Lisan - Prosiding Simposium Kimia Analisis Malaysia Ke-18, 2005

SKAM 18
SIMPOSIUM KIMIA ANALISIS MALAYSIA

KELAPAN BELAS
Sains Analisis Cemerlang Negara Terbilang
E-PROSIDING
12 – 14 September 2005
Hotel Hyatt Regency
Johor Bahru, Johor
anjuran bersama
Jabatan Kimia, Fakulti Sains, Universiti Teknologi Malaysia
dan
Persatuan Sains Analisis Malaysia (ANALIS)
dengan kerjasama
Institut Kimia Malaysia (Cawangan Selatan)
Prosiding Simposium Kimia Analisis Malaysia Ke-18, Johor Bahru
SESI KERTAS KERJA LISAN

1A Alam Sekitar I
Pengerusi: Prof. Madya Dr. Salihan Siais (UPM)
Bilik: Sri Mersing
1A-1 ANALISIS UNSUR DAN KERADIOAKTIFAN DALAM SAMPEL SEDIMEN
TASIK CHINI, PAHANG DARUL MAKMUR
AMRAN AB.MAJID (UKM)
1A-2 TRACE ELEMENTS IN OREOCHROMIS NILOTICUS (RED TILAPIA)
FROM FISH POND
ZAHARAH AIYUB (UM)
1A-3 THE CHARACTERISTICS OF GEOCHEMISTRY AND
SEDIMENTOLOGICAL IN PAKA RIVER SEDIMENT
ONG MENG CHUAN (KUSTEM).
1A-4 PYROLYSIS AND LIQUEFACTION OF ACETONE AND MIXED
ACETONE/TETRALIN PRE-SWELLED MUKAH BALINGIAN
MALAYSIAN SUB-BITUMINOUS COAL – THE EFFECT ON COAL
CONVERSION AND OIL YIELD
MOHD FAUZI ABDULLAH (UiTM)
1A-5 KANDUNGAN LOGAM BERAT DAN RADIONUKLID TABII DALAM
IKAN, AIR, TUMBUHAN DAN SEDIMEN DI BEKAS TASIK LOMBONG
MUHAMAD SAMUDI YASIR (UKM)
1B Kimia Tak Organik dan Pemangkinan I

Pengerusi: Prof. Madya Dr. Razali Ismail (UTM)
Bilik: Sri Pontian
1B-1 X-RAY CRYSTALLOGRAPHY AND BIOLOGICAL PROPERTIES OF
INDOLE-AROMATICHYDRAZONE AND THEIR METAL COMPLEXES
HAPIPAH MOHD ALI (UM)
1B-2 CHARACTERIZATION OF NOVEL ORGANOTIN(IV) COMPLEXES WITH

O, N, O – DONOR LIGAND DERIVED FROM CARBOHYDRAZIDE: X-RAY
CRYSTAL STRUCTURE OF [PH2SN(H2CBS)]
LIEW YEW ZION (UNIMAS)
1B-3 THE PREPARATION AND CHARACTERIZATION OF ISOMERIC SCHIFF
BASES AND THEIR TRANSITION METAL COMPLEXES STRUCTURE-
REACTIVITY RELATION
FIONA HOW NI FOONG (UPM)
1B-4 STUDY ON BIMETALLIC NI/CO CATALYST DOPED WITH
PRASEODYMIUM USING VARIOUS SUPPORT FOR CO2 / H2
METHANATION TOWARDS PURIFICATION OF NATURAL GAS
FARIDAH MOHD MARSIN (UTM)
1B-5 KAJIAN KRISTALOGRAFI SINAR-X BIS(N-(O-METILFENIL)-N’-(P-
METOKSIBENZOIL)-TIOUREA -κS)-DIIODOMERKURI(II) DAN BIS[µ-1-
(2-PIRIDIL-κN)ETANONE-4-FENILTIOSEMIKARBAZONATO-
κ2N1,S]BIS[IODOMERKURI(II)]
MOHD SUKERI MOHD YUSOF (KUSTEM)
1C Teknik Pemisahan I
Pengerusi: Prof. Dr. Mohd. Marsin Sanagi (UTM)
Bilik: Sri Muar
1C-1 MULTIRESIDUE ANALYSIS OF PESTICIDES IN FRUITS AND
VEGETABLES USING SOLID PHASE EXTRACTION AND GAS
ii

CHROMATOGRAPHY WITH AN ELECTRON CAPTURE DETECTOR
GUAN H. TAN (UM)
1C-2 SUBCRITICAL WATER EXTRACTION OF ESSENTIAL OIL FROM
CORIANDER (CORIANDRUM SATIVUM L.) SEEDS
NORASHIKIN SAIM (UiTM)
1C-3 PERKEMBANGAN SATU TEKNIK PENGEKSTRAKAN FASA PEPEJAL
BAGI ANALISIS ASID HALOASETIK (HAAs) DALAM AIR MINUM
MARINI AB. RAHMAN (UKM)
1C-4 ANALYSIS OF RESIDUAL TOLUENE IN FOOD PACKAGING
MATERIALS USING HEADSPACE GAS CHROMATOGRAPHY
LIM YING CHIN (UiTM)
1C-5 SYNTHESIS OF 3-O-SUCCINYL-BETULINIC ACID AS ANTI –CANCER
AGENTS AGAINST HUMAN MYELOID LEUKAEMIA AND HUMAN T-4
LYMPHOBLASTOID CELLS LINE.
MOHD. TAJUDIN MOHD. ALI (UiTM)
1D Organik/Bioteknologi I
Pengerusi: Prof. Madya Dr. Habsah Mohamad (KUSTEM)
Bilik: Ballroom
1D-1 KEKABU SEED OIL EXTRACTION AND PUFA ISOLATION
JUMAT SALIMON (UKM)
1D-2 TOXICITY AND ANTITERMITE ACTIVITIES OF THE ESSENTIAL OILS

FROM PIPER SARMENTOSUM
CHIENG TIONG CHIN (UNIMAS)
1D-3 MICROBIAL INFLUENCED CORROSION CAPABILITY OF SULFATE-
REDUCING BACTERIA ISOLATED FROM THE MALAYSIAN SHIPYARD
AND ENGINEERING HARBOURS, PASIR GUDANG
FATHUL KARIM SAHRANI (UTM)
1D-4 KAJIAN PENGOPTIMUMAN TINDAK BALAS SAPONIFIKASI MINYAK
KACANG SOYA
HASNISA BINTI HASHIM (UKM)
1D-5 ENZYMATIC SCALE-UP PRODUCTION OF PALM OIL WAX ESTERS −
OPTIMIZATION USING RESPONSE SURFACE METHODOLOGY
KENG PEI SIN (UPM)
2A Alam Sekitar II
Pengerusi: Prof. Madya Dr. Ahmad Saat (UiTM)
Bilik: Sri Mersing
2A-1 VOLATILE ORGANIC COMPOUNDS IN MALAYSIAN DRINKING
WATER : IS IT AN ISSUE?
MD. PAUZI ABDULLAH (UKM)
2A-2 REMOVAL OF HEAVY METAL IONS WITH ACID ACTIVATED
CARBONS DERIVED FROM OIL PALM AND COCONUT SHELLS
MOHAMMAD ADIL (UTM)
2A-3 KANDUNGAN TOLUENA DAN XILENA DALAM DEBU JALAN DI
SEKITAR KUALA LUMPUR
MOHD ROZALI OTHMAN (UKM)
2A-4 TABURAN 228Th, 230Th DAN 232Th DI DALAM AIR LAUT DAN SEDIMEN
PERMUKAAN DI PERAIRAN PANTAI TIMUR SEMENANJUNG
MALAYSIA
JALAL SHARIB@SARIP (MINT)
2A-5 210
Po/210Pb DALAM AIR HUJAN, AIR SUNGAI DAN AIR LAUT
iii

