
or down hole depending on the location and severity of the scaling risks, or periodic scale squeeze treatments
may be required to place the inhibitor in the reservoir matrix if the scaling occurs in the near wellbore region or at
the completion. Scale inhibitor chemistries used in the oil industry generally fall into three main types:
a. Inorganic Phosphates:
These compounds are both cheap and easily prepared. They are readily soluble in water, non-toxic and
effective at low treatment concentrations (typically 0.5 to 20 ppm), particularly in controlling calcium
carbonate scale. However, their oilfield applications are limited because, even at fairly modest temperatures,
they hydrolyze in water to form orthophosphates that have little scale inhibitor activity.
b. The use of Organophosphorous Compounds
Organophosphorous compounds are degradable organic compounds containing carbon–phosphorus bonds
(thus excluding phosphate and phosphite esters, which lack this kind of bonding). Therefore, they could be:
(i) Organic Phosphate Esters: Phosphate esters are more tolerant of acid conditions than polyphosphates.
Stable to temperatures of 150°F-160°F (65-71°C), they can withstand temperatures of 180°F (82°C)- 200°F
(93°C) for a few hours.
Within these temperature limitations, phosphate esters are generally very effective calcium carbonate (CaCO3)
and calcium sulphate (CaSO4) inhibitors. Except in acid environments (pH < 5.5), they also provide excellent
control of strontium sulphate (SrSO4) and barium sulphate (BaSO4) precipitation. In general, phosphate esters
are soluble in and compatible with high-calcium brines.
(ii) Phosphonates: Several different types of phosphonates are used as scale inhibitors. Each has different
characteristics of thermal stability, calcium tolerance and efficiency relative to scale type. A broad range of
characteristics are available because phosphonate scale inhibitors are supplied in acid form or with any portion of
the acidity neutralized by ammonia, amines or alkaline hydroxides. They are commonly deployed as scale
inhibitors for calcium, strontium and barium scales. They are mainly polydentate ligands that can form chelate
complexes with the metal ions in question, thus inhibiting their precipitation and effects.
Structures of some typical phosphonate scale inhibitors to treat carbonate and sulphate scales are shown
below. Most of these reagents are organic ligands that form metal complexes with available metal ions.
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c. Organic Polymers: Organic
polymers are chiefly crystal distorters although they also reduce precipitation in typical oilfield brines. By
modifying or distorting crystal shapes, organic polymers (primarily low molecular weight polyacrylics)
prevent scales from growing and adhering to equipment surfaces. Polycarboxylic acids are commonly used
in oilfield applications. Effective polymers tend to have a low molecular weight (typically 1000-5000) and
have regularly spaced ionizable groups. These compounds have excellent thermal and hydrolytic stabilities.
The most common classes of inhibitors include polyacrylates, polyphosphinocarboxylates, polymalates,
polyvinylsulphonates and polyacrylamides.
Stable to 204°C or higher, polymers are generally effective at very low concentrations for control of CaCO3 and
BaSO4 in waters containing low concentrations of scale-forming ions. They are also effective under acidic
conditions, particularly in the control of BaSO4. Polymers are often blended with other types of scale inhibitors to
obtain a single product with a broader range of applications.
Scale Inhibition mechanisms
There are many proposed mechanisms by which scale inhibitors operate. Generally they interfere with either
nucleation and/or with crystal growth. At the nucleation stage, threshold scale inhibitors bind with scale-
forming ions, but unlike chelants, the bound ions must be available to interact with their counter ions. This
disrupts the ion cluster at the early equilibrium stages of crystal formation, disrupting them before they reach the
critical size for nucleation. As a result, the ions dissociate, releasing the inhibitor to repeat the process. At the
growth stage, growth inhibitors slow the growth of the scale by blocking the active edges of the crystal.
Good crystal growth inhibitors have a strong affinity for the active growth sites, but should readily diffuse over
the crystal surface to other active sites as they form. Once the inhibitor has bound to the lattice, the crystal will
form much more slowly and be distorted. Often they are more rounded in shape, which makes them less likely to
adhere to surfaces and more easily be dispersed throughout the system. At the deposition stage, dispersants
prevent new crystals from coming together to form a large body of scale material. Dispersant-type inhibitors
interact with the surface and repulse other charged particles to prevent binding.
Scale inhibitor applications
Scale inhibitors should be used wherever a risk of scale damage is predicted (or known to exist from past
experience). For example, inhibitors are often incorporated into drilling muds, completion brines, and process
water used for sandwashing or desalting. Scale inhibitors have been used in injection water that is incompatible
with the formation brine present in the zones into which the water is being injected. Continuous injection of scale
inhibitors into production systems is commonly practised, and batch (squeeze) treatment of production wells is
now a routine operation.
ASPHALTENES
Asphaltenes are well known to be the most important components causing blockages in oil
pipelines and fouling in refineries which lead to enormous costs. They are also responsible for the formation
of tar mats in petroleum reservoirs, which can impede crude oil recovery (Wilhelms & Larter, 1994).
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Structure of Asphaltene molecule (wikimedia).
Asphaltene Inhibitor refers to chemical additives that help prevent asphaltene deposition when dosed at
very low concentrations into the oil (up to a few hundred parts per million by volume), unlike asphaltene
solvents that typically require much higher concentrations to work.
Although generally referred to as asphaltene inhibitors by the industry, this class of additives can actually
further divided as asphaltene inhibitors and asphaltene dispersants, depending on their mechanisms of
preventing asphaltene deposition. It has been reported that asphaltene inhibitor performance from
laboratory testing does not necessarily reflect the performance in the field (Montesi, et. al., 2011).
Mechanisms of inhibition by asphaltenes
In general asphaltene inhibitors are polymeric compounds with charged groups that are aimed to
function as artificial resins to keep the asphaltenes in solution. They provide real inhibition since they
prevent the aggregation of asphaltene molecules and therefore shift the asphaltene onset pressure. If an
asphaltene inhibitor contains long alkyl chains, it can help disperse any formed asphaltene aggregates as
well. Therefore, some asphaltene inhibitors can function as asphaltene dispersants.
A thermodynamic model has been proposed for asphaltene inhibition by treating asphaltenes as
micelles (Pan & Firoozabadi, 2000). The absorption interaction between an asphaltene and an
amphiphile molecule (natural resin or synthetic additive) is considered the most important parameter for
the stabilization of asphaltene micelles in crude oil.
DEMULSIFIERS
Demulsifiers, or emulsion breakers, are a class of special chemicals used to separate emulsions
(e.g. water in oil). They are commonly used in the processing of crude oil, which is typically produced along
with significant quantities of saline water. This water (and salt) must be removed from the crude oil prior to
refining. If the majority of the water and salt are not removed, significant corrosion problems can occur in the
refining process. Demulsifiers are typically based on the following chemistry:
·Acid catalysed phenol -formaldehyde resins
·Base catalysed phenol-formaldehyde resins
·Epoxy resins
·Polyamines
·Di-epoxides
·Polyols.
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The above are usually ethoxylated (and/or propoxylated) to provide the desired degree of water/oil solubility.
The addition of Ethylene oxide increases water solubility, while propylene oxide decreases it.
HYDROGEN SULPHIDE (H2S)
Hydrogen Sulphide (H2S) is a very toxic and pungent gas that causes problems in both the upstream and
downstream oil and gas industry. H2S in oil can be found as a component of formation gases dissolved in
water, hydrocarbons or even liquid sulphur. Thermal degradation of organic materials and sulphate reducing
bacteria (SRB) can create H2S along with other gases.
H2S is poisonous and risky to the health and safety of oil gas personnel. Besides, it causes corrosion problems to
drill strings, transport pipes, storage tanks. It also causes sulphide stress cracking, hydrogen embrittlement and
pitting corrosion in oil and gas operational structures.
The process of removing H2S is known as gas sweetening, by either iron sponge H2S scrubbers (forming iron
sulphide) or chemical scavengers. Typical H2S scavengers used in the oilfield are amine based chemicals. They
can be roughly categorized into regenerative and non-regenerative H2S scavengers.
Regenerative H2S scavengers Chemicals
In large production facilities, the most economic solution to remove H2S in the gas process stream is to install a
regenerative system for treating the sour gas. After absorbing the H2S, the chemical is then regenerated, usually
by heating and reused in the system. The separated H2S is treated by a modified Claus process to form elemental
sulphur.
Several types of amine solutions can be used as the absorbent depending on the sour gas specifications. Typical
amines are: monoethanolamine (MEA), diethanolamine (DEA), N-methyldiethanolamine (MDEA),
diisopropylamine and diglycolamine (DGA), also known as 2-(2-aminoethoxy)ethanolamine. These are ligands
and their chemical structures are as shown below:
H2NCH2CH2OH; Monoethanolamine (MEA)

HOCH2CH2NHCH2CH2OH; Diethanolamine (DEA)
HOCH2CH2N(CH3)CH2CH2OH; N-methyldiethanolamine (MDEA)
HOCH2CH2-O-CH2CH2NH2; Diglycolamine
CH3 CH
C NH C (Diisopropylamine)
CH3 CH
Amine gas treating can be utilized to remove H2S in the gas stream. Most modern amine gas sweetening processes
are MDEA-based, which typically only absorbs H2S. MEA, DEA and DGA typically absorb other acid gases as
well (i.e. CO2) besides H2S.
Non-regenerative H2S scavengers This type of
scavengers can be divided into the following categories:
(a) Triazine - alkaline and can cause carbonate scaling (b)
Solid scavengers (generally zinc or iron based materials) (c)
Oxidizing chemicals (e.g. NaClO2, NaBrO3, NaNO2, etc.) (d)
Metal carboxylates and chelates. Both water and oil soluble high valence metal chelates have been used
as H2S scavengers for treating drilling fluids and contaminated water and oil streams.
Triazine scavengers are the most commonly used (Al-Duailej, Al-Mutairi & Al-Humaidan, 2010). These
triazines are also chelating agents that could function via chelation.
383
N N N
N N
N N N
N
1,2,3- triazine 1,2,4- triazine 1,3,5- triazine
Application processes using solid scavengers.
H2S scavenging with solid scavengers, which includes a loading manway on the top and a media dump near the
bottom. The solid scavenger media is in the middle supported by a bed support system. Sour gas comes in from
the top and goes out at the bottom as sweet gas.
Non-generative H2S scavengers are typically applied via an in-line injection quail to finely disperse the liquid
chemical into the gas stream to maximize reaction.
Corrosion Inhibitors and Chemistry

A corrosion inhibitor is a chemical compound that, when added to a liquid or gas, decreases the corrosion
rates of a material, typically a metal or an alloy (Hubert et. al., 2002). The effectiveness of a corrosion inhibitor
depends on fluid composition, quantity of water, and flow regime. A common mechanism for inhibiting
corrosion involves formation of a coating, often a passivation layer, which prevents access of the corrosive
substance to the metal. Permanent treatments such as chrome-plating are not generally considered inhibitors,
however; instead corrosion inhibitors are additives to the fluids that surround the metal or related object.
Corrosion Inhibitors and Their Role

The nature of the corrosive agent depends on (i) the material being protected, which are most commonly metal
objects, and (ii) on the corrosive agent(s) to be neutralized. The corrosive agents are generally oxygen, hydrogen
sulphide, and carbon dioxide. Oxygen is generally removed by reductive inhibitors such as amines and
hydrazines (N2H4).
384
O2 + N2H4 → 2H2O + N2
In this example, hydrazine converts oxygen, a common corrosive agent, to water, which is generally benign.
Related inhibitors of oxygen corrosion are hexamine, phenylenediammine and dimethylethanolammine, and
their derivatives. Antioxidants such as sulphites and ascorbic acid are sometimes used. Some corrosion
inhibitors form a passivating coating on the surface by chemisorption. Benzotriazole is one such species used to
protect copper. For lubrication, zinc dithiophosphates are common - they deposit sulphide on surfaces.
N
Cu
N N
Benzotriazole, a ligand inhibits corrosion of copper by forming an inert layer (Cu-benzotriazole complex) of this
polymer on the metal's surface.
In the fuel industry, corrosion inhibitors are commonly added to coolants, fuels, hydraulic fluids, boiler
water, engine oil, and many other fluids used in the industry. For fuels, various corrosion inhibitors can be
used. Some components include zinc dithiophosphates (Finšgarand & Milošev, 2010). Zinc dithiophosphates
(ZDDP) are coordination complexes. A typical example is zinc dialkyldithiophosphate whose monomeric
structure is shown below.
RO S S OR
P Zn P
RO S S OR
Structure of a monomeric zinc dialkyldithiophosphate (ZDDP).
Zinc dialkyldithiophosphates (often referred to as ZDDP) are a family of coordination compounds invented by
Castrol that feature zinc bound to the anion of dithiophosphoric acid. These uncharged compounds are not salts.
They are soluble in non polar solvents, and the longer chain derivatives easily dissolve in mineral and synthetic
oils used as lubricants. In aftermarket oil additives, the percentage of ZDDP ranges approximately between 2-
15% (Barnes, Bartle & Thibo, 2001).
The alkyl groups can be branched and linear alkanes between 1-14 carbons length, 2-butyl, pentyl, hexyl, 1,3-
dimethylbutyl, heptyl, octyl, isooctyl (2-ethylhexyl), 6-methylheptyl, 1-methylpropyl, dodecylphenyl, and
others.These species are produced in two steps. First, phosphorus pentasulphide is treated with suitable alcohols
to give the dithiophosphoric acid. A wide variety of alcohols can be employed, which allows the lipophilicity of
the final zinc product to be fine-tuned.
PS5 + 4ROH 2(RO)2PS2H + H2S
The resulting dithiophosphate is then neutralized by adding zinc oxide:
2 (RO)2PS2H + ZnO → Zn[(S2P(OR)2]2 + H2O
385
In Zn[(S2P(OR)2]2, the zinc is tetrahedral. This monomeric compound also exists in equilibrium with dimers and
oligomers caused by opening of the four-membered ZnS2P ring. Partial hydrolysis gives the cluster
Zn4O[(S2P(OR)2]6, which adopts the structure seen for basic zinc acetate.
Zinc diethyldithiophosphate, Zn[(S2P(OEt)2]2, is a polymeric solid, consisting of infinite linear chains. Ball-
and stick- model of part of the chain in the crystal structure of zinc diethyldithiophosphate is as shown
below.
Applications
The main use of ZDDP is as an anti-wear additive to lubricants such as greases, gear oils, and motor oils, which
often contain less than 1% of this additive. It has been reported that zinc and phosphorus emissions which may
damage catalytic converters and standard formulations of lubricating oils for gasoline engines, now have reduced
amounts of the additive, though diesel engine oils remain at higher levels. Crankcase oils with reduced ZPPD
have been cited as causing damage to, or failure of, classic/collector car flat tappet camshafts and lifters which
undergo very high boundary layer pressures and/or shear forces at their contact faces, and in other regions such as
big-end/main bearings, and piston rings and pins. Roller camshafts are more commonly used to reduce camshaft
lobe friction in modern engines. The same ZDDP compounds serve also as corrosion inhibitors and antioxidants.
Ethylenediamminetetraacetic Acid (EDTA)

EDTA refers to the chelating agent that is widely used to sequester di- and trivalent metal ions. EDTA
features four carboxylic acid and two amine groups that can all bind to metals. EDTA forms specially strong
complexes with most metals including: Mn(II), Cu(II), Fe(III), and Co(III). The structure is as shown below.
CH2COOH CH2COOH
N CH2 CH2 N
CH2COOH CH2COOH
In Oil production, EDTA is added into the borehole to inhibit mineral precipitation coordination complex
formation with offending metal components.
Studies on the stimulation of sandstone and carbonate reservoirs using environmentally friendly
chelating agent, glutamic acid-N,N-diacetic acid (GLDA), has been reported (Mahmoud et al., 2011). The
objective of stimulation is to remove the damage caused to the production zone during the drilling or completion
process in sandstone reservoirs, and to create channels or wormholes in carbonate reservoirs. It was concluded
that GLDA showed a significant ability to chelate calcium, magnesium, iron, and aluminum ions from the
sandstone cores and in creating wormholes in calcite cores.
386
The use of polymer pendant catechol ligands in removal and recovery studies using arsenic compounds
that were characterized in Green River Formation oil shale and its pyrolysis products. The molecular
characterization of metallo-non-porphyrin compounds of vanadium and nickel in petroleums and their
asphaltenes was also addressed to clearly demonstrate the need to understand these complex mixtures and to use
this knowledge for exploration, catalyst poisoning and removal studies that will directly benefit the petroleum
industry (Annual report, 1984).
CONCLUSION
Proper understanding of the chemistry of reactions in oil and gas industry is very important in order to
minimize operational difficulties often encountered in the operation materials and processes. The success
of these processes have been attributed in part to the successful applications of the complex-forming
reagents whose chemical interactions with some materials in oil and gas industry usually lead to the
formation of stable coordination complexes, and consequently, to an optimum and sufficient production
of petroleum fractions and chemicals.
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York, pp. 64-131.
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review" Corrosion Science, 52, 2737-2749.
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Mahmoud, M. A., Nasr-El-Din, H. A., De Wolf, C. (2011). Evaluation of a New Environmentally
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Scale inhibitors Across a Wide Temperature Range", SPE International Symposium on Oilfield Scale,
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388
USING INDEX MODELS FOR HEAVY METAL POLLUTION ESTIMATION OF SEDIMENTS
FROM BOMU AND OGINIGBA RIVERS
Marcus, A. C. and Edori, O. S.
Department of Chemistry, Ignatius Ajuru University of Education, P.M.B. 5047, Rumuolumeni, Port
Harcourt, Nigeria. Email: onisogen.edori@yahoo.com
ABSTRACT
Sediments collected from Bomu and Oginigba Rivers were treated according to recommended standards and
subsequently analyzed for heavy metals and further tested with index models. The results showed that all the
metals examined were higher than the recommended upper limits in sediments by China and USEPA except lead
in the OginigbaRiver and cadmium in both rivers. In both rivers, all the metals, copper (Cu), nickel (Ni),
chromium (Cr) and zinc (Zn)fell within the category of heavily polluted, lead (Pb) in the sediment from the
Oginigba River was moderately polluted and cadmium (Cd) showed no indication of contamination/pollutionin
any of the sediments from the rivers. The order of the concentration of the heavy metals in the sediments wasZn>
Ni > Cr >Cu>Pb>Cd. Contamination factor (Cf) analysis showed that the sediments from both rivers were free
from Cd pollution, slightly polluted with Cu and Cr and moderately polluted with Ni, Zn and Pb. Pollution load
index (PI) results showed that Bomu River sediments were moderately polluted while Oginigba river sediments
were slightly polluted by the metals. Geo-accumulation index (I-geo) measurements showed that sediments
from both rivers are free from pollution by the metals. Ecological risk factor and ecological risk assessment of the
sediments from both rivers indicated that both aquatic environments are free from ecological risk. However, the
results suggested that there is the possibility of a near future or long run pollution. Therefore, adequate measures
should be put in place to monitor thes environments.
Keywords: Heavy metals, pollution, index models, Bomu River, Oginigba River, sediments
INTRODUCTION
The prevalence of pollution has become a matter of public debate all over the world. The increase in pollution
primarily resulted from human activities aimed at making life on earth worthwhile.Pollution is diverse and
therefore comes in different ways and forms. It has a fairly negative consequence on our health and thus affects
every other part of human existence. Among all the pollutions, heavy metal is one that should be given much
attention by individuals, cooperate bodies and the government. When adequate attention is paid to the
contamination of the environment, then possible ways of improving the environment through pollution
reduction can be put in place.
Industrialization and agricultural activities which are the routes for economic growth havediversified the
pathways through whichheavy metals are introduced into air, water, soils and sediments. Soilis as the ultimate
sink for heavy metals and other pollutants discharged into the environment [1]. Heavy metals, which may be
essential (useful) and non-essential (harmful)to human systemare very significant in the study of ecology and
toxicology. They are persistent in the environment and possess potential toxicity to living organisms in the
environment. The concentrations of heavy metalsin aquatic ecosystems are monitored by measuring their
concentration in water [2], sediments and life forms in the aquatic environment.
Sediment as an important component of the aquatic environment gives vital information on the pollution status
of the marine ecosystem [3]. The sediment which acts as the carriers and sinks for contaminants provides
information on the history of pollutionthe extent of pollution or contamination of the river concerned[4] and also
provides a data bank on that ecosystem which further studies could be compared with.In the aquatic
environment, there are three ways in which heavy metals are retained. These ways are water, sediment and
biological organisms (aquatic plants and animals)[5]. The analysis of the sediment of any aquatic system gives
information on the impact of human activities and natural processes that may have impacted on that environment
over time[6,7].
389
The accumulation of heavy metals in river sediments involvesadsorption processes (physical and
chemical)which depends on the nature of the sediment matrix (simple or complex) and the characteristics of the
compounds entrained in the sediments [8].
This study was therefore carried out to examine the effect of oil exploration activity at Bomu on the Bomu River
and the effect of industrial activity on the Oginigba River with reference to the concentration of heavy metals in
these rivers.
MATERIALS and METHODS

Sediment samples were collected at low tide from three locations from each river using Eckman Grab Sampler.
The samples were put in clean black polythene bags and then transported to the Chemistry Laboratory of the
Ignatius Ajuru University of Education Port Harcourt.Sediment samples were air-dried at ambient temperature
in the laboratory to constant weight. The dried samples were powdered in a ceramic mortar without crushing
stones and pebbles and other hard materials such as pieces of wood and leaves of vegetables. The powdered
samples were then then sieved with 0.5 mm sieving mesh.Two grams (2 g) of the air-dried and powdered
sediment samples were weighed using a high precision balance and was transferred into 100 ml conical flask.
0
The digestion process for sediment samples was done at 60 C for 3 hours to near dryness with a mixture of HCI,
HNO3 and H2SO4 acids in in the ratio of 1:3:1 in a water bath. Precisely 20 cm3 ofdeionized water were added after
which the digest was allowed to cool to room at temperature and was then filtered into a 50 ml volumetric flask
using Whatman No.1 filter paper. The digest was made up to mark of 25 ml with distilled water and the
concentrations of the trace metals were determined by a Buck Scientific model 200 Å Spectrophotometer
equipped with air-acetylene flame.
The data obtained were subjected to mathematical index models on pollution. These models are used in the
interpretation of the extent of contamination/pollution of the environment by heavy metals.
Contamination Factor/ Pollution Index
The contamination factor is expressed as:
Contamination Factor (Cf) = Cm/Cb
The Pollution Index (PI) is mathematically expressed as:
Pollution Index (PI) = n√(Cf1xCf2xCf3x…xCfn)
Where, CF = contamination factor, n = number of metals, Cm = metal concentration in polluted sediments,Cb =
background value of that metal.
The significance of intervals of contamination factor/ pollution index indicates that values < 1 indicates
contamination of the sediment while values > 1 is an indication of pollution[9].The world average concentration
valuesfor the metals are Cu (45 mg/Kg), Ni (68 mg/Kg), Cd (0.3 mg/Kg), Zn (95 mg/Kg), Cr (90 mg/Kg) and
Pb(20 mg/Kg) were used as the background value.
Geo-accumulation index (I-geo)
Geo-accumulation index was calculated according toequation given by[10].
Mathematically, I-geo = log 2 (Cn / 1.5 Bn)
Where, Cn = Concentration of heavy metal in the sediment.
Bn = Geochemical background value in average shale of the metal.
The factor 1.5 is used for the possible variations of the background data due to lithological variations.
The classification of the I-geo shows that values < 1 are not polluted, while values > than 1 ranges from slight
pollution to extreme pollution [10].
Ecological Risk Factor (Ef)/ Potential Ecological Risk(RI)
Ecological risk factor (Ef) was used to express the potential ecological risk of a given contaminant, while the
potential ecological risk assesses the effect of multiple metal pollutions in the sediments from the aquatic
390
environment [11].
Mathematically, Ef=Tr x Cfand
RI = ∑Ef
Where Tr = the toxic-response factor for a given element and
Cf= the contamination factor of the element.
The toxic response values of the heavy metals are: Cu = Ni = Pb = 5, Cd = 30, Cr =2 and Zn =1.
The terms used to describe the ecological risk factor (Ef)and potential ecological risk(RI) are given in table 1.
Table 1: Terminologies used for ecological risk factor (Ef) and potential ecological risk (RI)
Factor Low risk Moderate Considerable risk High risk Very high risk
Ef <40 40≤Ef<80 80≤Ef<160 160≤Ef<320 Ef≥320
RI <150 150≤RI< 300 300≤RI<600 - RI>600

RESULTS and DISCUSSION
Heavy Metal Concentration
The concentrations of the heavy metals in the sediments of Bomu and Oginigba Rivers are given in Table 1. All
the metals assessed in the sediments from the two rivers were higher than the world average value for each metal
and the limit value set by China except cadmium. Further examination of the concentrations of the heavy metals
in the sediments revealed that the sediments are heavily polluted by the individual metals except lead which was
moderately polluted in the Oginigba River and cadmium with no pollution threat.
Table 2: Heavy metal concentrations of sediments from Bomu and Oginigba
Metal Bomu Oginigba Standard deviation

± SD
Cu 66.301 ±38.152 74.610 ± 27.152 5.657
Ni 148.919±84.793 151.219 ± 34.473 1.627
Cd <0.001 <0.001 0.00
Cr 129.731 ±30.483 109.297 ± 21.843 14.449
Zn 203.513 ±63.101 213.315 ± 60.214 6.931
Pb 60.301 ±38.152 43.307 ± 21.521 12.017
Table 3: World average and standards Limits for trace metal contamination of sediments in mg/Kg
Metal World China USEPA
average Not polluted Moderately polluted Heavily polluted
Cu 45 35 < 25 25 -50 > 50
Ni 68 - < 20 20 -50 > 50
Cd 0.3 0.5 - - >6
Cr 90 - < 25 25 -75 > 75
Zn 95 150 < 90 90 -200 > 200
Pb 20 60 < 40 40- 60 > 60
391
The high values of heavy metals concentrations in sediment is traceable to anthropogenic activities or
interferences which in this case are the oil exploration and exploitation in Bomu in Ogoni and the industrial
activities in the Port Harcourt Trans Amadi Industrial layout of most of the industries discharge their effluents
treated and untreated into the river. This observation corroborates the findings of [12], who observed that toxic
metals are derived from waste arising from industries and from fuel sources used in these industries.
Accordingly, they argued that most of these metals are also derived from the disintegration or corrosion launches
abandoned by the factories along the river shore. This situation was observed in the Oginigba River where many
launch boats were lying unattended to by the shores of the river.
The variation of heavy metals concentrations in sediments depends on rate at which particles sediments, the rate
of heavy metals are deposited, the size of the particles generated and organic matter presence in the sediments
[13]. Other factors that contribute to heavy metal concentration in sediments includes the type of industry
present, the rate of discharge of effluents, the available quantity of particulate matter present in the river or the
aquatic environment and others such as the landscape which can facilitate runoffs into the aquatic environment.
Contamination Factor/Pollution Load Index of Heavy Metals
In the present study the contamination factor of Cd (0.00) in the sediments were found in the category of no
contamination. The sediments were slightly polluted with Cu and Cr whose values in the sediments ranged from
1.214-1.658 in the sediments, while the sediments were moderately polluted by Ni, Zn and Pb metals. The
pollution index status of the sediment showed that Bomu sediment was moderately polluted, while Oginigba
sediment was slightly polluted (Table 4).
Table 4:Contamination Factor/Pollution Load Index of heavy metals in sediments from Bomu and Oginigba
Rivers.
Station Metals Pollution
Cu Ni Cd Cr Zn Pb Index
Bomu 1.473 2.190 0.00 1.414 2.142 3.315 2.0048*
Oginigba 1.658 2.224 0.00 1.214 2.245 2.165 1.8515*
*Cadmium value not added in the pollution index calculation