CHE ABD RAHIM MOHAMED (UKM)
2A-6 PENGGUNAAN PENAPIS BIOLOGI TERAKTIF BAGI RAWATAN AIR
SISA
RAZALI ISMAIL (UTM)
2B Industri I
Pengerusi: Prof. Madya Dr. Wan aini Wan Ibrahim (UTM)
Bilik: Sri Pontian
2B-1 PENGHASILAN SERTA PENCIRIAN HITAM KARBON DAN KARBON
TERAKTIF YANG DIHASILKAN DARIPADA PELBAGAI SISA
LIGNOSELULOSA
AHMAD MD. NOOR (USM)
2B-2 SYNTHESIS AND CHARACTERISATION OF MOLYBDENUM-
VANADIUM BASED OXIDE PREPARED BY THE ANDERSON-TYPE
HETEROPOLYMOLYBDATES ROUTE
TAN YEE WEAN (UPM)
2B-3 AN ELECTROGENERATIVE PROCESS FOR THE RECOVERY OF GOLD
FROM CYANIDE SOLUTIONS
YAP CHIN YEAN (USM)
2B-4 KESAN SUHU PUNARAN DALAM MENAMPAKKAN SEMPADAN
BUTIRAN ALOI ALUMINIUM AA5083 DENGAN MENGGUNAKAN ASID
NITRIK
AZMAN JALAR (UKM)
2B-5 INFLUENCE OF PRECIPITATING AGENT ON COPPER(II) OXIDE
SYNTHESISED VIA PRECIPITATION METHOD
LAU HOOI HONG (UPM)
2B-6 LINKING STUDENTS’ LEARNING PREFERENCES WITH THEIR
CONCEPTUAL UNDERSTANDING IN BASIC CHEMISTRY
JAAFAR JANTAN (UITM)
2C Elektroanalisis/Penderia I
Pengerusi: Prof. Madya Dr. Md. Sani Ibrahim (USM)
Bilik: Sri Muar
2C-1 PHASE BOUNDARY HETEROGENEOUS TIO2/ZRO2 CATALYST FOR
EPOXIDATION
HALIMATON HAMDAN (UTM)
2C-2 TRIIODIDE RESPONSIVE MEMBRANE AND ITS APPLICATION FOR
THE SEQUENTIAL FLOW INJECTION DETERMINATION OF
CHLORINE SPECIES
WAN TATT WAI (USM)
2C-3 PENGGUNAAN 1-(2-HIDROKSIFENIL)ETHANON BENZOILHIDRAZON
(HPEBH) SEBAGAI REAGEN UNTUK PENENTUAN KUANTITATIF
AL(III) SECARA SPEKTROFOTOMETRI UL-NAMPAK
NURIAH MOHAMAD (UKM)
2C-4 BAHAN PENDERIA pH BERASASKAN REAGEN FENILFLUORON
TERPEGUN DALAM MATRIK HIBRID SOL-GEL - KITOSAN
ROSMAWANI MOHAMMAD (UKM)
2C-5 VOLTAMMETRIC STUDIES OF CADMIUM ION AT THE MERCURY
ELECTRODE IN THE PRESENCE OF GLUTATHIONE
M. ZIDAN (UPM)
2C-6 ADSORPTION OF CHROMATE ANION FROM AQUEOUS SOLUTION BY
iv

SURFACTANT MODIFIED ZEOLITE Y
NIK AHMAD NIZAM (UTM)
2D Organik/Bioteknologi II
Pengerusi: Prof. Madya Dr. Jumat Salimon (UKM)
Bilik: Ballroom
2D-1 DPPH FREE RADICAL SCAVENGING, ANTIMICROBIAL AND
ANTIAMOEBIC COMPOUNDS FROM ZIZYPHUS MAURITIANA
(RHAMNACEAE)
HABSAH MOHAMAD (KUSTEM)
2D-2 PRODUCTION OF WATER SOLUBLE AND LOW MOLECULAR WEIGHT
CHITOOLIGOSACCHARIDES USING COMMERCIAL MIXED-
GLUCANASE FROM ASPERGILLUS ACULEATUS
LEE LIH FEN (UKM)
2D-3 SYNTHESIS, CHARACTERIZATION AND BIOLOGICAL ACTIVITY OF A
NEW SCHIFF BASE AND ITS COMPLEXES DERIVED FROM 1-(2-
THIENYL)-1-PROPANONE
M. IBRAHIM M. TAHIR (UPM)
2D-4 PENGHASILAN SURFAKTAN TAK BERION BERASASKAN LAURIL
ALKOHOL DARIPADA MINYAK KELAPA SAWIT
NORHAFIPAH MOHAMAD (UKM)
2D-5 THE OPTIMIZATION OF ACIDIC HYDROLYSIS OF CHITIN BY
RESPONSE SURFACE METHODOLOGY
CHEE KAH LEONG (UKM)
2D-6 FRACTIONATION OF SUPERCRITICAL FLUID EXTRACT OF ZINGIBER
ZERUMBET AND IT’S SKIN WHITENING PROPERTIES
MAZITA MOHD. DIAH (SIRIM)
3A Alam Sekitar III

Pengerusi: Prof. Dr. Ibarhim Baba (UKM)
Bilik: Sri Mersing
3A-1 KAEDAH MENGURANGKAN COD DALAM AIR KUMBAHAN KILANG
PEMBERSIHAN GULA
SALIHAN BIN SIAIS (UPM)
3A-2 CHLORPYRIFOS AND MALATHION RESIDUES IN SOILS OF A
TERENGGANU GOLF COURSE: A CASE STUDY
JURRIFFAH ARIFFIN (KUSTEM)
3A-3 LONG-TERM STUDIES ON THE SALINITY (TDS) AND LEACHING
CHARACTERISTICS OF COAL FLY ASH FROM TNB POWER
GENERATION PLANT
KHOO KOK SIONG (UKM)
3A-4 PRELIMINARY TROPHIC STATE INDEX AND WATER QUALITY
ASSESSMENT RESULTS OF TASIK CHINI
MAKETAB MOHAMED (UTM)
3A-5 PENENTUAN KEPEKATAN UNSUR-UNSUR SURIH DALAM TANIH
SAWAH DI KAWASAN PERLIS SECARA ANALISIS INSTRUMENTASI
PENDARFLOUR SINAR-X (XRF)
PUI VUI MING (UKM)
3B Industri II
Pengerusi: Prof. Madya Dr. Ahmad Mohd. Nor (USM)
Bilik: Sri Pontian
v

3B-1 PENENTUAN KUALITI AIR TASIK KEJURUTERAAN UKM KAMPUS
BANGI: KE ARAH SISTEM PENGURUSAN SUMBER AIR BERSEPADU
MAZLIN MOKHTAR (UKM)
3B-2 KAJIAN PENGEMBANGAN PENGGUNAAN ZEOLIT DARIPADA SEKAM
PADI DALAM SISTEM PENAPISAN AIR MINUM
NORHASNI RAMLI (UKM)
3B-3 REMOVAL OF BASIC AND REACTIVE DYES: A COMPARISON OF
SORPTION AND PHOTODEGRADATION STUDY
ONG SIEW TENG (UPM)
3B-4 BINARY ION-EXCHANGE OF DIVALENT LEAD AND CADMIUM IN
AQUEOUS SOLUTION WITH THE INDIGENOUS IONS OF RICE HUSK
ASH-SYNTHESIZED ZEOLITE P
TAN SEE HUA (UTM)
3B-5 SYNTHESIS AND PROPERTIES OF ZINC-ALUMINIUM-
ANTHRANILATE-NANOCOMPOSITE
MAZIDAH MAMAT (UPM)
3C Elektroanalisis/Penderia II
Pengerusi: Prof. Madya Dr. Marcus Jopony (UMS)
Bilik: Sri Muar
3C-1 KAJIAN KECEKAPAN ANOD KORBANAN ALOI AL-5.5ZN-2.0MG-XSN DI
DALAM AIR LAUT TROPIKA
ABDUL RAZAK DAUD (UKM)
3C-2 DETERMINATION OF LEAD BY NEW OPTICAL CHROMOGENIC
SENSOR
ABDUSSALAM SALHIN (USM)
3C-3 Al(III) SENSING USING IMMOBILIZED PAN ENTRAPPED IN SOL GEL
FILM BASED ON FLUORESCENCE BEHAVIOR
Rita Sundari (UKM)
3C-4 SQUARE WAVE CATHODIC STRIPPING VOLTAMMETRIC TECHNIQUE
FOR DETERMINATION OF AFLATOXIN B1 IN GROUND NUT SAMPLE
MOHAMAD HADZRI YAACOB (USM)
3C-5 FABRICATION OF CATECHOL BIOSENSOR BASED ON IMMOBILIZED
MBTH AND LACCASE ENZYME
JAAFAR ABDULLAH (SIRIM)
3D Kimia Tak Organik/Pemangkinan II

Pengerusi: Prof. Madya Dr. Hapipah Mohd. Ali (UM)
Bilik: Ballroom
3D-1 SPECTROPHOTOMETRIC DETERMINATION OF ARSENIC (III) BASED
ON COMPLEX FORMATION WITH GALLOCYANINE
NOR AZAH YUSOF (UPM)
3D-2 X-RAY CRYSTALLOGRAPHY AND BIOLOGICAL PROPERTIES OF
ISATIN-2-THIOPHENECARBOXYLIC HYDRAZONE WITH ZINC,
NICKEL, CADMIUM, MANGANESE, COBALT AND COPPER
COMPLEXES.
SITI NADIAH ABDUL HALIM (UM)
3D-3 PHOTOLUMINESCENCE OF EUROPIUM-PICRATE-POLYETHERS
COMPLEXES
ENY KUSRINI (USM)
3D-4 TINDAK BALAS PENGKOMPLEKSAN ASID-3-(3-BENZOILTIOUREDO)
PROPIONIK DENGAN BIS[ASETATOTRIFENILFOSFINARGENTUM(I)]
DAN KOBALT(II)KLORIDA
vi

NURZIANA NGAH (UKM)
3D-5 PHOTODEGRADATION OF MDEA, PARAQUAT AND PHENOL USING
TiO2 AND ZnO BASED PHOTOCATALYSTS: COMPARISON STUDY
NORAIDURA AMIN (UTM)
4A Alam Sekitar IV
Pengerusi: Prof. Madya Dr. Mohd. Rozali Othman (UKM)
Bilik: Sri Mersing
4A-1 ASSESSMENT OF WATER QUALITY IN SEMENYIH WATER RESOURCE
AREA AND HULU LANGAT DOMESTIC WATER SUPPLY
ZAINI HAMZAH (UiTM)
4A-2 HEAVY METALS DISTRIBUTION IN URBAN SOIL OF SEBERANG
PERAI TENGAH, PULAU PINANG
POH SENG CHEE (KUSTEM)
4A-3 ASSESSMENT OF ELEMENTAL POLLUTION IN THE STRAIT OF
MELAKA
AWAD A. ALZAHRANY (UPM)
4A-4 ANALYSIS OF NATURAL OCCURRING RADIOACTIVE MATERIALS AT
AND AROUND KOTA TINGGI WATERFALL IN JOHOR
REDZUWAN YAHAYA (UKM)
4B Industri III
Pengerusi: Prof. Madya Dr. Musa Ahmad (UKM)
Bilik: Sri Pontian
4B-1 CHARACTERIZATION OF TWO DIFFERENT LIQUID SCINTILLATION
COUNTERS: EFFECTS ON AGE DETERMINATION IN RADIOCARBON
DATING SYSTEME
NORAISHAH OTHMAN (MINT)
4B-2 DECOLOURISATION AND DEGRADATION OF REACTIVE ORANGE 16
DYE IN AQUEOUS SOLUTIONS BY THE CUO/H2O2 PROCESS
WONG WAN YUAN (UPM)
4B-3 ANALYSIS OF POLYNUCLEAR AROMATIC HYDROCARBON (PAHs) IN
EDIBLE OILS
SITI NURDIANA JAAFAR (USM)
4B-4 PREPARATION AND MODIFICATION ACTIVATED CARBON WITH
NITRIC ACID (HNO3)
LOO LI YIN (UPM)
4C Analisis Am
Pengerusi: Prof. Madya Dr. Norhayati Mohd. Tahir (KUSTEM)
Bilik: Sri Muar
4C-1 MICROSCALE CHEMISTRY: AN ALTERNATIVE WAY FOR DOING
CHEMISTRY EXPERIMENTS?
MASHITA ABDULLAH (USM)
4C-2 CLASSIFICATION OF CHILLI SAUCES : MULTIVARIATE PATTERN
RECOGNITION USING SELECTED GCMS RETENTION TIME PEAKS OF
CHILLI SAUCE SAMPLES
LOW KAH HIN (UM)
4C-3 CORRELATING STUDENTS’ VIEWS ABOUT CHEMISTRY AND THEIR
CONCEPTUAL KNOWLEDGE IN FIRST YEAR CHEMISTRY
ZARILA MOHD SHARIFF (UiTM)
vii

4C-4 KEMAHIRAN PROSES SAINS – SATU WAHANA KE ARAH KEJITUAN
DAN KEPERSISSAN MEMBUAT ANALISIS
NAZIYAH KHALID (UPSI)
4D Organik/Bioteknologi III
Pengerusi: Prof. Madya Dr. Sharifuddin M. Zain (UM)
Bilik: Ballroom
4D-1 SYNTHESIS AND CHARACTERISATION OF WATER SOLUBLE
CHITOSAN DERIVATIVES
NOORRASYIDAH MOHD. SARMIN (SIRIM)
4D-2 PRODUCTION OF RESISTANT STARCH FROM HIGH-AMYLOSE
STARCH DERIVED FROM ENZYMATIC DEBRANCHING OF SAGO
STARCH
CHEN SIOW WOON (UKM)
4D-3 SYNTHESIS AND FLUORESCENCE CHARACTERISTIC OF 2-
SUBSTITUTED AND 6-SUBSTITUTED PURINES
MAIZATUL AKMAN A. BAKAR (UM)
4D-4 TINDAK BALAS PENGETOKSILAN ASID LAURIK BERASASKAN
MINYAK KELAPA SAWIT
ZARINA EDRIS (UKM)
5A Alam Sekitar V
Pengerusi: Prof. Madya Dr. Amran Abdul Majid (UKM)
Bilik: Sri Mersing
5A-1 WELL WATER QUALITY IN NORTH-EASTERN DISTRICTS OF
KELANTAN
AHMAD SAAT (UiTM)
5A-2 A PRELIMINARY INVESTIGATION OF INLAND AQUATIC
ACIDIFICATION ON SEMENYIH RESERVOIR
S.MARIAM SUMARI (UiTM)
5A-3 AKTIVITI 226Ra DAN 228Ra Di PERMUKAAN SEDIMEN KAWASAN
BAGAN LALANG, SELANGOR
NIOO SIEW YEW (UKM)
5A-4 REACTIONS OF HEAVY METALS (CU, ZN, FE AND MN) IN ACIDIC
METAL-RICH WATER WITH CALCAREOUS MUDSTONE
MARCUS JOPONY (UMS)
5A-5 KERADIOAKTIFAN TABII DAN KANDUNGAN LOGAM BERAT DALAM
IKAN DARI SUNGAI ATOK DAN SUNGAI TAHAN , TAMAN NEGARA
PAHANG
MUHAMAD BIN OTHMAN (UKM)
5B Industri IV
Pengerusi: Dr. Meor Yusoff Meor Sulaiman (MINT)
Bilik: Sri Pontian
5B-1 FABRIKASI SENSOR KIMIA OPTIK KAPSAISIN BERASASKAN KAEDAH
PENDARFLOUR
MUSA AHMAD (UKM)
5B-2 HYDROMETALLURGICAL PROCESS TO SEPARATE LANTHANIDES
MD FAZLUL BARI (KUKUM)
5B-3 SYNTHESIS, CHARACTERIZATION AND BIOLOGICAL ACTIVITIES OF
SUBSTITUTED SALICYLALDEHYDE-HYDRAZONE AND THEIR METAL
COMPLEXES
PUVANESWARY SUBRAMANIAM (UM)
viii

5B-4 The Effect of pH on the Formation of Zinc/Aluminium-4-Chlorophenoxy Acetate
Nanocomposite
SITI HALIMAH SARIJO (UPM)
5B-5 HPW ENTRAPPED IN MESOPOROUS SILICA VIA SOL GEL
TECHNIQUE FOR ESTERIFICATION REACTION
JUAN JOON CHING (UKM)
5C Tak Org/Pemangkinan III

Pengerusi: Prof. Madya Dr. Abdul Halim Abdullah (UPM)
Bilik: Sri Muar
5C-1 KETIDAKTERTIBAN DALAM PENYELESAIAN DAN PENGHALUSAN
STRUKTUR MOLEKUL SINAR-X
BOHARI M. YAMIN (UKM)
5C-2 POLYMER-ADDED TITANIUM DIOXIDE PHOTOCATALYSTS FOR THE
DEGRADATION OF BENZENE
AZLIYANA ABDUL RAHMAN (UTM)
5C-3 SYNTHESIS, STRUCTURAL CHARACTERIZATION AND BIOLOGICAL
ACTIVITY OF ORGANOTIN(IV) DERIVATIVES WITH
CARBOHYDRAZIDE -BIS(SALICYLALDEHYDE) (H4CBS): X-RAY
CRYSTAL STRUCTURE OF [Me2Sn(H2CBS)]
M. ABU AFFAN (UNIMAS)
5C-4 HYDRAZONE DERIVATIVE CHROMOGENIC REAGENT: SYNTHESIS,
CHARACTERIZATION AND ANALYTICAL APPLICATION
TEOH BOON SIEW (USM)
5C-5 Bi-Fe PROMOTED VANADIUM PHOSPHATE (VPO) CATALYST FOR

THE SELECTIVE OXIDATION OF n-BUTANE TO MALEIC ANHYDRIDE
GOH CHEE KEONG (UPM)
6A Alam Sekitar VI
Pengerusi: Prof. Madya Dr. Zaini Hamzah (UiTM)
Bilik: Sri Mersing
6A-1 POLYCYLIC AROMATIC HYDROCARBONS IN URBAN SOILS OF
KEMAMAN
NORHAYATI MOHD TAHIR (KUSTEM)
6A-2 PERLAKUAN ISOTOP URANIUM DI PERAIRAN PANTAI TIMUR
SEMENANJUNG MALAYSIA: PENENTUAN NISBAH KEAKTIFAN
234
U/238U
ZAL UYUN WAN MAHMOOD (MINT)
6A-3 ASSESSMENT OF NATURAL RADIOACTIVITY IN WATER AND
SEDIMENT FROM AMANG (TIN TAILING) PROCESSING PONDS
MOHSEN NASIRIAN (UKM)
6A-4 ASSESSMENT OF HYDROCARBON POLLUTIONS ON COASTAL
ENVIRONMENT BY PETROLEUM PROCESSING ACTIVITIES.
ABDUL NASIR HAJI SAMOH (USM)
6B Industri V
Pengerusi: Noorrasyidah Mohd Sarmin (SIRIM)
Bilik: Sri Pontian
6B-1 COATING THICKNESS MEASUREMENT FOR GOLD BY USING EDXRF
MEOR YUSOFF M.S. (MINT)
6B-2 PREPARATION AND CHARACTERIZATION OF CHEMICALLY
IMMOBILIZED CROWN ETHER TO MESOPOROUS SILICA FOR SOLID
ix