Contamination index is a quick tool used to examine the pollution status of the individual metal in water,
sediments and soils while Pollution load index examinesthe combined effects of all the individual metals
examined and its variation along the sites[14]. The pollution of these sediments is an indication of external
influence, which in this case is the industrial activity. The result of the contamination index and the pollution load
index based on the [9] contamination and pollution index is aconfirmation that the Bomu and OginigbaRiversare
facing environmental pollution problems which need urgent attention of the government and relevant agencies.
Geo-accumulation Index
The geo-accumulation index of the sediments by the metals from the two rivers all fall into the category or class 0,
Class 0 = Igeo ≤ 0, practically uncontaminated (Table 5). This is based on the classification proposed by [10].
Geo-accumulation is a measure of the extent of pollution of aquatic sediments which is categorized into seven
levels. From the results obtained in this work, it follows that the heavy metals in sediments from these rivers are
within the background level or little above the background value from world average in shale.
Table 5: Geo-accumulation Index (Igeo) ofheavy metals in sediments from Bomu and Oginigba Rivers.
392
Station Metals
Cu Ni Cd Cr Zn Pb
Bomu 0.294 0.440 0.00 0.289 0.430 0.665
Oginigba 0.333 0.446 0.00 0.244 0.451 0.435
Ecological Risk Factor (Ef) and Potential Ecological Risk Assessment (RI)
Based on the equation of ecological risk factor (Ef) and potential ecological risk assessment (Ri) the analysis of
factors were found to be in the category of low risk. All Ef results for the metals were far < 40 and Ri results were
all < 150 (Table 6). Based on the classification in Table 1 as proposed by [11], the metals in the sediments do not
show any form of risk to the aquatic environment.
Different guidelines are used to monitor the quality of sedimentsand protectaquatic organism (plants and
animals) from the dangerous, harmful and toxic effects that comes along with sediment bound contaminants
[15]. The reasons behind these guidelines is to examine the extent that the sediment-bound chemical status has
affected aquatic animals and plants and also to give adequate ranking of sediment pollution and also allow time
for further investigations and make necessary recommendations [16].
Table 6: Ecological Risk Factor(Ef) and Potential Ecological Risk Assessment (RI) ofheavy metals in
sediments from Bomu and Oginigba Rivers.
Station Metals RI
Cu Ni Cd Cr Zn Pb
Bomu 7.365 10.95 0.00 2.828 2.142 16.575 39.859
Oginigba 8.29 11.12 0.00 2.428 2.245 10.825 34.908
CONCLUSION
The activities of oil exploration and exploitation in Bomu in Ogoni and the industrial activities in the Trans
Amadi Industrial Layout in Port Harcourt all in the Rivers State Nigeria has slightly impacted negatively on the
rivers within the areas. The heavy metals assessed were all slightly higher than the recommended limits for heavy
metals in sediments by the relevant bodies cited. Even though the assessment by the different index models
showed little or no threat to the environment, yet consistent activity on these rivers for a long time can pose
detrimental effects on the later years.
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Should We Worry about Their Environmental Risks? Environmental Research, 97: 258–273.
2. M. Ebrahimpour, andI. Mushrifah, (2008). Heavy metal concentrations in water and sediments in
TasikChini, a freshwater lake, Malaysia. - Environmental Monitoring and Assessment 141: 297-307.
3. J. M. Casas, H. Rosas, M. Sole, and C. Lao, (2003). Heavy metals and metalloids in sediments from the
llobregat basin, Spain. Environmental Geology, 44(3), 325-332.
4. K. P.Singh, D. Mohan, V. K. Singh, and A. Malik, (2005). Studies on distribution and fractionation of
heavy metals in Gomti River sediments – a tributary of the Ganges, India. Journal of Hydrology, 312(1-
393
4), 14-27. 2005.
5. S. B. Saha, S. B Abhijit, and A. Choudhury, (2001). Status of sediment with special reference to heavy
metal pollution of a brackish water tidal ecosystem in northern Sundarbans of West Bengal. Tropical
Ecology, 42(1): 127-132.
6. A. Goorzadi, G. Vahabzadeh and A. R. Carbassi, (2009). Assessment of heavy metals
pollution in Tilehbon River sediments.Iran Journal of Applied Science, 9(6): 1190-1193.
7. F. E. Olubunmi, and O. E. Olorunsola, (2010). Evaluation of the status of heavy metal pollution of
sediment of Agbabu Bitumen deposit area, Nigeria. European Journal of Scientific Research, 41(3),
373-382.
8. G.T. Ankley, K. Lodge, D.J. Call, M.D. Balcer, and B. J. Smith, (1992). Heavy metal concentrations in
surface sediments in a near shore environment, Jurujuba Sound, Southeast Brazil. Environmental
Pollution, 109.
9. Lacatusu, R. (2000). Appraising Levels of Soil Contamination and Pollution with heavy Metals.
European Soil Bureau Research Report, 4: 393 - 402.
10. G. Muller, (1969). Index of geoaccumulation in sediments of the Rhine River. Geology Journal, 2:
109–118.
11. L. Håkanson, (1980). An Ecological Risk Index for Aquatic Pollution Control: A Sedimentological
Approach. Water Research, 14: 975–1001.
12. A. M. Rabee, Y. F. Al-Fatlawy, A. N. Abdown and M. Nameer (2011). Using Pollution Load Index
(PLI) and Geoaccumulation Index (I-Geo) for the Assessment of Heavy Metals Pollution in Tigris
River Sediment in Baghdad Region. Journal of Al-Nahrain University, 14 (4): 108-114.
13. J. Mwamburi, (2003). Variations in trace elements in bottom sediments of major rivers in Lake Victoria's
basin, Kenya Lakes & Reservoirs. Research and Management, 8: 5- 13.
14. K.O.Adebowale, F.O.Agunbide, and B. Olu-Owolabi, (2009). Trace metal concentration, site variations
and partitioning pattern in water and bottom sediments from coastal area: A case study of Ondo Coast,
Nigeria. Environmental Research Journal, 3(2): 46-59.
15. S. McCready, G. F. Birch, andE. R. Long (2006). Metallic and organic contaminants in sediments of
Sydney Harbour, Australia and vicinity-a chemical dataset for evaluating sediment quality guidelines.
Environment International, 32:455–65.
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Kosore, (2011). Assessment of the metal pollution, potential toxicity and speciation of sediment from
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394
EFFECT OF SEASONAL WATER FLUCTUATION OF A WATER BODY ON ANTIOXIDANT
ACTIVITY OF SELECTED PLANTS OF LOWER PHYLUM(A CASE STUDY OF NCHESTREAM)
1
Duru M.K.C; 2Akubugwo E.I; 2Chinyere G.C; 3Amadi B.A; 4Agomuo E.N; 5Alisa, C. O; 6NJoku V. O; and
4
Amadi P.U
1
Department of Chemical Sciences (Biochemistry Unit), Rhema University, Nigeria
2
D e p a r t m e n t o f B i o c h e m i s t r y, A b i a S t a t e U n i v e r s i t y, N i g e r i a
3
D e p a r t m e n t o f B i o c h e m i s t r y, U n i v e r s i t y o f P o r t H a r c o u r t , N i g e r i a
4
D e p a r t m e n t o f B i o c h e m i s t r y, I m o S t a t e U n i v e r s i t y, N i g e r i a
5
D e p a r t m e n t o f C h e m i s t r y, F e d e r a l U n i v e r s i t y o f Te c h n o l o g y, N i g e r i a
6
Department of Chemistry, Imo State University, Nigeria
ABSTRACT
Effect of seasonal water fluctuation of a water body on antioxidant activity of selected plants of lower phylum
using Nche stream as a case study was investigated using standard methods. Three plants of lower phylum
(watercress, moss plant, and spirogyra) were selected and studied for both enzymatic and non-enzymatic
antioxidants. Results obtained for levels of ascorbic acid (0.81-11.87 µmoles/g DW), glutathione (1.47-
3.01µmoles/g DW) and proline (1.27-3.01 g/100g) non-enzymatic antioxidants and those of superoxide
dismutase (289.19-615.85 µg/g protein), peroxidase (32.56-52.79 µg/g protein), and catalase (57.80-
73.20µmoles/g DW) of enzymatic antioxidants were higher in dry season against rain season. Since it has been
noted that a slit difference in these indicators could be as result of enormous stress. The reduction in volume of
water of the host stream in dry season, may have resulted in increased concentration of the pollutants of the water
body hence inducing the plants to absorb more of the pollutants. This may have triggered more stress on the
plants, which reflected on the levels of the observed stress indicatorswhen compared tothe indicators as observed
in rainy season. This study has shown the seasonal water fluctuation of a water body on antioxidant activity of
selected plants of lower phylum.
Keywords: Antioxidant, lower phylum, seasonal fluctuation, stress enzymes,water body,
INTRODUCTION
According to Dominika and Barbara [1], biological systems stress can be defined as an adverse force, effect, or
influence that trends to inhibit normal systems from functioning. It has been noted that a wide range of
unfavourable environmental conditions may induce stresses in plants, which can alter plant growth,
development, metabolism, and even may lead to plant death[1]. Plants react to environmental stresses on various
levels including biochemical, cellular and morphological scales depending on type of species or population [2-
5]. Abiotic stress is defined as the negative impact of non-living factors on the living organisms in a specific
environment [6]. Basically, reaction of plants to abiotic stresses depends on type of plant species having
fundamental differences in development and anatomy as well as environmental limiting factors [7].
Water stress is among the abiotic stresses that affect plants. This type of stress could be generated by drought
orflooding [6]. According to Bartels and Souer [8], water deficit, caused by “lack of water” has been among the
problems for agriculture, affecting virtually every aspect of plant physiology and metabolism.The mechanisms
of abiotic stress effect on plants have been reported by different authors [9-10], and have been linked to
- -
generation of reactive oxygen species (ROS), which include O2 , H2O2, and OH . These molecules are highly
reactive and can alert normal cellular metabolism through oxidative damage to membranes, proteins and nucleic
acids [11]. Reactive oxygen species can also cause lipid peroxidation, protein denaturation and DNA mutation
[12].
395
Generally, to prevent the damage inherent from reactive oxygen species (ROS) on cellular components, plants
have developed a complex antioxidant systems[11]. According to Rahimizadehet al. [11],the primary
components of this system include carotenoids, ascorbate, glutathione and tocopherols. Others are superoxide
dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), perioxidase (POX; EC 1.11.1.7) and glutathione
reductase (GR; EC 1.1.4.2) [12]. The antioxidant enzymes play key role in the defense against oxidative damage
[9, 11-12].
Nche stream is among the natural waterbodies that supply domestic water to the people of Umunchi. Umunchi is
among the communities found in IsialaMbano L.G.A of Imo State, Nigeria. Water level of the stream is known to
increase in rainy season, and decrease in during dry season. Apart from domestic water supply, the stream houses
other organisms that depend on it.An earlier study on the water by Duruet al. [13], ascertained theseasonal water
quality assessment of the water body. There is need to extend the study on the water body to accommodate lower
organisms that depend on water from the water body.
Different studies have investigated the effect of water stress on plants, but most of the studies were centered on
drought stress [14-19]through isolated observations. The present study investigated the seasonal water
fluctuation of a water body on antioxidant activity of selected plants of lower phylum, with a view to ascertain the
stress induced by such fluctuation, using Nche steam as a case study.
Location of Nche Stream
IsialaMbano L.G.A lies within latitude 5°40´ 3.6´´ (5.6677°) north and longitude 7 ° 12´ 22.2´´ (7.2034°) east
with an average elevation of 149 meters (about 489 feet). Within these latitude and longitude lies Umunchi
community, and hence Nche stream.
Collection and Preparation of Samples
Three plants of lower phylum were used for this study. The plants were spirogyra(seaweed),
watercress(Nasturtium sp.) and mosses. Spirogyra (seaweed) sampling was done using the method described by
Okafor[20]for phytoplankton. Coned-shaped, silk plankton net were employed. At free-flowing part of the water
body, a net was used by sinking and drawing it against the water current. Those found on hard surfaces such as
the walls built by local population to safe-guide the stream were collected with the help of sterile scraper.
Watercress (Nasturtium sp.) samples were collected from banks of the stream. Moss plants used in this study
were collected from the walls as built by the local population to protect the stream, making sure that they receive
water from the stream both at high tide and low tide.The samples were transported to the laboratory in a cold
condition (container packed with ice). At the laboratory, the sampled plants were prepared for further studies.
The sampling was done at peak of the two seasons.
DETERMINATION OF NON-ENZYMATIC ANTIOXIDANTS
The titrimetric method described by Conklin [21]was used for determined ascorbic acid. Glutathione and phenol
were determined as described by Murugan and Harish [22]. Carotene was determined by Dereet al. [23].
Speckmanet al. [24] method was used for proline while flavonoid was determined using AOAC [25].
ESTIMATION OF ENZYMATIC ANTIOXIDANTS
The method as described by Lowry et al. [26] was used for protein assays. Measurement of superoxide dismutase
enzyme activity was done using the method Misera and Fridorich[27].Peroxidaseenzyme activity was estimated
using the method as described by AOAC[25]. Catalase activity was measured using Paglia and Valentine
[28].Activity of Glutathione reductase (GR) was estimated by the method of Foyer and Halliwell [29] modified
by Rao [30].
RESULTS AND DISCUSSIONS
396
Fig. 1: Ascorbic acid levels of the plants. Fig. 2: Glutathione level of the plants.
Fig 3. Phenol level of the plants . Fig. 4: Carotene level of the plants.
Fig. 5: Flavonoids level of the plants. Fig. 6: Prolinelevel of the plants.
Figures 1-6 shows the presence of non-enzymatic antioxidants found in the studied plants. Ascorbic acid, also
known as vitamin Cis the most abundant, powerful and water soluble antioxidant which minimizes or prevents
damage caused by reactive oxygen species (ROS) in plants. Ascorbic acid (AA) is one of the most studied and
has been detected in majority of plant cell types, organelles and appoplast [31]. Wu [32] noted that ascorbic acid
-
reacts not only with H2O2, but also with O2 , OH and lipid hydroperoxidases.Ascorbic acid can also directly
-
scavenge 1O2, O2 and HO. and regenerate tocopherol from tocopheroxyl radicals providing membrane
protection [1]. Smirnoff and Wheeler [31] summarized that ascorbic acid reacts non-enzymatically with
superoxide, hydrogen peroxide and singlet oxygen. Ascorbic acid levels of the studied plants were between
10.12-11.87 µmoles/g DW in watercress; 1.02-1.32 µmoles/g DW in moss plant and 0.81-0.84 µmoles/g DW in
spirogyra (Figure 1). Glutathione (GSH)is a tripeptide (á-glutamyl-cysteinyl-glycine), which is considered as
the most important intracellular defense against ROS-induced oxidative damage [1]. Glutathione is important in
plant chloroplasts because it helps to protect the photosynthetic apparatus from oxidative damage [33].
Glutathione levels of the present study ranged from 1.15-1.20 µmoles/g DW in watercress; 1.79-1.88 µmoles/g
DW moss plant; and 2.58-2.60 µmoles/g DW in spirogyra (Figure 2).It has been noted that phenols are aromatic
secondary metabolites broadly distributed in plant kingdom [34]. They are essential to the physiology and
397
cellular metabolism. Phenolic compounds play a role of protection against insects and other animals to the
plants.The antioxidant activity of phenolic compounds depends largely on their chemical structures. Among the
phenolic compounds with known antioxidant activity are flavonoids, tannins chalcones and coumarins as well as
phenolic acids [35-36]. According to Daiand Mumper[37], phenolics have been considered as great antioxidants
and proved to be more effective than Vitamin C, E and carotenoids in recent times. Phenolic levels of the present
study were between 1.47-1.58 µ equ. of Gal/g in watercress; 2.58-3.01 µ equ. of Gal/g in moss plants; and 1.79-1
88 µ equ. of Gal/g in spirogyra (Figure 3).Carotenoidsare lipid soluble antioxidants pigments that play
multitude of function in plant metabolism including oxidative stress tolerance [1]. Carotenoids observed in the
present study were between 660.10 -671.60 µg/100g FW in watercress; 1870.13 to 1920.24 µg/100g FW in moss
plant; and 2310.00-2370.15 µg/100g FW in spirogyra as presented in Figure 4.Flavonoids are widely distributed
in plants; and come in four classes depending on their structure; flavonols, flavones, isoflavones and
anthocyanines. Flavonoides[1]. Flavonoids belong to one of the most reactive secondary metabolites
of plants [37], and play important role as reactive oxygen species (ROS) scavenger by locating and
neutralizing radicals before they damage cell structure [1]. It has been proved that they are involved in plant
responses to both, biotic or abiotic stresses such as wounding, drought and metal toxicity [38]. Flavonoids level
of the present study ranged from 0.53-0.58 mg/100g in watercress; 0.29-0.35 mg/100g in moss plant; and 1.03-
1.18 in spirogyra (Figure 5).Prolineas á-amino acid, is an antioxidant and potential inhibitor of programmed cell
death[1]. It has been suggested that free proline acts as osmoprotectant, a protein stabilizer, a metal chelator, an
inhibitor of lipid peroxidation and OH· and 1O2 scavenger [1].Proline is not only an important signaling
molecule, but also an effective reactive oxygen species (ROS) quencher [1, 39]. Levels of proline in the present
study were between 2.26-3.01 g/100g protein in watercress; 1.27-1.40 in moss plant; and 1.79-1.85 in spirogyra
(Figure 6). Trovatoet al. [40] reported increased proline accumulation during abiotic stresses. It has been found
that the important role of proiline is in potentiating pentose-phosphate pathway activity as important component
of antioxidative defense mechanism [39].Flavonoids are widely distributed in plants leaves, floral part and
pollens. They often accumulate in the plant vacuole as glycosides or as exudates on the leaves surface and other
aerial part of the plant. There are four flavonoid classes depending on their structure: flavonols, flavones,
isoflavones and anthocyanines. Flavonoides belong to one of the most reactive secondary metabolites of plants
[41]. Flavonoids play important role as ROS scavenger by locating and neutralizing radicals before they damage
cell structure. Flavonoides have function as flowers, fruits and seed pigmentation, they play protective role
before UV light, drought and cold and defense against pathogens. Flavonoids play an important role in plant
fertility and germination of pollen. They are involved in plant signaling with interaction with plant microbes [41-
42]. It has been proved that they are involved in plant responses to both, biotic or abiotic stresses such as
wounding, drought and metal toxicity [38].Proline, á-amino acid is an antioxidant and potential inhibitor of
programmed cell death. It has been suggested that free proline act as osmoprotectant, a protein stabilizer, a metal
chelator, an inhibitor of lipid peroxidation and OH· and 1O2 scavenger. Increased proline accumulation appears
especially during salt, drought and metal stresses [40]. Therefore proline is not only an important signaling
molecule, but also an effective ROS quencher. It has been found that the important role of proline is in
potentiating pentose-phosphatase pathway activity as important component of antioxidative defense mechanism
[39].
398
Fig. 7: Superoxide dismutase level of the plants. Fig. 8: Peroxidase level of the plants.
Fig. 9: Catalase level of the plants. Fig. 10: Glutathione reductase level of the plants.
According to Dominika and Barbara [1], superoxide dismutase (SOD) constitutes the first line of defense against
reactive oxygen specie (ROS). SODs are metalloproteins and basedon their metal cofactor, they are
classified into three known types; the copper/zinc (Cu/Zn-SOD), the manganese (Mn-SOD) and the
iron (Fe-SOD) that are localized in differentcellular compartment [43].Dominika and Barbara [1] noted
- -
that SODs remove O2 by catalyzing its dismutation, one O2 is reduced to H2O2 and another to O2.
Superoxide dismutase of the present study ranged from 506.94-508.10 µg/g protein in watercress; 289.19-
369.02 µg/g protein in moss plant; and 612.09-615.85 µg/g protein in spirogyra (Figure 7).Peroxidase could be
in the form of glutathione peroxidase, ascorbate peroxidase or guaiacolperoxidase [1]. According to Abdollahet
al. [9], the activity of peroxidases depends on plant species and stress condition.Peroxidase levels of the studied
plants ranged from 32.56-34.67 µg/g protein in watercress; 50.89-60.87 µg/g protein in moss plant; and 46.38-
48.29 µg/g protein in spirogyra (Figure 8). Catalase is a light-sensitive protein that has a high rate of
turnover and environmental stresses which reduce the rate of protein turnover, such as salinity, heat
shock or cold, cause the depletion of catalase activity [44,45].Dominika and Barbara [1] noted that it
remains unclear whether variability in catalase response to different unfavourable conditions may be
ofimportance in plant stress tolerance level. Observed catalase levels as presented in Figure 9,
ranged from 57.80-59.21 µg/g protein in watercress; 58.90-60.87 µg/g protein in moss plant; and 73.20-76.14
µg/g protein in spirogyra. Glutathione reductase is a flavor-protein oxidoreductasethat is thought to play an
essential role in defense system against reactive oxygen species [42, 46]. Glutathione reductase levels of the
present study ranged from 7.14-9.04 µg/g protein in watercress; 6.20-8.18 µg/g protein in moss plant; and 6.16-
8.10 µg/g protein in spirogyra.
All the antioxidants both the non-enzymatic and enzymaticof the investigated plants, increased in dry season
than rainy season.
399
CONCLUSION
Since it has been noted that a slit difference in antioxidant levels could be as a result of enormous stress. It
therefore follows that the observed increase in antioxidant levels of the investigated plants in dry season, could
be as a result of increased concentration of pollutants found in the host stream due to reduction in volume (low
tide) of the water body in dry season. The plants may have absorbed more of the pollutants in dry season, which
may have stressed the plants hence leading to increased levels of the antioxidants in dry season against those of
the rainy season where dilution of the pollutants due to increase in level of water of the water body, and their
reduced absorption may have induced a reduced stress on the plants. This study has shown the effect of seasonal
water fluctuation of a water body on antioxidant activity of selected plants of lowerphylum.
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402
CHARACTERIZATION OF VIOLET TREE (Securidaca longepedunculata) ROOT
POWDER
1 ? 1 1 2
Donatus, R.B ., Barminas, J.T., Maitera,O.N., Riki, .E.Y
1
Department of Chemistry, Modibbo Adama University of Technology, P.M.B 2076
Yola- Nigeria.
2
Department of Basic Science, College of Agriculture, P.M.B 1025 Jalingo-Nigeria
Abstract
The characteristic proximate analysis was examined of the modified and unmodified Violet Tree Root Powder
(VTRP) samples as done on moisture content, ash content, volatile content, fixed carbon, and bulk density.
Others include porosity, specific gravity and swellability. The values for modified and unmodified VTRP
compared were moisture content 6.9 ±0.01 % - 10.14 ±0.03 %, ash content 8.86 ±0.01 % - 9.56 ±0.02 %, volatile
content 40.94 ±0.02 % - 18.07 ±0.02 %, fixed carbon 67.92 ±0.01 % - 71.80 ±0.02 % and bulk density 1.10 ±0.01
g/cm3 – 1.33 ±0.01 g/cm3. The porosity gave 97.02 ±0.01 % - 93.8 ±0.01 %, specific gravity 0.6909 ±0.04 –
0.9906 ±0.01, swellability 220.02 ±0.02 % - 304.7 ±0.02 % accordingly. The improved physico-chemical values
were indications of better hydrophobic behaviour of the modified VTRP as a sorbent.
Keywords: Characterization, Modified, Unmodified, Violet Tree Root Powder.
Introduction
Increasing environmental concern especially after several hazardous incidents in the past decades when large
quantities of oil were spilled into the sea renewed the interest for natural fibers (Radetic et al., 2003). The
chemical modifications of wood with anhydride reagents have been the subject of research for many decades
(Hill et al., 1998; Rowell et al., 1987 and Rowell, 1983).
Recent research and development on sorbent, is focused on the improvement of known products and synthesis
paths as well as on new derivatives and alternatives synthesis concepts (Heinze, 2005).
Application of agro-industrial residues in industries on the other hand provides alternative substrate and also
helps in solving pollution problems, which their disposal may otherwise cause (Sun et al., 2004). In addition,
with decreasing wood resources, work has increased regarding the use of no Acetylation (Ethanoylation in
IUPAC nomenclature) describes a reaction that introduces an acetyl (CH3CO) functional group into a chemical
Parameter UMVTRP MVTRP

Mean with S. D Mean with S. D
Ash content (%) 9.56 ± 0.02 8.86 ± 0.01

Moisture content (%) 10.14 ± 0.03 6.95 ± 0.01
Volatile content (%) 18.07 ± 0.02 40.94 ± 0.02
Bulk density (g/cm2) 1.33 ± 0.01 1.10 ± 0.01
Fixed carbon (%) 71.80 ± 0.01 67.92 ± 0.01
Specific gravity (g/cm3) 0.9966 ± 0.01 0.6909 ± 0.04
Porosity (%) 93.8 ± 0.11 97.02 ± 0.01
Swelling ability (%) 304.7 ± 0.02 220.02 ± 0.02
403
UMVTRP – unmodified violet tree root powder, MVTRP – modified violet tree root powder.
The results of the physical properties of UMVTRP and MVTRP showed that during the course of modification,
the ash content which is a reflection of the inorganic composition of the VTRP samples decreased from
9.56±0.02 % - 8.86±0.01 %. In general, ash content of fibrous raw materials has been reported to be between 1 %
- 20 % according to Ruben and Bokelman (1987). It showed that modification also reduced the moisture content
from 10.14±0.03 % - 6.95±0.01 %. Hence the MVTRP will have low water uptake and become more
hydrophobic. The volatile matter and porosity increased from 18.07±0.02 % - 40.94± 0.02 % and 93.8±0.11 % –
97.02±0.01 % respectively during modification. This is an indication of effective acetylation derived from the
use of cetyltrimethylammonium bromide (CTAB) as catalyst, whose great advantage is removal of hydroxyl
component of hemi-cellulose and lignin of the cellulose (Hill et al., 2000). The increased porosity of the fiber is
an indication that expected MVTRP is to have high sorption capacity for oil due to the introduced acetyl group
present.
3
The determined bulk density, fixed carbon and specific gravity gave reductions from 1.33±0.01 g/cm –
3 3 3
1.10±0.01 g/cm ; 71.80±0.01 % - 67.92±0.01 % and 0.9966±0.01 g/cm – 0.6909±0.04 g/cm respectively.
The bulk density and specific gravity are indication of efficient buoyancy expected upon application of
MVTRP in water, however, the fixed carbon and ash content material
Conclusion
In this study, the proximate analyses of UMVTRP and MVTRP were investigated, alongside the sorption
behaviours of the respective samples type evaluated in oil and water. The characterization revealed decreased
properties in ash content, moisture content, bulk density, specific gravity and fixed carbon as 9.56±0.02 % -
3 3 3
8.86±0.01 %, 10.14±0.03 % - 6.95 ±0.01 %, 1.33±0.01 g/cm - 1.10±0.01 g/cm , 0.9966±0.01 g/cm -
0.6909±0.04 g/cm3 and 71.80±0.01 % - 67.92±0.01 % by unmodified to modified samples respectively. The
volatile content and porosity observed increased values from 18.07±0.02 % - 40.94±0.02 %, 93.8±0.11 % -
97.02±0.01 %, accordingly. The swelling ability decreased from 304.7±0.02 % -220.02±0.02 % at 24 h.
404
DETERMINATION OF THE SACCHARIN CONTENT IN SOME ICE CREAMS CONSUMED IN
PORT HARCOURT
Dibofori-Orji Amalo Ndu & Didia Lucky Ejikeme
DEPARTMENT OF CHEMISTRY
IGNATIUS AJURU UNIVERSITY OF EDUCATION
RUMUOLUMENI, PORT HARCOURT
jc_orji@yahoo.com GSM: 08039572370
Place of Research: Department of Chemistry, Rivers State University of Science &
Technology, Nkpolu Port Harcourt
ABSTRACT
The scope of this study covered six different ice creams purchased outside the gates of some primary and
secondary schools in Port Harcourt. Each category of ice cream was assayed for saccharin (n=3). Standard
deviation was calculated to ascertain the level of deviation from the mean. A total of 18 samples (6x3) were
analyzed. The Saccharin concentration was determined quantitatively at a wave length of 425nm using ultra
violet spectrophotometric method. Some conventional ice creams and locally prepared ice creams were
compared in this study. The calibration data obtained were satisfactory with a correlation coefficient of 0.996.
The concentration of saccharin in these samples ranged between 25.0mg/Kg – 69.0mg/Kg. Both conventional
and locally prepared ice creams showed similar trend in saccharin concentration. Two samples (a conventional
ice cream and a locally prepared ice cream) gave highest concentration of 69.0mg/Kg saccharin respectively,
while the lowest concentration (25.0mg/Kg) was assayed in one of the locally prepared ice creams. Both locally
prepared and conventional ice creams showed very high levels of saccharin when compared with the World
Health Organization (WHO ) permissible level of 5mg/Kg/day. From the findings of this work, continuous
consumption of these ice creams may result in adverse health conditions.
KEY WORDS: Saccharin, Ice Cream, Sweeteners, UV Spec.
INTRODUCTION
Saccharin with an IUPAC name 1,1–dioxo–1, 2–benzoisothiazol-3-one is a sweet tasting synthetic
compound. It is a non-caloric sweetening agent which has been used to weeten foods and beverages.
Etymologically, its name is derived from a Latin word “Saccharium” meaning sugar. Saccharin was discovered
accidentally in 1879 by Constantine Feldberg, a graduate student at John Hopkins University1. The use of
saccharin became wide spread during the sugar shortages of World War 1. Its popularity further increased
during the 1960s and 1970s among dieters as a result of its calorie-free status. The excellent stability of saccharin
under food processing makes it ideally suited in many different products. It is used in a wide range of cases
where heat processing is required (e.g. jams, canned products). However, at low pH (pH 2.5), it can be slowly
2
hydrolyzed to 2-sufobenzoic acid and 2-sulfoamylobenzoic acid .
In the past, saccharin was used in a variety of applications. It was first used as an antiseptic and
preservative to retard fermentation in food. Later on, saccharin was used in the plastic industry as an antistatic
3
agent and as a brightener in nickel plated automobile bumpers . Today, saccharin is used in a wide variety of food
products including baked goods, beverages, soft drinks, sugar preserves and confectionery, alcoholic drinks,
vinegar, pickles and sauces other food products. It is also used in cosmetic industries and in manufacturing of
toothpaste.
ICE-CREAM: is a smooth, sweet, cold food prepared from a frozen mixture of milk products and
flavourings, containing a minimum of 10 percent milk fat and eaten as a snack or desert. Ice cream is derived
405
from earlier “iced cream” or “cream ice” that was similar to “iced tea”. The name was later abbreviated to “ice
cream” the name we know today. It is a refreshing food that is liked by both children and adults. Ice cream is rich
in calcium and phosphorus and also contains many vitamins (natural and fortified), including vitamin A, C, D
and E as well as thiamin, riboflavin, niacin, folate, vitamins B-6 and B-12.
Arising from some controversies surrounding the toxic effects of saccharin to the body physiology,
several studies have been conducted using different analytical procedures to establish this fact. The effect of lose
of sodium saccharin on the induction of rat urinary bladder proliferation was investigated using autoradiography
and scanning electron microscopy4. A study was conducted on the Rapid Spectrophotometric Determination of
5
saccharin in soft drinks and pharmaceuticals using Azure B. as reagent . The Rapid Determination of Saccharin
6
sodium in ice cream by High performance liquid chromatography (HPLC) was also investigated . A radial
compression (18 column, 100mm x 8mm i.d; and ultraviolet detector at 421nm were applied in the
determination without extraction.
Another researcher studied the estimation of saccharin in soda beverages syrups, kulfies and candies. Saccharin
contents of these different food products were measured quantitatively by gravimetric method, using acetic acid
and lead acetate as solvent 7. Some others workers, investigated saccharin analysis in pharmaceutical and
cosmetics preparations as follows: by derivative ultraviolet spectrophotometry8; the determination of saccharin
9
in low calorie products using flow through spectrophotometric sensor ; the use of a simplified
10
spectrophotometric method for routine analysis of saccharin in commercial non-caloric sweetners ; the
spectrophotometric method for the determination of saccharin in food and pharmaceutical products11 and the
determination of saccharin in preserved fruits by High Performance liquid chromatography12.
Ice cream comes in different brands. We have “conventional ice creams” with well known brand names.
There are also several “locally prepared ice creams” with unbranded names in the market. This study was aimed
at determining the level of saccharin content in some commonly consumed ice creams in Port Harcourt
metropolis. The assay covered both the conventional and locally manufactured ice creams.
MATERIALS / METHOD
The materials and method used in this study were all of analytical standard and principles. Ice cream
samples were purchased at the gates of some Primary and Post Primary schools within Port Harcourt metropolis.
13
Method of analysis of saccharin was by UV Spectrophotometer (UV – 2500) at 425nm .
3.4 Sample Preparation/Extraction
25grams of each sample were weighed and transferred to 100ml volumetric flask with small amount of
water. Enough boiling water was added to make up 75ml; the mixture was allowed to stand for one hour shaking
occasionally. Then 3ml ethanoic acid was added and mixed thoroughly; followed by excess (5ml) of 20%
neutral lead (II) acetate solution. Cold water was added to mark, stirred to complete homogenization and the
mixture was allowed to stand for 20mins and filtered using Whatman 25mm filter paper. 25ml of filtrate was
transferred into a separator, followed by 3ml HCl. Saccharin in the filtrate was extracted using 1:1 (v/v) of
Ethoxyethane and Petroleum Ether . The extract was concentrated and the absorbance measured in the UV Spec
at 425nm using Nessler's reagent for colour development. Saccharin forms blue colour complex with Nessler's
reagent in a slightly acidic medium of Lead (II) Acetate - Ethanoic Acid buffer. Prior to sample analysis, a
calibration curve was developed from absorbance readings of serial dilutions of a standard solution of saccharin.
Formula for Calculation of Concentration of Serial Dilutions of Stock Standard Solution:
Conc. of Std. Soln. x Req. Vol. (Std) = Req. Conc. x Desired Soln. Vol.
Req. Vol (Std) = Req. Conc. x Desired Vol.
Conc. of Std. Soln.
Table 1: Volume of 1000ppm standard Stock solution required to prepare series of dilute standard
solutions.
406
Volume of 1000ppm Stock Total volume of solution n Concentration of standard
solution (ml) (ml) solution ppm
2.5 100ml 25
5.0 100ml 50
7.5 100ml 75
10.0 100ml 100
12.5 100ml 125
Table 2: Absorbance Readings of Different Concentrations of Standard Solution.

Concentration of Standard Absorbance
Solution (ppm)
25 0.011
50 0.026
75 0.040
100 0.058
125 0.070
RESULT ANDDISCUSSION
Fig. 1: Calibration Curve for Saccharin

Table 2: The Absorbance and Concentration of Saccharin in Different Ice Cream Samples
407
Samples 1st 2nd 3rd Mean S.D Conc.
Absorbance Absorbance Absorbance Absorbance (Mg/Kg)
C1 0.039 0.036 0.035 0.037 0.002 69.0
C2 0.025 0.025 0.024 0.024 0.001 48.0
C3 0.014 0.016 0.015 0.015 0.001 32.0
UB1 0.015 0.016 0.016 0.016 0.007 34.0
UB2 0.013 0.011 0.011 0.011 0.001 25.0
UB3 0.037 0.033 0.037 0.037 0.001 69.0
The standardized linear concentration ranges between 25mg/kg – 125mg/kg with absorbance range of 0.011 –
0.070. This was found to obey the Beer-Lambert's law which relates absorption to light path length, and also
absorption to the concentration of absorbing species in the material. The general Beer-Lambert's law is usually
expressed as “A = abc”14.
Using the calibration curve, the concentrations of saccharin in the various ice cream samples were determined.
C-1, C-2 and C-3 represent three brands of conventional ice creams respectively while UB-1, UB-2 and UB-3
are three different unbranded ice creams. The amounts of saccharin contents determined from the six samples of
ice cream indicate different levels of saccharin, except for C1 and UB3 which had the same saccharin content
(69.0mg/Kg). The saccharin content in these ice creams ranged between 25.0mg/kg – 69.0mg/kg. Interestingly,
a locally prepared ice cream, UB2 recorded the lowest concentration of saccharin (25.0mg/Kg).
When the saccharin concentrations in the studied ice creams were compared to the World Health
Organization15 permissible dose of 5mg/kg/day, they all exceeded it. The implication of this finding is that, both
local ice creams and conventional ice creams are prepared without considering safe level of saccharin
consumption. Children are most likely the largest consumers of ice cream especially the unbranded and
affordable ones. This may increase the susceptibility of life hazards, as reported in a study of bladder cancer
patients, where a possible link between the disease and use of saccharin has been established16.
CONCLUSION AND RECOMMENDATION