PHASE EXTRACTION (SPE) APPLICATIONS
MARDIANA SAAID (USM)
6B-3 SYNTHESIS AND CHARACTERISATION OF MOLYBDENUM OXIDE
CATALYSTS FROM CONTROLLED AGEING PROCESS
SITI MURNI BINTI M. ZAWAWI (UPM)
6B-4 MOLECULAR RECOGNITION OF A CHROMOGENIC HYDRAZONE
DERIVATIVE TOWARDS AMINES
ABDASSALAM ABDELHAFIZ (USM)
6C Teknik Pemisahan II
Pengerusi: Prof. Madya Dr. Mohd. Basyaruddin (UPM)
Bilik: Sri Muar
6C-1 DETERMINATION OF CAROTENE, TOCOPHEROLS AND
TOCOTRIENOLS IN RESIDUE OIL FROM PALM PRESSED-FIBER
USING PRESSURIZED LIQUID EXTRACTION-NORMAL PHASE LIQUID
CHROMATOGRAPHY
M. MARSIN SANAGI (UTM)
6C-2 APPLICATION OF SOLID-PHASE MICROEXTRACTION FOR THE
DETERMINATION OF PESTICIDES IN VEGETABLE SAMPLES BY GAS
CHROMATOGRAPHY WITH AN ELECTRON CAPTURE DETECTOR
CHAI MEE KIN (UNITEN)
6C-3 APPLICATION OF SOLID PHASE MICROEXTRACTION (SPME) IN
PROFILING HYDROCARBONS IN OIL SPILL CASES
ZURAIDAH ABDULLAH MUNIR (UiTM)
6C-4 ISOLATION AND PURIFICATION OF LINOLEIC ACID FROM CEIBA
PENTANDRA SEED OIL
KHAIRUL ASMAK ABD. KADIR (UKM)
x
ANALISIS UNSUR DAN KERADIOAKTIFAN DALAM SAMPEL SEDIMEN TASIK CHINI,

PAHANG DARUL MAKMUR
Amran Ab.Majid, Siti Rahimah Umar, Redzuwan Yahaya, Muhamad Samudi Yasir &
Mohd Suhaimi Othman
Program Sains Nuklear, Fakulti Sains & Teknologi, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor
Darul Ehsan
Email:amran@pkrisc.cc.ukm.my
Abstrak Kajian taburan unsur dan keradioakifan dalam sampel sedimen dari 12 lokasi Tasik Chini telah dilakukan
menggunakan teknik spektrometri gama dan analisis pendarfluor sinar-X (XRF). Sebanyak dua belas unsur surih iaitu As,
Ba, Co, Cr, Cu, Ni, Pb, Rb, Sr, V, Zn, Zr, tiga unsur radionuklid tabii iaitu 40K, 238U, 232Th dan satu unsur radionuklid buatan
iaitu 137Cs telah dapat dikesan dalam sampel sedimen. Keputusan yang diperolehi menunjukkan nilai kepekatan unsur surih
adalah berbeza mengikut lokasi persampelan dan ketiga-tiga radionuklid tabii menunjukkan hampir semua stesen
mempunyai kepekatan melebihi had purata dunia dalam tanah. Radionuklid 137Cs telah ditemui di lima stesen tetapi secara
kualitatif sahaja. Secara umumnya, kepekatan unsur disetiap stesen berkait rapat dengan aktiviti yang terdapat di stesen
masing-masing. Kajian menunjukkan bahawa stesen Laut Jemberau yang terletak berhampiran dengan bekas kawasan
perlombongan dan pelbagai gunatanah menunjukkan lokasi yang mengandungi kepekatan unsur As, Co, Cr, Cu, Pb, V dan
Zn tertinggi berbanding lokasi lain.
Keywords: Sedimen, radionuklid tabii, logam berat.
Pengenalan
Tasik Chini merupakan tasik semulajadi yang kedua terbesar di Malaysia, di mana ia berada
dalam persekitaran asal. Ianya terletak di Mukin Penyur, Pekan, Pahang Darul Makmur pada
kedudukan 3o26’–102o55’. Jaraknya dari Kuantan adalah lebih kurang 100 km. Tasik Chini
merupakan tasik yang berpola dendritik (berbentuk ranting) dengan keluasan 150 hektar atau
12 km2. Sekitaran Tasik Chini ini dikelilingi oleh hutan tropika. Namun kebanyakan kawasan
telah dibalak dan kini ia telah menjadi hutan sekunder dan terdapat juga banyak berlakunya
gunatanah di kawasan sekitar Tasik Chini seperti sebagai pusat pelancongan dan pertanian
komersil. Malahan terdapat juga bekas kawasan perlombongan di sekitar Tasik Chini. Tasik
Chini dijadikan lokasi kajian kerana ia merupakan tasik semulajadi yang menjadi kawasan
penempatan penduduk orang asli di mana kawasan tersebut menjadi sumber bekalan air
[1&2].
Oleh yang demikian, terdapat satu inisiatif untuk mengkaji kesan aktiviti yang berlaku ini
dengan menganalisis unsur dan keradioaktifan di kawasan sekitar Tasik Chini. Ia bertujuan
melihat samada pembangunan yang dijalankan ini boleh mengakibatkan pencemaran di
kawasan sekitar Tasik Chini atau pun tidak. Jadi dengan menganalisis sedimen dapat
memberi gambaran tahap pencemaran air yang boleh mengakibatkan pencemaran logam
berat dan dapat mengkaji perkaitan antara kepekatan unsur dengan lokasi persampelan.
Sedimen merupakan dasar atau tapak bagi sesebuah tasik di mana ia merupakan perhentian
terakhir unsur dan radionuklid serta pemberi “maklumat” terbaik mengenai lokasi kajian.
Dalam kajian ini, sedimen Tasik Chini dari pelbagai lokasi telah dianalisis kandungan unsur
dan keradioaktifannya menggunakan dua teknik analisis iaitu analisis pendarfluor sinar-X
(XRF) dan spektroskopi sinar gama.
Eksperimen
Lokasi Kajian
Sebanyak 15 sampel sedimen telah diambil dari 12 lokasi persampelan di sekitar Tasik Chini
pada bulan Mei 2004 seperti yang ditunjukkan dalam Rajah 1.
1
Pengumpulan Sampel
Sampel sedimen di setiap stesen diambil menggunakan pensampel sediment secara kaut
(Ekman). Alat pensampel sediment diturunkan perlahan-lahan menggunakan tali dari bot
dengan mulut pengautnya terbuka. Apabila sampai didasar tasik, satu alat penghentak
dilurutkan turun melalui tali dan menyebabkan mulut pengaut ini tertutup dan memerangkap
sedimen di dasar.
Sg. Datang
Sg. Chini
4a
Laut Genting Teratai
Laut Tanjung Jerangking 4b
Laut Chenanan
Sg. Gumun
5
Laut Mempiteh 3 Laut Pulau Besar
1a
2 Laut Gumun
6
1b
Laut Kenawar Pulau Besar
Laut Batu Busuk

7 Pulau Balai
8 Laut Labun
10
11
12
Laut Serodong Laut Jemberau

Laut Melai 9b
9a Ke Segamat
Ke Segamat
PETA TASIK CHINI
RAJAH 1 : Peta lokasi persampelan sediment Tasik Chini
Penyediaan dan Rawatan Sampel.
Semua sampel dimasukkan ke dalam dulang dan dibersihkan dari bendasing yang nyata dan
berat basahnya dicatatkan. Sampel sediment kemudian dikeringkan pada suhu 105oC
semalaman dan setelah disejukkan berat keringnya ditentukan. Sampel kemudiaannya
dihancurkan dengan menggunakan mortar dan ditapis menggunakan penapis bersaiz 0.5 mm.
Penyediaan Sampel untuk Teknik Pendarfluor Sinar-X
Semua sampel sedimen yang telah dihancurkan dan ditapis, perlu ditumbuk halus dan
dihomogenkan. Sebanyak 1.00 g sampel sedimen dimasukkan ke dalam radas pembuat pelet
2
dengan ditambahkan asid borik sebanyak 6.00 g sebagai pengikat pelet. Setelah itu, sampel
bersama asid borik yang dicampurkan tadi diberi tekanan sebanyak 5 tons dan 15 tons untuk
mendapatkan palet tertekan yang baik. Pada bahagian belakang palet dilabelkan mengikut
stesen masing-masing. Palet seterusnya dimasukkan ke dalam alat pendarfluor sinar-X iaitu
Spektrometer PW 1480 (Philips) untuk dianalisis kandungan unsurnya. Bahan piawai yang
digunakan adalah basalt.
Penyediaan Sampel untuk Teknik Analisis Keradioaktifan.
Sebanyak 15 sampel yang diketahui berat masing-masing dimasukkan kedalam botol