The UV spectrophotometric analytic procedure for the quantification of saccharin is a good method. The
concentration of saccharin in all the ice cream samples is alarmingly high including the branded ones. While the
association between saccharin consumption and bladder cancer risk is still controversial, many health groups
still believe that its used should be limited in infants, children and pregnant women. This is due to the possibility
of allergic reactions. Saccharin belongs to a class of compounds known as sulfonamides, which can cause
allergic reactions in some individuals. These reactions may include: headaches, breathing difficulties, and
diarrhoea and skin problems.
It is therefore recommended that, if saccharin must be used as an artificial sweetening agent in foods,
recommended dietary standards should be complied with. Different regulatory bodies in the country should
work in synergy to enforce and ensure compliance with existing national and international standards.
References
1. V. M. Sardesai and T. H. Waldshan (1991) Natural and synthetic Intense sweeteners. J. Nutr. Biochem.
2:236-44.
2. R. L. Pearson (1991). Alternative sweeteners. Ed. Lyn O' Brien Nabors, 147-65. US: Marker Dekker.
408
3. D. L. Arnold; D. Krewski and I. C Munro (1983). Saccharin: A toxicological and h i s t o r i c a l
perspective. Toxicol. 27: 179-256.
4. G. Murasaki and S. M. Cohen (1981). Effect of dose of sodium saccharin on the inductionn of rat urinary
bladder proliferation. Cancer Res. 41(3): 942-4.
5. P. G. Ramappa and N. A. Nayak (1983). Rapid Spectrophotometric determination of saccharin in soft
drinks and pharmaceuticals using Azure B as reagent. Analyst , 108, 966-970.
6. X. Zhang, W. Zhang and J. Dai (1999). Rapid determination of saccharin sodium in ice cream
by high performance liquid chromatography (HPLC). Se Pu 17(4) 408-9. Article in Chinese.
7. J. Masuma; I. Najma; A. Sarfraz and A. Iftikhar (2001). Estimation of saccharin in s o d a
beverages, syrups, kulfies and candies. Pakistan Journal of Applied Science 1 (4): 486-488,
8. C.D. Vianna-soares, and J. L. Seferin-Martins (2002). Saccharin analysis in Pharmaceutical and
cosmetic preparations by derivative ultra violet spectrophotometry. Revista Brasileira de ciencia
farmaceuticas DOI: 10.1590.
9. L F. Capitan valley, M.C. Valencuia, and E. Arana Nicolas (2005). Flow through spectrophitimetric sensor
for the determination of saccharin in low-calorie products. E-18071, Granada, Spain.
10. P. Los Weinert, H. R. Pezza de Oliverira, J. E. and L. Pezza (2004). A simplified S p e c t r o p h o t o m e t r i c
method for rountine analsysis of saccharin in commercial non caloric sweetener. Journal
Agricultural Food Chemistry, 29:52(26) 2288-92.
11. S. B. Matthew; A. K. Pillai and V. K.Gupta (2006) Spectrophotometric method for the determination of
saccharin in food and pharmateutical products. Indian journal of pharmaceutical science; Vol. 68
p821.
12. K. Pakasit, and P. Sirirat (2009). Determination of saccharin in preserved fruits by high performance
liquid chromatography. Journal of chemistry and chemical engineering's Vol. 3 Issue 12, p50.
13. AOAC (1990). Official methods of analysis. JAOAC: 56,162.
14. J. D. How; A. B. Burch; B. Miller; P. Morrison: L. Cordon; L. Weldon; C. Chambers Fordor and G.M.
Winsor (1977). Artificial sweetener and human bladder Cancer Lancet no. 8038:528.
st
15. A. S. Marcus (2013). Applied Spectroscopy (A Functional Approach). 1 ed., Enugu, University of
Nigeria Press Ltd. 57-61.
16. World Health Oganisation (1993). Evaluation of Certain Food Additives and Contaminants (WHO
Technical Report Series 837), Geneva. 17-19, 46.
409
EVALUATION OF CHEMICAL COMPOSITION OF Murraya koenigii (Linn) SPRENG LEAF
1 2 3 1 1
C. E. IGARA, D. A. OMOBOYOWA, A. A. AHUCHAOGU, N. U. ORJI AND M. K. NDUKWE
1
Chemistry Research Unit, 2Biochemistry Research Unit, Department of Science Laboratory
Technology, Akanu Ibiam Federal Polytechnic Unwana, Ebonyi State, Nigeria
2
Michael Okpara University of Agriculture, Umudike, Umuahia, Abia State, Nigeria.
ABSTRACT
MurrayaKoenigii (Linn) Spreng has been used for flavouring and spicing of food since ancient time. Its
medicinal value has also been identified. This work investigated the phytochemical screening, proximate
composition, vitamin and mineral contents of the plant leaves using standard procedures. The result of the
phytochemical screening revealed the presence of the bioactive constituents comprising flavonoid 7.43 ± 0.03%,
phenols 4. 25 ± 0.04%, saponins 2.50 ± 0.01%, alkaloids 1.90 ± 0.01%, tannins 0.86 ± 0.02% and glycosides 0.11
± 0.01%. The proximate composition was found to be as follows; carbohydrate 39.44 ± 0.04%, moisture content
23.42 ± 0.10%, crude fibre 6.30 ± 0.05%, ash content15.60 ± 0.21%, fats 6.48 ± 0.22% and protein 8.38 ± 0.02%.
The analysis of the vitamin content showed the presence of vitamin A (â-carotene) 6.04 ± 0.02 mg/100g, vitamin
C (ascorbic acid) 0.04 ± 0.002 mg/100g, thiamin 0.89 ± 0.01 mg/100g, riboflavin 0.09 ± 0.001mg/100g, niacin
2.73 ± 0.02 mg/100g and vitamin E 0.03mg/100g. The plant leaf was found to contain some important minerals;
Calcium 19.75 mg/100g, magnesium 49.06 ± 0.02 mg/100g, sodium 16.50 ± 0.21 mg/100g, potassium and zinc
0.04 ± 0.001 mg/100g each. Curry plant leaves contain some substantial amount of important phytochemicals
which possess anti-oxidant properties and some nutritive vitamins and minerals thus supporting its use as
medicinal plant and as food flavoring and spicing condiment.
Keywords: Phytochemicals, Minerals, Vitamins, Curry leaf, Anti-oxidant
INTRODUCTION
Humans have relied mostly on plants for nutritional and medicinal needs, Herbal plants provide most of the
medicinal needs. Important herbal products include spices, herbal teas, functional foods, medicinal raw
1
materials, essential oils,flavouring and dietary supplements . The medicinal use of plant is as a result of the
phyto-constituents present in them.Some of these chemicals are bioactive and produce definite physiological
and biochemical actions in humans and animals. They are known as secondary metabolites or
phytochemicals and comprises alkaloids,flavonoids, tannins, phenolics,saponin, steroids,
2
glycoside,terpenesetc . The use of plants in traditional medicine is of global interest.Traditional and herbal
medicines have been promoted as a source of less expensive and comprehensive medicare especially in
developing countries 3. Some of these medicines are easily available, cheaper and safer than these modern
synthetic drugs4. This led to the resurgence in the use and demand for medicinal plants as they play roles in
prevention and management of some health ailments such as diabetes , cancer, arthritis , degenerative
3
disorders like Parkinson and Alzheimer's diseases . The medicinal value of many plants remain unexploited.
Investigations are ongoing to discover novel drugs or templates for the development of new therapeutic
agents.
Murraya koenigii commonly known as curry plant belongs to the family Rutaceae. The plant is a medicinal
1
plant and native to India,Sri Lanka and other South Asian countries . It is usually found in tropical and sub-
tropical region and cultivated in China, Australia, Nigeria etc.The plant is highly valued for its leaves which
is used for flavouring and spicing of food.The curry leaf is believed to have several medicinal properties
such as anti-diabetic, anti-oxidant ,antimicrobial, anti-inflammatory, anticarcinogenic and hepato-protective
410
properties5. Curry leaves have the richest source of carbazole alkaloids such as koenigine, mahanimbine and
mueoninine extracted from the leaves which have been found to demonstrate anti-cancer and anti-oxidant
5
properties .The medicinal values of Murraya koenigii are numerous and beneficial to humans, hence this
work seeks to evaluate the phytochemical and proximate compositions; vitamin content and minerals
elements present in the plant.
SAMPLE COLLECTION:The curry plant leaves were purchased from New Market Ngwa Road , Aba in
Aba South L.G.A. Abia state. It was authenticated in the Department of Agricultural Technology, Akanu
Ibiam Federal Polytechnic, Unwana,Ebonyi state and deposited in the herbarium with voucher specimen
no. MK 00342.
SAMPLE PREPARATION: The plant sample was washed and dried on laboratory bench for 3 weeks. It
was ground to coarse particles using an electric blender and weighed to be 920g. The sample was later stored
until require for analysis. The analyses were carried out in the Chemistry Laboratory of Akanu Ibiam Federal
Polytechnic, Unwana. Ebonyi State,
PHYTOCHEMICAL DETERMINATIONS.
Two grams of sample was defatted with 100ml of diethyl ether.
Phytochemical determination was carried out using method of Harbone 6. The phytochemical include
alkaloids, saponins, flavonoids, saponin, tanins glycoside and phenols
Proximate Analysis:Proximate composition of the plant leaf was carried out according to the method
of(A.O.A. C. )7. Those analysed for include; carbohydrate, moisture content, ash content, protein , fat and crude
fibre.
Vitamin content determination: Vitamins A, C, E, and B-complex vitamins were determined using the method
of (Okwu and Josiah ;A.O A.C)8,7
Mineral elements analysis:The sample was first digested. Five gram of sample was digested with perchloric.
Then 50mls of nitric acid and 10mls of concentrated sulphuric acid were added . The mixture was then heated
until a clear solution was obtained and the mixture reduced to half of its original volume with the aid of hot plate.
The mixture was poured into 250ml volumetric flask and made up to mark with distilled water. The digested
sample was then analysed using Atomic Absorption Spectrophotometer with standards for each mineral
element.Sodium and potassium were determined using flame photometer.
RESULTS AND DISCUSSIONS
Table 1: Result of Phytochemical composition of Murraya koenigii plant leaf
Phytochemicals Values (%)
Alkaloids 1.90±0.01
Saponins 2,50±0.01
Flavonoids 7.43±0.03
Tanins 0.86±0.02
Phenols 4.25±0.04
Glycosides 0.11±0.01\
Table 2: Result of Proximate composition of Murraya koenigii plant leaf
Proximate composition Values (%)
Moisture content 23.4±0.10
Protein 8.38±0.02
411
Carbohydrate 39.44±0.04
Fats 6.48±0.22
Ash content 15.60±0.21
Crude fibre 6.30±0.05
Table 3: Result of vitamin content Murraya koenigii plant leaf
Vitamins Values (mg/100g)
Vitamin A (B-carotene) 6.04±0.02
Vitamin C (ascorbic acid) 0.04±0.002
Vitamin E (tocopherol) 0,03±001
Vitamin Bi (thiamin) 0.89±0.01
Vitamin B2 (riboflavin) 0.09±0.002
Vitamin B3 (niacin ) 2,73±0.02
Values are triplicate determination ±std deviation
Table 4: Result of Mineral elements of Murrayakoenigii plant leaf

Minerals Values (mg/100g
Calcium 19.73±0.02
Iron 0.16±0.01
Magnesium 49.06±0.02
Sodium 16.50±0.21
Zinc 0.04±0.001
Potassium 0.04±0.001
The phytochemical composition of the Murraya koenigii leaf is shown in Table 1.Flavonoid content was found
to be the highest7.43±0.03% whileglycoside was detected to be the least. Flavonoids have been shown to possess
9
anti-fungal and anti-bacterial activity . Flavonoids have strong anti-oxidant and anti-inflammatory properties.
10
They have ability to scavenge hydroxyl radicals, superoxide anions and lipid peroxy radicals . Thesefree
radicalshave been implicated in causing some age-long diseases such as diabetes, cardiovascular, Parkinson and
Alzheimer's diseases.The presence of flavonoids in the curry leaf may account for its use in treatment of these
diseases. Saponin was detected in an appreciable amount 2.50±0.01%. Saponins are known as immune booster.
Plants rich in saponins have been shown to demonstrate anti-inflammatory,cholesterol lowering and ant-cancer
properties11. Alkaloid was found to be 1.90±0.01% in the curry leaf. Alkaloids have anti-microbial properties
owing to their ability to intercalate with DNA of the micro-organisms12. Phenol was found to be 4.25±0.04%.
13
Urquiaga and Leighton reported that phenols and phenolics have anti-tumour and antioxidant effects . They
also have anti-inflammatory and anti-carcinogenic properties and play roles in scavenging H2O2 by donating
electrons to the
14
peroxide thus neutralizing them to water .
Table 2 reveals the proximate composition of Murraya koenigii leaf. The plant leaves contain a substantial
amount of carbohydrate 39.44±0.04% and moisture 23.42±0.10%. This shows that the leaf is a good source of
carbohydrate while the moisture content is high indicating a low shelf –life,so the plant material can be preserved
by drying it to retain other useful components. The low amount of fat indicates that the vegetable is not a good
15
source of lipid accumulation which can cause artherosclerosis and aging . The proteinsin the leaf can make fair
contributions to protein diet as proteins are involved in formationof hormones, enzymes and structural
412
membranes.M. koenigii contains a good amount of dietary fibre. Dietary fibre lowers cholesterol level, risk of
16
coronary heart disease, diabetes and cancer . Vitamin content of the M.koenigii is shown in Table 3. Vitamins
are found in trace amounts. Niacin vitamin B3 was found to be high 2.73±0.02mg/100g compared to other
vitamins.Vitamins A and C were found to be low 6.04±0.02mg/100gand 0.04±0.02 respectively. Niacin helps to
17
lower and regulate cholesterol level and helps in maintaining good blood circulation . Vitamin A helps to
provide good vision and healthy immune system. It also fights cancer by inhibiting the production of DNA in
cancerous cells18. Vitamins C and E are strong anti-oxidants. They are good in fighting cancer through their
scavenging of free radicals. Other B-groups vitamins B1 thiamine, B2riboflavine were also detected. Riboflavine
helps in production of red blood cells and is important for growth and healthy body. The presence of these
vitamins in the plant leaf has shown that the leaf possesses strong medicinal and nutritive values.
Table 3 gives the mineral content of the curry plant leaf. Magnesium was found to be the highest
49.06±0.02mg/100g followed by calcium 19.75±0.02mg/100g while zinc and potassium were found to be low
0.04±0.001mg/100g each. Mineral and trace elements play important roles in immune function and health.The
high content of magnesium proves that the curry leaf can be a good source of magnesium which is an activator of
many enzyme systems and maintain electrical potential in nerves19. Calcium was detected in an appreciable
amount. The M. koenigiileaf can serve as food supplement for calcium. Calcium combines with phosphate to
20
enhance strong bone and teeth formation . Sodium content was also found to be high 16.50±0.21mg/100g.
Sodium and potassium are needed in body fluids and nerves for transport and balance. The plant leaf contains
zinc ,though in small amount, zinc is important in the body as it enhances immunity, cell growth and insulin
regulation. It is used in restoring dehydration and stoppage of diarrhea. The presence of the mineral elements in
the plant leaf shows that consumption of M.koenigii can be used as a supplement for these essential nutrients.
CONCLUSION: This study has shown that Murraya Koenigii (curry) leaf used as spice and flavouringagent in
food contains substantial amount of phytochemicals and phytonutrients. The proximate analysis showed high
content of carbohydrate, protein ,fibre and fat. The presence of the vitamins and mineral elements in the plant
leaves showed that it could be consumed to supplement these scarce nutrients. With its content of these anti-
oxidants flavonoids, phenols,vitamins E and C, curry leaf possess anti-cancer and cardio-protective agents
supporting its use as medicinal plant.
REFERENCES
1) Harish, K.H., Pandith, A. &Shruthi, S.D. (2012). A review on MurrayakoenigiiMultipotential
medicinal plant. Asian Journal of Pharmaceutical and Clinical Research 5(4): 5-14.
2) Ujuwundu, C. O. ,Okafor O.E, Agha, N.C.,Nwaogu, L.A, Igwe, K.O. &Igwe, C.U. (2010).
Phytochemical and chemical composition of Combretumzenteri leaves, Journal of Medicinal Plant
Research4 (10): 965-968.
3) Kaur, G. Daftardar, S. &Barve , K. H. (2014). Modifying anti-inflammatory effect of Diclofenac
with Murrayakoenigii: Recent patent on inflammation and allergy. Drug Discovery8(1): 77-81.
4) Summan, S., More, P.K. &Sandhya, M.M. (2014).Curry leaves (Murrayakeonigii LinnSpreng) a
miracle plant. Indian Journal of Science Research, 4 (1): 46-52.
5) Kirupa, S.L.S.&Kariitha, R. (2015).Anti-oxidant enhancing property of curry leaf powder M.
keonigii in type ll diabetes mellitus. Intl Journal of Phamacy&Biosciencs 6 (1)
507-514.
6) Harbone, J.B.(1973). Phytochemical method, Chapman and Hall, London, Pp 110-113.
7) A.O.AC.(1994). Official method of analysis 15ed , Association of Official Analytical Chemist,
413
Benjamin Franklion Station Washington D. C. U.S.A.
8) Okwu, D. E. & Josiah, C. (2006): Evaluation of chemical composition of two Nigerian medicinal
plants. African Journal of Biotechnology5(4): 357-361.
9) Galeoti, F., Barile, E., Curir, P., Dolci, M &Lanzotti, V. (2008). Flavonids from Dianthus
Caryophyllus and their anti-fungal activity, Phytochem. Lett1: 44
10) Allan, L.& Miller, N. D. (1996). Anti-oxidant flavonoids, function ,structure and clinical usage. Alt
Med. Rev. 1: 103-111.
11) Sauvauire, Y. ,Baissac,Y., Leconte, O. &Ribes, G. (1996): Steroid saponinns from Fencegreek and
some of their biological properties In: Saponins used in food and Agriculture. Advances in Experimental
Medicine and Biology405: 37-46.
12) Kasolo, J. N., Gabriel, L.O. Biamenya, O,J. &Ogwal-Okeng, J. W.(2010). Phytochemicals and uses
ofMoringaOleifera leaves in Uganda rural communities. J.Med, Plant Res. 4 (9): 753-757.
13) Urquiaga, I & Leighton, F. (2000). Plant polyphernol anti-oxidants and oxidative stress, Biological
Research33; 159-160.
14) Mikail, P., Madirad, S.,Molougdizargari, M. &Sarahroodi, S. (2013). Therapeutic uses and
pharmacological properties of garlic, shallot and their biological active components, Iran Journal of
Basic Med. 16 (10): 1031-1048.
15) Bamishaiye, E.I., olayeme, F.F. &Bamishaiye , O. M. (2011) Proximate and phytochemical
composition of Moringaoleifera leaves at three stages of maturation, Advance Journal of Food
andTechnology3(4): 233-237.
16) Ishida, H. ,Suzuno, H., Sugiyama, N., innami, S., Tadokoro, T., &Maekawa, A.(2000). Nutritive
evaluation chemical components of leaves, stalks and stems of sweet potatoes (Ipomoea batatapoir).
Food Chem, 68: 359-368
17) Okwu, D. E. (2004).Phytochemicals and vitamin content of indigenous spices of South Eastern
Nigeria. J Sustain. Agric. Environ. 6(1): 30-37.
18) Brett, J. (2013). Health/vitamins and supplements facts. www.howstuffworks.com Retrieved
14/03/16.
19) Olusanya, J.O.(2008).The food nutrients In: Essentials of Food and Nutrition, Apex book Limited
Lagos xxi
20) Uwakwe, A. A.&Ayalogu, E. O. (1998). Nutritional composition of Food and some Nigerian fruits.
Journal of Food Technology 6(1) 30-37.
414
PHYTOCHEMICAL STUDIES AND APHRODISIAC ACTIVITY OF Byrsocarpus coccineus
ROOT EXRACT
By
*1
Balarabe M. M., Mann A., Ndamitso M. M. Adeyemi O. Y. H. and 3Nma N. Y.
2 2
*Corresponding Author: balamah99@yahoo.com

08067570013, 08050602652
1
Department of Physical Sciences, Niger State Polytechnic, Zungeru.
2
Chemistry Department, Federal University of Technology, Minna.
3
Department of Science Laboratory Technology, Federal Polytechnic Bida.
Being a Paper Presented at the 39th Annual International Conference of Chemical Society of Nigeria
(CSN)
th rd
Date: 18 – 23 September, 2016
Venue: Law Faculty, Rivers State University of Science and Technology (RSUST), Port Harcourt,
Nigeria.
Abstract
Sexual dysfunction have becomes a global issues for male and female due to disease condition, medication
and even stress. Plants have served as primary healthcare for the rural dwellers for several centuries. The
aim and objectives of this study is isolate bioactive constituents which have serve as stimulant for sexual
drive in man, from Byrsocarpus coccinues (short pod) root extract. Phytochemical screening was carried out
on the crude and the ethyl acetate fraction of the root extract and all the parameters were significant in both
except for phlobatannins and reducing sugar in the fraction. Thin-layer chromatography revealed four
different spots with their respective Rf values. An aphrodisiac activity of both the crude extract and the ethyl
acetate fraction was carried out on experimental wistar albino rats. Both shows activity, but ethyl acetate
fraction has the highest significant in the entire dose. This is an indication that the ethyl acetate fraction
contains relatively more the phytoconstituents responsible for sexual activity than the crude extract.
Therefore, Brysocarpus coccinues root extract could be recommended for the treatment of low libido and
sexual dysfunction in human beings as claimed by the traditional healers.
Keywords: Phytochemicals, Aphrodisiac activity, Wistar albino rats, Byrsocarpus coccinues, Thin-layer
chromatography, GC-MS.
Introduction
Byrsocarpus coccinues shrub up to 2m high with light brown stem and very fine lenticel. Leaves are pinnate,
[1]
petiole 1-2cm long with rachis and 6 pairs of oval glabrous. Fruits are capsular, glabrous, oblong and
yellow about 15mm long by 7mm broad. Slightly convex at one side, and showing a black seed with orange
aril. [2] The other names for Byrsocarpus coccineus are as follows; English (Short pod), Nupe (Darabagi),
Yoruba (Kanti-kanti), Hausa (Kimbar maharba), Fulani (Wangarabubi), Igbo (Oke abolo or Mybo-
[1,2]
apepea) and Bassange (Kogi).
415
Plants have been the source of medicinal agents since earliest times, and today have continued to play a dominant
role in the local and primary health care of about 80% of the world's population [3]. In Nigeria, more than 70% of
the estimated 140 million people are rural dwellers, who depend largely on indigenous herbal medicines as
[2,4]
source of their health care .
Ethno-medicinal studies are very important in order to understand the social, cultural and economic factors
influencing ideas and actions concerning health and illness and also to get information on types of diseases and
health problems prevalent among the people of a particular locality. Such studies may help to provide the basic
health-care services to the greater part of the rural resources and poor populations in an effective way, provided
that such studies are conducted hand-in-hand with phytochemical, pharmacological and perhaps clinical studies
[5]
Natural stimulants are types of stimulants that increase energy, reduced appetite or improve mental focus and
clarity naturally. The most common natural stimulants include caffeine, cocaine and nicotine but many other
natural stimulants also exist. There are a variety of natural stimulants found throughout the world, and these are
typically referred to as herbal stimulants. Some of these herbal stimulants include ginseng, tyrosine, and coconut
oil. Most of the stimulants are roots, herbs or foods such as: guarana, ginsing, L-tyrosine, gotukula, B-vitamins,
[5]
chlorophyll, coconut oil and taurine .
Collection of Plant Material; Roots sample of Byrsocarpus coccineus was collected in a bush opposite the
Central Gate of Niger State Polytechnic, Zungeru, Wushishi Local Government Area of Niger State, Nigeria in
August 2014. The root sample was collected by digging, properly washed and kept in plastic sealable bag which
was taken to the laboratory and dried under laboratory condition for six (6) weeks. The dried root sample was
grounded to coarse powder using pestle and mortar, and preserved in a dry plastic bag for extraction.
Methods; The extraction was done using cold maceration method, in which 500 g of the dried powdered root
3 3
materials was accurately weighed into a clean plastic rubber containing 700cm methanol and 300cm distilled
water (70:30 solvent), covered and allowed to stay for 72 hours[9]. The phytochemical screening was done using
standard methods described by [9,10,11,12]. The partitioning of the crude extract into four solvent soluble fractions
[11,12]
and column chromatography were determined by methods described by . Wistar albino rats were obtained
from the animal house of the Department of Biological Sciences, Niger State Polytechnic, Zungeru and the
aphrodisiac screening was conducted in the same school using methods described by [13]. The GC-MS analysis of
the ethylacetate fraction was carried out at Chemical Engineering Department of the University of Ilorin, Nigeria
[14,15]
using a GC-MS machine (QP-2010; Shimadzu, India) according to .
Results
Table 1: Phytochemical screening of Byrsocarpuscuccinues
Secondary metabolites Tests Crude extract Ethyl acetate fraction
Flavonoids Shinoda test + +

NaOH test + +
Tannins Ferric chloride test + +
Lead acetate test + +
Saponins Froth test + +
Kede test + +
Alkaloids Mayers test + +
Dradendroff’s test + +
Steriods/tripterpenes Liebermann-burchard test + +
Salkowski test + +
Cardiac glycoside Keller-kiliani test + +
416
Anthraquinone Borntragers test + +
Combined test + +
Phlobatannins HCl test - -
Reducing sugar Molisch test + -
Fehlings test + -
Keys; + = present, - = absent
Table 2: Preparative Thin Layer Chromatography of Ethyl acetate fraction with Solvent mixture of
(9:1).
Spots Colour of components RF Values
1 yellow 0.15
Aphrodisiac Screening of both the crude extract and the ethyl acetate fraction
Table 3. Activity of crude root extract of Byrsocapus cuccineus on male wistar albino rats for three days
after administration.
Group Concentration (mg/kg) Mount Frequencies

Day 1 Day 2 Day 7
A Negative control 0.0 1.0 0.0
B Positive control 12.0 7.0 2.0
C 50 0.0 1.0 0.0
D 100 1.0 1.0 0.0
E 200 5.0 1.0 1.0
F 400 9.0 5.0 2.0
Table 4. Activity of ethyl acetate fraction of Byrsocapus coccineus on male albino rats for three days after
administration.
Group Concentration (mg/kg) Mount Frequencies

Day 1 Day 2 Day 7
A Negative control 0.0 0.0 1.0
B Positive control 13.0 5.0 3.0
C 50 0.0 2.0 1.0
D 100 2.0 2.0 1.0
E 200 5.0 2.0 1.0
F 400 10.0 6.0 2.0
Table 5: Result of GC -MS Showing Molecular Masses, Percentage Areas and Major Peaks of
Some of the Constituents of the Ethylacetate Soluble Fraction
417
Line Compound Mol. Mass Area (%) Peaks (m/z)
3 4-cyclopentene-41,3-dione 96 6.86 (96), 68, 54.
2 Aziridine 112 6.18 (112), 98, 82, 70,

55.
3 n-butyric acid 172 3.44 (172), 139, 112, 83,

71, 55.
4 Pantalactone 130 30.60 (130), 85, 76, 71,
57.
5 2-cyclohexen-1-one 161 5.20 (161), 122, 94, 66.
6 7-tridecanone 198 4.41 (198), 155, 128,

133, 71, 58.
5 Phenol, 2-methoxy 124 4.88 (124), 109, 81, 53.
Discussion
This research was carried out to evaluate the traditional claim of aphrodisiac and sexual stimulant activities of
byrsocarpus coccinues using wistar albino rats. Phytochemical screening was observed on both the crude extract
and the ethyl acetate fraction using standard methods. The results revealed that both the crude and ethyl acetate
fraction have almost the same significance except for phlobatannins absent in the crude extract, while
phlobatannins and reducing sugar absent in the case of ethyl acetate fraction. These results agreed with the
reports of [13], and it was also reported by [14] that extract of Brysocarpus coccinues have been used for the treatment
of gonorrhea, pile, fever, diabetes, hypertension and even ear problem.
The ethyl acetate fraction gave four (4) spots with RF values of 0.15, 0.63, 0.77 and 0.83 respectively. Each spot
revealing different colour as shown table 2 above, and it is an indication that the ethylacetate fraction contains
combined phytochemicals which are responsible for aphrodisiac activities and other bioactivity such as
[15]
treatment of sexual dysfunction. This also have a close agreement with the result of using Asparagus
racemosus.
The mating behavior study of the experimental groups revealed that the dose or concentration of the prepared
extract significantly increased mount frequency and intromission frequency, when compared with the control
[15]
groups and it agreed with . Although its significance is less than that of positive control but higher than the
negative control standard. The negative control shows very low significant, while the positive control standard
shows the highest degree of mount frequency, intromission and ejaculation latencies when compared with the
test or prepared extract. This is an indication that libido and potency were enhanced by the prepared
phytoconstituents. The enhancement of sexual behavior on the tested animals was more prominent with increase
in dose of administration of the tested plant extract.
However, hormonal modulations have been reported to be attributed to the presence of steroidal saponins
protodiocin in the plant root extract. Protodioscin has been observed to show improvement in libido, sexual
[13,14]
activity and intracavernous pressure in experimental animals .
Conclusion
418
The study concluded that the cumulative dose of the phytoconstituents can enhance overall sexual performance
in male rats increasing spermatozoa and testosterone concentration. The results of this research work suggest that
the tested extract could serve as alternative to the orthodox drug and could be used to improve sex life.
Recommendations
1. More pharmacological studies should be embark on the whole plant to discover its medicinal potentials
2. Toxicological studies should also be done on plant extract
3. Isolation and characterization of the active phytoconstituent responsible for sexual drive be carried out.
4. Research be carried out on its nutritional and other medicinal properties.
5. The general public should be educated on the potentials of Byrsocarpus coccinues root extract.
Acknowledgement
To my mentor Dr A. Mann Department of Chemistry Federal University of Technology Minna, for his advice and
support towards achieving the set goal. I also want to acknowledge Dr M.T. Yakubu, University of Illori and Dr
O. H. Y. Adeyemo Department of Biochemistry, Federal University of Technology Minna.
References
1. C.H. Wasis, J.A. Anuka, S.Y. Timothy, A.U. Zezi, & I.M. Hussaini (2013). Preliminary
Phytochemical Screening and Invitro Effects of Leaf Extracts of Byrsocarpus CoccineusShum &
Thonn on Pregnant Rat Uterus. International Journal of Pharmacy Teaching & P r a c t i c e s 4 ( 2 ) ,
649-654.
st
2. A. Mann, M. Gbate, & A. Nda-umar (2003). Medicinal and Economic Plants of NupeLand 1 E d i t i o n
Jube Evans Books & Publications P. 85.
3. A.A. Ahmadu, I.M. Akpulu, H.S. Hassan, M.I. Sule & U.U. Pateh (2006). Preliminary Phytochemical
and Antimicrobial Screening of the Leaves of Byrsocarpus coccineusSchum and Thonn. (Connaraceae).
Journal of Pharmacy and Bioresources 3(2), 107-110.
4. A.J. Akindele & O.O. Adeyemi (2007). Anti-inflammatory Activity of the Aqueous Leaf Extract of
Byrsocarpus coccineus. Fitoterapia, 78, 25–28.
5. I.O. Lawal, N.E. Uzokwe, A.B.I. Igboanugo, A.F. Adio, E.A. Awosan, J.O. Nwogwugwu, B. Faloye,
B.P. Olatunji & A.A. Adesoga (2010). Ethnomedicinal Information on Collation and Identification of
some Medicinal plants in Research Institutes of South-West Nigeria. African Journal of Pharmacy and
Pharmacology 4(1), 001-007.
6. R.H. Howland (2008). "Lisdexam fetamine: A Prodrug Stimulant for ADHD". Journal of Psychosocial
Nursing and Mental Health Services 46 (8), 19–22.
7. T.J. Spencer, A. Brown, L.J. Seidman, E.M. Valera, N. Makris, A. Lomedico, S.V. Faraone & J.
Biederman (2013). "Effect of Psychostimulants on Brain Structure and Function in ADHD: A Qualitative
Literature Review of Magnetic Resonance Imaging-based Neuroimaging Studies". J. Clin. Psychiatry
74 (9), 902–917.
8. K.A. Montgomery (2008). "Sexual Desire Disorders". Psychiatry (Edgmont) 5 (6), 50–55.
9. J.B. Harborne (1998). Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. 3rd
Edition Chapman and Hall Co, New York.1-302.
10. A. Sofowora (1993). Medicinal Plants and Traditional Medicine in Africa. Spectrum Books L t d . ,
Ibadan. Pp. 95-238.
11. A. Mann (2007). Survey of Ethnomedicine for the Treatment of Tuberculosis: Chemistry Perspective.
Ayanwola Printing Works, ISBN 978-978-085-536-9, Minna, Niger State, Nigeria. 117.
419
12. G.E. Trease and W.C. Evans (2009). Pharmacognosy.10th Edition, ELBS/BailliereTindall, London, Pp.
185-187, 340-346.
13. A.W. Javeed, N.A. Rajeshwara & R.K. Nema (2011). Phytochemical Screening and Aphrodisiac
Activities of Asparagus racemosus. International Journal of Pharmaceutical Sciences and Drug
Research, 3(2), 112-115.
14. S. Amos, L. Binda, O.F. Kunle, C. Wambebe & K. Gamaniel (2002). Uterotonic Properties of the
Ethanolic extract of Byrsocarpus coccineus. Pharmaceutical Biology, 40, 33-38.
15. O.O. Adeyemi, A.J. Akindele & K.I. Nwumeh (2010). Acute and Subchronic Toxicological
Assessment of Byrsocarpus coccineus Schum and Thonn. (Connaraceae) Aqueous Leaf Extract.
Journal of Biochemistry 3 (2), 1-11.
420
GC-MS analysis of leaf, stem-back and root extracts of Alstonia boonei
1 1 2
Babatunde O*. ; Godwin R. E. ; Wahab N.O.
1
Department of Chemical Sciences, Ajayi Crowther University, Oyo, Nigeria.
2
Tai Solarin University of Education, Ijagun, Ijebu-Ode, Ogun state, Nigeria
Corresponding author: toyeenbab@yahoo.com

ABSTRACT
Medicinal plants have had a crucial role in human culture and civilization. Alstonia boonei De Wild is a
medicinal plant commonly found in West African and is popularly known as God's tree. The plant parts have been
traditionally used for painkiller, antimicrobial, antimalarial and antidiabetic which have also been proved
scientifically. Previous studies revealed little information on phytochemcial components ofAlstonia boonei. The
present work aims at investigating and comparing the chemical components of the leaf, the stem and the root of
the plant in order to provide sufficient baseline information for future work and for commercial exploitation.
Leaf, stem and root extracts of Alstonia boonei were prepared by maceration using 1:1 EtOAc/MeOH and
100% DCM as extraction solvents. Gas chromatography and Mass spectroscopy studies were performed
to identify the phytochemical constituent of the plant.
The GC-MS analysis of DCM extract of the leaf revealed ten chemical components with Eugenol as major
component (54.58%); DCM extract of the stem showed forty one components with alpha-amyrin (32.25%)while
DCM extract of the root revealed twenty components with 1,2-benzenedicarboxylic acid (49.2%) as major
component.
This study shows that the A. boonei extracts of the leaf, stem-back and root consist of different types of
compounds with few compounds common to two of the parts. The most identified compound by GC/MS
were Eugenol, benzenedicarboxylic acid and alpha-amyrin
Keywords:Phytochemicals, Alstonia boonei, Gas chromatography/Mass Spectroscopic Studies
Introduction
Alstonia. Alstonia boonei De Wild belongs to the family Apocynaceae. It'sa herbalmedicinal plant ofWest
African origin, popularly known as cheese wood and known locally among Yoruba in Nigeria as Ahun.All the
parts of the plant are very useful but the thick bark cut from the matured tree is the part that is most commonly
used for therapeutic purposes.Therapeutically, the bark has been found to possess antirheumatic, anti-
inflammatory (Abbiw 1990), analgesic, antimalaria, antidiabetic, antihelminthic, antimicrobial and antibiotic
properties (Hadi and Bremner, 2001; Fakae et al., 2000; Kam et al., 1997).A wide array of chemical compounds
has been isolated from Alstonia boonei. These include alkaloids, tannins, iridoids, and triterpenoids (Ayiku,
1992). The alkaloids isolated from the plant include echitamine,echitamidine and voacangine(Ayiku, 1992;
Kucera et al., 1972; Croquelois et al.,1972;Oguakwa, 1984). Echitamine possess a battery of pharmacological
and autonomic activities (Ojewole,1984 and Ojewole,1983) including anticancer activities (Chandrasekaran
and Nagarajan ,1983;Saraswathi et al.,1997;Saraswathi et al.,1998a;Saraswathi et al.,1998b;Saraswathi et
al.,1999;Viswanathanet al .,1997;Kamarajanet al.,1991; Keawpradubet al., 1997)
Iridoids isolated from Alstonia boonei include boonein and loganin. Loganin is a key intermediate in the
biosynthesis of indole alkaloids.The triterpenoids isolated from Alstonia boonei include lupeol, ursolic acid and
â-amyrin.
Material and Methods
Sample preparation
421
Alstonia booneileaf, stem-back and root were collected at a farmland in Akinmarin village, Oyo and were
identified at the Department of Botany, University of Ibadan, Ibadan. The air-dried plant materials: (204.7 g)
leaves, (282.5 g) stem-bark, and (102.9 g) root of, Alstonia boonei, exhaustively extracted separately with 1:1
EtOAc:MeOH for 3days. The extracts were filtered separately, concentrated on rotary evaporator to about 50
mL and evaporated to dryness under vacuum. Each extract wasmarcerated with DCM to obtain DCM fraction
of the three parts. Obtained fractions were evaporated to dryness and stored in a refrigerator at +4°C until use.
Gas Chromatography (GC)–Mass Spectrometer (MS) Analysis