pembilang kedap udara dan disimpan selama 30 hari bagi mencapai keseimbangan sekular.
Analisis keradioaktifan dalam sampel sedimen ditentukan dengan menggunakan sistem
spektrometri gama yang terdiri daripada pembilang gama HPGe (Tennelec) dan penganalisis
multisalurann berasaskan komputer peribadi untuk analisis keradioaktifan. kandungan
keradioaktifan dalam sampel dikrikan secara kaedah bandingan menggunakan Bahan
Rujukan Piawai (SRM) yang digunakan ialah Soil-375 (IAEA) sebagai piawai. Masa
pembilang sampel dan piawai adalah selama 12 jam.
Hasil Dan Perbincangan
Keputusan Analisis Unsur
Keputusan analisis kandungan unsur dalam sampel sedimen Tasik Chini menggunakan
teknik pendarfluor sinar-X ditunjukkan dalam Jadual 1. Daripada keputusan analisis,
sebanyak 12 unsur telah dikesan dalam sampel sedimen Tasik Chini yang terdiri daripada
As, Ba, Co, Cr, Cu, Ni, Pb, Rb, Sr, V, Zn dan Zr. Keputusan kajian menunjukkan bahawa
setiap unsur mempunyai kepekatan yang berbeza bagi setiap lokasi persampelan. Secara
amnya keputusan mendapati Laut Jemberau mempunyai kepekatan yang paling tinggi bagi
unsur As, Co, Cr, Cu, Pb, V dan Zn dengan kepekatan masing-masing unsure sebesar 50, 46,
113, 68, 156, 156 dan 1679 ppm. Dua bekas lombong besi dan barit serta kedudukanyan yang
berdekatan dengan Laut Gumun dan terdapatnya kawasan pelancongan Tasik Chini,
pembukaan FELDA, penempatan dan yang terbaru ialah Pusat Latihan Khidmat Negara
dipercayai menyumbang terhadap kandungan unsur yang tinggi ini. Hal ini menunjukkan
sememangnya perlombongan dan pembangunan yang terdapat di sekitar lokasi persampelan
mempengaruhi kepekatan unsur dan keputusan ini mempamerkan sedimen boleh bertindak
sebagai sinki kepada pencemaran akuatik sebagaimana yang dilaporkan oleh Mouhi et.al
(2003) [3].
Kajian para penyelidik Kanada di Tasik Killarney, Canada mendapati sampel dari kawasan
perindustrian mempunyai unsur-unsur kadmium, kuprum, plumbum, nikel dan zink manakala
pada kawasan yang masih lagi terpelihara terdapat unsur-unsur ferum, mangan, arsenik dan
kobalt [4]. Kajian lain pula melaporkan arsenik biasanya ditemua dalam sedimen tasik
dengan kepekatan 1 - 15 ppm [5]. Menurut Fytianos dan Laurantou (2004) [6] dalam
kajiannya terhadap sampel sedimen di Tasik Volvi dan Koronia di Greece terdapat tujuh
unsur yang dapat dikesan iaitu kadmium, plumbum, kromium, kuprum, mangan, zink dan
ferum. Sampel ini diambil di dua keadaan cuaca yang berbeza dan kedua-dua tasik ini
merupakan tasik yang masih belum tercemar.
3
Krumgalz dan Fainshtein (1991) [7] pula melaporkan sedimen bagi kawasan Haifa Bay yang
berdekatan dengan kawasan perindustrian dan perbandaran menunjukkan terdapat
pengumpulan merkuri, plumbum, zink, kadmium, ferum dan kuprum. Kuprum, zink,
plumbum dan kadmium juga ditemui dalam sedimen Kenyan Coast. Kajian oleh Mouhi et.al
(2003) [3] juga mendapati kawasan persampelan sediment Makupa yang berdekatan dengan
kawasan perindustrian dan pelupusan sampah tinggi kandungan unsur. Sebagai contoh
mereka mendapati kepekatan masing-masing kupram, zink, plumbum dan kadmium adalah
setinggi 102.0 ± 46.0, 1017.0 ± 840.0, 103.0 ± 35.8, dan 51.0 ± 14.3 manakala kepekatan
unsur kawasan Port Reitz Creek yang kurang tercemar adalah kuprum, 21.6 ± 7.1; zink, 57.1
± 17.9; plumbum, 26.2 ± 11.6 dan kadmium, 1.38 ± 14.3.
Menurut Kamaruzzaman et. al (2004) [8], kepekatan logam berat dalam sedimen dipengaruhi
oleh pelbagai faktor seperti ciri sedimen, jenis, kualiti bahan organik dan saiz partikel
sedimen itu sendiri. Kajian oleh Suhaimi-Othman & Tan (2004) [9], pula saiz butiran
sedimen memainkan peranan penting dalam menentukan keupayaan menjerap logam di mana
pengurangan saiz sedimen didapati meningkatkan keupayaan memerangkap logam.
Analisis keradioaktifan tabii
Selain kandungan unsur, analisis keradioaktifan juga telah ditentukan menggunakan

spektrometri gama seperti yang diberikan dalam Jadual 2. Keputusan analisis menunjukkan
kepekatan radionuklid tabii bagi masing-masing uranium-238, torium-232 dan kalium-40
pada hampir semua stesen mencatatkan bacaan melebihi had purata dunia. Kajian
menunjukkan purata kepekatan uranium-238 adalah 191.0 ± 94.7 Bq/kg (14.5 ± 7.2 ppm)
manakala bagi torium-232 pula ialah 160.3 ± 32.8 Bq/kg (39.5 ± 8.9 ppm) iaitu 5 atau 6 kali
lebih tinggi berbanding kepekatan purata dunia. Walau pun kepekatan Th-232 dan U-238
lebih tinggi, tetapi nisbah Th:U masih lagi 3:1. Ini menunjukkan pertambahan U-238 dan Th-
232 berpunca dari sumber tabii. Bagi kawasan kajian Tasek Chini ini, aktiviti
perlombongan serta jenis batuan yang terdapat disini dijangka banyak mempengaruhi nilai
keradioaktifan tabii yang tinggi ini. Peningkatan kepekatan uranium-238, torium-232 dan
kalium-40 di kerak bumi secara umumnya semakin tinggi apabila kandungan SiO2 dan K2O
dalam kerak bumi semakin tinggi [10,11].
Keputusan kajian ini juga menemukan radionuklid buatan manusia iaitu Cs-137 di
lima stesen secara kualitatif iaitu Laut Gumun (1a,1b), Laut Mempiteh (6), Laut Melai (9b)
dan Laut Labun (11). Radionuklid buatan Cs-137 memang boleh ditemui dalam sampel yang
tidak terganggu kerana ianya berpunca dari radionuklid guguran. Kajian oleh Muhamad
Omar [12] melaporkan kehadiran Cs-137 antara 0.2 – 10.0 Bg/kg bagi beberapa sampel
tanah di Malaysia.
Kesimpulan
Kajian penentuan unsur dalam sampel sedimen Tasik Chini menggunakan teknik pendarfluor
sinar-X telah dapat mengesan 12 unsur. Unsur-unsur tersebut ialah As, Ba, Co, Cr, Cu, Ni,
Pb, Rb, Sr, V, Zn dan Zr. Kesemua unsur ini mempunyai kepekatan yang berbeza di setiap
lokasi kajian iaitu As (11-50 ppm), Ba (553-944 ppm), Co (8-46 ppm), Cr (82-113 ppm), Cu
(11-68 ppm), Ni (67-193 ppm), Pb (37-156 ppm), Rb (105-149 ppm), Sr (18-127 ppm), V
(93-156 ppm), Zn (147-1679 ppm) dan Zr (190-290 ppm). Walaupun persekitaran Tasik
Chini dikatakan masih lagi terpelihara, namun terdapat juga kepekatan unsur yang tinggi
terutamanya di Laut Jemberau. Sebanyak tujuh jenis unsur di laut ini mempunyai kepekatan
4
paling tinggi jika dibandingkan dengan lokasi kajian yang lain. Unsur tersebut ialah As (50
ppm), Co (46 ppm), Cr (113 ppm), Cu (68 ppm), Pb (156 ppm), V (156 ppm) dan Zn (1679
ppm). Kawasan Laut Jemberau ini dikelilingi oleh banyak aktiviti manusia seperti berdekatan
dengan kawasan carigali barit, pembalakan dan pembukaan tanah untuk pembinaan FELDA
dan kawasan rekreasi serta berdekatan dengan kawasan pelancongan Tasik Chini di Laut
Gumun dan lombong besi di bahagian selatan Laut Jemberau.
Kajian juga menemukan kepekatan radionuklid tabii uranium-238, torium-232 dan kalium-40
dalam sediment Tasik Chini melebihi nilai purata kepekatan dunia tetapi bersumberkan
secara tabii. Selain itu radionuklid guguran Cs-137 turut ditemui di lima lokasi kajian secara
kualitatif sahaja.
Rujukan:
1. ERINCO. 1992. Kuala Sungai Chini Gateway Development Plan Vol 1. Petaling Jaya: ERINCO.
2. ERINCO. 1993. Tasik Chini Eco Tourism Development Study Environment Impact Assesment Vol II.
Petaling Jaya: ERINCO.
3. Muohi, A.W., Mavuti, K.M., Omondi, J.G. & Onyari, J.M. 2003. Heavy metals in sediments from
Makupa and Port-Reitz Creek systems: Kenyan Coast. Environment International. 28: 639-647.
4. Nelson , B., Chen, Y. W., Gunn, J. M. & Dixit, S. S. 2004. Sediment Trace Metal Profiles in Lakes of
Killarney Park, Canada from Regional to Continental Influence. (atas talian)
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi (14 September 2004)
5. Lollar, B. S. 2004. Treatise on Geochemistry Environmental Geochemistry. Ed. ke-9.Oxford: Elsevier
Pergamon.
6. Fytianos, K. & Lourantou, A. 2004. Speciation of elements in sediment samples collected at lakes
Volvi and Koronia, N.Greece. Environment International. 30(1):11-17.
7. Krumgalz, B. S. & Fainshtein, G. 1991. Trace Metal and Organic Matter in Nearshore Sediment Cores
from the Eastern Mediterranean (Haifa Bay of Israel). Marine Environmental Research. 31: 1-15.
8. Kamaruzzaman, B. Y., Ong, M. C. & Willison, K. Y. S. 2004. Taburan Kepekatan Elemen-elemen Kimia
di dalam Teras Sedimen di Hutan Paya Bakau Paka, Terengganu. Prosiding Simposium Kimia Analisis
Malaysia Ke-17.
9. Suhaimi-Othman, M. & Tan, B. F. 2004. Kajian Kandungan Logam Berat (Cu, Cd, Zn dan Pb) di
dalam Air, Sedimen dan Udang Air Tawar Macrobrachium Ianchesteri di Sungai Langat. Prosiding
Simposium Kimia Analisis Malaysia Ke-17.
10. International Atomic Energy Agency (IAEA). 1990. The Use of Gamma Ray Data To Define The Natural
Radiation In Environment. IAEA-TECDOC_566, Vienna: IAEA.
11. Khursyid Alam Butt, Amanat Ali & Aziz Ahmad Qureshi. 1998. Estimation of Environmental Gama
Background Radiation Levels in Pakistan. Health Physics. 75(1): 63-66.
12. Muhamat Omar (1991), Environmental Radiation and Its Relation with Man. Nuclear Buletin of
Malaysia. 1(2). 16-18.
5
Jadual 1 : Keputusan analisis unsur bagi setiap stesen persampelan
Stesen Kepekatan (ppm)
As Ba Co Cr Cu Ni Pb Rb Sr V Zn Zr
1a: Laut Gumun 21 571 20 92 43 125 67 119 75 134 326 226
1b : Laut Gumun 14 888 26 87 57 128 50 136 18 134 1145 290
2: Laut P. Balai 15 557 Lod 90 60 157 76 132 101 130 147 208
3: Laut Chenanan 13 553 Lod 99 66 78 65 105 59 142 260 190
4a: L. Tanjung 12 714 22 95 11 143 48 128 99 139 693 225
Jerangking
4b: L. Tanjung 17 603 26 87 44 149 39 133 100 130 329 235
Jerangkin
5: Laut Genting 15 630 Lod 96 60 101 38 119 78 127 321 201
Teratai
6: Laut Mempiteh 13 714 15 96 60 141 44 131 93 147 645 227
7: Laut Senawar 11 824 26 96 20 189 37 138 113 126 655 249
8: Laut Serodong 13 944 24 98 49 145 46 129 98 148 948 274
9a: Laut Melai 17 582 Lod 85 68 186 56 149 127 117 488 230
9b: Laut Melai 15 831 35 82 Lod 151 61 120 88 117 1130 284
10: Laut Batu 13 651 25 83 55 146 83 131 92 105 1097 211
Busuk
11: Laut Labun 18 555 8 87 54 193 59 146 104 93 1032 209
12: Laut Jemberau 50 782 46 113 68 67 156 109 23 156 1679 221
Min 17.1 693.3 18.2 92.4 47.7 140.0 61.7 128.3 84.5 129.7 726.3 232.0
± ± ± ± ± ± ± ± ± ± ± ±
9.5 132.1 14.1 8.0 21.1 36.8 29.5 12.3 30.5 16.7 433.5 30.0
Had pengesan 0.5 2 1 2 1 1 1 0.5 0.5 1 1 0.5
(Lod)
Lod – di bawah had pengesanan
Jadual 2: Kepekatan radionuklid tabii sampelsediemn Tasik Chini