GC-MS: Hewlett-Packard 5890 gas chromatograph,combined with a Jeol JMS-HX 110 mass spectrometer
with sourceat 270°C at 70 eV. Injector was set at 270°C with splitting ratio1:30. The analysis was performed on
the aforementionedprogramme on equivalent column HP-5 (25 m × 0.22 mm and 0.25ìmdf). A mass spectral
survey was performed using the NISTmass spectral search program.
GC-MS analyses of DCM fraction of the leaf, the stem-back and the root column of Alstonia boonei are
presented in Table 1. Constituents were listed in order of elution from HP-5 capillary column (Table 1). Gas
chromatography-mass spectrometry analysis of the leaf extract resulted in the identification of ten (10)
compounds, forty one (41) compounds in stem-back extract and twenty (20) compounds in root extract.
Table 1: Chemical composition of leaf, stem-back and root of Alstonia boonei
S/N Constituent % composition
Leaf Stem-back Root
1 Eugenol 54.58(9.730) 0.14(9.722) -
2 4-Tetradecene - - 1.39(10.185)
3 Caryophyllene 6.99(10.631) - -
4 Phenol 2.01(12.148) - 0.56(11.970)
5 1H-2-Benzopyran-1-one - 0.54(12.434) -
6 Cyclododecane - 0.28(12.943) -
7 Dichloroacetic acid - - 4.83(12.943)
8 Hexadecane - 0.16(13.033) 1.32(113.029)
9 2H-Pyran-2-one - 1.54(13.161) -
10 2-Ethyl-3-methoxypyrazine - 0.34(14.334) -
11 1-Octadecene - 0.37(15.324) 6.59(15.323)
12 Octadecane - 0.15(15.398) 1.35(15.392)
13 Cyclononasiloxane - 0.20(15.713) -
14 Bicyclo(3.1.1)heptane 2.61(15.833) - -
15 1,2-Benzenedicarboxylic acid - - 49.20(16.210)
16 Phthalic acid - 0.15(16.216) -
17 7,9-Di-tert-butyl-1-oxaspiro(4,5) deca-6,9- - - 1.13(16.777)
diene-2,8-dione
18 Hexadecanoic acid 2.13(16.765) 0.67(16.777) -
19 n-Hexadecanoic acid 5.40(17.183) 1.40(17.303) -
20 Dibutyl phthalate - - 2.61(17.200)
21 6H-Furo[2',3':4,5]oxazolo[3,2-a]pyrimidin- - 0.24(17.338) -
6-one
22 Heptafluorobutyric acid - 0.25(17.458) -
23 n-Nonadecanol-1 - 0.28(18.391) 5.90(17.452)
24 Eicosane - 0.16(20.365) 1.34(17.509)
25 Ethanone - - 1.30(17.767)
26 1-Heneicosanol 2.88(17.950) - -
27 Behenic alcohol - - 3.21(18.373)
28 9,12-Octadecadienoic acid - 0.34(18.488) -
422
29 11-Octadecenoic acid - 0.26(18.545) -
30 Phytol 1.63(18.665) - -
31 3-Isopropoxy-1,1,1,7,7,7-hexamethyl-3,5,5- - 0.33(18.820) -
tris(trimethylsiloxy)tetrasiloxane
32 Oleic Acid - 0.46(18.986) -
33 1-Octadecene - 0.19(19.192) -
34 1-Docosene 13.28(22.585) - 3.65(19.392)
35 2-Eicosanol - 0.14(19.398) -
36 Docosane - - 0.66(19.438)
37 6-Acetamido-2-methylbenzothiazole - 0.78(20.090) -
38 Cyclotetracosane - - 2.56(21.337)
39 Hexasiloxane - 0.40(21.686) -
40 2(3H)-Furanone - - 0.82(22.430)
41 1-Heptadecene - - 0.92(22.659)
42 Tetratetracontane - 0.95(22.693) -
43 Oxirane - 0.27(23.186) -
44 Olean-12-ene - 0.63(24.948) -
45 1S,6R,9S)-5,5,9,10- - 1.29(25.245) -
A comparative analysis of chemical profiles of the leaf and stem-backshowed thatHexadecanoic acid and n-
Hexadecanoic acid were present in the two parts but in higher proportion in the leaf while phenol and 1-Docosene
were present in the leaf and the root. Hexadecane, 1-Octadecene, Octadecane, n-Nonadecanol-1 and Eicosane
were present in both stem-back and root.
The results revealed that Eugenol(54.58), 1-Docosene(13.28),2,6,10,14,18,22-Tetracosahexaene(8.50),
Caryophyllene(6.99) and n-Hexadecanoic acid (5.40) were found as the five major components in the leaf
extract, the minor compound was Phytol. The major constituents of stem-back extract werealpha-amyrin
(32.25), 2(1H) Naphthalenone (10.32),9,19-Cycloergost-24(28)-en-3-ol (9.13)and Urs-12-en-3-ol (7.76) while
the major component found in root were 1,2-Benzenedicarboxylic acid (49.20), 1-Octadecene(6.59) and n-
Nonadecanol-1(5.90).
Conclusion
The present study represents the comprehensive analysis of phytochemical constituents ofAlstonia booneileaf,
stem-back, and root DCM extracts. The presence of various phytochemicals contributes to the medicinal activity
of the plant. The results will form the basis for selection of plant species for further investigation in the potential
discovery of new natural bioactive compounds.
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Plants.Kew,London:Intermediate Technology Publications, Royal Botanical Garden.
Croquelois G, Kunesch N, Debray M, Poisson J.(1972) Alstoniaboonei alkaloids. Medicinal Plants and
Phytotherapy.;6(2):122–127.
Hadi S, Bremner JB. (2001); Initial studies on alkaloids from Lombok medicinal plants.
Molecules. 6(2):117–129.
Fakae B.B, Campbell AM, Barrett J, et al. (2000); Inhibition of glutathione S-transferases (GSTs)
from parasitic nematodes by extracts from traditional Nigerian medicinal plants.
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Kam TS, Nyeoh KT, Sim KM, Yoganathan K. (1997) Alkaloids from Alstoniascholaris.
Phytochemistry.;45(6):1303–1305.
423
Ayiku MNB. Ghana Herbal Pharmacopoeia. Technology Transfer Centre; 1992.
Kucera M, Marquis VO, Kucerova H. (1972); Nigerian medicinal plants. I. TLC thin-layer
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Phytochemistry.;23(11):2708–2709.
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of Alstoniaboonei L. (Apocynaceae) International Journal of Crude Drug
Research.;22(3):121–143.
Ojewole JAO(1983). Autonomic pharmacology of echitamine, an alkaloid from Alstoniaboonei De Wild.
Fitoterapia.;54(3):99–113.
Viswanathan S, Ramamurthy N, Subramanian S, Mathuram V, Govindasamy S. (1997)
Enhancement of the cytotoxic effects of Echitamine chloride by vitamin A: an in vitro
study on Ehrlich ascites carcinoma cell culture. Indian Journal of
Pharmacology.;29(4):244–249.
Saraswathi V, Ramamoorthy N, Subramaniam S, Mathuram V, Gunasekaran P, Govindasamy S.

(1998) Inhibition of glycolysis and respiration of sarcoma-180 cells by echitamine
chloride. Chemotherapy. ;44(3):198–205.
Saraswathi V, Subramanian S, Ramamoorthy N, Mathuram V, Govindasamy S.(1997) In vitro
cytotoxicity of echitamine chloride and adriamycin on Ehrlich ascites carcinoma cell
cultures. Medical Science Research.;25(3):167–170.
Saraswathi V, Shyamala AC, Subramanian S, Govindasamy S. (1998) Studies on the effects of
echitamine chloride on serum glycoproteins and lysosomal hydrolases of sarcoma-180
induced mice. Fitoterapia.;69(1):73–75.
Saraswathi V, Mathuram V, Subramanian S, Govindasamy S(1999). Modulation of the impaired
drug metabolism in sarcoma-180 bearing mice by echitamine chloride. Cancer
Biochemistry Biophysics.;17(1-2):79–88.
Kamarajan P, Sekar N, Mathuram V, Govindasamy S. (1991) Antitumor effect of echitamine
chloride on methylcholonthrene induced fibrosarcoma in rats. Biochemistry International.
;25(3):491–498.
Keawpradub N, Houghton PJ, Eno-Amooquaye E, Burke PJ. (1997); Activity of extracts and
alkaloids of Thai Alstonia species against human lung cancer cell lines. Planta Medica.
63(2):97–101.
424
Isolation and Characterization of 2, 4 - Dihydroxycinnamic Acid from the Stem Bark
of Adina microcephala Delile
1,2
R.G. Ayo, J.D. Habila, J. I. Achika and 2O.O. Akinwande
2 2 1
Samaru
College of Agriculture, Ahmadu Bello University, Zaria
2
Department of Chemistry, Ahmadu Bello University, Zaria
*Corresponding author's email: gbekeayo@yahoo.com
ABSTRACT
Adina microcephala which belongs to the family Rubiaceae is widely distributed in Africa. It is used in
African traditional medicine for the treatment of diarrhoea and other stomach or digestive tract problems. A
microcephala stem bark was collected, identified, dried and pulverized. The pulverized plant material was
subjected to Soxhlet extraction using petroleum ether (60-80 °C), chloroform, ethyl acetate and methanol.
The ethyl acetate extract was subjected to further purification by chromatographic techniques to yield a
compound labeled R1 which was then fully characterized as 2, 4-dihydroxycinnamic acid using IR, 1D and
2D NMR spectroscopic techniques.
Keywords: 2,4 dihydroxycinnamic acid, Adina microcephala, Rubiaceae, stem bark

INTRODUCTION
Adina microcephala Delile is an evergreen plant and a small to large tree of up to 40 m tall, with slender branches
near the base. Its synonymous are: Breonadia salicina, Breonadia microcephala and Nauclea microcephala [1].
The root decoctions of Adina microcephala are drunk as a purgative in Tanzania and against tachycardia in South
Africa. An infusion of the powdered stem is given to breast-fed children in Ethiopia for the treatment of diarrhoea
and vomiting. In Nigeria the plant is locally called hakoren babauniya or káïányàr kaúrmìì or káïányàr ràfíí (
Hausa) and emido in Yoruba. It is commonly known as Africa teak or water shea-nut [2]. The bark of A.
microcephala is rich in tannins [3]. Leaf extracts of Breonadia salicina have also been found to reduce activity of
both Gram-negative and Gram-positive bacteria [4]. It is believed that these same anti-bacterial properties of
Breonadia salicina can be used for the preservation of foods [5]. Antifungal compounds isolated and
characterised from the leaves of Breonadia salicina protect oranges from infection [6]. Cinnamic acids are used
as precursor for the synthesis of commercially important cinnamic esters, which are obtained from various plant
sources and find application in flavourings, synthetic indigo, perfumery, cosmetic industries and in
pharmaceutics [7]. A major use of cinnamic acids is in the manufacturing of the methyl, ethyl, and benzyl for the
perfume industry [8]. Cinnamic acid is also a precursor to the sweetener, aspartame via enzyme-catalyzed
amination to phenylalanine [9]. Cinnamic acid derivatives are also naturally occurring substances found in
fruits, vegetables, flowers and are consumed as dietary phenolic compounds, which play a vital role in the
formation of commercially important intermediate molecules used for the production of different
pharmaceutical ingredients [7].
The aim of the study was to isolate and characterize chemical constituent(s) present in the ethyl acetate extract of
the stem bark of Adina microcephala.
Plant sample
The stem bark of A. microcephala was collected from Kundingi area, Samaru- Zaria (11° 04/N, 07° 42/ E),
located in the Northern Guinea Savannah zone of Nigeria in May, 2015. The plant was identified and
authenticated by the curator of the Herbarium, Department of Biological Sciences, Ahmadu Bello University,
Zaria, and a Voucher Specimen Number 345 was deposited there. The stem bark was air dried, powdered, and
stored in air tight container before use.
425
Extraction and Isolation
The dried pulverized stem bark (148 g) of A. microcephala was extracted exhaustively and successively with
petroleum ether (60-80°C), chloroform, ethyl acetate and methanol, respectively to afford the four extracts upon
evaporation of the solvent at reduced pressure using rotary evaporator. The ethyl acetate extract (7 g) was
chromatographed on a silica gel column and eluted with gradient elution technique using a solvent systems of
petroleum ether:ethyl acetate mixtures 95:05, 90:10, 85:15 and 80:20, respectively to yield 25 fractions.
Compound R1 (5 mg) was obtained from fraction Fr-2 (eluate of petroleum ether:ethyl acetate = 80:20), purified
further using preparative thin layer chromatography and a solvent mixture of petroleum ether /ethyl acetate
75:25.
Equipment and Chemicals
Thin layer chromatography analysis, silica gel 60 F254 (Merck) was used, column chromatography was
performed using Merck silica gel (60 – 120) mesh, while spots on thin layer chromatography plates were
visualized by spraying with 10% H2SO4, followed by heating at 100°C for 5 minutes. The IR spectrum was
measured on a Shimadzu FT-IR8 400S Fourier transform infrared spectrophotometer. Nuclear magnetic
resonance (NMR)-spectra were recorded on a Bruker Avance spectrometer (400 MHz) for 1H- and (100 MHz) for
13
C-NMR. Internal standard was residual solvent signal with methanol as a solvent.
RESULTS O
6 7
5
1 9
OH
8
4 2
HO OH
3
Figure 1: 2,4-dihydroxycinnamic acid

Table 1: NMR spectral data for compound 1 (CD3OD, 400 MHZ)
No äC äH DEPT
1 113.17 C
2 163.17 - C
3 103.44 6.72(H, d, J =2.12 Hz) CH
4 157.27 - C
5 114.5 6.82(H, dd, J = 2.24, 852 Hz) CH
6 130.69 7.47 (H, d, J = 8.52 Hz) CH
7 146.10 7.86 (H, d, J = 9.48 Hz) CH
8 112.33 6.82 (H, dd, J = 2.24, 8.52 Hz) CH
9 163.76 C
DISCUSSION
The IR spectrum indicated the presence of carbonyl (1735 cm- 1), hydroxyl (3361 cm- 1) and aromatic groups
(1550–1480 cm- 1). The aromatic region of the proton NMR spectrum of compound R1 revealed the presence of 5
proton signals at äH 6.20(H, d, J = 9.44 Hz) 6.72(H, d J =2.12 Hz), 6.82 (H,dd, J = 2.24, 8.52 Hz), 7.47 (H, d, J =
13
8.52 Hz) and 7.86 (H, d, J = 9.48 Hz). The C spectrum of R1 revealed the presence of 9 carbon atoms, of which
one carbon atom was assigned to a carboxylic acid carbonyl at äC 163.76; six unsaturated carbons at äC 103.44,
113.17, 114.56, 130.69, 157.27 and 160.17. Three of the six unsaturated carbons are methine carbons (103.44,
114.56, and 130.69), and two oxymethine carbons äC 157.25 and 163.17; and two olefinic carbons at äC 146 and
112.33. Considering that the proton NMR spectrum revealed five aromatic protons. The six unsaturated
426
(olefinic) carbons could be assigned to a mono-substituted benzene ring, joined to a three carbon straight olefinic
carboxylic acid chain, with two significantly deshielded carbons being oxygenated, while three of the remaining
up-field aromatic carbon atoms bore single aromatic proton [10].
The 1H and 13C assignments were further supported by the two-dimensional (2D) NMR data including1H-1H
COSY, HSQC and heteronuclear multiple-bond correlation (HMBC) experiments. In the five carbon
1 1
polyhydroxy carbon chain, H- H COSY experiment indicated the following connectivity: on the aromatic ring
H-5'/H-6', suggesting that the two aromatic protons are adjacent to each other and ortho-couple to each other. The
H-1H COSY experiment showed connectivity between H-3'/H-5', H-3'/H-4' suggesting that the aromatic protons
1 1
are meta couple to each other. Furthermore, H-7 on the carboxylic side chain showed H- H COSY with H-8' of
the olefinic carbon. The connectivity of the carboxylic side chain was supported by the HMBC correlations of H-
7'/C-3, H-7'/C-1', H-7'/C-6 H-7'/C-4, H-7'/C-2 and H-7'/C-9. From the above 1D and 2D NMR spectra analyses,
the compound isolated from the ethyl acetate extract of the stem bark of Adina microcephala is 2, 4
dihydroxycinnamic acid. Dihydroxycinnamics acids are found to possess significant pharmacological activity
and are used as a precursor of in pharmaceutical industries [7].
REFERENCES
1. M.A. Hyde, B.T. Worsen, P. Billing and M.P. Coates (2016). Flora of Zimbabwe. Species information,
Breonadia salicina: Pp. 245 – 246.
2. H.M. Burkill (1985). The Useful Plants of West Tropical Africa. Second Edition, Vol 1. Royal Botanic
Gardens, Kew, UK, 510 pp.
3. H.D. Neuwinger (1994). African Ethnobotany: Poisons and Drugs: Chemistry, Pharmacology,
Toxicology. Stuttgart, Germany: Chapman & Hall.
4. S.M. Mahlo (2010). Characterisation and Biological Activity of Antifungal Compounds Present in
Breonadia salicina (Rubiaceae) Leaves. PhD Thesis, University of Pretoria, Pretoria, South Africa, 120
pp.
5. F. Al-Qurainy, G. Abdel-Rahman, S. Khan, M. Nadeem, M. Tarroum, A, Alaklabi, J. Thomas (2013).
Antibacterial activity of leaf extract of Breonadia salicina (Rubiaceae), an endangered medicinal plant
of Saudi Arabia. Genetics and Molecular Research, 12(3): 3212 – 3219.
6. S.M. Mahlo (2010). Antifungal activity of leaf extracts from South African trees against plant
pathogens. Crop Protection, 29(12): 1529 – 1533.
7. P. Sharma (2011). Cinnamic acid derivatives: A new chapter of various pharmacological activities.
Journal of Chemical and Pharmaceutical Research, 3(2): 403 – 423.
8. S. Budavari (1996). The Merck Index: An Encyclopaedia of Chemicals, Drugs and Biologicals. Merck,
th
12 edn.
9. D. Garbe (2002). Cinnamic Acid in Ullmann's Encyclopaedia of Industrial Chemistry, Wiley-VCH,
Weinheim 022.
10. M. A. Ponce, M. J. Bompadre, J. M. Scervino, J. A. Ocampo, E. J. Chaneton, A. M. Godeas (2009).
Flavonoids, benzoic acids and cinnamic acids isolated from shoots and roots of Italian rye grass (Lolium
multiflorum Lam.) with and without endophyte association and Arbuscular mycorrhizal fungus.
Biochemical Systematics and Ecology, 37: 245 – 253.
427
GROUNDWATER QUALITY IN OKITIPUPA TOWNSHIP OF ONDO STATE, NIGERIA
?1Ayedun, H. and Ediagbonya T. F
Department of Chemical Sciences, Ondo State University of Science and Technology, P. M. B 353,
Okitipupa, Ondo State, Nigeria.
?Author of Correspondence
E-mail: hayedun2000@yahoo.com; ht.ayedun@osustech.edu.ng
Tel.: +234 – 803 2952831
Abstract
This study was conducted to assess the quality of groundwater in Okitipupa (Okpa), the head quarter of
Okitipupa local government area of Ondo State, Nigeria. Physico-chemical parameters were determined using
standard methods while metals were determined using Atomic Absorption Spectrophotometer. The pH range
-1
(Mean ± SD) are 8.5 – 11.2 (10.1 ± 0.9), the Electrical Conductivity range are 1095 – 1932 (1503 ± 211) µSCm ,
the TDS range are 657 – 1159 (902 ± 126) mgL-1, the chloride concentration range are 220- 299 (276 ± 18.5) mgL-
1
, the Sulphate range are 80.7 – 260 (201 ± 53.5) mgL-1 and Nitrate range 10.1-92 (35.1-19.6) mgL-1. The metal
index (MI) was used to assess the level of metal contamination in comparison with WHO maximum acceptable
concentration (MAC). The calculated MI values of metals (Fe, Cr, Pb, Cu and Ni) ranged from 1.47 – 46.3. The
results show that the overall quality of water decreases as MI >1 is a threshold of warning. Improved sanitation
and treatment of groundwater meant for drinking is necessary.
Keyword: groundwater, Okitipupa,. Metal Index, anions, Ondo State
Introduction
Groundwater contamination occurs when man made products such as gasoline, rock salts and chemicals get into
groundwater and cause it to become unsafe and unfit for human use [1]. Water is an excellent solvent that
contains a lot of dissolved chemicals which moves through rocks and subsurface soil. It has a lot of opportunity to
dissolve substances as it moves through rocks and subsurface soil. For that reason groundwater will often have
more dissolved substances than surface water. Even though the ground is an excellent mechanism for filtering
out particulate matter such as leaves, soil still dissolved chemicals and gases can still occur in a large enough
concentration in groundwater to cause problems. In developing countries, the provision of safe and clean
drinking water supply schemes is considered to be the key responsibilities of the government because people
have limited access to safe drinking water [2]. Many inorganic contaminants are toxic and pose a great health and
environmental concern in quite low concentrations.
Heavy metals are considered as environmental contaminants because of its persistence, toxicity and bio-
accumulative nature in the environments [3]. Water containing several types of toxic metals may cause different
types of health problems like respiratory and cancers in humans [4].The metal Index (MI) was defined as the sum
of metal concentration divided by maximum acceptable concentration (MAC) [5]. The higher the concentration
of a metal compared to its MAC value, the worse the quality of the water. The objective of the present study is to
assess the physicochemical parameters of groundwater in Okitipupa township and compare MI values to
ascertain its suitability for human consumption.

The Study Area: Okitipupa town lies between Latitude 5.125oN Longitude 4.008oE and Latitude 8.042oN
o
Longitude 6.011 E. It is the headquarter of Okitipupa local government of Ondo State, Nigeria. It has an area
428
of 63 square kilometer and a population of 233,565 at 2006 census [6]. It is bounded in the North by Odigbo
Local government, East by Irele Local government, South by Ilaje Ese Odo Local government and West
by Ogun State. The inhabitants are mainly Yoruba of Ikale ethinic group while agriculture is the main
occupation.
Sample Collection and Preservation:
Well water samples were collected from twenty one locations across Okitipupa. Water samples were collected
into a cleaned polythene bottles of one liter capacity which were rinsed with water to be sampled before
collection. A cleaned polyvinyl chloride (PVC) rope was attached to a drawer made up of high density
polyethylene (HDPE) bucket which was used to draw water from the wells.
Sample Analysis:
The physico-chemical parameters such as pH, Temperature, Electrical Conductivity (EC) and position of the
wells were measured on the field using the following instruments. Hand-held 2000VWR Scientific meter for pH,
Thermo Orion model conductivity meter (USA made) was used to measure the conductivity and TDS of the
water. Samples meant for metal analysis were preserved with 5 mL concentrated nitric acid per liter. Samples for
anions were kept in ice, transferred to the laboratory and analyzed immediately. The concentration of anions
were determined using standard methods (APHA, 1998) [7]. The salicylate method was used for nitrate
determination, turbidimetry method was used for sulphate determination, Vanado-molybdo-phosphoric acid
method was used for phosphate determination and Indophenol colorimetric method was used for ammonium
concentration determination. Cecil UV-Visible Spectrometer was used for the measurement. The chloride and
alkalinity were determined by Mohr's method and titration methods respectively. Samples were digested in aqua-
regia and analyzed for trace elements using a Perkin Elmer Atomic Absorption Spectrophotometer.
Chemicals and Reagents: All glassware and polyethylene bottles were washed, rinsed with doubly distilled
water and soaked in 10% HNO3 (v/v) for 72 h. The glass wares were rinsed again three times with doubly
deionized water and finally dried in oven before use. All reagents were of analytical grade and the water used was
doubly deionized water. Nitric acid (65% Suprapur, Merck, Darmstadt, Germany) was used for cleaning
glassware and for preservation of water samples.
Statistical Analysis: Microsoft Excel was used for data entry and descriptive statistics (range, mean, standard
deviation). SPSS 18.0 was used to generate the Pearson correlation data.
The range of physicochemical parameters and metallic elements in groundwater of Okpa were presented in
Table1while the concentration of metallic element from different locations were presented in Table 2. The metal
Index were presented in Table 3 while Pearson correlation of metals were presented in Table 4.
Physicochemical Parameters:
The range of pH are 8.5 – 11.2 in groundwater of Okitipupa township of Ondo State (Table 1). Ten percent (10 %)
of the samples were found to be very close to 8.5, the maximum recommended by World Health Organization
(WHO) [8].
Table 1: Range of physicochemical parameters and metallic elements in groundwater of Okpa
429
Elements/Parameters Min Max Mean WHO
pH 8.5 11.2 10.1±0.900 6.50-8.50
Conductivity (µSCm-1) 1095 1932 1503±211 250
TDS(mgL-1) 657 1159 902±126 250
Cl-(mgL-1) 220 299 276±18.5 250
PO43-(mgL-1) 3.5 41 7.40±7.79 -
SO42-(mgL-1) 80.7 260 201± 53.5 250
NO3-(mgL-1) 10.1 92 35.1±19.6 0.05
Cd(mgL-1) 0.001 0.035 0.0098±0.002 0.003
Fe(mgL-1) 0.012 0.315 0.112±0.103 0.300
Cr(mgL-1) 0.003 0.044 0.0208±0.013 0.050
Cu(mgL-1) 0.071 0.681 0.4201±0.224 2.00
Pb(mgL-1) 0.001 0.006 0.004±0.003 0.01
Ni(mgL-1) 0.005 0.057 0.028±0.014 0.020
This is in variance to 6.8 – 7.7 that was reported in Punjab Pakistan [9]. The Electrical Conductivity (EC) in
Okitipupa groundwater ranged from 1095 - 1932 µScm-1 with the mean value of 1503 ± 211 µScm-1. The WHO
recommended EC value in drinking water was 250 µScm-1. The high value recorded may be due to dissolved
ions of the salty nature of the water. Total Dissolve Solid (TDS) from Okitipupa groundwater ranged from 657 -
1159 mgL-1 with the mean value 902 ± 126 mgL-1. Based on TDS content, groundwater are classified into
desirable for drinking (up to 500 mgL-1), permissible for drinking (500 – 1000 mgL-1), useful for agricultural
-1 -1
purposes (up to 3000 mgL ) and unfit for drinking and irrigation (above 3000 mgL ) [10]. Eighty one percent
(81%) of groundwater samples collected from Okitipupa are within permissible for drinking level while 19 % are
useful for agricultural purposes.
Chloride concentration in Okitipupa groundwater ranged from 220 – 299 mgL-1 with the mean value of 276 ±
-1 -1
18.5mgL . The WHO recommended limit of 250 mgL in drinking water were exceeded by 86 % of the samples.
-1
Phosphate concentration in groundwater samples from Okitipupa ranged from 3.5 – 41mgL with a mean value
of 7.4 ± 2.1 mgL-1. Sulphate concentration in groundwater samples from Okitipupa town ranged from 80.7 - 260
mgL-1 with a mean value of 201 ± 53.5 mgL-1. . Ten percent (10%) of the groundwater samples collected has
-1 2-
sulfate concentration greater than WHO limit of 250 mgL . The high level of SO4 may be due to run off from
agricultural land where fertilizers is applied to the land which might have contaminated the groundwater. Nitrate
concentration in groundwater samples from Ore ranged from 10.1 - 92 mgL-1 with a mean value of 35.1 ± 19.6
mgL-1. The results indicated that none of the samples exceeded the WHO limit of 50 mgL-1 in drinking water.
Previous studies reported similar nitrate concentration and was correlated it with land use pattern [11]. Nitrate in
groundwater is of great concern because of it causes met-hemoglobinemia in infants while the primary sources of
groundwater nitrate are domestic sewage disposal and fertilizer.
Metallic Elements
-1
Cadmium concentration in Okpa groundwater ranged from 0.001 – 0.035 mgL with the mean value of 0.0098 ±
-1 -1
0.002 mgL . Twenty nine percent (29 %) of the samples exceeded WHO limit of 0.003mgL . The highest Cd
concentration value were recorded from Idepe area (Okpa 5) (Table 2).
Table 2: Metallic elements in groundwater of Okpa
430
Locations Cd(mgL-1) Fe(mgL-1) Cr(mgL-1) Cu(mgL-1) Pb(mgL-1) Ni(mgL-1)
Okpa1 n=4 0.001±0.001 0.212±0.001 0.0133±0.001 0.611±0.01 0.0103±0.003 0.027±0.002
Okpa 2 n=4 0.005±0.001 0.309±0.004 0.029±0.004 0.534±0.009 0.0013±0.00 0.026±0.001
Okpa3 n=4 0.002±0.00 0.068±0.002 0.0037±0.00 0.652±0.017 0.0047±0.00 0.024±0.00
Okpa 4 n=4 0.003±0.001 0.015±0.00 0.0243±0.00 0.074±0.002 0.0083±0.00 0.04±0.00
Okpa 5 n=4 0.021±0.001 0.0533±0.022 0.031±0.00 0.152±0.0312 0.011±0.00 0.021±0.00
Okpa 6 n=4 0.003±0.002 0.101±0.037 0.04±0.004 0.614±0.00 0.002±0.00 0.007±0.001
Okpa 7 n=4 0.016±0.001 0.028±0.014 0.004±0.00 0.305±0.027 0.0067±0.00 0.054±0.002
Key: Okpa1- garage, Okpa2- Bagudu str, Okpa 3-Ayonio str, Okpa 4-ofedepe, Okpa 5-Idepe, Okpa 6-Igbodigbo,
Okpa 7-Erinje/ NDDC quarters
High Cd concentration may be due to discharge from waste or by leaching from sewage laden landfills. It may
also be attributed large use of polyvinyl plastics, nickel-cadmium batteries, insecticides, motor oil, and disposal
of slugde in dumpsites. Chronic exposure to Cd lead to anemia, anosmia, cardiovascular diseases and renal
failure [12]. High values were recorded from two locations (Okpa 5 and Okpa7) with the mean values of
-1 -1
0.021±0.001 mgL and 0.016±0.001 mgL respectively (Table 2). Results from previous studies shows Cd
concentration range in groundwater of Punjab which were lower than the range obtained from the present study
[9].
-1
Iron concentration in Okpa groundwater ranged from 0.012 – 0.315 mgL with the mean value of 0.112 ± 0.103
-1
mgL . Previous study reported iron concentration in the Sahelian region of Northern Ghana, West Africa with
higher range but lower in mean value than the results from the present study [13]. Fourteen % of the Fe
concentration in well water samples from Okpa exceeded WHO limit of 0.3mgL-1. In most wells, rusty well cover
used could also be a possible source of iron in these well. Turbidity, taste, discoloration, deposits and growth of
iron-bacteria could be widespread in wells with high iron concentration.
Chromium concentration in Okpa groundwater ranged from 0.003 – 0.044 mgL-1 with the mean value of 0.021 ±
0.013 mgL-1. None of the samples exceeded WHO limit of 0.05mgL-1. The results from the present study is
similar to what was reported in previous study of groundwater in Riyadh region of Saudi Arabia [14].
-1
Copper concentration in Okpa groundwater ranged from 0.071 – 0.681 mgL with the mean value of 0.420 ±
0.224 mgL-1. None of the samples exceeded WHO limit of 2.0 mgL-1. This is similar to results obtained in
groundwater of Iwo and Ikonifin of Osun State, Nigeria [15].
-1
The Pb concentration in Okpa groundwater ranged from 0.001 – 0.006 mgL with the mean value of 0.004±
0.003 mgL-1. Three percent (38 %) exceeded the WHO limit of 0.01 mgL-1. The difference is due to improper
waste disposal which eventually pollute groundwater resources. This results is in agreement with the study that
reported lead as one of the elements in the groundwater samples of Kwahu west district of Ghana that is above the
safety limits recommended by WHO [16]. Lead can initiate hypertension, tiredness, irritability, anemia
behavioral changes and impairment of intellectual functions in affected population that consume those
groundwater samples.
-1
Nickel concentration in groundwater of Ore ranged from 0.005 – 0.057 mgL with the mean value of 0.028 ±
-1 -1
0.014 mgL . Fourteen percent (14%) of the groundwater samples exceeded WHO limit of 0.02 mgL . Ni
concentration in groundwater up to 0.20mgL-1 was reported in the past [21]. Nawab et al (2013) reported highest
431
-1
Ni concentration in groundwater of Shangla, Pakistan to be 0.064 mgL [2]. Combustion of fuel as well as
municipal waste are possible sources of Ni in groundwater.
Metal Index: The metal index for drinking water was preliminary defined as MI = ?Ci/MACi, where Ci is the
concentration of each element in solution and MAC is the maximum allowable concentration [5]. The MI in
Okpa groundwater for Cd, Fe, Cr, Cu, Pb and Ni are 46.3, 2.59, 2.9, 1.47, 4.4 and 9.95 respectively (Table 3).
Table 3: Metal Index (MI) in Okpa groundwater
Elements/Parameters ?i MACi MI = ?i/ MACi
Cd(mgL-1) 0.139 0.003 46.3
Fe(mgL-1) 0.779 0.300 2.59
Cr(mgL-1) 0.145 0.050 2.9
Cu(mgL-1) 2.94 2.00 1.47
Pb(mgL-1) 0.044 0.01 4.4
Ni(mgL-1) 0.199 0.020 9.95
If the concentration of a certain element is higher than the respective MAC value, the water cannot be used. The
presence of several elements with concentrations smaller than but close to the respective MAC values will also
decrease overall quality of water because of additive effect. Thus MI greater >1 is a threshold of warning even in
the case where Ci < MACi for all elements.
Pearson Correlation of metals: Significant positive correlation P < 0.05 was recorded between Cd and Cr (r =
0.813), Fe and Cu (r = 0.558) (Table 4).
Table 4: Correlation of metals in groundwater of Okpa
Cd Fe Cr Ni Pb Cu
Cd 1
Fe .033 1
Cr .813** .227 1
Ni -.306 -.305 -.631** 1
Pb -.158 -.384 -.218 .278 1
Cu -.248 .558** -.127 -.492* -.542* 1
Negative correlation was recorded between Cr and Ni (r = -0.631), between Pb and Cu (r = -0.542). Inter-element
positive correlation matrix of metals indicated a common source of contamination, however, negative correlation
shows that the sources are different.
Conclusion
Groundwater in Okpa contain dissolved ions and most especially chloride level was very high. The metal index
showed that cumulative effect endanger human health as MI >1 is a threshold of warning. Improved sanitation
may reduce the danger but treatment of groundwater meant for drinking is necessary.
REFERENCES
1 . G . F. G r o u n d w a t e r F o u n d a t i o n ( 2 0 0 8 ) . S o u r c e s o f g r o u n d w a t e r c o n t a m i n a t i o n .
Wwwgroundwater.org/gi/sources of gwcontain.htm. Accessed on 15th Feb. 2009..
2. J. Nawab, S. Ali, A. Rauf, U.U. Rehman, A. Khan, M. Sajjad and W. Khan (2015), Health risk associated
with heavy metals via consumption of surface and groundwater in District Shangla, Pakistan. Journal of
Himalayan Earth Sciences. 48(2), 62-73.
3.H. Pekey, D. Karaka and M. Bakoglu (2004), Source apportionment of heavy metals in surface waters of a
432
polluted stream using multivariate statistical analyses. Marine Pollution Bulletin, 49, 809-818.
4. P. Kavcar, A. Sofuoglu and S. C. Sofuoglu (2009), A health risk assessment for exposure to trace metals via
drinking water ingestion pathway. International Journal of Hygiene and Environmental Health, 212(2),
216–227.
5.T. Gabriella and C. Renso (2004). Heavy metals in drinking waters from mount Aminata (Tuscany Italy).
Possible risk from Arsenic for public health province of Sirena. Sci Tot. Env. 327, 41-51.
6. NPC ( 2006). National Population Commision Official Gazette.
7. APHA.(1998). American Public Health Association Standard methods for the examination of
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water and waste water, 19 Edition. APHA, AWWA, WEF. Washington DC. p 4-67, 99-144.
8. WHO (2008).World health Organisation. Guidelines for drinking water quality (3rd Ed incorporating the 1st and
2nd Addenda) volume 1 recommendation .Geneva http://
. Accessed on 19th August 2009.
9. M. T. Mahar, M. Y. Khuhama, M. A. Baloch and T. M.Tahangir. (2013). Health risk assessment of heavy
metals in groundwater, the effect of evaporation ponds on distillery spent wash: A case study of Southern
Punjab Pakistan. World Appllied Science Journal. 28 (11),1748-1756.
10. S.N. Davis and R. J.M De-Wiest (1963), Hydrogeology. New York. Wiley. pp 463.
11..K.K. Gardner and R.M. Vogel (2005), Predicting Ground Water Nitrate Concentration
from Land Use. Ground Water, 43(3), 343–352
12. W. H. Holl, C. H. Bartosch, X. Zhao and S. He (2002), Elimination of trace heavy metals
from drinking water using weakly basic anion exchange. Jahrgang Heft, 3, 5-20.
13. S. J. Cobbina, F.K. Nyame and S. Obiri (2012), Groundwater Quality in the Sahelian Region
of Northern Ghana, West Africa Research Journal of Environmental and Earth Sciences, 4(4):
482-491.
14. A. Abdulrahman, I. A. Abdullah and A. K. Mujahid (2011), Assessment of Trace Metals in Groundwater
Sources Used for Drinking Purposes in Riyadh Region. International Journal of Water Resources and Arid
Environments, 1(1), 05-09.
15. A. M. Gbadebo, H. Ayedun and A. I. Moses (2015), Hydrogeochemical assessment of groundwater in Iwo,
Ikonifin and Ife-Odan, Osun and Oyo states, Southwest Nigeria, Environ Earth Sci, 73, 3633–3642
16. M. A. Nkansah, N. O. Boadi and M. Badu (2010), Assessment of quality of water from hand dug wells in
Ghana. Environmental Health Insights. 4,7-12.
433
PREPARATION AND CHARACTERIZATION OF ACTIVATED CARBON FROM Hura Crepitans
Linn SEED SHELLS
E. W. NSI1 , AND A. E. AKPAKPAN2* U. D. AKPABIO3
1,2
Department of Chemistry Akwa Ibom State University, Nigeria
3
Department of Chemistry University of Uyo, Uyo, Akwa Ibom State, Nigeria
ani4sucess@yahoo.com
ABSTRACT
Activated carbons were thermally prepared from Hura Crepitan L. seed shells. Zinc chloride (ZnCl2) and
Phosphoric acid (H3PO4) were separately used as the activating agents. The activated carbons obtained were
characterized by determining the percentage yield, moisture content, ash content and percentage fixed carbon.
The adsorption of methylene blue by the activated carbon was done using 0.1 to 0.5g of the activated carbon. The
results revealed that the percentage yield and ash content of H3PO4 impregnated activated carbon was higher than
ZnCl2 impregnated activated carbon. On the other hand ZnCl2 impregnated activated carbon had higher moisture
content and percentage fixed carbon. It was also revealed that ZnCl2 impregnated activated carbon had greater
adsorption capacity than H3PO4 impregnated activated carbon. However it was found that the higher the
adsorbent (activated carbon) dosage, the higher the adsorption capacity.
Keywords: Hura Crepitan L., activation, activated carbon, Zinc chloride and Phosphoric acid
INTRODUCTION
Activated Carbons are carbon of highly micro-porous form with both high internal surface area and porosity, and
commercially, the most common adsorbents used for the removal of organic pollutant from air, water and
industrial products (Tsai et al., 2001).
Food and pharmaceutical industries uses activated carbon in the removal of compounds that are responsible for
undesirable colour, odour and taste in their products. Activated carbon is also used in gas cleaning applications
where it is extensively used in air filters at industrial level as well as in air condition equipment (Vigayan et al.,
2012).
Commercial activated carbon is produced from bituminous or lignite coal. Because of the environmental
problems and increasing high cost associated with petroleum products, research have been reported on the
production of activated carbon from wood which is environmentally friendly and cost effective. However,
because of the environmental problem associated with deforestation, seed shells have been used to replace wood
for the production of activated carbon. These products are highly cost effective as most of the seed shells are
discarded as waste materials, and the activated carbon produced from them have similar potential in industrial
application.
The production of activated carbon from coconut shells, palm kernel shells, Cananium scheqinflurthi nut shell,
corn cob and other seed shells have been reported (Olawale andAjayi, 2007; Diya et al., 2008; Chang et al, 2001
and Vijayan et al., 2012).
Hura crepitans L., the sand box tree also known as possum-wood, is an evergreen tree of the spurge family
Euphorbiacae. The wood is used for furniture under the name “Hura”. The oil extracted from the seed is used in
the production of alkyl resin (Ezeh et al., 2012) and metallic soap (Umoren et al., 2013). The seed pods (shells)
are not utilized for any industrial purposes; hence they are waste and thereby pollute the environment. These
shells are hard and therefore suitable for the production of activated carbon which can be used for the treatment of
434
waste water from textile industries and can also be used in food and pharmaceutical industries to remove
pollutants/contaminants.
Therefore, the aim and objectives of this work is to prepare activated carbon from Hura crepitans L. shells,
characterize and apply it in the removal of methylene blue from contaminated water.
2.0 MATERIAL AND METHODS