Stesen Kepekatan radionuklid Nisbah
K-40 Th-232 Th-232 U-238 U-238 Th:U
(Bq/kg) (Bq/kg) (ppm) (Bq/kg) (ppm)
Laut Gumun: 1a 599 126.8 32.0 215.6 16.0 2:1
Laut Gumun: 1b 757 109.8 28.0 130.7 10.0 3:1
L. Pulai Balai: 2 549 189.7 48.0 225.8 17.0 3:1
Laut Chenanam: 3 674 206.7 52.0 268.4 20.0 3:1
L. Tanj. Jerangking: 4a 713 139.3 35.0 142.1 11.0 3:1
L. Tanj. Jerangking:4b 594 143.7 36.0 114.0 9.0 4:1
L. Genting Teratai: 5 690 178.8 45.0 170.2 13.0 4:1
Laut Mempiteh: 6 656 152.2 39.0 135.9 10.0 4:1
LautKenawar: 7 760 152.1 39.0 90.5 7.0 6:1
Laut Serodong: 8 818 105.9 27.0 151.3 12.0 2:1
Laut Melai: 9a 537 160.3 41.0 54.5 4.0 10 : 1
Laut Melai: 9b 741 170.5 27.0 153.5 12.0 2:1
L. Batu Busuk: 10 435 168.3 43.0 400.5 31.0 1:1
Laut Labun: 11 475 221.4 56.0 292.8 22.0 3:1
Laut Jemberau: 12 277 178.8 45.0 319.3 24.0 2:1
Min 618.3 160.3 39.5 191.0 14.5 3:1
± 1454 ± 32.8 ± 8.9 ± 94.7 ± 7.2
UNSCEAR (tanah) [14]. 400 - 6.3 - 2.0 3:1
6
PYROLYSIS AND LIQUEFACTION OF ACETONE AND MIXED

ACETONE/TETRALIN SWELLED MUKAH BALINGIAN MALAYSIAN SUB-
BITUMINOUS COAL – THE EFFECT ON COAL CONVERSION AND OIL YIELD
Mohd Fauzi Abdullah*, Mohd Azlan Mohd Ishak and Khudzir Ismail
Fuel Combustion Research Laboratory