2.1 Sample Collection
The seed shells of Hura crepitans L. also known as sand box were collected from the University of Uyo Town
Campus. The samples were cut into chips of workable size.
Activation-Carbonization
Activation of the sample was carried out in a muffle furnace using the solution of zinc chloride (ZnCl2) and
phosphoric acid (H3PO4) separately as the activating agents. 400g of the sample chips were charred in the furnace
at 500oC for 2 hours. 30g of the charred sample was weighed into a beaker containing 300cm3 of 1M solution of
each activating chemical. The contents of each beaker was thoroughly mixed and heated until all the activating
liquor was absorbed by the sample.
o
The samples were then transferred into the crucible and dried in the oven at 100 C for 12 hours. The crucible was
then placed in the furnace and fired at 650oC for 2 hours.
The sample was then removed from the furnace and cooled in the desiccator, it was then washed with 0.5M
o
NaOH followed by distilled water to a neutral pH. The activated carbons obtained were dried in the oven at 100 C
to a constant weight.
2.2 Characterization of the Activated Carbon.
· Percentage Yield
The yield of the activated carbon was calculated using;
Percentage yield = M2 x 100%
M1
Where M1 = Weight of the charred sample
M2 = Weight of the activated carbon
· Moisture content
Moisture content of the activating carbon was calculated using :
Percentage moisture content = M1-M2 X 100%
M1
Where M1 = Weight of the activated carbon before drying in the oven
M2 = Weight of the activated carbon after drying in the oven
· Determination of Ash content and percentage fixed carbon

Ash content of the activated carbon was determined as follows: 2g of each activated carbon sample was put in a
o
crucible and fired at 900 C in a muffle furnace for 3 hour. The resultant ash was removed and allowed to cool in
the desiccator.
Ash content was calculated using:
% Ash content = W2 x 100%
W1
435
Where W2 = Weight of the ash
W1 = Weight of the activated carbon
Percentage fixed carbon was calculated from the ash content in accordance with the method reported by
Ogbonnaya (1992) and Duff and Ross, (1988).
Adsorption Study
The adsorption potentials of the activated carbon sample obtained from different activating agents were
determined according to the method reported by Odebunmi and Okelola (2001). In this method, batch sorption
experiments were carried out at room temperature. 0.1 to 0.5g of the activated carbon was added into a beaker
containing 100ml of 250mg/l methylene blue solution. The mixture was thoroughly mixed and shaken for 1 hour
using a mechanical shaker. After shaking, the suspensions were filtered and remaining concentration was
determined spectrophotometrically at ënm of 250nm using UV/V spectrophotometer. The amount of methylene
blue adsorbed from each solution was calculated as shown below.
Percentage methylene blue adsorbed = (Co – C) X 100

Co
Where:
Co (mg/L) = Initial concentration of methylene blue
C (mg/L) = Post-adsorption concentration of methylene blue

The percentage yield, ash content, moisture content and percentage fixed carbon of the activated carbon
are presented in Table 1. These parameters are factors which could influence the adsorption capacity of the
activated carbon (Okeola et al., 2011).
Result revealed that the percentage yield of H3PO4 activated carbon was 44.90% and that of ZnCl2
activated carbon was 31.94%. The decrease in the yield of ZnCl2 activated carbon may be due to the reactions
between ZnCl2 and the sample.
The ash content and percentage fixed carbon compare favourable with those for the commercial
activated carbon. Also the ash content of H3PO4 activated carbon compare favourable with that of H3PO4, ZnCl2
and KOH activated carbon from maize cob (Odebunnin and Okeola, 2001) and NaCl activated carbon of
Jatropha curcas seed coats (Okeola et al., 2011). The ash content obtained for ZnCl2 activated carbon compare
favourably with the value obtained Coconut shell (Odebunmi and Okeola, 2001). However, ZnCl2 impregnated
activated carbon has greater percentage fixed carbon than H3PO4 impregnated activated. There is a correlation
between percentage fixed carbon and adsorption capacity. Sample with higher percentage fixed carbon are
expected to have high adsorption capacities (Ogbonnaya, 1992). The moisture content of ZnCl2 activated carbon
was higher than that of H3PO4 activated carbon, implying that ZnCl2 activated carbon has greater pore size for
absorption than H3PO4 activated carbon.
Table 1: Physical parameters of the activated carbon
436
Activating Agent H3PO4 ZnCl2
Yield (%) 44.90 31.94
Moisture content (%) 5 17
Ash content (%) 28 16.5
Percentage fixed carbon (%) 72 83.8
Adsorption Studies
Methylene blue was chosen in this study because of its recognized usefulness in characterizing
adsorptive material (Itodo et al., 2010). The size of methylene blue molecule is much larger and its adsorption is
restricted mainly to the mesopores adsorbent. However a small portion is also found in larger micropores
(Cleiton et al., 2011).
The results of the methylene blue adsorption by the activated carbons are presented in Table 2 and 3. The
results revealed that the quantity of the adsorbent has greater effect on the adsorption of methylene blue from
aqueous solution. The result shows that the adsorption efficiency increased with increase in mass of the
adsorbent. The increase in adsorbent mass increases the contact surface area of the adsorbent particles which
means that it become more probable for solute molecules to be adsorbed on the adsorption sites and thereby
increasing adsorption efficiency (Okeola et al., 2012).
Effect of Activating Agents
With all the adsorbent dosages, ZnCl2 impregnated activated carbon sample has greater adsorption
efficiency than the one prepared with H3PO4. This is due to the fact that samples with high percentage fixed
carbon always have high adsorption capacities (Ogbonnaya, 1992). This result also suggests that ZnCl2
impregnated activated carbon sample have well developed mesoporosity favorably for adsorption of larger
molecules compared to H3PO4 impregnated activated carbon sample. However, the percentage adsorptions of
methylene blue by these activated carbons are low, this may be due to the fact that the size of the methylene blue
molecule is much larger and its adsorption is restricted mainly to the mesopores activated carbon (Cleiton et al.,
2011).
Table 2: Adsorption of Methylene Blue by H3PO4 activated Carbon
AC dosage (mg) Post Adsorption Conc. of Adsorption efficiency (%)

MB (mg/l)
100 201.67 19.33
200 197.54 20.98
300 180.64 27.74
400 167.36 33.05
500 140.58 43.77
437
Table 2: Adsorption of Methylene Blue by ZnCl2 activated Carbon
AC dosage (mg) Post Adsorption Conc. of Adsorption Efficiency (%)
MB (mg/l)
100 195.84 21.66
200 186.29 25.48
300 178.64 28.54
400 145.06 41.98
500 122.77 50.89
CONCLUSION
Hura Crepitan L. seed shells were successfully converted into activated carbon using H3PO4 and ZnCl2 as the
activating agents. The results of the physical properties of the activated carbon compared favourably with the
commercial activated carbon and with other activated carbons prepared from seed shells and cob. The adsorption
of methylene blue by ZnCl2 impregnated activated carbon was higher than H3PO4 impregnated activated carbon.
However H3PO4 impregnated activated carbon is recommended for used in food and pharmaceutical industrials
since it is non toxic activating chemical as it does not contain heavy metal while ZnCl2 impregnated activated
carbon can be apply for the treatment of waste water from textile and allied industries.
REFERENCES
Ajayi, O.A., and Olawale, A. S. (2009). A comparative study of thermal and chemical activation of Canarium
Schweinfurthii Nutshell. J. Appl. sci. Res., 5(12): 2148-2152.
Cleiton Nunes A., Guerreiru Mario C., (2011). Estimation of surface area and pore volume of activated carbons
by methylene blue and iodine numbers, Quim. Nova, 34(3) 472-476.
Diya udden, B. H., mohammed,I. A. Ahmed, A.S. and Jibril, B. Y. (2008). Production of activated carbon from
corncobs and its utilization in crude oil spillage clean up. Agricultural Engineering International: The
CIGR E-Journal. Vol. x: 1-9.
Duff, O.G. and Ross, S.M. (1988). Adsorption on activated charcoal. J of chem. Educ. 65(3) 815
Ezeh, I.E., Umoren, S.A., Essien , E.E. and Udoh. A.P. (2012). Studies on the utilization of Hura Crepitans
L. seed oil in the preparation of alkyl resins, industrial crops and products 36:94-99,
Itodo, A. U., Abdulrahman, F. W., Hassan. L. G., Maigandi, S. A. and Itodo, H. U. (2010). Application of
Methylene blue and iodine Adsorption in the measurement of specific area by four acid and salt treated
activated carbons. New York Science Journal, 3(5): 25-33.
Joshi, S., Adhikari, M., Pokharel, B. P. and Pradhanga, R. R. (2013). Effect of Activating agents on the Activated
carbons prepared from Lapsi seed stone. Research Journal of Chemical Sciences, 3(5): 19-24.
Odebunmi, E. O. and Okeola, O. F. (2001). Preparation and characterization of activated carbon from waste
material. J. Chem. Soc. Nigeria, 26(2): 149-155.
Ogbonnaya, O. (1992). Preparation and comparative characterization of activated carbon from Nigerian sub-
bituminous coal, palm kernel and cow bone. J. Chem. Soc. Nigerian, 17,11.
438
Okeola, O. F., Odebunmi, E. O. and Ameen, O. M. (2012). Comparison of Sorption capacity and surface area of
Activated carbon prepared from Jatropha curcas fruit pericarp and seed coat. Bull. Chem. Soc. Ethiop,
26(2) 171-180.
Olawale, A. S. and Ajayi, O. A. (2009). Thermal activation of Canarium Schweinfhurthi nutshell. Australian
Journal of Basic and Applied Sciences, 3(4): 3801-3807.
Tsai, W., Chang, C., Wang, Y., Chang C., Chien, F., Sun, H.(2001). Preparation of activated carbons from corn
cob catalyzed by potassium saltz and subsequent gasification with CO2. Bioresource technology 78(2) ;
203-208
Umoren. S.A., umoren, I. U. Johnson A.S.(2013). Metal soaps of Hura crepitans seed oil: potential stabilizer
for unplasticised polyvinyl chloride against thermal degradation. Journal of chemical Acta, 2:92-94
Vijayan, S. N., Makeshkumar, M. and Sridhar, K. (2012). Physical and chemical Analysis of Activated carbon
prepared from coconut shell charcoal and usage-A case study. International Journal of Advanced
Scientific Research and Technology, 2(3): 168-175.
439
CHARACTERIZATION OF BEACH WATER AND ITS EFFECT ON NEARBY GROUNDWATER
QUALITY IN ETI-OSA AND IBEJU-LEKKI LOCAL GOVERNMENT AREAS, OF LAGOS STATE,
NIGERIA.
V. E. *Agbazue and B. U. Ukazu
Centre for Environmental management and control (CEMAC),
University of Nigeria, Enugu Campus.
Abstract
Groundwater in Eti-Osa and Ibeju-Lekki local government areas of Lagos state were investigated for the effects
of beach water influence on adjacent aquifers. Beach water and Borehole samples at distances of 40-50m and
100-150m from adjacent beaches in the study areas were taken and analysed for pH, Temperature, Electrical
2+ 2+ + + - - - 2-
Conductivity, Total Dissolved Solids, Salinity, Ca , Mg , K , Na , HCO3 , NO3 , Cl ,SO4 , Total Hardness and
Dissolved Oxygen. The results were compared with World Health Organisation (WHO) and National Agency for
Food and Drug Administration and Control (NAFDAC) standards for water quality. Piper diagram was used to
delineate the hydrochemicalcalfacies of the aquifer and to identify hydrogeochemical changes. The results
-
showed that Cl dominated in boreholes of both Ibeju-Lekki and Eti-Osa. Sampled boreholes in Eti-Osa showed
prevailing dominance of major seawater ions ( Na+> Ca2+> K+> : Cl- > SO4-) at levels far beyond permissible
limits of potable water quality set by WHO and NAFDAC and showed a Ca-SO4 and NaCl water type for the
boreholes 40-50m and 100-15 from the coastline line respectively. Water quality requirements of WHO and
NAFDAC are within permissible limits for boreholes in Ibeju-Lekki. Analysed Water samples from Ibeju-
Lekkiare characterised as Ca-SO4 water type.Groundwater resources of studied boreholes in Ibeju-Lekki are less
influenced by saltwater intrusion from nearby beach, as opposed to Eti-Osa. This is can be attributed to overtly
unsustainable exploitation of groundwater resources in Eti-Osa, Which makes groundwater in Eti-Osa
unsuitable for domestic purposes.
Keywords: Characterization, Groundwater, Seawater, Intrusion, Standard.
* Corresponding author: e-mail: agbazuestars05@yahoo.com

Phone: 08068283058
INTRODUCTION
Water resources permeate every aspect of life, thus requiring utmost attention. . Effective water resource
management meets the needs for potable water and domestic usage, agriculture, transport and for other public
and industrial usage [1]. It also provides for the natural course of existence of water bodies, as home to many
aquatic organisms, plants and animals and a huge ecological capital. Improvement of information in water
quality and quantity with the Protection and restoration of the quality and function of ecological systems within
the coastal zones are key policy focus to improve the efficient use of water resources particularly in fragile
environments [1]. Lagos state, Nigeria in this context can be defined as a fragile environment following
recurrent reports of residents in the island areas abandoning boreholes, flooding, building collapses, land
encroachment by the ocean (due to ocean water level rise), changing coastal aquifer dynamics etc, which has
made these areas vulnerable to climate change hazards. To manage water resources adequately, monitoring and
investigative studies are essential; as a noble earth friendly task and responsibility.
Seawater intrusion is part of the changing dynamics with most coastal environments globally, increasing
populations in coastal environments which today are highly priced and sought after destinations for habitation
comes with disturbance to ecological balance as a consequence.
Researchers have lamented on the increasing and excessive abstraction of groundwater resources at rates faster
than natural recharge processes, owing to exploding population.
440
Unsustainable withdrawals from non-renewable aquifers, the unreliability of water availability, and the current
level of water demand exceeds the sustainable water supply in many places with serious long-term implications
and this is detrimental to the ecosystem and as well as the quality of water available.
Lagos, located on the southwest coast of Nigeria is made up of island and mainland areas. With an area of 1,341
2
square miles (3,400km ) of which half is water as creeks, lagoons etc and bordered by Atlantic ocean [2].
2
Population is estimated at 24.3million people [2]. Average population density is over 20,000 persons / Km [3].
The specific study areas; Alpha beach in Eti-Osa Local Government Areas (LGA) and Eleko beach in Ibeju-
Lekki LGA of Lagos state are highly vulnerable to climate change hazards [4]. These coastal environments are
densely and moderately populated respectively and the natural state of the coastal environment is likely retained
in the later unlike the former which some form of reconstructive engineering was ongoing as at the time of this
research, to stem the tide of coastal encroachment.
Knowledge gaps exists in understanding seawater intrusion (SI) process.In particular associated transient SI
processes and timeframes, characterization and prediction of saltwater influences on adjacent groundwaterover
regional scales and in highly heterogeneous and dynamic settings [5] like Lagos is a hard nut to crack. Oteri and
Atolagber (2003) posited that there is “Lack of adequate knowledge of the nature of salt water intrusion” [6].
Saline water intrusion has been identified as a major pollution issue, especially in areas that are close to the ocean
and lagoon.
Sadly there is no monitoring programme for saline water intrusion in Nigeria [6].
The occasion of excessive extraction of underground water beyond its restorative limit leads to pressure and
density variations that cause salt water intrusion in order to maintain ecological equilibrium, as the system of the
earth abhors vacuum. [7]have expressed that Intensive exploitation of groundwater aquifer disturbs the natural
equilibrium between fresh and saline water and results in the increase of groundwater salinity and the
hydrochemical complexities of freshwater-seawater contact. They further observed that the freshwater –
seawater mixing creates diversity in the geochemical process, and this manifests in compositional changes away
from the groundwater theoretical composition line.
According to Ohwoghere-Asuma (2014) groundwater is an important resource and consequently, its
management is only appropriate and necessary for its sustainability [8]. He suggested that the best way to
manage coastal aquifers is dependent on the ability to prevent re-occurrence of saline water intrusion by way of
early detection which is achieved by unrelenting monitoring of these aquifers. It is then vital and strategic to
understand the groundwater chemistry changes that may have occurred in Eti-Osa and Ibeju-Lekki coastal
environment as a result of saline water intrusion.
Hence this study is expected to provide findings to support early warning measures on the migration of the fresh
water/sea water intrusion into groundwater and its effects on groundwater quality. These findings will add a
voice to the clamour for a saline water intrusion monitoring and assessment plan and policy measures for coastal
environments in Nigeria.

Two borehole were purposively chosen in each study area:Ibeju-Lekki LGA (Eleko beach area) and Eti-Osa
LGA (Alpha beach area), following the criteria of (1) heavily/ publicly used borehole, where pumping is carried
out daily; (2) boreholes sited at distance of 40-50m and 100-150m from the coastline for the first and second
boreholes respectively. Groundwater and beach water samples were taken weekly for four weeks in the month
of November, 2015.A total of 48 samples were collected and analysed. Triplicate samples were collected per
time of sampling at each point after which a mean value was calculated for each parameter measured and a
standard deviation (SD) was used as a precision indicator for each measured parameter [9]. Sample points were
441
marked using a Global positioning system (GPS) instrument. Water samples were collected in plastic bottles that
had been washed and cleaned with hydrochloric acid (1:1) and then rinsed severally at the point of sampling with
the water to be sampled. Groundwater was pumped and allowed to rush out for 10–15 minutes before sample
fractions were collected [9].The beach water was collected with a sterilized bucket rinsed severally with the
beach water before sample collection. The water was scooped from three different side points of about 500m
apart along the coastline. Water from the bucket was poured into the bottles carefully, avoiding contact with the
bottle opening
These samples after collection were transported in a cooler of ice to the laboratory for analysis within 24 hours of
sample collection. in-situ measurements, pH and temperature, were carried out using portable digital meter and
platinum bulb thermometer respectively. Samples were analysed for physiochemical parameters which
included, salinity, Electrical Conductivity (EC), total dissolved solids (TDS), major seawater cations and anions
2+ 2+ + + - - - 2-
(Ca , Mg , K , Na , HCO3 , NO3 , Cl , SO4 ), total Hardness (TH) and dissolved oxygen (DO) using standard
methods [10].
The reliability check of the quality of the laboratory analysis and data obtained was done by the Error on ionic
balance (%). The analysis is considered as reliable for the error on the ion balance is smaller than 5 % [10].
However an error of ≤ ±10 % is within the acceptable range. The statistical package for social sciences (SPSS)
Version 16 and piper plot or diagram was employed to analyse laboratory results obtained.

Results
The location of the borehole, and beaches are as shown on table 1. Table 2 shows the mean values of
physicochemical parameters of borehole samples.
TABLE 1: LOCATION COORDINATES OF SAMPLE POINTS AND THEIR DISTANCE FROM
COASTLINE.
BOREHOLE CODES AREA LOCATION LATITIUDE LONGITUDE DISTANCE

FROM THE COAST
0 0
BH 1 ELEKO – IBEJU LEKKI 6.4394 N 3.85423 E 50m
0 0
BH 2 ELEKO – IBEJU LEKKI 6.44113 N 3.85825 E 100-150m
0 0
BH 3 ALPHA BEACH – ETI OSA 6.42514 N 3.52306 E 50m
0 0
BH 4 ALPHA BEACH – ETI OSA 6.42687 N 3.52708 E 100-150m
442
TABLE 2: MEAN VALUES OF PHYSIOCHEMICAL PARAMETERS OF BOREHOLE SAMPLES AND CORRESPONDING BEACH WATER .
ETI-OSA
IBEJU-LEKKI
S/N PARAMETER BH1 BH2 ELEK BH3 BH4 ALPHA
S O BEACH
MOV SD MOV SD BEAC MOV SD MOV SD
H
1. pH 7.825 0.377 7.750 0.479 7.0 7.375 0.350 7.525 0.512 7.3
2. Temperature, 0C 31.075 1.719 31.815 0.850 31 30.165 0.579 30.500 0.577 30
3. EC (µScm-1) 267.090 16.074 208.620 9.480 32440 3282.83 261.84 2036.340 231.580 60,520
4. TDS(mg/l) 186.500 13.127 145.860 6.688 22,720 2307.91 190.786 1427.100 162.830 42623.3
5. SAL (mg/l) 0.0225 0.005 0.020 .000 6.19 0.832 0.038 0.875 0.246 18.13
6. Ca2+ (mg/l) 11.740 1.788 10.750 0.957 700.0 167.850 11.888 111.680 34.250 1,837.0
443
7. Mg2+ (mg/l) 4.750 0.957 3.500 1.000 100.0 39.750 8.770 27.500 10.660 470.0
8. K+ (mg/l) 5.000 1.154 6.500 1.000 141.0 37.250 2.629 54.00 13.880 443.3
9. Na+ (mg/l) 9.832 1.373 10.500 1.000 863.3 158.16 13.379 265.92 195.080 6,870
10. HCO3- (mg/l) 0.022 0.0170 0.020 0.017 ND 0.022 0.015 0.022 0.017 0.01
11. NO3- (mg/l) 2.190 0.181 1.825 0.085 26.51 28.470 2.670 16.747 2.287 62.3
12. CL- (mg/l) 13.835 3.627 12.332 1.155 3,523.3 481.580 7.410 524.580 140.745 12,526
13. SO42- (mg/l) 8.665 1.300 6.750 0.500 125.0 33.167 13.989 22.875 11.862 141.0
14. TH (mg/l) 16.165 1.370 15.000 2.581 803.3 218.330 24.120 152.50 38.622 2,003.3
15. DO (mg/l) 5.805 0.066 6.007 0.304 5.1 5.150 0.336 5.624 0.309 5.6
Key to Tables 1 and 2:
NAFDAC: National Food Drug Administration and Control
SON: Standard Organisation of Nigeria.
STD: standard
MOV: mean of observed values
SD: standard deviation of mean value
SDF WHO: Significance difference from WHO standard
SDF WHO: Significance difference from NAFDAC/SON standard
BH1: borehole 1, Ibeju-Lekki LGA, 50m from coastline
BH2: borehole 2, Ibeju-Lekki LGA, 100-150m from coastline
BH3: borehole 3, Eti-Osa LGA, 50m from coastline
BH4: borehole 4, Eti-Osa LGA, 100- 150m from coastline
-1 0
UNITS: EC = (µScm ), Temperature = C, All other parameters = mg/l
Discussion
The pH followed an increasing trend i.e. from neutral to more alkaline every other week from the first week to the
fourth week for all boreholes
This is indicative of a reduced carbonic acid process (i.e. a decreased dissolution of CO2 in water). The coastal
aquifer in Eti-Osa showed to have higher mean values of electrical conductivity (EC) at 3282.83 (µScm-1) and
-1
2036.340(µScm ); TDS values of 2307.91 (mg/l) and 1427.1 (mg/l ) for bore holes 50m and 150 m respectively
away from the coast, both figures being far above the standard limits. This is indicative of high enrichment of salts
inthe aquifer, a pointer to saltwater intrusion (as a result of a pumping operation) [7] and contamination from
industrial activities in that it is densely habited and industrialized. It can be seen in general manner that the high
values of electrical conductivity (EC) and TDS are related to saltwater intrusion, as supported by corresponding
+ -
high levels of Na and Cl observed in Eti-Osa, but not in Ibeju-Lekki where no domination of any cation or anion
was observed.
The high Cl- and Na+ ion concentration is indicative of saltwater impact on the groundwater of Eti-Osa[7],
whereas that is not the case in Ibeju-Lekki, where values of measured parameters are within the permissible limits
as prescribed by WHO and NAFDAC.
I. Soluble Cations and Anions
Generally, the chemical composition of groundwater is primarily dependent on the type of geochemical
processes and chemical reactions taking place within the groundwater system. The composition of ions in Eti-
Osa and Ibeju-Lekki is dependent on the variant geochemical processes and resultant reactions that take
place in both locations.
Analytical result presents the abundance of these ions in the following order: Na+> Ca2+> K+> Mg2+ = Cl- > SO4->
- - 2+ + + 2+ -
NO3 > HCO3 for Eti-Osa and Ibeju-Lekki showed relative abundance in the order; Ca > Na > K > Mg = Cl >
- - -
SO4 > NO3 > HCO3 . Comparing this trend with the compositional make up for seawater (table 2) and fresh
groundwater shows similarities, which is indicative of saltwater intrusion, giving the closeness between the
saltwater nature of the beach and the adjacent groundwater.
Sampling points at Eti-Osa had the highest groundwater salinity, the nearest well to the shorelines showed 0.853
measure of salinity and the later which is about 100-150m had a salinity of 0.875. Although compared to the
salinity of the beach, this is only about 4.82% of Alpha beach's (Eti-Osa LGA) salinity. In view of this, consider
444
the assumption of the Environment Canterbury Regional Council (2012), that “If groundwater became
contaminated by 2% sea water this would result in chloride concentrations above the guideline values for
drinking water” [11]. This assumption supports a case for intrusion of saltwater in the groundwater of Eti-Osa as
opposed to the situation in Ibeju-Lekki where the percentage of salinity comparable to Eleko beach is 0.004%.
Comparison with WHO and NAFDAC Water Quality standards.