Faculty of Applied Sciences
University Technology MARA
Arau Campus, 02600, Arau, Perlis
*e-mail: mohdfauz_678@yahoo.com
Abstract. The effect of swelling on Mukah Balingian (MB) Malaysian sub-bituminous coal
macrostructure was observed by pyrolysing the swelled coal via thermogravimetry under
nitrogen at ambient pressure. The DTG curves of the pyrolysed swelled coal samples show
the presence of evolution peaks at temperature ranging from 235 – 295 °C that are due to
releasing of light molecular weight hydrocarbons. These peaks, however, were not presence
in the untreated coal, indicating some changes in the coal macrostructure has occurred in the
swelled coal samples. The global pyrolysis kinetics for coal that follows the first-order
decomposition reaction was used to evaluate the activation energy of the pyrolysed untreated
and swelled coal samples. The results thus far have shown that the activation energy for the
acetone and mixed acetone/tetralin-swelled coal samples exhibit lower values than untreated
coal, indicating less energy is required during the pyrolysis process due to the weakening of
the coal-coal macromolecular interaction network Moreover, liquefaction on the these
swelled coal samples that was carried out at temperatures ranging from 360 to 450 °C at 4
MPa of nitrogen pressure showed the enhancement of the coal conversion and oil yield at
temperature of 420 °C, with retrogressive reaction started to dominate at higher temperature
as indicated by decreased and increased in oil yield and high molecular weight pre-
asphaltene, respectively. These observations suggest that the solvent swelling pre-treatment
using acetone and mixed acetone/tetralin can improve the coal conversion and oil yields at
less severe liquefaction condition.
Keywords: Swelling, liquefaction, pyrolysis, coal conversion, oil yield.
Introduction
Coal liquefaction is usually carried out at temperatures higher than 400 °C and at relatively
high pressure. Many attempts have been made to develop methods of dissolving coal at less
severe conditions in order to increase coal liquefaction efficiency [1-4] and to reduce capital
and operation costs [5]. It is known that, by lowering reaction severity, coal conversion
reaction rates and liquid product yields will also be reduced, unless the intrinsic coal
reactivity can be sufficiently enhanced [4]. Joseph [1] reported that pre-swelled US
bituminous and lower rank coals (i.e. sub-bituminous and lignite) with tetrabutylammonium
hydroxide, tetrahydrofuran and methanol prior to liquefaction at 400 °C and at 7.6 MPa
enhanced coal conversion and product quality. He found that the enhancement depends on the
coal rank and the type of swelling agent being used and suggested that the beneficial effect of
swelling might be due to the expansion of the coal macromolecular structure, making it more
accessible to the hydrogen donor solvent during liquefaction. In another work, Simsek et al.
7
[2] studied the effect of pre-swelling with various organic solvents and acid-demineralised
(pre-treatment) Turkish coals on the supercritical toluene extract yield at 350 °C and at 7
MPa. They found that the highest improved liquid yields for each coal was obtained by the
combined effects of pre-treatment and pre-swelling of coals. Although the combination
effects proved to increase liquid yields, no simple trend between reactivity improvements
with properties of the coals was observed.
In our recent study [6], we carried out the solvent swelling pre-treatment using
hydrogen and non-hydrogen-bonding solvents on low-rank Malaysian coal. The pyrolysis of
the untreated and swelled coal using thermogravimetric analyzer (TGA) under nitrogen
atmosphere thus far has shown that the activation energy barrier in the swelled coals was
lower with an increase in the volatile yield in comparison to the untreated sample. Moreover,
the results of the treated samples also show the increase of coal reactivity as revealed by DTG
results. Thus, these preliminary results might provide useful information for coal extraction
and liquefaction processes. Hence, this study investigates the effect of acetone and mixed
acetone/tetralin-swelled, respectively on the pyrolysis behaviours using TGA and the
conversion and product yields from the liquefaction using 1-liter high-pressure high-
temperature batch-wise reactor system with tetralin as the hydrogen-donor solvent.
Materials And Methods
Coal Preparation
The sample used in this study is Mukah Balingian (MB) originating from Sarawak, Malaysia.
The procedure for coal preparation has been reported earlier [7]. Table 1 represents the
ultimate, proximate and petrography analyses of the untreated MB coal sample.
Solvent Swelling Procedure
The volumetric method described by Onal et al. [8] was adopted in the measurement of
swelling ratio without density correction. Acetone (analytical grade solvent from Fisher
Chemicals) is used as swelling solvent. For the mixed solvent, which consists of acetone and
tetralin, the volume mixed ratio was 80:20. The swelling ratio of the coal samples was
measured at room temperature (30 °C) and at ambient pressure. After the swelling procedure,
the swelled coal was filtered, dried in vacuum-oven at 80 °C for overnight and finally stored
in bottle prior to liquefaction experiments. The determination of activation energy and
volatile yield of the coal samples have been reported earlier [6]. The results of swelling ratio,
activation energy and pyrolysis volatile yields of the coal samples are given in Table 2.
Coal Liquefaction Experiments
All liquefaction experiments were carried out in a 1-liter magnetically stirred (at 500 rpm)
high-pressure high-temperature batch-wise reactor (Parr, 4571 model) fitted with stainless
steel tubing condenser cooled with ice to ensure maximum capture of the volatile materials.
The procedure of coal liquefaction has been reported earlier [7]. Briefly, 20 g of untreated or
acetone-swelled coal and 200 g of tetralin were mixed in the reactor and the liquefaction was
carried out at temperature ranging from 360 – 450 °C and at 4 MPa nitrogen pressure.
However, for mixed acetone/tetralin coal sample, the reaction temperature used was at 420
°C. After reaching at the particular temperature, the reaction was held for 30 minutes before
8
the system was cooled to ambient temperature. Coal conversion and the product yields of all
the samples were finally evaluated [7] and are shown in Table 3.
Sample Analyses
The procedure of ultimate and proximate analyses has been reported earlier [7]. The ultimate
analyses were carried out using Elemental Analyser Leco 932 model. The proximate analyses
were done using Thermogravimetric analyzer DTA/DSC TA model SDTQ600 under nitrogen
gas atmosphere with heating condition follows the ASTM D2974 [9].
Table 1: Characteristics of MB coal sample. Table 2: Results of swelling ratio (Q),
activation energy (Ei, kJ/mol) and

Ultimate Proximate pyrolysis volatile yield (VY) of MB
coal.
analysis (wt% daf) analysis (wt% db)
Sample Q Ei VY
(%)
Carbon 63.9 Volatile matter 44.7
Hydrogen 5.1 Fixed carbon 51.1 Untreated 1.0 312.2 37.6
Nitrogen 1.9 Ash content 4.2 Tet-sw 1.0 246.3 41.4
Sulphur 0.5 Ace-sw 1.3 230.5 40.4

Oxygen* 28.6 Ace+Tet-sw 1.4 131.6 49.8
Calorific value = 24.60 MJ/kg Tet-sw = Tetralin-swelled;

Ace-sw = Acetone-swelled;
*Calculated by difference. Ace+Tet-sw = Acetone/Tetralin-
swelled.
Results And Discussion
Effect of Swelling On Coal by Pure and Mixed Solvents During Pyrolysis
Table 2 shows the results of swelling ratio, activation energy and volatile yield of untreated,
acetone-swelled and mixed acetone/tetralin-swelled of MB coal samples. From the table, it
can be seen that the swelling ratios of the coal in acetone and mixed acetone/tetralin are
higher than that of tetralin. This observation indicates the interaction of nucleophile in the H-
bonding solvent (i.e. acetone) with the reactive sites such as hydroxyl, carboxyl and carbonyl
that are presence in the coal forming hydrogen bondings. The increase in solvent-coal
interactions enhanced the bonds strength, thus weakened the coal-coal macromolecular
interactions, and promotes the swelling process of the coal. Tetralin, which is a non-H-
bonding solvent, however, did not exhibit any swelling activity with the swelling ratio value
closed to that of untreated coal [6]. Interestingly, when tetralin was mixed with acetone, the
swelling ratio of the coal increased to a value of 1.4, which is slightly higher than the pure
acetone. This minor increment indicates an additional interaction occurred between the coal
macromolecular network and the solvents mixture. The effect of swelling by the mixed
solvents seem to weakens the coal macromolecular network rigidity and promotes the
9
diffusion of the solvent mixtures, though the exact mechanism is still not well understood.
Further, in order to prove this effect, the solvent pre-swelled and untreated coal samples were
subjected to pyrolysis via thermogravimetric analysis and the activation energy of the process
was evaluated. The global pyrolysis kinetics for coal that follows the first-order
decomposition reaction was used to determine the activation energy of the process [10].
Indeed, the mixed solvent swelled coal exhibits the lowest activation energy with an
increased in volatile yield, indicating less energy was required during the pyrolysis process
due to the weakening of the coal-coal macromolecular interaction network that occurred
during swelling process. Thus, this observation indicates the presence of synergistic effect of
the mixed solvent towards swelling activity of the coal that enhanced the lowering of the
activation energy with an increased in volatile yield during pyrolysis.
Figure 1 shows the DTG curves of untreated, acetone-swelled and mixed
acetone/tetralin-swelled coal during pyrolysis. Apparently, both the pyrolysed acetone and
mixed acetone/tetralin-swelled coal samples show the presence of evolution peaks at
temperatures ranging from 175 to 395 °C that are due to the releasing of the light molecular
weight hydrocarbons [10]. These peaks however, were absence in the untreated coal.
Another point of interest is that for the mixed acetone/tetralin-swelled coal sample there
appears to be another evolution peaks at high temperature ranging from 500 to 650 °C that
are due to the released of heavy hydrocarbons [10], and were not presence apparently in both
untreated and acetone pre-swelled coal samples. Hence, these observations further proves
that some changes in the coal macrostructure have occurred especially in the mixed solvents
swelled coal sample that promotes the released of light and heavy molecular weight
hydrocarbons during pyrolysis. Hence, this shows that the swelling pre-treatment on coal
would possibly increase the liquefaction performance by increasing coal conversion and
product yields (especially percent oil) at less severe conditions.
Figure 1: The DTG curves of untreated, acetone-swelled and mixed

acetone/tetralin-swelled of MB coal samples.
10
Effect of Swelling on Coal Conversion and Oil Yield
Table 3 shows the percent of coal conversion and product yields of liquefaction on the
untreated, acetone-swelled and mixed acetone/tetralin-swelled coal samples.
Table 3: Liquefaction results of untreated, acetone- and mixed acetone/tetralin-swelled coal.
Product yields
Temp. (°C) % Coal conversion % Oil+gas % Asphaltene %

Preasphaltene
Unt. Ace. Mix. Unt. Ace. Mix. Unt. Ace. Mix. Unt. Ace
Mix
360 42 35 - 28 24 - 8 5 - 6 6
-
380 57 43 - 44 29 - 8 3 - 5 11
-
400 66 62 - 50 46 - 10 6 - 6 10
-
420 84 86 86 67 70 78 10 6 1 7 10
7
450 90 83 - 80 75 - 8 4 - 2 4
-
Unt. = Untreated; Ace. = Acetone-swelled; Mix. = Mixed acetone/tetralin-swelled.