Groundwater from BH1 and BH2 from Ibeju-Lekki were found to meet all WHO and NAFDAC portable water
quality requirements, however groundwater samples from Eti-Osa LGA (BH3 and BH4), did not meet all WHO
and NAFDAC set standards for water quality. Table 3 shows mean parameter values of groundwater samples
investigated in comparison with standards of WHO and NAFDAC as discussed onwards.
445
TABLE 3: COMPARISONS WITH WHO AND NAFDAC STANDARDS FOR WATER QUALITY.
ARAMET WHO NAFD IBEJU-LEKKI ETI-0SA
ERS STD AC
STD
BH2 BH2 BH3 BH4
MOV SDF SDF MOV SDF SDF MOV SDF SDF MOV SDF SDF
WHO NAFD WHO NAFD WHO NAFD WHO NAFD
AC AC AC AC
pH 7.5 (6.5 – 7.5 (6 -5 7.825 0.184 0.184 7.750 0.374 0.374 7.375 0.527 0.527 7.525 0.928 0.928
8.5) -8.5)
Temperatur - - 31.075 - - 31.815 - - 30.165 - - 30.500 - -

e, 0C
EC (µScm - 1200 1000 267.090 .000 .000 208.620 .000 .000 3282.83 0.001 .000 2,036.34 0.005 0.003
1
) 0
TDS(mg/l) 1500 500 186.500 .000 .000 145.860 .000 .000 2,307.91 0.003 .000 1,427.10 0.436 0.001
0
446
SAL (mg/l) - - 0.0225 - - 0.020 - - 0.832 - - 0.875 - -
Ca2+ (mg/l) - 75 11.740 - .000 10.750 - .000 167.850 - 0.001 111.680 - 0.122
2+
Mg (mg/l) 20 20 4.750 .000 .000 3.500 .000 .000 39.750 0.200 0.200 27.500 0.254 0.245
K+ (mg/l) - 10 5.000 - 0.003 6.500 - 0.006 37.250 - .000 54.00 - 0.088
+
Na (mg/l) 200 - 9.832 .000 - 10.500 .000 - 158.16 0.009 - 265.92 0.548 -
HCO3- - - 0.022 - - 0.020 - - 0.022 - - 0.022 - -
(mg/l)
NO3- (mg/l) 10 10 2.190 .000 .000 1.825 .000 .000 28.470 0.001 0.001 16.747 0.010 0.010
-
CL (mg/l) 250 100 13.835 .000 .000 12.332 .000 .000 481.580 .000 .000 524.580 0.030 0.009
SO42- (mg/l) 500 100 8.665 .000 0.002 6.750 .000 0.001 33.167 .000 .000 22.875 .000 .000
TH (mg/l) 500 100 16.165 .000 .000 15.000 .000 .000 218.330 .000 0.002 152.50 .000 0.073
DO (mg/l) - - 5.805 - - 6.007 - - 5.150 - - 5.624 - -

Ibeju-Lekki LGA
Based on SPSS analysis it was observed that peculiar interrelationships exists between the parameters which
mirror the groundwater condition in the area.
In BH1, pH has very high correlation with HCO3– (r = 0.969, p = 0.031). pH indicates the strength of the water
–
reacting as an acid or base, a very strong correlation with HCO3 supports the dissolution of CO2 into carbonic
acid characteristic of weathering process involving a calcite substrata.
EC has a very high positive correlation with Ca2+, Mg2+ and SO42– at r values greater than 0.9, P < 0.05. TDS has a
very high positive correlations with Mg2+ (r = 0.968, p = 0.032). This implies that EC and TDS are controlled by
2+ –
these ions in a linear relationship. Mg has a very high positive correlations with NO3 (r = 0.960, p = 0.04),
+ -
whereas TH have a very high positive correlation with Na (r = 0.961, p = 0.039) and Cl (r = 0.988, p = 0.12).
In BH2, pH has very high correlation with EC, TDS, NO3, Cl- and TH at values of r ≥ 9.0, p < 0.05. EC has very
high correlations with NO3- and TH at values of r = 0.99, p = 0.001. TDS have very high correlations with HCO3–
–
(r = 0.971, p = 0. 021), NO3 (r = 1.000, p = 0.000) and TH (r = 0.998, 0.002). This implies that EC and TDS are
controlled by these ions.
K+ has very high positive correlation with Na+ ( r = 1.000, p = 0.000) and Cl- (r = 0.962, p = 0.38). Na+ has very
high positive correlation with Cl- (r = 0.962, p = 0.38). HCO3– have very high positive correlations with NO3- of (r
= 0.979, p = 0.029) and high positive correlation with TH (r = 0.983, p = 0.017).
Eti-OsaLGA
– -
In BH3, EC has a high positive correlation with NO3 (r = 0.988, p = 0.012), Cl (r = 0.970, p = 0.030). TDS has a
– - 2+
high positive correlation with NO3 (r = 0.990, p = 0.010), Cl (r = 0.971, p = 0.029) and Mg (r = 0.966, p =
0.034). This implies that NO3– and Cl– are the controlling factors of the concentration of EC and TDS.
Ca2+ as well has a high positive correlation with Na+ (r = 0.994, p = 0.006), NO3– (r = 0.979, p = 0.021), and Cl- (r
+ – -
= 0.971, p = 0.029). Na has a high positive correlation with NO3 (r = 0.961, p = 0.039), and Cl (r = 0.984, p =
– -
0.016). NO3 has a high positive correlation with Cl (r = 0.995, p = 0.005) and TH (r = 0.952, p = 0.048). The
strong correlations with the major sea water ions with NO3- and it suggests impact of contamination from sewage
waste and other anthropogenic activities, not neglecting the influence of seawater intrusion into the aquifer. This
assertion holds frail as wells built in areas proximate to the coast are often times not built very deep far below the
direct impact of sewage pits.
In BH4, salinity has a very high positive correlation with Na+ (r = 0.99, p = 0.02) and has a very high negative
correlation with DO (r = - 0.966, p = 0.034). Also K+ has a very high positive correlation with Cl– and SO42– at r
–
values > 0.9 and p values < 0.05, but has a high negative correlation with HCO3 (r = - 0.998, p = 0.02) and DO (r
- 2–
= - 0.993, p = 0.07). Cl as well have a high positive correlation with SO4 (r = 1.000. p = 0.00). These correlations
support the case for saltwater intrusion in the sample point, as salinity is influenced in a linear relationship by
sodium ions concentration. This as well is the case between K+ ,Cl- and SO42–. It can also be deduced that these
–
parameters originate from a common source (Al-Khatib and Al-Najar, 2011). On the other hand, HCO3 has high
- 2– – -
negative correlations with Cl and SO4 , similarly, it can be deduced that HCO3 is not of same source as Cl and
So42–.
447
Figure 1: mean values of weekly physiochemical measurements of boreholes in both Ibeju-Lekki (1, 2) and
Eti-Osa (3, 4)
Figure 1 is a clustered column plot of mean values of water characteristics for the four boreholes under
investigation.
Groundwater classification by piper diagram

To follow on the likely conformational changes to the aquifer water that due to saltwater intrusion a plot of
chemical data on diamond shaped trilinear diagram (Figure 2) reveals groundwater hydrochemicalfacies
classification of BH1 and BH2 from Ibeju-Lekki, which fall in the fields of Ca-Mg-Cl-SO4 and thus
characterized as Ca-SO4 waters - typical of gypsum ground waters and mine drainage water type. In Eti-Osa LGA
the borehole proximate to the coastline i.e. BH3 which is about 50m to the coastline was found to be of Ca-SO4
water type. It is important to note that it is often times difficult to identify the hydrogeochemical facie of shallow
groundwater due to excess infiltration from secondary recharge from irrigation water [12] or precipitation as
well as influxes from anthropogenic activities .
However, BH4 in Eti-Osa was found to have Na-Cl water type, characterized by high EC, TDS and salinity, and
has been in used for over a year; however the borehole is located 100-150m from the coastline. This suggests that
the continuous exploitation of groundwater has led to saltwater intrusion [13]. Figure 3 shows the water quality
situation or changes in Eti-Osa is characterised by TDS, this depicts influence from a source of dissolvable
solids.
448
Figure 2: Hydrochemicalfacies of groundwater in Eti-Osa and Ibeju-Lekki LGA
Figure 3: Hydrochemical Characteristics in Proportion to TDS in Eti-Osa and Ibeju-Lekki LGAs
Satellite imges of the study areas.
Conclusion
Groundwater sample from coastal environments with varying tendencies to climate change hazards were
studied, Eti-Osa and Ibeju-LekkiLocal Government coastal communities with different patterns in land usage,
population densities and industrial activities having groundwater which are influenced differently by nearby
coast were focused. Water samples from two beaches in each Local Government were analysed for
physiochemical parameters (pH, Temperature, EC, TDS, Salinity, Ca2+ , Mg2+ , K+ , Na+, HCO3- , NO3- , CL- ,SO42- ,
TH, DO). The results were used in assessing water quality and the effects of seawater intrusion on groundwater
which was analyzed.
Considering hydrochemical changes in groundwater chemistry, Eti-Osa aquifers were compared to Ibeju-Lekki
with less pumping pressure. Thus, saltwater intrusion into the aquifer was established in Eti-osa, making water
samples from Eti-Osa to be of low quality, as it failed to meet requirements of potable water quality set by WHO
and NAFDAC. However water samples from Ibeju-Lekki met all requirements for portable water quality set by
WHO and NAFDAC.
The groundwaters were characterised using piper diagram. Ibeju-Lekki water samples showed
hydrochemicalfacies of Ca-SO4 water type showing early signs of oncoming intrusion, whereas Eti-Osa aquifers
449
showed Ca-SO4 and Na-Cl water type, which confirmed salt water intrusion in the area.
Recommendations
Considering the findings from this research it is recommended as follows:
1. Establishment of an environmental risk assessment network which would initiate a hydrogeological
monitoring and management plan to address seawater intrusion issues in Lagos state.
2. Encouragement of sustainability practices through awareness creation and public environmental health
and resource management education is highly recommended.
REFERENCES
1. UNEP/ROLAC (2012), Global Environmental Outlook (GEO 5): policy options for Latin America and
the Caribbean , UNEP, Panama. Chapter 12
2. J. A.Jideonwo, (2014). Ensuring Sustainable Water Supply in Lagos, Nigeria, University of
Pennsylvania, ScholarlyCommons; Master of Environmental Studies Capstone Projects Department
of Earth and Environmental Science, University of Pennsylvania, USA
http://repository.upenn.edu/mes_capstones/58 accessed 21/01/2015
3. LSG Lagos State Government (2011).Population.Retrieved from
www.lagosstate.gov.ng/pagelinks.php?p=6
4. J. Omenai, & D. Ayodele, (2014). The Vulnerability of Eti-Osa and Ibeju-Lekki Coastal
Communities in Lagos State Nigeria to Climate Change Hazards, Research on Humanities and
Social Sciences, 4(27).
5. A. D. Werner, M. Bakker, V. E. A. Post, A. Vandenbohede, L. Chunhui, B. Ataie-Ashtiani, C. T.
Simmons, & D. A. Barry (2012). Seawater intrusion processes, investigation and management:
Recent advances and future challenges.Advances in Water Resource, 1.
6. A. U. Oteri, & F. P. Atolagbe, (2003). Saltwater Intrusion into Coastal Aquifers in Nigeria.The
Second International Conference on Saltwater Intrusion and Coastal Aquifers — Monitoring, Modeling,
and Management. Mérida, Yucatán, México, March 30 - April 2, 2003
7. A. Z. Aris, M. H. Abdullah, K. W. Kim, & S. M. Praveena, (2009). Hydrochemical changes in a small

tropical island's aquifer: Manukanisland, Sabah, Malaysia. Environmental Geology, 56:1721–1732.
DOI 10.1007/s00254-008-1275-3
8. O. Ohwoghere- Asuma, I. A. Akpoborie, E. G. Akpokodje, (2014). Investigation of Saltwater
Intrusion in Warri –Effurun Shallow Groundwater Aquifer from 2D Electrical Resistivity
Imaging and Hydraulic Gradient Data.New York Science Journal 7(12):20-29.
http://www.sciencepub.net/newyork
9. N. M. Isa, A. Z.Aris, W.N. Azmin, &W. Sulaiman, (2012). Extent and severity of groundwater
contamination based on hydrochemistry mechanism of sandy tropical coastal aquifer. Science of the
Total Environment, 8(69): 414–425. DOI: 10.1016/j.scitotenv.2012.08.069
10. American Public Health Association (APHA), (1998).American Water Works Association and Water
Pollution Control Federation-Standard methods for the examination of water and wastewater.
Washington DC. 20: 143- 146, 248-252.
11. ECRC Environment Canterbury Regional Council, (2012).Seawater intrusion network review,
Environment Canterbury Technical Report, September 2012, Report No.R12/35,Canterbury,
England. Authors: Marta Scott & Nicola Wilson
450
12. E. Dano, August, 2010. Identification of geochemical facies through major ion data and additional
parameters from shallow groundwater utilizing a comparison of geomathematics and traditional
methods in Las Vegas Valley, Nevada.UNLVTheses/Dissertations/Professional
Papers/Capstones.Paper 849.Retrieved from .
13. N.U.Kura,M.F. Ramli,W.N.A. Sulaiman,S. Ibrahim,A.S.Aris, & A. Mustapha, (2013). Evaluation
of Factors Influencing the Groundwater Chemistry in a Small Tropical Island of Malaysia,
International Journal of Environmental Research andPublic Health, 10: 1861-1881;
doi:10.3390/ijerph10051861
451
COMPARATIVE STUDIES OF THE INHIBITIVE POTENTIALS OF RED PEANUT SKIN
EXTRACT AND LASIANTHERA AFRICANA P. BEAUV. (NKANKA LEAF) MUCILAGE ON MILD
STEEL CORROSION IN SULPHURIC ACID SOLUTION.
a b c
A.O. James, Kalu Nnennaand Akaranta O.
a, b & C
Department of Pure and Industrial Chemistry, University of Port Harcourt,
P.M.B. 5323, Port Harcourt
a
Email address:abosede.james@uniport.edu.ng
b
Email address: nellyebony@gmail.com
c
Email address: akaranta.onyewuchi@uniport.edu.ng
a
Phone Number: 08057314332
b
c
Abstract:
The inhibition efficiency of red peanut skin extract and lasianthera africana P. beauv. mucilage on mild steel
deterioration (corrosion) in 2M sulphuric acid was studied by the method of weight loss. The extracts were
prepared by immersing the red peanut skin and nkanka leaves in hot water (1000g/l and 100g/0.5l of water
0 0 0
respectively). The inhibition efficiencies were studied at various temperatures: 30 C, 40 C and 50 C. Results
obtained indicated that mild steel corrosion in sulphuric acid was mitigated significantly by the extracts. As the
inhibitor concentration in the acid increased, the inhibition efficiency also increased while temperature increase
resulted to a decline in the inhibition efficiency. The difference in the inhibition potency of the two extracts is
explained in terms of the difference in the molecular structures of their active component. The active component
in red peanut skin extract is catechin while that of lasianthera africana p. beauv. (nkanka leaf) mucilage is pectin.
Keywords:Corrosion; Red peanut skin; Mild steel; Mucillage; Weight loss.
INTRODUCTION
Corrosion is the weakening in the structure of a material due to interactiob with its surroundings. It can also be
described as the destructive attack of materials mostly metals, by electrochemical reactions (Natty, 2010).
Corrosion processes are redox reactions involving oxidation and reduction. (Lebo, et al., 2001).
Mild steel is exposed to the action of acid in industrial processes in which acids have important functions, for
example in oil well acidification, acid pickling, acid cleaning, and acid descaling. Corrosion inhibition of mild
steel in acid solutions has become one of the most urgent and severe challenges in acid pickling process (Shukla
and Qurarishi, 2010; Aljourani et al., 2009).
Corrosion inhibitors are substances added in small quantities to an environment to prevent corrosion of metals
(Quarishi, 1999).The use of inhibitors is one of the best options of protecting metals and alloys against corrosion.
The selection of inhibitor is controlled by its economic availability, its efficiency to inhibit the substrate material
and its environmental side effects.
Most of the efficient acid inhibitors are organic compounds containing nitrogen, sulphur and/or oxygen atoms in
their molecule (Sudhish and Quraishi, 2010; Eddy and Ebenso, 2008; Sharma et al., 2008). The toxicity of
organic and inorganic corrosion inhibitors to the environment has prompted the search for safer corrosion
inhibitors such as green corrosion inhibitors as they are biodegradable and do not contain heavy metals or other
toxic compounds. In addition to being environmentally friendly and ecologically acceptable, plant products are
452
inexpensive, readily available and renewable (Eddy and Ebenso, 2008).
In this study, the inhibition efficiency of mild steel corrosion in red peanut skin extract and lasianthera africana
mucilage in sulphuric acid at different concentrations and temperatures wasinvestigated by the method of weight
loss.
EXPERIMENTAL PROCEDURE
Preparation of the Metal Coupons

The metal coupons were purchased at steel market, Mile 1, Port Harcourt. They were mechanically press-cut into
4 x 3cmcoupons with a hole, 2mm in diameter at the tip for suspending the metal into the acid solution. The mild
steel coupons were not polished any further but was degreased in absolute ethanol and dried in acetone; then
stored in a dessicator to prevent any form of contamination before the experiment.
Preparation of Various Concentrations of Red Peanut Skin Extract

The red skin of roasted peanut was obtained locally from a groundnut seller at Choba junction in Port Harcourt,
Rivers State. 1000g of the red peanut skin was boiled in 1litre of water for about 3 hours. Filtration was done after
cooling of the mixture. The acid solution (2M H2SO4) was added to varied amounts of the extract solution in %v/v
using the following procedure:
10%v/v concentration was prepared by adding 90ml of H2SO4 to 10ml of the extract in a 250ml beaker; same
procedure was followed to prepare other concentrations: 20%v/v, 30%v/v, 40%v/v and50%v/v. Another beaker
containing 100ml H2SO4 was used as blank for the weight loss experiment.
Preparation of Mucilage From Nkanka Leaves (Lasianthera Africana)
Nkanka leaves (Lasianthera africana) were obtained locally from a farmland in Etchie Local Government of
Rivers State. The leaves were washed, dried, chopped and weighed. 100g of the leaves were completely
o
immersed in 0.5litres of hot water (100 C) for 3 hours till the water turned cold. The mucilage (slimy liquid) was
squeezed out of the leaf and filtered with a sieve to remove tiny leaf particles. 2M H2SO4 solution was added to
varied amounts of the extract solution in %v/v.
Weight Loss Determination

The various concentrations (blank, 10%v/v, 20%v/v, 30%v/v, 40%v/v, and 50%v/v) of both extracts of red
peanut skin and mucilage in 2M H2SO4 were maintained at 300C. The mild steel coupons which had been weighed
were placed in the corrodent-inhibitor solutions. Each coupon was recovered from these solutions every 24hours
consistently for 168hours (7 days). Each retrieved coupon was immediately dropped into distilled water to stop
the corrosion reaction, washed, dried and reweighed. The coupon weight difference was recorded as the weight
loss. The temperature was raised to 40oC and 50oC and the experiment was performed again in a thermostatic
water bath.
Efficiency of the inhibition (%E) was computed from the outlined equation below:
%E = ? WB – ? Wi X 100------------------- (1)
? WB 1
Where ? WBand ? Wi are the weight loss values of the mild steel coupons in the absence
(blank) and presence of an additive respectively.
453
The results obtained from the weight loss experiment with inferences drawn from them are discussed below:
Inhibitory Effect of Red Peanut Skin Extract on Mild Steel Corrosion
Fig. 1: Variation of Weight Loss with Time for Mild Steel Coupons in 2.0M H2S04 and various concentrations of
red peanut skin extract solution at 300C.
From the variation of weight loss with time of exposure of mild steel in 2M sulphuric acid (blank) at 300C (Fig. 1)
compared with those containing the additives, there is a remarkable decrease in weight loss signifying corrosion
inhibition.
The same observations were made at 40oC and 50oC. This shows that the red peanut skin extract is very
economical since it could inhibit corrosion greatly even at small amounts.
Effect of Temperature on the Inhibition Efficiency of Red Peanut Skin Extract
454
Fig. 2: Variation of inhibition efficiency with inhibitor concentration of red peanut skin extract for mild steel
corrosion in 2M H2SO4 solution at different temperatures.
From fig. 2, it can be seen that as the temperature of the system was increased from 300C to 500C, the inhibition
0
efficiency decreased. But even at this temperature (50 C), the red peanut skin extract still inhibited corrosion
significantly.
Effect of lasianthera africana mucilage on mild steel corrosion in 2M H2SO4 solution:

The figures below show the effect of lasianthera africana mucilage on the weight loss of mild steel in
sulphuric acid solution at the temperatures studied.
Fig. 3: Variation of Weight Loss with Time for Mild Steel Coupons in 2.0M H2S04 solution containing
different concentrations of the mucilage extract from lasianthera africana (nkanka leaves) at 300C.
Fig. 3 shows the effect of the addition of the mucilage the acid, it was observed that as the additive concentration
increased, the metal weight loss decreased significantly, with 10%v/v showing the highest weight loss while the
highest additive concentration 50%v/v showed the least weight loss at the different temperatures studied. This
o
signifies inhibition of the metal dissolution in sulphuric acid.Same behaviour was recorded at 40 and 50 C.
Effect of Temperature on the Inhibition Efficiency of lasianthera africana mucilage
455
Fig. 4: Variation of inhibition efficiency with inhibitor concentration of lasianthera africana mucilage for mild
steel corrosion in 2M H2SO4 solution at different temperatures.
From fig. 4, it can be seen that as the temperature of the system was increased from 30 to 500C, the inhibition
efficiency decreased. But even at this temperature (500C), the mucilage still inhibited corrosion significantly.
The active components in the extracts and mucilage.

The active component in red peanut skin extract is catechin while that of lasianthera africana p. beauv. (nkanka
leaf) mucilage is pectin. Both possess electron-rich sites for their adsorption unto the metal surface leading
corrosion inhibition.
OH
OH
HO O
OH
OH
Figure 5: The structure of Catechin (Red Peanut skin).
456
Figure 6: The structure of Pectin (lasianthera africana mucilage)
Comparison of the corrosion inhibition potentials of the extract and the mucilage.
Table 1: The Inhibition Efficiency (%)for mild steel corrosion in 2M H2SO4 solutionby lasianthera
africana mucilage and red peanut skin extract at the temperatures studied
Inhibitor Inhibition Efficiency (%)

Concentration
(% v/v) Lasianthera africana mucilage Red peanut skin extract
30oC 40oC 50oC o
30 C 40oC 50oC
10 79.71 75.11 65.62 74.66 70.04 61.54
20 86.07 81.45 69.95 80.73 72.73 68.85
30 90.39 86.30 80.65 85.86 80.62 79.01
40 93.55 90.28 85.91 89.45 84.11 83.63
50 96.49 94.13 91.68 93.73 90.07 88.98
The inhibitive effect of the extract and the mucilage may be explained by considering the adsorption of their
active compounds through the electron rich oxygen atoms in the molecules on the corroding metal surface. The
active ingredient in lasianthera africana mucilage is Pectin while that of red peanut skin extract is catechin.It is
evident that the inhibition efficiency of the two compoundsdepends mainly on the molecular size of their
structures, the charge density on the adsorption sites,electron clouds, mode of interaction with the metal surface
and formation of metallic complexes. Adsorption is expected to take place primarily through the oxygen atoms
on the molecules. Considering the structures of the compounds (figs.5-6), there are numerous electron-rich
oxygen atoms in them, hence the high percentage inhibition efficiencies recorded (Table 1).
A comparative study of the two extracts shows that lasianthera africana mucilage recorded higher inhibition
efficiency than red peanut skinextracts (Table 1) at all the temperature studied. This may be due to the fact that
the oxygen atoms in pectin are more than those in catechin. Thus creating more adsorption sites and increased
inhibitive potency.
457
CONCLUSION
The following conclusions were made from this study:
§ Red peanut skin extract and lasianthera africana mucilageare good corrosion inhibitors for mild steel in
2M H2SO4 solution.
§ Inhibition efficiency decreased with increased temperature.
§ In both extracts, increase in inhibitor concentration led to an increase in the inhibition efficiency.
§ The mucilage extract exhibited greater inhibition efficiencies (max. = 96.49%) than the red peanut skin
extract (max. = 93.73) under the same conditions. This may be attributed to the size and nature of their
active components; larger structures with larger number of heteroatoms always give a higher inhibition.
Red peanut skin extract and lasianthera africana mucilage should be employed as eco-friendly corrosion
inhibitors and be used to replace toxic organic corrosion-inhibitors.
REFERENCES
1. Aljourani, J., Raeissi, K., Golozar, M.A. (2009). Benzimidazole and its derivatives as corrosion
inhibitors for mild steel in 1M HCl solution. Corros. Sci. Journal, 51:1836-1843.
2. Eddy, N.O., and Ebenso, E.E. (2008). Inhibitive and adsorption properties of ethanol extract of Musa
sapientum peels as a green corrosion inhibitor for mild steel. Afr. J. Pure Appl. Chem. 2(6):46-54.
3. James, A.O., Akaranta, O. and Awatefe, K. J. (2011). Red peanut skin: an excellent green inhibitor for
mild steel dissolution in hydrochloric acid solution. Alfa Universal, An International Journal of
Chemistry, 2(2): 72 – 78.
4. Lebo, S. E., Gargulak, J. D. and McNally, T. J. (2001). Encyclopedia of Chemical Technology; 7th ed.
John Wiley & Sons Inc. NewYork. pp. 420 – 424.
5. Natty, K. (2010). Corrosion and its Mitigation in the Oil and Gas Industry; Leonardo Journal of
Science, 23(6): 12, 19.
6. Quraishi, M. A. (1999). “Investigation of some green compounds as corrosion and scale inhibitors for
cooling systems,” Corrosion Journal,55(5) 493–497.
7. Shukla, S.K., and Quraishi, M.A. (2010). The effects of pharmaceutically active compound
doxycycline on the corrosion of mild steel in hydrochloric acid solution. Corros. Sci. 52(1): 314-321.
8. Sudhish, K.S., and Quraishi, M.A. (2010). Cefalexin drug: A new and efficient corrosion inhibitor for
mild steel in hydrochloric acid solution. Mater. Chem. Phys.120:142-147.
458
ASSESSMENT OF PHYSICOCHEMICAL PROPERTIES OF CLAY SAMPLES OBTAINED FROM
ASHAKA, POTISKUM AND TANGO (NIGERIA) AND A COMMERCIAL BENTONITE
OBTAINED FROM KOFAR RUWA MARKET.
1 2
Abdullahi S. L and A.A Audu
1
Kano State Polytechmic, Kano Nigeria
2
Department of Pure and Applied Chemistry, Bayero University Kano, Nigeria.
Correspondence author: slabdullahi39@gmail.com
Abstract
Samples of clays suspected to be bentonite were obtained from Ashaka, Tongo and Potiskum, Nigeria and a
commercial bentonite obtained from Kofar Ruwa market, (Kano State) were investigated for some of their
physicochemical properties. These determinations were carried out on the wet beneficiated clay samples.
The results were compared with results obtained from other studies on bentonite clays. The moisture content,
pH, density and swelling power were between 4-7%, 7.78-9.89,1.21-1.82g/cm3 and 10-13ml/2g respectively.
The CEC was between 61-72meq/100g, Ca2+ and Na+ were the main exchangeable cations (20.87-39.752 and
26.55-33.18meq/100g respectuvely) with calcium ion as the dominant. The chemical composition of the
clays indicates high percentages of CaO 1.155-14.850wt%. The silica and alumina contents of the samples
were in the ranges of 40.705-49.873 wt% and 14.856-16.744 wt% respectively. Fe2O3 content of the samples
was in the range of 4.802 – 5.606 wt%. . The results of the physicochemical analysis of the samples when
compared with the results from other studies on bentonite may suggest that the samples are bentonite clays.
Key words: Bentonite, montmorillonite, smectite, CEC, swelling power, chemical composition.
Introduction c
Moisture content is not a relevant parameter or indicator of bentonite performance. Preliminary
limiting (maximum) value for water (moisture content) was set to 13 % (Trauger, 1994). The moisture
contents of the four samples obtained in this study were in between 4-7%. These values fall within the range
reported by Ahonen et al., (2008). The pH values of the clays were similar to the pH reported by Nweke et
al., (2015) Shah et al., (2013); James et al., (2008) and were within the range given for both Na and Ca
bentonite( 8.5-10.5 and < 8.5) respectively.
The density values for the samples were higher than the value obtained by James et al, (2008), Table
1, but similar to the values reported by Nweke et al., (2015), Table 2.
A moderately swelling bentonite will produce 15-20ml/g gel, a good variety 25ml and an excellent
grade will produce 30ml or more (Ahonen et al., 2008; Shah et al.,2013). All the samples in this study have
lower swelling index <15ml/2g, but higher than the value reported by Shah et al., (2013) of 7ml/2g for
Ca-bentonite. The weak swelling properties are thought to buttress the presence of low expansive
montmorillonite, most probably the calcium variety. Variation in exchangeable cations affects the maximum
amount of water uptake and swelling.
The Na, K, Ca and Mg concentrations in the extracts were determined using flame emission
spectrometer/ atomic absorption spectrometer (FES/AAS). The CEC was calculated as sum of exchangeable
+ + 2+ 2+
elements (Na , K , Mg and Ca ) and given as cmol+/kg (centimoles of charge per kilogram of material,
which corresponds to the also commonly used CEC meq/100 g). The result in Table 1for the four samples
revealed Ca2+ and Na+ as the main exchangeable cations with calcium ion as the dominant cation. Calcium
was higher in three of the samples (GT, GA and P) and also higher than the values reported by Ahonen et al,
(2008), but lower than those reported by Nweke et al, (2015) [Table 3]. The Calcium content reflects both
459
the interlayer cation and the calcite content of the samples. The other sample (KR) has higher sodium content
which also reflects the interlayer cation. The values of Mg2+among the exchangeable cations in all the
samples were the lowest. The values obtained in Table 1 for Na+, Ca2+, K+, Mg2+ and CEC when compared
2+
with the values obtained by James et al, (2008), they were found to be higher except Mg values which were
+
lower. The K cations present in the samples was in the range 6.45-9.2 meq/100g which was higher than the
values obtained by Nweke et al, (2015) and Ahonen et al, (2008). Potassium content is associated with
smectites as an exchangeable cation and may also be considered as an indication of the presence of feldspar
or micas in the samples. Natural clays rich in montmorillonite generally contain a mixture of exchangeable
2+ + 3+ + 2+
cations, including Mg , K and Fe , although Na and Ca predominate but in varied proportions
The typical CEC range of pure smectite is from 80 meq/100 g to 150 meq/100 g (Grim, 1968).
According to Gomes, (1988), the CEC of the montmorillonite clays is between 40 and 150 meq/100 g of
clay. The CEC values in this study ranged from 61-72meq/100g, sample KR with the highest value. CEC
values published for MX-80 purified sample are 76 meq/100g (Madsen, 1998); 102-140 meq/100g; 88-110
meq/100g (Pusch, 1999; 2001); 97 meq/100g (Neaman et al., 2003); 84-109 cmol+/kg (Carlson, 2004). CEC
values for other studied samples are: Friedland clay, 60 meq/100g; 40 meq/100g (Pusch, 1999; 2001) Rokle
bentonite, 62 meq/100 g (Pusch, 2001). The lower values are in good accordance with values received in this
study.
The results for the chemical analysis of the samples are presented as percentage of oxides. The silica
and alumina contents of the samples were in the ranges of 40.705 - 49.873 wt% and 14.856 - 16.744 wt%
respectively, the values were close to the reported values by Ahonen et al., (2008); Nweke et al., (2015) for
Wyoming bentonite (45 and 17wt% respectively). Ahmad et al., (2012) reported a literature value of
13.33wt% alumina content for Ca-bentonite. The iron content of the samples in this study was in the range of
4.802 – 5.606 wt% which falls within the range reported from other studies (Table). The method does not
differentiate between divalent and trivalent iron, therefore the values of Fe2O3 obtained for each sample
represent its total iron content. Sample P and KR have values higher than values obtained from reported
studies (14.692 and 8.850 wt %), samples GT and GA have values similar to reported values by James et al.,
(2008); Ahonen et al., (2008); Shah et al., (2013) and Nweke et al., (2015).
Conclusion
This study assed the physicochemical properties of some clay samples suspected to be bentonite
from Ashaka, Tango, Potiskum and commercial bentonite clay. All the results of the analysis in this study
were similar with the results obtained from other studies on bentonite samples although there were some
variations. Physicochemical properties of bentonite clays typically vary both within and between deposits
due to differences in the degree of substitution within the smectite structure and the nature of the
exchangeable cations present and also due to type and amount of impurities present, hence the variations
observed as such. Therefore the samples could be concluded as bentonite clays.
The quality and grade of bentonites are related to the physicochemical properties and smectite content of the
clay sample and its measure of likely industrial application. This study did not report on the mineralogical
and geotechnical properties of the samples, therefore further analyses such as FTIR, XRD, SEM, TEM,
TGA, liquid limit, plastic limit, shrinkage limit, viscosity, porosity, particle size etc would be required in
order to obtain more information on the mineral content and quality of the samples. Usually the quality of
bentonite can be improved by activation with Na2CO3 in order to convert the Ca-bentonite into Na-bentonite.
Table 1: Physical characteristics of the samples
460
Parameter GA GT KR P James et
al.,
(2008)
Moisture 6.7 6.9 4.7 5.4 NA

(%)
pH 8.78 7.89 9.89 7.78 8.82
Density 1.26 1.35 1.21 1.82 0.961
(g/cm3)
Swelling 11 13 10 13 NA
Test
(ml/2g)
NA: not available
Table 2: The physical characteristics of the samples compared with other studies in a range.
Parameter Nweke et Ahonen et

al., (2015) al., (2008)
This study
Moisture
(%) 4-7 NA 6.0-16.0
pH 7.89-9.89 7.8-9.3 NA
Density
(g/cm3)
1.21-1.82 1.45-1.68 NA
Swelling
Test (ml/2g)
10.0-13.0 NA 23-30
NA: not available
Table 3: Exchangeable cations and cation exchange capacity of the samples. CEC was calculated as
sum of exchangeable cations (Meq/100g).
Parameter GA GT KR P James et
al.,
(2008)
K+ 8.1 9.2 6.45 7.025 1.26
+
Na 30.169 20.87 39.752 21.643 1.37
Ca2+ 33.183 32.378 26.546 33.183 27.86
Mg2+ 0.028 0.037 0.019 0.037 1.08
CEC 71.480 62.485 72.767 61.888 31.57
Table 4: Exchangeable cations and cation exchange capacity (Meq/100g) of the samples in a range
compared with other studies.
461
Nweke et al.,
Ahonen et
(2015)
Parameter This study al., (2008)
0.5-2.6
K+ 6.45-9.2 1.49-3.66
46.5-81.3
Na+ 20.87-39.752 11.1-16.6
26.546-33.183 12.2-28.7
Ca2+ 28.8-38.6
7.2-13.2
Mg2+ 0.019-0.037 4.3-6.7
CEC 61.89-72.77 45.69-65.56 85-104.3
Table 5: Chemical composition (wt-%) of bentonite samples presented as oxide; analyzed by XRF.
Sample (wt%)
GA GT KR PK Shah et James et
al., (2013) al.,
chemical (2008)
oxide
1.655 1.426 2.269 1.551 0.24 0.14
Na2O
MgO 2.075 2.077 5.257 8.561 6.71 1
14.856 14.981 15.93 16.744 17.99 13.58
Al2O3
48.164 49.873 43.717 40.705 59.59 58.79
SiO2
1.061 1.014 1.201 1.201 NA 0.04
P2O5
1.595 1.759 1.411 1.394 1.02 0.94
K2O
CaO 1.155 1.806 8.692 14.85 1.59 1.11
0.937 0.874 0.826 0.77 0.32 1.36
TiO 2
4.802 5.124 5.192 5.606 2.9 7.06
Fe2O3
NA: not available
Table 6: Chemical composition (wt-%) of bentonite samples presented as oxide; analyzed by XRF
compared with other studies in a range.
Nweke et al.,
Chemical Ahonen et al.,
(2015)
oxide This study (2008)
0.68-1.98 1.9-3.0
Na2O 1.426-2.269
MgO 2.075-8.561 0.32-2.02 2.3-3.8
19.90-25.08 14.1-18.7
Al2O3 14.856-16.744
50.10-58.96 43.1-56.6
SiO2 40.705-49.873
ND 0.04-0.79
P2O5 1.014-1.201
0.52-1.40 0.1-1.1
K2O 1.394-1.759
CaO 1.155-14.850 1.00-5.42 1.3-5.6
1.10-2.10 0.16-1.82
TiO 2 0.77-0.937
3.80-4.67 3.5-11.4
Fe2O3 4.802-5.606
ND: not detected
462
Reference
Ahmed A.S, Salahudeen, N., Ajinomoh, C.S., Hamza, H. and Ohikere, A. (2012); Studies on the Mineral
and Chemical Characteristics of Pindiga Bentonitic Clay. Petroleum Technology Development Journal
(ISSN 1595-9104). An International Journal. 1: 1-8
Ahonen, L., Korkeakoski, P., Tiljander, M., Kivikoski, H. and Rainer L.(2008); Quality Assurance of
the Bentonite Material. POSIVA OY Working Report 33
Apugo-Nwosu, T.U., Mohammed-Dabo, I.A., Ahmed, A.S., Abubakar, G., Alkali, A.S. and Ayilara, S.I.
(2011). Studies on the Suitability of Ubakala Bentonitic Clay for Oil Well Drilling Mud Formation. British J.
Appl. Sci. Technol. 1(4):152-171.
Arabi A.S, Ibrahim AA, Muhammad MA, Kwaya MY, Mustapha S (2011). Comparative Evaluation of
Rheological Properties of Standard Commercial Bentonite and a Locally Beneficiated Bentonitic Clay from a
Marine Deposit in Upper Benue Basin, Nigeria. British J. Appl. Sci. Technol. 1(4):211-221.1.
Asad, A., Shantanu, K., Mohammad, A.D. and Raquibul, H. (2013); Suitability of Bentonite Clay: An
Analytical Approach. International Journal of Earth Science, 2(3): 88-95.
Bergaya, F., Theng, B.K.G and Lagaly (2006); Modified Clay and Clay Minerals. In: Handbook of Clay
Science.pp 261-422. Elsevier, Oxford, united Kingdom.
Carlson, L. (2004); Bentonite Mineralogy. Part 1: Methods of Investigations – A Literature
Review; Part 2: Mineralogical Research of Selected Bentonites. Posiva Oy, Working
Report 02.
Donald S. R. and Quirine K. (2011); Recommended Methods for Determining Soil Cation Exchange
Capacity, Chapter 9: Recommended Soil Testing Procedures for the Northeastern United States.
Cooperative Bulletin No. 493
2+
Emmerich, K., Plotze, M. & Kahr, G. (2001) Reversible collapse and Mg release of de- and rehydroxylated
homoionic cis-vacant montmorillonites. Applied Clay Science, 19:143-154.
nd
Grim, R.E. (1968); Clay Mineralogy. 2 Edition, McGraw- Hill, New York, 596 pp.
Holtzer, M., Bobrowski, A. and Grabowska, B. (2011); Montmorillonite: A Comparison of Methods for
its Determination in Foundry Bentonites, Metabk 50(2):119-122
James O. O., Adediran, M. M., Adekola, F. A., Odebunmi, E. O. and Adekeye, J. I. D. (2008); Beneficiation
and Characterisation of a Bentonite from North-Eastern Nigeria. Journal of the North Carolina Academy of
Science, 124(4):154–158.
M adsen, F. T., (199 8). Clay Mineralogical Investigations Related to Nuclear Waste Disposal. Clay Minerals
33:109-129.
463
Neaman, A., Pelletier, M. and Villieras, F., (2003); The Effects of Exchanged Cation, Compression, Heating
and Hydration on Textural Properties of Bulk Bentonite and its Corresponding Purified Montmorillonite.
Applied Clay Science, 22:153-168.
Nweke, O. M., Igwe, E. O. and Nnabo, P. N. 2015): Comparative Evaluation of Clays from Abakaliki
Formation with Commercial Bentonite Clays for Use as Drilling Mud. African Journal of
Environmental Science and Technology. 9(6):508-518
Pusch, R., (1999); Is Montmorillonite-rich Clay of MX-80 Type the Ideal Buffer for Isolation of HLW? SKB
Technical Report TR-99-33.
Pusch, R., (200); The Buffer and Backfill Handbook. Part 2: Materials and techniques.
SKB Technical Report TR-02-12, 198 pp.
RMRDC Raw Materials Research and Development Council (2007), Technical Brief on Mineral Raw
Materials in Nigeria – Bentonite, Revised edn., Abuja.
Shah, L.A., Khattak, N.S., Valenzuela, M.G.S., Manan, A.and Valenzuela di´az, F.R. (2013); Preparation and
Characterization of Purified Na-activated Bentonite from Karak (Pakistan) for Pharmaceutical Use. Clay
Minerals, 48:595–603.
Trauger, R.L (1994); The Structure, Properties and Analysis of Bentonite in Geosynthetic Clay Liners.
Geosynthetic Resins, Formulation and Manufacturing. Proceedings of 8th GRI Conference.
464
EXTRACTION AND CHARACTERIZATION OF MAGNESIUM CHLORIDE FROM DIFFERENT
BRACKISH WATER SOURCES IN RIVERS STATE
1 2 1
Konne, J.L., Ujile, A.A.and Ogolo, J.J.
1
Department of Chemistry, Faculty of Science, Rivers State University of Science and Technology,
Nkpolu-Oroworukwo, P.M.B. 5080, PortHarcourt, Nigeria. Email: konne.joshua@ust.edu.ng
2
Department of Petroleum Engineering, Faculty of Engineering, Rivers State University of Science and
Technology, Nkpolu-Oroworukwo, P.M.B. 5080PortHarcourt, Nigeria
ABSTRACT
Magnesium chloride, an important source of magnesium, chlorine and other related products was extractedfrom
brackish water obtained from Kaa, Eagle Island, Opobo and Iwofewater fronts in Rivers State using
conventional methods. This involved the concentration of the sea water from 10L to below 1L before reacting
with CaO to precipitate Mg(OH)2.The precipitates were washed, filtered and dissolved in 200 ml of 1.0M HCl to
form MgCl2 solution.The solution was evaporated to dryness in order to crystallize MgCl2.6H2O salt. The
different yields of the salts obtained were 0.79, 0.73, 0.4 and 0.2 g/L for the Kaa, Eagle Island, Opobo and
Iwofebrackish waters, respectively. Physicochemical parameters such as pH, conductivity and total salinityof
the salts obtained followed the same trend of Kaa> Eagle Island>Opobo>Iwofe. The salts were characterized
using XRD and XRF. The XRD gave major reflectionsof hydrated magnesium chloride salt while the XRF
showed the elemental compositions indicating the presence of magnesium as the major component of the salt.In
addition, high sulphur content as well as significant presence of Sn, Sb, Al, Si, P, Fe, Cu, Zn, W and Mo were also
observed on the XRFcharts of all the four salts.
Key words: MgCl2.6H2O, Rivers State, Brackish waters (Kaa, Eagle Island, Opobo, Iwofe),XRD and XRF.
INTRODUCTION
The exploitation of seawater for mineralsappears to have garnered lessattentionin comparison to efforts
put in search for crude oil in various on- and offshore finds around the world. However, the sea in addition to
supporting marine flora and faunais also a sink to numerous minerals or salts of economic value from
weathering. Minerals found in seawater (brines) include salts of Na, Mg, Ca, Ba, Li, K, Al, S, Si, Fe, Sn, Mn, Mo,
Zn, Ni, Co, Cr, Cu, V, Ti, Cd, Pb, Au, Th, U, etc. (Al Mutaz and Wagialia, 1990; Bardi, 2010).
Magnesium and calcium chlorides rank second and third respectively to sodium chloride in terms of
relative abundance in seawater. Ca and Mgchlorides find applications in road construction as dust control agents,
road de-icing, drying agents (hygroscopic), etc. However, there is renewed interest in MgCl2because of its
applicationsas catalyst support for Ziegler Natta catalyst (used in the petrochemicals/plastic industries for the
production of Polyethylene or polypropylene products), Grignard reagents, borates and carnallites.This
translates toaddedcommercial value (Tripp, 2009; Rojanotaikul, 2012; Podder, et al., 2013; Derun, 2013;
Kipcak, et al., 2013).
Brines currently serve as major commercial sourcesof magnesium and its products.Forinstance,major
exporters like Norway, United States of America and Union of Soviet Socialist Republic, mined Mg from
brines(Al Mutaz and Wagialia, 1990; WTIC, 1997; Maddan, 2001).Hydrated magnesium chloride,
MgCl2.xH2O,exists as MgCl2.12H2O in seawater ( 0.55%) and in otherhydrated forms,MgCl2.x (H2O), where x
=1,2,4,6,8 and 12 due to its hygroscopic nature (Kipouros and Sadoway, 2001). The hexahydrate, MgCl2.6H2O,
on the other hand is the stable form at room temperature. Production of anhydrous or high purity grades and
crystal modifications of magnesium chloride have also been reported (Rojanotainul, et al., 2011; Zhang et al.,
2012).
465
The conventional method used for the extraction of MgCl2from seawater is by the Dow method.This
method exploits the low solubility ofthe precursor, magnesium hydroxide compared to that of calcium hydroxide
since solubility of the salts of binegative ions increases down the group. The precipitated magnesium hydroxide
reacts with HCl to form the hydrated chloride salt according to equation 1
Mg(OH)2(s) + 2HCl → MgCl2(aq) + 2H2O(l) (1)
In general, a litre of seawater is said to contain over 1.0g of magnesium ions (Bhatti, et al., 1984; Atkins and
Shriver, 2009).However, it could be less with brackish water due to possible river-brackish water mix.
There is currently no available report on the magnesium chloride content of Nigerian waters despite
similar report in North Africa (Behij, et al., 2013). This project therefore investigates the presence, yield and
characterization of crude magnesium chloride extract obtained from some brackish waters (Kaa-BoriOgoni,
Eagle IslandRumueme-Oroakwo, Opobo and Iwofe) in Rivers state of Nigeria. This study would also provide
some useful information on the elemental composition of unrefined salts from brackish waters in River State.
Experimental
Collection and concentration of brackish (brine) water
Brackish water samples from four selected locations namely:-Kaa-BoriOgoni, Eagle Island Rumueme-
Oroakwo, Opobo and Iwofe in Rivers state of Nigeria were collectedin a 10 litredry plastic containereach,
prewashed with detergent andrinsed with deionise water.All samples were collected at high tide at each location.
This was done because salinity is usually higher at high tides than at low tides. The 10 L was heatedat 100 ºC in a
clean 20 L steel pot andlater transferred to a 2 L beaker where it was finally reduced to 1 L saturated salt solution
viaevaporation.
Precipitation of Mg(OH)2 and crystallization of MgCl2.6H2O
The precipitation of Mg(OH)2 precipitates was done by the reaction of200 mL of a 11.2g (0.2M) CaOwith 200
mL of the saturated brackish (brine) water to form white gelatinous precipitates of Mg(OH)2.The reaction was
allowed for a day for proper aging before separation by decantation of the clear supernatant. The precipitate was
further heated to remove any remaining supernatant liquid present (but not to dryness). The
recoveredMg(OH)2precipitate was reacted with (dissolved in) 200mL of 1M HCl to form the magnesium
chloride solution. Evaporation of the resultant solution crystallized the hydrated magnesium chloride salt,
MgCl2.6H2O.
The triple function conductivity meter was used to record the pH, conductivity and salinity of the salt
solutions. A 5 min flux of nitrogen gas into a second magnesium chloride solution was done in order to
investigate possible reaction of the Mg2+ ions in solution with free nitrogen for confirmation.The sample with the
highest yield was chosen for X-ray diffraction characterization (on a Siemens D5000 Diffractometer) while X-
ray fluorescence analyses were donefor only two samples due to cost.
Result and Discussion
The XRD pattern of the extracted salt is shown in Figure 1a. The three major peaks (27.7, 32.05 and 46.25º)
reflect at 2-theta angles reported previously for MgCl2 (Zhang, et al., 2014).The particle size estimated using
Scherrer's formula was 63.23 nm. The unlabeled minor peaks could be those of associated impurity phases not
indexed due to insufficient data from available literature. This was confirmed by the X-ray Fluoresce (XRF) data
which indicated the presence of impurities like S (Major), Sn, Sb, Al, Si, Fe, P, Ni, Cu, Zn, W, Mo, V, Rb and Nb.
See Figure 1b,c and d for the XRF charts of MgCl2.6H2O salts from Kaa,Nitrogenated (Kaa) and Opobo brackish
waters,respectively.
The high concentration of sulphur suggested that most of the impurity phases were sulphates or that the salt
was a mixture of Magnesium chloride and sulphates but confirmation will be done in further studies. S counts on
466
the XRF of MgCl2.6H2O from Kaaand Opobo (Figure 1b and d respectively) were slightly higher than those of
magnesium. However, the Mg peak on the nitrogenated MgCl2.6H2O chart diminished to less than half of its
2+
original peak before nitrogenation. This could bedue to possible reaction ofMg ions with free nitrogen to form
Mg3N2 but would be confirmed in future studies. However, the reaction confirmed the Mg peak being one of the
characteristic reactions of Mg2+ ions.
The XRF also confirmed significant presence of toxic elements likePb,Sn and Sb in all the samples. This
report would help to createpublicawareness ofthe dangers of using brackish water salts as table salts.The
common occurrence of these toxic elements in both areas investigated reflects the impact of heavy
environmental pollution due to oil spills. The metals including sulphurare inorganic components of crude oil.
Figure 1(a) XRD patternof MgCl2.6H2O (Kaa), (b) XRF of MgCl2.6H2O (Kaa), (c) XRF of MgCl2.6H2O (Kaa)
fluxed with N2, and (d) XRF of MgCl2.6H2O (Opobo)extracted from the respective brackish waters.
The yields in grams of MgCl2.6H2O extracted from the four brackish waters were 0.79, 0.73, 0.40 and
0.20 g/L for Kaa, Eagle Island, Opobo and Iwofe respectively. Similarly, the physicochemical properties like pH,
conductivity and salinity also followed the same trend as shown in Table 1. The lower salinity of Opobo and
Iwofe brackish waters could be due to dilutions from river water unlike Kaa that empties into the Bonny high sea.
Table 1 Showing the brackish water location, Yield, pH, Conductivity and Salinity of each solution of
extracted MgCl2.6H2O
467
S/N Location Yield (g/L) PH Conductivity (mS/cm) Salinity (ppt)
1 KAA 0.79 9.75 101.10 67.20
2 ISLAND 0.73 9.64 79.40 49.30
3 OPOBO 0.40 9.62 69.20 43.60
4 IWOFE 0.20 9.62 56.50 34.20
Conclusion
The extraction and characterization of magnesium chloride,MgCl2.6H2O salt obtained from brackish water
obtained ofKaa, Eagle Island, Opobo and Iwofe in Rivers State has been reported for the first time. The XRD
showed three major reflections of MgCl2.6H2O salt while the XRF indicated the presence (higher counts) of
Mgas well as Swith lower counts forSn, Sb, Al, Si, P, Fe, Cu, Zn, W and Mo on the XRF charts of all the four salts.
However lower counts were also recorded for Mg on the XRF chart of the pre-nitrogenatedMgCl2.6H2O sample
indicating possible conversion of the Mg to nitride. The yields of the salts were 0.79, 0.73, 0.4 and 0.2 g/L for the
Kaa, Eagle Island, Opobo and Iwofebrackish waters, respectively. This report recommends that the use of
extracted salt from brackish waters for food seasoning should be discouraged due to the possible presence of
toxic elements in suchsalt.
REFERENCE
Bhatti, A.S., Dollimore, D. and Dyer, R. (1984). Magnesia from Sea Water: A Review.Clay Minerals, 19:865-
875.
Al Mutaz, I.S. and Wagiala, K.M.(1990). Production of magnesium from desalination brines.Resour. Conserv.
Recycl., 3:231-239.
Kipouros, G.J. and Sadoway, D.R. (2001). A thermochemical analysis of the production of anhydrous MgCl2.J.
Light. Met., 1:111-117.
Wisconsin Transportation Information Centre (WTIC). (1997).Wisconsin Transport Bulletin, 13:1-6.
Downloaded on the 21st of Sept. 2016.
Maddan, O.L. (2001). Apparatus and Method for producing magnesium from sea water.US Patent,
US6267854B1.
Tripp, T.G. (2009). Production of Magnesium from Great Salt Lake, Utah USA.Nat. Resour. Env. Iss.,15: 55-61
Bardi, U. (2010). Extracting Minerals from Seawater:An Energy Analysis.Sustainability, 2:980-992.
Rojanotaikul, P., Ayudha, S.K.N., Charoenchaidet, S., Faungnawakij, K. and Soottitantawat, A. (2012).
Preparation of porous anhydrous magnesium chloride particles by spray drying process. Engineering
Journal, 16:109-114.
Kipcak, A.S., Gurses, P., Kunt, K., MoroydorDerun, M,E. and Piskin, S.(2013). Magnesium Borate Synthesis by
Microwave Method using MgCl2.6H2O and H3BO3. International Journal of Chemical, Molecular,
Nuclear, Materials and Metallurgical Engineering, 7: 290-295.
Derun, M.E., Gurses, P., Yildirim, M., Kipcak, A.S., Ibroska, T. and Piskin, S. (2014). The Effect of the Reaction
Time on the Microwave Synthesis of Magnesium Borates from MgCl2.6H2O, MgO and
H3BO3.International Journal of Chemical, Molecular, Nuclear, Materials and Metallurgical Engineering,
8: 565-569.
Behij, S., Hammi, H., Hamzaoui, A.H. and M'Nif, A. (2013). Magnesium Salts as Compounds of the Preparation
of Magnesium Oxide from Tunisian Natural Brines. Chemical Industry & Chemical Engineering
Quarterly, 19:263-271.
468
Podder, J., Gao, S., Evitts, R.W., Besant, R.W. and Matthews, D. (2013). Crystallization of carnallite from KCl-
MgCl2 brine solutions by solvent evaporation process and its structural and mechanical characterization.
Journal of Metals, Materials and Minerals, 23: 37-42.
Athkins, P. and Shriver, B. (2009). Occurrence of magnesium, properties and uses; inorganic chemistry, 312-
313.
Zhang, Z., Lu, X., Pan, F., Wang, Y. and Yang, S.(2012). Preparation of anhydrous magnesium chloride from
magnesium hexahydrate.Metallurgical and material tranction B, 9777-9782.
Zhang, Z., Lu, X., Wang, T., Yan, Y. and Pan, F.(2014). The dehydration of MgCl2.6H2O-KCl-NH4Cl
system.Journal of Analytical and Applied Pyrolysis, 110:248-253.
469
Neutron Activation Analysis (NAA) and X-ray Fluorescence Analysis (EDXRF) of Corchorus tridens Linn
1
Umar S. Gwarzo, 2Gimba, C.E., 3Adeyemo, D.J. and 2Paul, E.D.
1
Chemistry Department, Federal College of Education, Kano.
2
ChemistryDepartment, Ahmadu Bello University, Zaria, Nigeria.
3
Centre for Energy Research and Training, Ahmadu Bello University, Zaria, Nigeria.
E-mail of the corresponding author:
Abstract Analysis of
Corchorus tridens Linn was carried out for its elemental composition using neutron activation analysis (NAA)
and energy dispersive x-ray fluorescence analysis (EDXRF). A total of nineteen (19) elements (Al, Ba, Br, Ca,
Co, Cr, Cu, Fe, Hf, K, La, Mn, Na Rb, Sm, Sc, Th, V and Zn) were detected and determined while ten (10) others
(As, Cs, Dy, Eu, Lu, Mg, Ta, Ti, U and Yb ) were detected at BDL levels. Variations of their concentrations from
the two techniques and the different parts of the plant were compared using student's t – test and ANOVA test.
Translocation factor of the elements and their concentrations in the different parts of the plant were compared
with normal concentrations in plants and RDA values. It was found that for some of the elements, pattern of
distribution is in the order of roots > stems > leaves while for some there is no regular pattern of distribution.
Also, amounts of some essential elements in C. tridens Linn shows that it can serve as a means of
supplementation of required minerals for man and his animals.
Key words: NAA, EDXRF, C. tridens Linn, translocation and bioaccumulation
Introduction
The plant Corchorus tridens Linn belong to the family Tiliaceae. Its common names include turgunnuwa, or Lalo
– Hausa/Fulani (Dalziel, 1955). The leaves and young shoots of the plant are used as a vegetable and soup herb,
and are also a good fodder for camels and other domestic stock. It yields a good fiber used for fishing lines in
northern Nigeria and elsewhere. When long, the stems are used for horizontal ties of conical hut roofs (Dalziel,
1955).
Phytochemical analysis of the corchorus species to which C. tridens L. belongs indicates the presence of cardiac
glycosides, triterpenes, ionones, phenolics, sterols, coumarins, steroids and fatty acids (Khan et.al, 2006).
The contribution of medicinal plants in the traditional system of medicine for curing diseases has been
documented. Nowadays increased scientific interest and consumer demand have promoted the development of
herbal products as dietary supplements (Khan et.al, 2006). In view of renewed interest, oriental herbal medicines
have a prominent role to play in the pharmaceutical and health markets of the 21st century (CSIR, 1950). It has
been reported that whatever is taken as food could cause metabolic disturbance subject to the allowed upper and
lower limits of trace metals (Chopra et.al, 1958). Both the deficiency and excess of essential micronutrients and
trace of toxic metals may cause serious effects on human health (Satyavati et.al, 1976 and Wahid and Siddiqui,
1961).The use of medicinal plants in therapeutics or as dietary supplements goes back beyond recorded history,
but has increased substantially in the last decades (Bhatt et.al, 2003 and Sen, 1930). However, the safety of their
use has recently been questioned due to the reports of illness and fatalities. Medicinal herbs are easily
contaminated during growth, development and processing. After collection and transformation into dosage form
the heavy metals confined in plants finally enter the human body and may disturb the normal functions of central
nervous system, liver, lungs, heart, kidney and brain, leading to hypertension, abdominal pain, skin eruptions,
intestinal ulcer and different types of cancers.
With this background this researchers embarked on the comprehensive elemental analysis of C. tridens L. which
is to our knowledge is the first of its kind.
470
The plant in this work Corchorus tridens Linn) were identified at the Department of Biological Sciences of
Ahmadu Bello University (ABU) Zaria and voucher specimens are kept there. Samples of the plant materials
(roots, stems, fruits and leaves) were collected from Karkari village, Gwarzo Local Government Area of Western
part of Kano State, Nigeria and transported in polythene bags.
Sample treatment
After collection, the samples were washed twice with tap water and rinsed with deionised water. They were first
O
air dried and then further dried in an oven at 60 C. After drying, the samples were ground using pestle and mortar
and passed through 125µm mesh sieve.
Neutron activation analysis measurement
The ground sample was pulverized and approximately 150 mg sub-sample was weighed and wrapped in
polyethylene films. For the elements leading to short-lived activation products, the sample was packed and
3
sealed in 7 cm rabbit capsules. Irradiation with thermal neutrons was carried out at an outer irradiator channels
(i.e., B4) of the Nigeria Research Reactor-1 (NIRR-1) operating at a thermal neutron flux setting of 2×10 n/cm2s,
which corresponds to a neutron flux of 1×10 n/cm2s in the outer channels. After the irradiation, a PC- based
gamma ray spectrometry set-up performed measurement of induced radionuclide. This consists of a HPGG
detector coupled to a computer based Multi-Channel Analyzer (MCA) via electronic modules. The relative
efficiency of detector is 10 % and an energy resolution of 1.95 KeV at gamma-ray energy of 1332 KeV belonging
to 60Co. Through appropriate choice of cooling time, detector's dead time was controlled to be less than 10 %.
Identification of gamma ray of product radionuclides through their energies and quantitative analysis of their
concentrations was achieved using the gamma ray spectrum analysis software, WINSPAN - 2004. The certified
reference material IAEA – soil - 7 was used as the standard, while other two certified reference materials, GSD -
²² and GSR-5 were used as analytical quality control materials to validate the procedure for all the elements (Dim
et.al, 2004, Jonah et.al, 2006, Gwarzo et.al, 2014).
Energy Dispersive X-ray Flourescence Analysis Measurement
Approximately 0.5g of each sample was measured and then poured in to pelletiser (without a binder). A pressure
-2
of about 10tonnes (204081NM ) was applied to the sample for pelletisation using the SPECAC hydraulic press.
The pellets produced have dimensions of about 2mm thick and 25mm in diameter. Each pellet was placed on the
annular cadmium 109 isotopic excitation source which sits directly on the lithium drifted selenium detector (Se
(Li)) with a barium window. Underneath the detector is the liquid Nitrogen Dewar, which produces cooling
effect on the detector.
As soon as the source is exciting the sample, signals are sent to the preamplifier, the first stage of which a field
effect transistor (FET) is mounted adjacent to the crystal detector. Bias voltage is maintained at 500v and is
computer controlled. In the preamplifier, the signals are amplified, and moved to the main amplifier, which is the
spectroscopy amplifier where a voltage of 240v is applied for further amplification. With the aid of MAESTERO
computer software, these signals were acquired in 3000seconds, which is the counting time. The acquired spectra
was saved and copied to AXIL software for quantification. Before quantification, the AXIL software converted
the spectra to AXIL form which was fitted using a defined model. The fitting was between chi – square of 1 – 5
and the spectra was saved for each element (Gwarzo et.al a and b 2006, and Gwarzo et.al, 2011).
Table 1: Concentrations of the elements determined in the leaves, stems and roots of C. tridens L. using
INAA an EDXRF techniques.
471
Sample Leaves (CTLL) Stem (CTLS) Root (CTLR)
NAA EDXRF NAA EDXRF NAA EDXRF
Al 1453±119† Nd 209±19.0† Nd 334±29† Nd
As BDL Nd BDL Nd BDL Nd
a a
Ba? 138±19 † 8.825±2.1 111.0±18.0‡ Nd BDL Nd
Br 5.0±1.0† Nd 3.5±0.6† Nd BDL Nd
a a
Ca? 14750±2139 ‡ 17765±374 12350±1803‡ Nd 3788±591‡ Nd
Co 0.46±0.11† Nd 0.36±0.12† Nd BDL Nd
a a
Cr? 3.48±1.02 60.361±9.6 ‡ BDL 78.585±9.12‡ BDL 79.742±18.4‡
Cs BDL Nd BDL Nd BDL Nd
Cu Nd 114.885±11.2† Nd 32.274±2.14† Nd 22.457±13.7†
Dy BDL Nd BDL Nd BDL Nd
Eu BDL Nd BDL Nd Nd Nd
a
Fe? 113O±73 8945±89.34 ‡ BDL 8005±167‡ 376±45 8410±194.6a‡
a
Hf 1.9±0.1 Nd BDL Nd Nd Nd
a a a a a
K? 43120±517 21590±184.3 ‡ 54290±326 13210±349.5a‡ 21110±1457 8985±158.5 ‡
La 2.72±0.04 Nd 0.034±0.002 Nd 1.7±0.1† Nd
Lu BDL Nd BDL Nd BDL Nd
a a a a a
Mn? 62.3±2.6 496.224±39.3 ‡ 32.0±1.0 234.76±15.7 ‡ 23.0±1.0 196.92±25.2a‡
Na 416.0±3.0‡ Nd 198.0±2.0‡ Nd 315.0±180‡ Nd
Rb 23.0±2.0† Nd 20.0±3.0† Nd 3.9±0.8† Nd
Sb BDL Nd BDL Nd BDL Nd
Sc 0.33±0.01 Nd 0.043±0.008‡ Nd 0.11±0.01‡ Nd
Sm 0.39±0.01† Nd 0.056±0.006† Nd BDL Nd
Ta BDL Nd BDL Nd Nd Nd
Th 1.1±0.1† Nd BDL Nd BDL Nd
Ti BDL Nd BDL Nd BDL Nd
U BDL Nd BDL Nd BDL Nd
a a
V? 1.9±0.30 64.489±4.9 ‡ BDL 88.545±7.34‡ 0.7±0.2 73.309±5.3a‡
Yb BDL Nd BDL Nd BDL Nd
Zn 51.0±4.0 Nd 22.0±3.0† Nd 14.0±3.0† Nd
a
BDL: Below detection limit; Nd: Not determined; ?elements determined by INAA and EDXRF; superscript , are
not significantly different at p > 0.05. † Elements determined in the parts of C. tridens L. are not significantly
different while ‡ are significantly different at p > 0.05.
Table 2: Translocation factors of some of the elements determined between the roots to the stems (TS/R) and
stems to the leaves (TL/S) of C. tridens Linn.
Translocation factor Al Ca K La Mn Na Rb Sc Zn
TS/R 1.59 0.31 0.39 50.0 0.72 1.59 0.20 2.56 0.64
TL/S 0.14 0.84 1.26 0.01 0.51 0.48 0.87 0.31 2.32
Table 1 above indicates the results of the elemental analysis of different parts C. tridens L. by NAA and EDXRF
techniques. Statistical analysis (t – test and ANOVA) on the results indicated that, for some of the elements (Ba,
Ca, Fe, K, Mn and V) there was no significant difference in their concentrations using the two techniques and in
the different parts of the plant for (Al, Br, Co, La, Rb, Sm, Th and Zn). Significant differences were observed at p
472
≤ 0.5 for Ba, Ca, Cr, Sc, Na and V in the different parts of the plant analyzed. This is attributed to the capability of
the plant to bioaccumulate some of these elements in its different organs (while differences in the concentrations
of some the metals in the same samples and measured by the different methods may be attributed by the
accuracy, precision and capabilities of the two techniques used (Goulden, 1978).
Though the concentration of major elements including K and Ca known to be in high concentrations in tissues of
plants because of the roles they play as components of simple salts and complexes performing various functions
(Yalwa, 2002), their concentrations in the analyzed plants samples (8985 – 54290 ppm and 3788 – 17765 ppm)
are relatively higher than normal concentrations found in most plants (10 – 100 ppm and 50 – 60 ppm) (Allen et
al., 1974).
For microelements such as Ba, Fe, Mn, Zn, Rb and Co, their concentrations are also high and in fact some values
of the elements such as Fe (376 – 8945 ppm), 8.825 – 138.0 ppm (Ba), 23.0 – 496.224 ppm (Mn), 14.0 – 51.0 ppm
(Zn), 3.9 – 23.0 ppm (Rb), and 0.00 – 0.41 ppm (Co) may prompt the researcher to declare that C. tridens L. is an
indispensable sources of micronutrients to man and his animals when consumed. This is because the normal
concentrations of these metals in plants are Fe (40.0 – 500.0 ppm), Ba (9.2 – 131.9 ppm in Sedge and Nutgrass),
Mn (50.0 – 356.0 ppm), Zn (15.0 – 100.0 ppm), Rb (0.2 – 194.0 ppm), and Co (0.1 – 0.60 ppm) and the above
values of the metals determined are within normal range.
Moreover, for the ultra-trace elements; Sc, Sm, Th, Br, V, La, Cu, Sb, Lu and Eu, their concentrations are
relatively within normal levels. This is evident when concentrations of these elements in this work are compared
to normal levels in plants. For example, the range of Sc concentration C. tridens is 0.03 – 0.95 ppm while normal
range in vegetables is 5.00 ppb and 70.0 ppb in grasses, Sm in this work is 0.03 – 0.06ppm while 0.11– 0.12 ppm
-3 -3
was found by Zhang et al., 2013. Also Th in this work is 1.1ppm while normal range in plants is 2.0x10 – 3.6x10
Ci/g, V in this work is 0.7 – 73.309 ppm while normal range in plants is 96.0 – 112.0 ppm which signifies low
concentration of the element in the analyzed plant. For lanthanum however, the range is 0.034 – 2.72 ppm while
normal range in plants is generally low (Wang et al., 1997). In addition to that, the concentration range of copper
in this work is 22.457 – 114.885 ppm which is higher than normal concentrations (10.0ppm) of the element in
plants.
Since the motive for elemental analysis of green plants, food and feed samples is mainly aimed to investigate
nutritional quality or contamination, one of the indices for such is transfer factor (TF). In this work the TF has
been categorized as either TFShoot/Root (TFS/R) or TFShoot/Leaf (TFS/L) and most plant species have been found to have TF
>1. For Al, the TFS/R for C. tridens Linn (1.59), which signifies good transfer of the element from roots to shoots
while TFL/S was found to be <1 which indicates accumulation of the element in the stems of the plant.
However, the TF values for Ba, Ca, K, La, Mn, Na, Rb and Zn in C. tridens L. varied like that of Al discussed.
TFS/R values of the above elements that are >1 ranged 1.59 – 50.0 while those that are <1 ranged 0.31 – 0.64 as
depicted in Table 2 above. TFL/S values for the above elements are all <1 except for K and Zn as indicated in table
2.
Conclusion
This analysis was conducted in order to ascertain the metal composition in the different parts of C. tridens L. and
their transfer factors. Essential elements were found to be at high concentrations and compares well with the
exotic vegetables. This indicates that the plant can contribute to the diet of individuals that may consume it. The
presence of toxic metals at low concentrations or BDL levels in the plant shall strengthen this claim. Also the
values of the translocation factor (mostly < 1) indicates that C. tridens L. the plant cannot be a hyper accumulator
of any of the determined elements probably with the exception of La (TS/R = 50) which hyper – accumulate the
473
metal in its stems.
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475
SYNTHESIS AND EVALUATION OF SUBSTITUTED BENZIMIDAZOLE MOTIFS FOR
PRELIMINARY ANTIMICROBIAL DRUG TARGET
Olayinka O. Ajani , Damilola V. Aderohunmu1, Shade J. Olorunshola2, Bamidele M. Durodola1 and
1,*
1
Olatunde M. Ogunleye
1
Department of Chemistry, School of Natural and Applied Sciences, Covenant University, Canaanland, Km
10 Idiroko Road, P.M.B. 1023, Ota, Ogun State, Nigeria.
2
Department of Biological Sciences, School of Natural and Applied Sciences, Covenant University,
Canaanland, Km 10 Idiroko Road, P.M.B. 1023, Ota, Ogun State, Nigeria.
*Corresponding Author: E-Mail Address: ola.ajani@covenantuniversity.edu.ng Telephone Number:
+2348061670254.
ABSTRACT
Benzimidazole scaffolds are structural isosteres of naturally occurring nucleotides which allows them to interact
with the biopolymers and enzymatic sites of living systems. The aim of this present work is to synthesize selected
2-alkanamino-benzimidazole derivatives in order to investigate their antimicrobial efficiency for possible future
drug development. The series of targeted compounds were conventionally synthesized in good to excellent
yields via [4+1]-cyclo-addition of o-phenylenediamine with some L-amino acid and purified by recrystallization
or column chromatography where necessary. The chemical structures were confirmed by physico-chemical and
1 13
spectral data which include UV, IR, H- and C-NMR. In addition, the antimicrobial properties of the synthesized
benzimidazole derivatives were determined on six bacteria isolates alongside with gentamycin standard drug
using agar diffusion method. The synthesized compounds showed broader activities spectrum than gentamycin
and (1H-benzo[d]imidazol-2-yl)methanamine, 10a emerged as the most potent. Based on the versatility of the
synthetic pathway and improved antibacterial activity, these compounds are recommended as good candidates
for further studies in terms of MIC test, toxicity profile as some of them might pave way in the pharmaceutical
research for future drug design．
Key words: benzimidazole, cyclo-addition, zone of inhibition, spectroscopy, drug design.
INTRODUCTION
Benzimidazole is nitrogen containing heterocyclic scaffold with pKa of 12.75 while its conjugate acid has a pKa
1
of 5.68, which is less basic than imidazole . Among the large variety of heterocyclic systems developed, the
ortho-fused bicyclic moiety benzimidazole appears to be a particularly effective heterocyclic system
considering its interaction with the AT1 receptor2 and important enzyme sites. Numerous compounds containing
benzimidazole moieties have been reported to exhibit diverse biological and pharmacological properties
3 4 5 6 7 8 9
including analgesic , antimicrobial , anticancer , anti-HCV , anti-HIV , anthelmintic , antitumor ，
antitubercular10，antimalarial11，antihypertensive12, antioxidant13 and anti-inflammatory14 among others.
Conventionally, a drug is designated by its dominant or by its first recognized function; hence, benzimidazole
nucleus is the core structure of commercially available drugs such as anthelmintics (albendazole, 2;
thiabendazole, 3), antifungal agents (benomyl, 4), anticancer (bendamustine 5), anti-ulcer drug (rabeprazole, 6),
receptor antagonist drug (astemizole, 7), antihypertensive (telmisartan 8) and anti-inflammatory and anti-ulcer
drug (omeprazole 9)15．We have herein investigated the efficient synthesis of 2-substituted benzimidazole
derivatives in order to evaluate their antimicrobial potential for future drug development.
(a) Physico-Chemical and Spectra Study: Solvents used were of analytical grade and were used directly
without further purification. Melting points were determined in open capillary tubes on Stuart melting point
apparatus and were uncorrected. The IR spectra were run in solid state using the Bruker FT-IR while UV analyses
1 13
of all the samples were run in ethanol, using UV-Genesys. The H-NMR and C-NMR of the compounds were
run on Bruker NMR machine at 400 MHz and 100 MHz respectively using DMSO-d6.
(b) Antimicrobial activity: All the synthesized benzimidazole templates 10a-i and gentamycin standard drug
were screened for antimicrobial activity on the targeted six organisms using agar well diffusion method as
16
described by Russell and Furr Method .
General procedure for synthesis of 2-alkanaminobenzimidazole derivatives, 10a-i
o-Phenylenediamine (1.08 g, 10 mmol) and corresponding amino acids a-h or synthon i (10 mmol) was added
sequentially to 10 mL of toluene in a quickfit flask. The reacting mixture was heated under reflux for 9 h under the
influence of a magnetic stirrer to obtain coloured solution which was allowed to cool overnight. The crystals
formed were filtered and air-dried to afford the 2-alkanaminobenzimidazoles, 10a-i in excellent yields.
Synthesis of (1H-benzo[d]imidazol-2-yl)methanamine, 10a. When a = glycine, yield 97.3%, m.p. > 200 oC.
-1
ëmax in nm (log åmax): 236 (1.7782), 290 (1.324), 407 (0.8541). í in cm (functionality responsible): 3384, 3363 (N-
H of NH2, 2 bands), 3245 (N-H), 3021 (C-H aromatic), 2930 (C-H aliphatic), 1605 (C=C), 1580 (C=N), 741 (Ar-
H).
1-(1H-benzo[d]imidazol-2-yl)-2-methylpropan-1-amine, 10b. When b = L-valine, yield 85.1%, m.p. > 200
o -1
C. ëmax in nm (log åmax): 209 (3.3653), 236 (2.893), 293 (2.5185). í in cm (functionality): 3384, 3363 (N-H of
NH2, 2bands), 3117 (N-H), 2962 (C-H aliph. of CH3), 1603 (C=C arom.), 1591 (C=N), 775 (Ar-H).
4-amino-4-(1H-benzo[d]imidazol-2-yl)butanamide, 10c. When c = L-glutamine, yield 84.3%, m.p. > 200 oC.
-1
ëmax in nm (log åmax): 209 (3.2842), 236 (3.1562), 293 (2.3054). í in cm (functionality responsible): 3384, 3364
(N-H of NH2, two bands), 3235 (N-H), 3200 (N-H), 2927 (C-H aliphatic of CH2), 2803 (C-H aliphatic), 1685
(C=O of amide), 1605 (C=C aromatic), 1575 (C=N), 748 (Ar-H).
2-(pyrrolidin-2-yl)-1H-benzo[d]imidazole, 5d. When d = L-proline, yield 89.0%, m.p. > 200 oC. ëmax in nm (log
-1
åmax): 209 (3.3520), 239 (2.6522), 293 (2.2810). í in cm (functionality): 3322 (N-H), 3124 (N-H aliph.), 3027
(C-H arom.), 2928 (C-H aliphatic of CH2), 2857 (C-H aliph.), 1600 (C=C arom.), 1577 (C=N), 744.
4-(2-amino-2-(1H-benzo[d]imidazol-2-yl)ethyl)phenol, 5e. When e = L-tyrosine, yield 83.3%, m.p. > 200 oC.
ëmax in nm (log åmax): 209 (3.3677), 236 (2.796), 293 (2.4698). í in cm-1 (functionality responsible): 3384, 3363 (N-
H of NH2, two bands), 3200 (N-H), 3117 (O-H, broad), 2929 (C-H aliphatic of CH2), 2876, 2827 (C-H aliphatic),
1600 (C=C aromatic), 1584 (C=N), 739 (Ar-H).
1-(1H-benzo[d]imidazol-2-yl)-2-phenylethanamine, 5f. When f = L-phenylalanine, yield 84.0%, m.p. > 200
o
C. ëmax in nm (log åmax): 212 (3.4050), 236 (2.9974), 293 (2.6857). í in cm-1 (functionality responsible): 3384,
3363 (N-H of NH2, two bands), 3209 (N-H), 3022 (C=H aromatic), 2889 (C-H aliphatic), 1615 (C=C aromatic),
1575 (C=N), 749 (Ar-H).
1-(1H-benzo[d]imidazol-2-yl)-3-(methylthio)propan-1-amine, 5g. When g = L-methi- onine, yield 97.1%,
m.p. > 200 oC. ëmax in nm (log åmax): 209 (3.3181), 236 (2.6064), 293 (2.2553). í in cm-1 (functionality responsible):
3384, 3363 (N-H of NH2, two bands), 3276 (N-H), 3173 (N-H), 3029 (C-H aromatic), 2921 (C-H aliphatic of
CH2), 2885 (C-H aliphatic), 1600 (C=C), 1575 (C=N), 745 (Ar-H).
1-(1H-benzo[d]imidazol-2-yl)pentane-1,5-diamine, 5h. When h = L-lysine, yield 88.2%, m.p. > 200 oC. ëmax in
nm (log åmax): 206 (2.6222), 236 (1.5911), 296 (1.6021). í in cm-1 (functionality responsible): 3383, 3365 (N-H of
NH2, two bands), 3276 (N-H), 3173 (N-H), 3026 (C-H aromatic), 2929 (C-H aliphatic of CH2), 2845 (C-H
aliphatic), 1603 (C=C aromatic), 1583 (C=N), 742 (Ar-H).
2-(1H-benzo[d]imidazol-2-yl)aniline, 5i. When i: anthranilic acid, yield 83.4%, m.p. > 200 oC. ëmax in nm (log
åmax): 209 (3.1553), 242 (2.4914), 335 (2.0899). í in cm-1 (functionality responsible): 3384, 3366 (N-H of NH2,
two bands), 3201 (N-H), 3031 (C-H aromatic), 1576 (C=N), 748 (Ar-H).
Benzimidazole derivatives are crucial structural scaffolds found in diverse library of biologically active
compounds which are therapeutically useful agents in drug discovery and medicinal chemistry research. Hence,
in the continuation of our effort on the efficient synthesis of benzimidazole17 and investigation of their
antimicrobial potential, we have herein synthesized some 2-substituted benzimidazole derivatives via [4+1]-
cyclo-addition of o-phenylenediamine and nine COOH donors. The direct condensation of the -COOH
functionality of eight amino acids a-h and anthranilic acid synthon i with the NH2 groups of the o-
phenylenediamine was employed in this category in order to get the alkanamino- bearing benzimidazole
derivatives 10a-i (Scheme 1), because it was envisaged that the amino acid insertion in benzimidazole
framework at 2-position might create a synergistic effect to boost the biological activity of the titled compounds.
In detail, the reaction optimization study was conducted by comparing the synthesis in the presence of HCl: H2O
mixture (Method A) to that of simple reflux in catalyst-free medium using toluene solvent (Method B) as shown
in Fig 1. The reaction of the equimolar amount of o-phenylenediamine with nine different amino acids in aqueous
HCl, (hydrochloric acid) was successfully achieved by heating the mixture under reflux at a carefully controlled
temperature for 9 h to afford alkanamino- bearing benzimidazole derivatives 10a-i． Method B involved the use
of the same stoichiometry in toluene solvent without any catalyst. Method B was discovered to be eco-friendly,
atom-economical, afforded the products via easier work-up and in higher yields (Fig. 1).
N R 10 a: R = H
10 b: R = CH (C H3 ) 2
N N H2 10 c: R = CH 2 CH 2C ON H2
H
10a-c
H2 N
N H
N N
(i) a- c
N ( i) N
H 10i i (i ) H
10d
NH2 d
NH2
NH 2 (i )
( i) e -f N R'
h
N (i) g N N H2
H
N N H2 S 10e-f 10 e: R ' = O Ts
H 10 h 10 f: R' = H
N
N N H2
H 10g
Am ino a cid: (a) Gly c ine, (b) L-Valin e, ( c) L-Gluta mine , (d ) L -Pr oline, (e) L- Ty rosin e, ( f) L- Ph enyla lanin e,
(g) L-Me thion ine, (h) L -Ly s ine, (i) Anthranilic ac id. Re act io n c on ditio n: (i) Meth od A: EtOH , H2 O: H Cl
(1 m l), reflu x fo r 9 h. Met hod B: r ef lux in toluene for 9 h.
Scheme 1: Synthetic pathways for the preparation of targeted benzimidazole motifs 10a-I.
Fig. 1: Comparative study of two methods for optimization study.
From the spectroscopic point of view, the uv transition with the lowest wavelengths for the compounds 10a-i
observed at 206 to 212 nm, were as a result of the ð→ ð* transition of C=C which depicted the presence of
benzene ring in all the compounds. Bathochromic shifts led to the presence of other peaks at higher wavelengths
ranging from 236 to 406 nm. Some of these were as a result of ð→ n transition which may be ascribed to the
18
auxochromic C=N group; characteristic of K bands of C=N functional group . The infrared spectra of the
compounds 10a-i showed absorption bands due to the stretching vibrations of N-H, C-H aromatic, C-H aliphatic,
C=C and C=N at 3384-3201, 3031-3021, 2962-2885, 1615-1600 and 1591-1575 cm-1 respectively. An additional
band was noticed in 10c at 1685 cm-1 which depicted the presence of C=O of amide in 10c alone.
The result of antimicrobial activity in vitro was as shown in the Table 1. It was discovered that gentamycin was
least active against K. pneumonia and E. coli whereas 10a-i exhibited high to excellent activities on these two
organisms. Also, gentamycin was moderately active on S. aureus, S. faecalis, P. vulgaris and P. aeruginosa while
all benzimidazoles possessed high to excellent activities on these four organisms except in the case of 10e, 10d
and 10g against P. vulgaris and 10b against P. aeruginosa where moderate activities were noticed (Table 1).
Table 1: Result of in vitro antimicrobial screening with zones of inhibition in (mm).
Organism→ Staphylococcus Streptococcus Klebsiella Proteus Pseudomonas Escherichia
aureus faecalis pneumonia vulgaris aeruginosa coli
Comp No↓
10a ++++ ++++ ++++ ++++ +++ ++++
10b ++++ ++++ ++++ +++ ++ ++++
10c +++ +++ ++++ +++ ++++ ++++
10d ++++ ++++ ++++ ++ ++++ ++++
10e +++ +++ +++ ++ +++ +++
10f +++ +++ +++ +++ +++ +++
10g +++ +++ +++ ++ +++ +++
10h +++ +++ +++ +++ +++ +++
10i +++ +++ +++ +++ +++ +++
Gentamycin ++ ++ + ++ ++ +
+ = Least activity (1 – 10 mm); ++ = Moderate activity (11 – 20 mm); +++ = High activity (21 – 30
mm), ++++ = Excellent activity (>30 mm).
CONCLUSION
The alkanamino-benzimidazoles were successfully achieved in a higher yield, easier work-up and highly
economical procedure using toluene solvent as compared with former conventional method in aqueous acid as
catalyst. The compounds were screened in vitro against six bacterial isolates most of which were associated with
the gastrointestinal tract damage in man and animal. All the compounds exhibited better activity than the
gentamycin standard drug. Hence, they are good candidates for further study as the new replacement for
gentamycin might emerge from this series.
ACKNOWLEDGEMENT
OOA acknowledges The World Academy of Sciences for the sponsorship of this project under the TWAS
Research Grant for Individual Programme (Grant No: 14-069 RG/CHE/AF/AC_1; UNESCO FR: 324028564).
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1. T. Eicher, S. Hauptmann & A. Speicher, (2004), The chemistry of heterocycles: Structure, reactions,
syntheses, and applications. Wiley-VCH GmbH & Co. KGaA, Germany.
2. V.K. Vyas, & M. Ghate, (2010)，Substituted benzimidazole derivatives as Angioten- sin II-AT1
receptor antagonists: a review．Mini Rev. Med. Chem., 10, 1366–1384.
3. P.A. Datar & S.A. Limaye, (2015), Design and synthesis of mannich bases as benzimidazole
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benzimidazole-derived chalcones containing 1,3,4-oxadiazole moiety. Chem. Heterocycl. Comp., 51,
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5. P. Singla, V. Luxami & K. Paul, (2015), Triazine-benzimidazole hybrids: anticancer activity, DNA
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6. F. Zhao, N. Liu, P. Zhan, X. Jiang & X. Liu, (2015). Discovery of HCV NS5B thumb site I
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7. T. Pan, X. He, B. Chen, H. Chen, G. Geng, H. Luo, H. Zhang & C. Bai, (2015), Development of
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8. B. Munguía, M. Michelena, E. Melian, J. Saldaña, X. Ures, E. Manta & L. Domínguez, (2015),
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9. B. Chu, F. Liu, L. Li, C. Ding, K. Chen, Q. Sun, Z. Shen, Y. Tan, C. Tan & Y. Jiang, (2015), A
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Shackleford, M. Kaiser, C. Yeates, S.A. Charman & K. Chibale, (2011), Antimalarial pyrido[1,2-
a]benzimidazoles. J. Med. Chem., 54, 4581–4589.
12. Y. Zhang, J. Xu, Y. Li, H. Yao & X. Wu, (2015), Design, synthesis and pharmacological
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HETEROGENEOUS CATALYTIC EFFICIENCY OF SILICA SULFURIC ACID TOWARD THE
SYNTHESIS OF SUBSTITUTED PYRIMIDIN-2(1H)-ONE DERIVATIVES
Olayinka O. Ajani*, Taiwo F. Owoeye, Ifedolapo O. Olanrewaju, Adebusayo E. Adedapo and Christiana O.
Ajanaku
Departmentof Chemistry, College of Science and Technology, Covenant University, Canaanland, Km 10,
Idiroko Road, P.M.B. 1023, Ota, Ogun State, Nigeria.
*
Corresponding Author.E-mail: ola.ajani@covenantuniversity.edu.ng; Tel: +2348061670254
ABSTRACT
Pyrimidine template is a highly privileged motif for the development of molecules of biological and
pharmaceutical interest due to its prebiotic nature to life. This present study deals with the synthesis of pyrimidin-
2(1H)-one derivative from chalcones by the action of silica supported sulfuric acid (SSA) or conventional
refluxed in concentrated hydrochloric acid. The chemical structures were confirmed by analytical data and
1 13
spectroscopic means such as UV, IR, mass spectra, H and C NMR. SSA was found to be efficient method for the
quantitative transformation to pyrimidine frame work. It can be re-used after simple washing with chloroform
thereby rendering this procedure more economical.
Key Words:Spectroscopic means, chalcones,pyrimidine, 4-phenylbut-3-en-2-one.
INTRODUCTION
Over the years,tremendous amount of literature have be accumulated on pyrimidine heterocycle, owing to its
widespread application in medicinal research and occurrence in many biological entities valuable to life1. For
instance, pyrimidine moiety is the core structure in biomolecules such asnucleic acids components (uracil,
thymine and cytosine) and vitamin B1, and is an important constituentof numerous drug molecules in many
therapeutic areas2.The successful application of pyrimidine derivatives in many ways, their utility in applied
chemistry and in more fundamental and theoretical studies has made the literature of the subject to be
correspondingly vast. Many commercially available drugs are pyrimidine-based some of which are
3
pyrimethamine 1, l trimethoprim 2, amicetin3,idoxuridine 4, hexitidine 5 and phenobarbital 6 (Fig. 1) . Diverse
methods have been reported for the synthesis of substituted pyrimidines, the commonest being the reaction of 1,3-
dielectrophiles with nitrogen donors such as urea4, thiourea5, guanidine6, amidine7, benzamidine8and
9
formamidine .Thus, it is conceivable to develop aseries of pyrimidinones using SSA catalyst technique and also
compare it with the traditionalmethod heating approach in conc. Hcl．
OH
Cl
NH2 H3CO OCH3
H2N N NH2 O
H
N H3CO HO N
N N
H3C HO
N NH2 H3CO O
O N
pyrimethamine, 1 pyrimethamine, 2 amicetin, 3
(antimalarial) (antibacterial) (antibioitc)
CH3
OH
CH3
OH O
I CH3
N O NH NH
H3C
CH3
O N O N O N O
H idoxuridine, 4 H3C H H
(antiviral drug) hexitidine, 5 (antifungal) phenobarbital, 6 (anti-epilectic)
Fig. 1: Selected pyrimidine-based drugs commercially available in the market.
MATERIALS AND METHOD
General Condition: Melting points were determined in open capillary tubes on a Stuart melting point apparatus
and were uncorrected. Infrared spectra were recorded on a Schimadzu Spectrometer. The Ultraviolet spectra
were run on a Genesys Spectrometer using acetone solvent. 1H and 13C NMR were run on JEOL-JNM-GX-
300spectrometer at 300 MHz and 75 MHz respectively using DMSO-d6. Mass spectra were run on Finnigan
MAT 312 machine. All compounds were routinely checked by TLC on silical gel G plates using CHCl3:CH3OH
(9:1, v/v) solvent system.The elemental analysis (C, H, N) of compounds were performed using a Carlo Erba-
1108 elemental analyzer.
General procedure for chalcones (7a-g). To a solution of sodium hydroxide (2.5g) in water (20 mL), was added
ethanol (10 mL) with continuous stirring until it cools down to room temperature. To this solution was added a
mixture of appropriate ketone (14.15 mmol) and benzaldehyde (14.15 mmol or 28.30 mmol*) drop-wisely with
continuous stirring at room temperature for 30 min. The resulting solution formed colored precipitate which was
filtered by suction, washed and recrystallized from ethanol to afford7a-g.Where * = double molar equiv. of
benzaldehyde.
General procedure for synthesis of pyrimidinone derivatives(8a-g)
Method I: A mixture of any of chalcones 7a-g (10 mmol) and urea (1.30 g, 21 mmol) was ground in mortar and
quantitatively transferred into a 250 mL round-bottomed flask containing ethanol (30 mL). Later, concentrated
hydrochloric acid (10 mL) was drop-wisely added with continuous stirring and the reaction mixture was refluxed
for appropriate time and reduced by evaporation to half of it original volume. It was then cooled to room
temperature and neutralized with 30% sodium hydroxide and left in the freezer chest over night. The solid
product obtained was recrystallized from ethanol to afford the corresponding pyrimidinone 8a-g in moderate to
good yields.
Method II: To a mixture of any of chalcones 7a-g (10 mmol), urea (1.30g, 21 mmol) and ethanol (20 mL), was
added a catalytic amount of SSA (100 mg, 0.26 mmol), and the reaction mixturewas reflux for appropriate time.
The SSA catalyst was extracted with chloroform (20 mL) and removed from the entire solution. The remaining
solution was reduced to half its volume and cooled to room temperature. It was neutralized with 30% sodium
hydroxide and left in the freezer chest over night. The solid product obtained was recrystallized from ethanol to
afford the pyrimidinone8a-g in good to excellent yields.
4-Methyl-6-phenyl-5,6-dihydropyrimidin-2(1H)-one (8a): UV-VIS{ëmax(Log)}: 325 (3.96), 274 (3.33), 244
-1
(3.78), 226 (3.44), 202 (3.13). IR [, cm ,KBr]: 3241 (N-H), 2928 (CH aliphatic), 1685 (C=O), 1612 (C=C), 1575
(C=N). 1H NMR (300 Hz, ä ppm, DMSO-d6): 8.0 (s, 1H, NH, D2O exchangeable), 7.26-7.40 (m, 5H, Ar-H), 4.90
(t, 1H, CH,J = 7.0 Hz), 1.94 (s, 3H, CH3), 1.91-1.66 (m, 2H, CH2,J = 7.0 Hz). 13C NMR (75 Hz, ä ppm, DMSO-d6):
180.1 (C=O), 160.2, 143.5, 128.7, 128.5, 128.5, 126.9, 126.9, 126.7, 47.7, 40.0, 22.1 (CH3).
4-(4-Ethylphenyl)-6-phenyl-5,6-dihydropyrimidin-2(1H)-one (8b): UV-VIS{ëmax(Log)}: 310 (3.68), 265
(3.86), 230 (3.97), 215(3.77). IR [, cm-1,KBr]: 3133 (N-H), 1685 (C=O), 1570 (C=N). 1H NMR (300 Hz, ä ppm,
DMSO-d6): 8.0 (s, 1H, NH, D2O exchangeable), 7.27-7.40 (m, 7H, 2×Ar-H), 7.78 (d, 2H, Ar-H,J = 7.5 Hz), 4.90
(t, 1H, CH,J = 7.0 Hz), 1.91-1.66 (m, 2H, CH2,J = 7.0 Hz). 2.60 (q, 2H, CH2,J = 8.0 Hz), 1.25 (t, 3H, CH3,J = 8.0
Hz). 13C NMR (75 Hz, ä ppm, DMSO-d6): 164.6 (C=O), 160.1, 146.7, 143.5, 137.8, 128.5, 128.5, 127.8, 127.8,
127.0, 127.0, 126.9, 126.9, 126.7, 47.3 (CH), 42.7 (CH2), 28.2 (CH2), 14.5 (CH3).
4-Phenyl-3,4,4a,5,6,7-hexahydro-2H-cyclopenta[d]pyrimidin-2-one (8c):UV-VIS{ëmax(Log )}: 328 (4.12),
-1
274 (3.39), 247 (3.41), 208 (4.02). IR [, cm ,KBr]: 3295 (NH), 2928 (CH aliphatic), 1690 (C=O), 1600 (C=C),
1565 (C=N). 1H NMR (300 Hz, ä ppm, DMSO-d6): 8.01 (s, 1H, NH, D2O exchangeable), 7.25-7.41 (m, 5H, Ar-
H), 4.92 (d, 1H, CH), 2.67-2.84 (m, 5H, Cp-H), 1.22-1.41 (m, 4H, 2×CH2,J = 7.1 Hz). 13C NMR (75 Hz, ä ppm,
DMSO-d6): 208.4 (C=O), 150.0, 146.1, 142.9, 135.0, 135.0, 128.1, 128.1, 115.0, 115.0, 39.1(CH2), 23.8(CH2),
+ + +
20.4(CH2).MS m/z 214 [M , 12.5%], 137 [M – Ph, 75%], 109 [M – Ph – CO, 100%].
7-Benzylidene-4-phenyl-3,4,4a,5,6,7-hexahydro-2H-cyclopenta[d]pyrimidin-2-one (8d):UV-
VIS{ëmax(Log )}: 330 (3.98), 208 (4.14). IR [, cm-1,KBr]: 3387 (NH), 1685 (C=O), 1612 (C=C), 1575 (C=N). 1H
NMR (300 Hz, ä ppm, DMSO-d6): 8.0 (s, 1H, NH, D2O exchangeable), 7.27-7.60 (m, 10H, 2×Ar-H), 6.34 (s, 1H,
13
CH), 4.91 (d, 1H, CH,J = 7.0 Hz), 2.69 (t, 1H, CH,J = 7.0 Hz), 1.22-2.02 (m, 4H, 2×CH2,J = 7.1 Hz). C NMR (75
Hz, ä ppm, DMSO-d6): 163.0 (C=O), 160.1, 141.5, 137.1, 135.2, 130.8, 128.6, 128.6, 128.5 (four times), 128.1,
128.1, 127.9, 125.9, 49.9, 45.3, 33.6 (CH2), 31.3 (CH2).
7-(3-Methoxybenzylidene)-4-(3-methoxyphenyl)-3,4,4a,5,6,7-hexahydro-2H-cyclopent- a[d]pyrimidin-
2-one (8e):UV-VIS{ëmax(Log )}: 366 (3.98), 345 (3.77), 210 (4.14). IR [, cm-1,KBr]: 3387 (NH), 1685 (C=O),
1612 (C=C), 1575 (C=N). 1H NMR (300 Hz, ä ppm, DMSO-d6): 8.0(s, 1H, NH, D2O exchangeable), 6.82-7.59
(m, 8H, 2×Ar-H), 6.35 (s, 1H, CH), 4.90 (d, 1H, CH,J = 7.0 Hz), 3.84 (s, 6H, 2×CH3,J = 7.0 Hz), 1.81-2.32 (m,
5H, Cp-H,J = 7.1 Hz).
4-Phenyl-4,4a,5,6,7,8-hexahydroquinazolin-2(3H)-one (8f):UV-VIS{ëmax(Log )}: 375 (3.69), 344 (3.87), 210
(4.02). IR [, cm-1,KBr]: 3387 (NH), 1685 (C=O), 1600 (C=C), 1573 (C=N). 1H NMR (300 Hz, ä ppm, DMSO-
d6): 8.0 (s, 1H, NH, D2O exchangeable), 7.27-7.41 (m, 5H, Ar-H), 4.91 (d, 1H, CH,J = 7.0 Hz), 2.19 (q, 1H, CH,J
13
= 7.0 Hz), 1.19-1.41 (m, 8H, 4×CH2,J = 7.1 Hz). C NMR (75 Hz, ä ppm, DMSO-d6): 164.7 (C=O), 160.1, 137.1,
128.5, 128.5, 128.1, 128.1, 125.9, 49.8, 41.9, 33.8, 27.0, 24.8, 24.2.

In continuation of our recent works concerned with the synthesisof a variety of heterocyclic systems for
biological evaluation10, we reporthere on the facile synthesis of substituted pyrimidine motifs under the influence
of silica sulfuric acid (SSA) as heterogeneous catalyst.The synthetic route of the precursor, á,-unsaturated
carbonyls, otherwise known as chalcones, was as illustrated in Scheme 1 wherein á,-unsaturated carbonyls (7a-
g) were synthesized via condensation of benzaldehyde with ketones in basic medium.Compounds 7a-g were
formed in good yields via a continuous stirring and agitation at room temperature. They were subsequently
reacted with urea under two different conditions which include the common technique in the presence of
concentrated HCl (Method I) and the proposed technique using solid support catalyst, Silica Sulfuric Acid (SSA)
(Method II), to afford pyrimidin-2(1H)-one derivatives (8a-g).
As a case study, the condensation of an equimolar mixture of benzaldehyde andacetoneafforded4-
phenylbut-3-en-2-one,7a. The reactive intermediate chalcone 7a was subsequently treated with urea in
ethanol in the presence of either concentrated hydrochloric acid (Method I) or Silica Sulfuric Acid, SSA
(Method II) under reflux at 140 C to afford 4-methyl-6-phenyl-5,6-dihydropyrimidin-2(1H)-one,8a. This
procedure was repeated for the chemical transformation of other chalcones to their corresponding
pyrimidinone derivatives8b-g (Scheme 2). In Method I, upon completion (TLC), the reaction was worked up
to afford 8a in low yield 44% after refluxing for 8 h. However, in Method II, where conc. HCl was replaced
with solid support catalyst SSA, the reaction time did not only reduced drastically to 3 h but also led to the
formation of the product 8a at a higher yield, 77% (Fig. 2). The SSA catalyst was recovered with chloroform
(20 mL).
O
C
H
3
7
a
O
O
(a
)
C
H
2
5
C
H
O
(
b
)
7
b
(g
)
7
g
(
c
)
f(
)
O
O
(d
-e
)
O
7
c
7
f
R d
7
:
R
=
H
R
=
H
f
o
r
a
ll
c
o
m
p
o
u
n
d
s
e
x
c
e
p
t
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e
R
7
e
:
R
=
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C
H
*
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R
e
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c
t
io
n
C
o
n
d
it
io
n
s
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r
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h
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lc
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e
7
a
-g
:
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th
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w
it
h
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e
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r
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e
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n
e
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s
e
d
:
(a
)
a
c
e
to
n
e
,
(
b
)
4
-
e
th
y
l
a
c
e
to
p
h
e
n
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n
e
,
(c
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)
c
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n
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n
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,
(
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y
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,
(
g
)
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*
=
d
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u
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e
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iv
a
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n
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n
z
a
ld
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,
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a
ll
e
x
c
e
p
t
in
7
e
w
h
e
re
R
=
O
C
H
3
Scheme 1: The pathway for the synthesis of á,â-unsaturated carbonyl, chalcones, 7a-g.
O
HN N
C H3 O
O
8a
N NH H N N
( 7a )
CH 3
(7 b ) 8b
( 7g )
8g O
H 2N NH2
O (7 f ) U REA ( 7c )
O
N N H ( 7d - e)
N N H
O
N NH
8f 8c
8d: R = H R
R 8 e : R = O C H 3*
R e a c t io n C o n d i ti o n f o r 8 a - g : M e th o d I : R e f l u x in e t h a n o l u s in g co n c . H C l. M e th o d I I : R e f lu x
in e th a n o l u s in g s ili ca s u p p o rt e d s u lf u r ic a c i d , S S A . R = H in a ll e x c e p t in 8 e w h e r e R = O C H 3
Scheme 2: The pathway for the synthesis of substituted pyrimidin-2(1H)-ones, 8a-g.

The resulting filtrate was reduced to half its volume and cooled. It was neutralized with ammonium hydroxide
and filtered by suction to afford 8a. In a nutshell, it was observed that SSA did not only emerge as an efficient
catalyst in this study but also afforded the pyrimidinone products 8a-g in higher yields (75-95%) within smaller
reaction time (3-4 h) compared with concentrated hydrochloric acid which gave smaller yields (45-71%) at
higher reaction time of 8-9 h (Fig. 2). The physico-chemical parameters and analytical data were as shown in
Table 1 and were consistent with the proposed structures of 8a-g.
Fig. 2: The comparative study of efficiency of tradition method using conc. HCl (Method I) to that of new approach
using SSA (Method II) (a) considering % yields factor. (b). considering reaction time (h) factor.
Table 1: Physico-chemical properties of synthesized pyrimidin-2(1H)-ones (8a-g)
Comp Molecular Mol. M.P. Rf a Colour Elemental Analysis %Calcd. (%Found)
Code Formula Wt. (°C) CH N
8a C11H12N2O 188 124-127 0.77 Yellow 70.21(70.73) 6.38(6.09) 14.89(15.01)

8b C18H18N2O 278 211-213 0.56 white 77.70(77.31) 6.47(6.72) 10.07(9.85)
8c C13H14N2O 214 184-186 0.85 Green 72.90(72.71) 5.04(5.52) 13.08(13.53)
8d C20H18N2O 302 227-229 0.49 green 79.47(79.95) 5.98(5.82) 9.27(9.45)
8e C22H22N2O3 362 240-242 0.69 green 72.93(73.04) 6.08(5.97) 7.73(7.55)
8f C14H16N2O 228 198-200 0.76 Orange 73.68(73.81) 7.02(7.16) 12.28(12.33)
8g C21H20N2O 316 >320 0.63 black 79.75(79.11) 6.33(6.72) 8.86(8.43)
a
CHCl3:CH3OH (9:1, v/v), Mol, Wt. = Molecular Weight, M.P. = Melting Point
CONCLUSION
The pyrimidine derivatives were successful achieved by synthetic modification of the various chalcone
precursors via Silica sulfuric acid (SSA) heterogeneous catalytic approach. SSA was found to be a mild, efficient
and reusable solid catalyst for the reaction of á,-unsaturated carbonyl with urea to furnish the corresponding
pyrimidinone derivatives in good to excellent yield.The interesting behaviour of SSA lies in the fact that it can be
re-used after simple washing with chloroform thereby rendering this procedure more economical compared with
concentrated HCl method.
ACKNOWLEDGEMENT
OOA acknowledges The World Academy of Sciences for the sponsorship of this project under the TWAS
Research Grant for Individual Programme (Grant No: 14-069 RG/CHE/AF/AC_1; UNESCO FR: 324028564).
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Table of Content

S/N Topics and Authors Page

58 Phytochemical Studies And Aphrodisiac Activity Of Byrsocarpus Coccineus Root Exract

Materials and Methods

Determination of Minimum Inhibitory Concentration (MIC)

RESULTS AND DISCUSION

Table 1: Susceptibility test of the extracts of Flabellaria paniculata

Test organism Zone of inhibition (mm)

Test organism Zone of inhibition (mm)

Table 3: MIC of P2a and P2b isolates of Flabellariapaniculata

Compound Concentration (µg/ml) Test organism

MBC value;-, No growth;+, Light growth

OGODO AEMURERHIMEN DICKSON (Snr.) (Chartered Chemist)

Methodology (Experimental) (Materials and Method)

1.2: Ambient Air Monitoring

1.3: Methods for Analysis of Ambient Air Pollutants

1.4.1: Distance Mapping of Sampling Point

1.4.2: Heat Radiation

1.4.3: Noise Levels

1.4.4: Relative Smoke Density

1.4.5: Wind Speed and Direction

1.4.7: Air Quality

(b) Hydrocarbons/Volatile Organic Compounds (CxHy/VOC), Nitrogen Oxides (NOx), Sulphur

1.4.8: Suspended Particulate Matter (SPM)

3.4.10 Data Analysis

Table 3: Ringlemann Number/Percent Light Transmission and Smoke Density.

WIND SPEED AND DIRECTION

AMBIENT AIR QUALITY

Table 4: Nigerian Ambient Air Quality Standard.

Fig. 5A: Radial Graph Representation of SPDC Amukpe – Sapele

Table 5B: Comparison of WRPC With DPR Limits

Fig. 5B: Radial Graph Representation of WRPC in Ekpan (Uvwie)

Fig. 5C: Radial Graph Representation of NGC in Ekpan (Uvwie)

Table 5D: Comparison Of Ogini Isoko North with DPR Limits

Table 5E: Comparison o Ugbekoko with DPR Limits

Fig. 5E: Radial Graph Representation of SPDC in Ugbekoko (Sapele West)

Fig. 5F: Radial Graph Representation of Ogwashi -Uku (Reference/Control Station)

Table 5G: Comparison of Issele-Uku with DPR Limits

Air Turbulence and Wind Direction

Ambient Air Quality

VOLATILE ORGANIC CARBON

Air Pollution Abatement

3.3 Physico-Chemical properties of the Adsorbed: The data table is as follows;

% Removal of metal ions, Qe

% Removal of metal ions

Degree of % Removal of metal ions, Qe

Pb2+ Cd2+ Ni2+

Percentage inhibition efficiency at different concentrations of inhibitor

DLE 61.1 65.3 86.3 89.3 90.5 92.5

ASSESSMENT OF SOME METALS IN ROADSIDE DUST FROM DAMATURU, YOBE STATE,

*Isah Yinusa, Woli Ajoke Balkees and Hassan Adam Adeiza

Keyword: Anti-Oxidant, Sarcocephalus, Elemental, Fruit, Activity, Scavenging, Radical, Diet

The IC50of S. latifoliuswas obtained by linear regression equations: y = 4.3145x + 43.445

Element Concentration Standard Mean

* E. I. Uwah and E. A. Moses

Parameters Composition (%)

Mineral Elements Composition (mg/100g)

Anti-nutrients Composition (mg/100g)

Phytochemical Methanol Aqueous extract

Key: (+) = present, (-) = absent

Table 2: Quantitative analysis of phytochemical constituents of soy flour

Parameters Values FAO/WHO guidelines

Results and Discussion