Liquefaction conditions: 4 MPa, 1:10 coal:solvent, 30 min reaction time and stirred at 500rpm.
Apparently, the results thus far showed that the percent of coal conversion and oil
yield obtained during liquefaction of acetone-swelled coal was lower than the untreated coal
with preasphaltene predominantly increased at temperature below 400 °C. The fact that as the
liquefaction temperature increased at above 350 °C, the coal starts to soften and eventually
undergo fragmentation forming preasphaltene and asphaltene [7]. However, with the swelled
coal sample, much smaller molecules are being formed due to the weakening of the coal-coal
macromolecular interaction as revealed by the DTG curves. The hydrogen donor solvent
should cap these smaller radical molecules rapidly in order to prevent repolymerisation
reaction. The fact that high formation of preasphaltene suggests that repolymerisation
reaction starts to dominate due to the inability of the donor solvent to cap these smaller
molecules at temperature below 400 °C. However, as the liquefaction temperature increased
beyond 400 °C, a high amount of coal conversion and oil yield being obtained with decreased
amount of preasphaltene formation.
Obviously, the effect of mixed solvent swelled coal during liquefaction can be seen
by the high percent of oil yield of 78 % at 420 °C, that is higher than that achieved in the
acetone- swelled and untreated coal samples. The increase in oil yield corresponds to the
decrease in asphaltene and preasphaltene. These observations seem to agree with the
pyrolysis which resulting in the lowest activation energy with the greater increased of volatile
yield that indicate the high reactivity of the mixed acetone/tetralin-swelled coal. Thus, these
phenomenons proved the beneficial effect of mixed solvent-swelled coal in enhancing coal
conversion and oil yield prior to liquefaction.
11
Conclusion
The effect of acetone and mixed acetone/tetralin-swelled on the coal pyrolysis using TGA
and the liquefaction products using 1-liter high-pressure high-temperature batch-wise reactor
system with tetralin as the hydrogen-donor solvent were studied. The results thus far have
shown that the activation energy for the acetone and mixed acetone/tetralin-swelled coal
samples exhibit lower values than the untreated coal, indicating less energy is required during
the pyrolysis process due to the weakening of the coal-coal macromolecular interaction
network. Moreover, liquefaction on the swelled coal samples showed the enhancement of the
coal conversion and oil yield at temperature of 420 °C, where the oil yield was greatly
increased for mixed acetone/tetralin-swelled with further decreased in asphaltene and
preasphaltene, respectively. Thus, these observations proved the beneficial effect of swelling
pre-treatment on liquefaction at less severe condition.
Acknowledgements
The authors would like to thank the Ministry of Science Technology & Innovation, Malaysia
for funding the research (grant no.: 02-02-01-0017-EA0017) and University Technology
MARA for supporting the research work.
References
1. Joseph, J.T., “Liquefaction behaviour of solvent-swollen coals”, Fuel, vol. 70, pp. 139-
144, 1991.
2. Simsek, E.H., “The effect of pre-swelling and/or pre-treatment of some Turkish coals
on the supercritical fluid extract yield”, Fuel, vol. 81, pp. 503-506, 2002.
3. Hu, H., Sha, G. and Chen, G., “Effect of solvent swelling on liquefaction of Xinglong
coal at less severe conditions”, Fuel, vol. 68, pp. 33-43, 2000.
4. Artok, L., Davis, A., Mitchell, G.D. and Schobert, H.H., “Swelling pre-treatment of
coals for improved catalytic liquefaction”, Fuel, vol. 71, pp. 981-991, 1992.
5. Shams, K.G., Miller, R.L. and Baldwin, R.M., “Enhancing low severity coal
liquefaction reactivity using mild chemical pre-treatment”, Fuel, vol. 71, pp. 1015-
1023, 1992.
6. Abdullah, M.F., Ismail, K. and Ishak, M.A.M., “Investigation of pyrolysis behaviours
on solvent swelled Mukah Balingian coal using TGA analysis”, ACGC Chemical
Research Commun., in print.
7. Ishak, M.A.M., Ismail, K., Abdullah, M.F., Kadir, M.O.A., Mohamed, A.R. and
Abdullah, W.H., “Liquefaction studies of low-rank Malaysian coal using high-pressure
high-temperature batch-wise reactor system”, Coal Preparation Jour., in print.
8. Onal, Y. and Akol, S., “Influence of pre-treatment on solvent-swelling and extraction of
some Turkish lignites”, Fuel, vol. 82, pp. 1297-1304, 2003.
9. ASTM D2974, Annual Book of ASTM Standards, vol. 5.05, The American Society for
Testing and Materials (ASTM), Philadelphia, PA.
10. Probstein, R.F. and Hicks, R.E., ‘Synthetic fuels’, McGraw-Hill, Inc., 1982.
12
KANDUNGAN LOGAM BERAT DAN RADIONUKLID TABII DALAM IKAN, AIR,

TUMBUHAN DAN SEDIMEN DI BEKAS TASIK LOMBONG
Muhamad Samudi Yasir, Norlaili bt Ahmad Kabir, Redzuwan Yahaya & Amran Ab Majid
Pusat Pengajian Fizik Gunaan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor.
msyasir@pkrisc.cc.ukm.my
Abstrak: Tasik bekas lombong bijih timah di Malaysia kini giat ditebusguna untuk kegiatan pertanian, akuakultur, kawasan
rekreasi atau dijadikan sebagai kawasan perumahan dan perindustrian. Kesan aktiviti perlombongan boleh menyebabkan
peningkatan kepekatan atau pengkayaan radionuklid tabii dan logam berat terhadap ekosistem. Maka, kajian telah dilakukan
untuk menentukan kandungan radionuklid tabii dan 12 unsur logam berat (Hf, Zr, Mn, Cu, Zn, As, Cd, Sn, Sb, Ba, Hg dan
Pb) di dalam ikan, air, tumbuhan dan sedimen di tiga tasik bekas lombong sekitar Puchong dan Sepang, Selangor Darul
Ehsan. Analisis dilakukan menggunakan ICP-MS. Secara keseluruhannya kepekatan logam berat adalah tinggi dalam
sedimen serta tumbuhan berbanding dengan dalam ikan dan air. Barium mencatatkan kepekatan yang tinggi dikesan di
dalam sedimen dan air, manakala zink dan mangan paling tinggi dikesan masing-masing dalam ikan dan tumbuhan.
Kesemua unsur logam berat dalam ikan berada di bawah paras maksimum cemaran logam yang dibenarkan oleh Akta
Makanan (Akta 281) dan Peraturan-Peraturan 1983 kecuali bagi kepekatan merkuri bagi ikan di lombong kedua iaitu 0.53 ±
0.20 mg/kg) yang melebihi had yang dibenarkan iaitu 0.5 mg/kg. Aktiviti radionuklid thorium (Th-232) dan uranium (U-
238) dalam sedimen adalah tinggi berbanding dalam ikan, air dan tumbuhan dengan masing-masing berada dalam julat
(30.76 ± 2.71 – 35.34 ± 0.27 Bq/kg) dan (9.37 ± 2.30 - 26.32 ± 3.01 Bq/kg). Manakala aktiviti kalium (K-40) dalam
tumbuhan dan ikan didapati lebih tinggi berbanding di dalam air dan sedimen.
Katakunci: Logam berat, radionuklid tabii, tasik bekas lombong, ikan, sedimen
Abstract: Malaysia aggressively reclaimed most of their disused tin-mining pool especially for agricultural activities,
freshwater fish farming area, recreational area, houses area and even as an industrial area. Past mining activities might
induced the concentration of naturally occurring radionuclide (NORM) and heavy metal at the disused tin-mining pool
ecosystem. A study has been conducted on the status of heavy metal (Hf, Zr, Mn, Cu, Zn, As, Cd, Sn, Sb, Ba, Hg and Pb)
concentration and naturally occurring radionuclide activity in fish, water, plants and sediments at three different disused tin-
mining pool near by Sepang and Puchong, Selangor Darul Ehsan. Sample of fish, water, plant and sediment being analyze
using ICP-MS. The concentrations of heavy metal in sediment and plant are higher than its concentrations in fish and
followed by water. The highest concentration of heavy metal in sediment and water is barium, whereas the highest
concentration of heavy metal in fish and plant is zinc and manganese. The result also showed that only mercury level in fish
collected in second disused tin-mining pool (0.53 ± 0.20 mg/kg) is exceed the maximum limit (0.5 mg/kg) prescribe by the
Malaysian Food Act (Act 281). The activity of U-238 and Th-232 in sediment was found to be relatively higher than its
activity in fish, plant or water (30.76 ± 2.71 – 35.34 ± 0.27 Bq/kg) and (9.37 ± 2.30 -18.86 ± 2.60 Bq/kg). The determination
of K-40 activity showed that it is highly contained in plant and fish than in sediment or water.
Keywords: Heavy metal, natural radiobuclides, ex-mining lake, fish, sedimen
Pendahuluan.
Malaysia satu ketika dahulu merupakan salah sebuah negara pengeluar utama timah dunia. Malah
pembangunan Lembah Kelang dan Lembah Kinta mempunyai kaitan yang rapat dengan penemuan
timah. Walaupun kini tidak banyak lagi lombong timah yang masih aktif beroperasi, ujud pula
industri sampingan seperti pengekstrakan pelbagai mineral berharga dari amang (zirkon, ilmenit dan
struverit). Salah satu peninggalan industri perlombongan timah ialah bekas tasik lombong yang
dikategorikan sebagai tasik buatan. Kini tasik dan kawasan bekas lombong ini telah banyak ditebus
guna untuk dimajukan sebagai kawasan perumahan dan perindustrian, ditanami rumput untuk tujuan
penternakan, akuakultur atau pun dijadikan kawasan rekreasi (Amran Ab Majid et al. [1]). Selain
bijih timah, perlombongan turut sama mengeluarkan pelbagai jenis mineral lain dari perut bumi.
Sekiranya mineral sampingan ini terdiri daripada logam berat, dikhuatiri sisanya masih tertinggal di
kawasan bekas lombong yang kini telah dijadikan kawasan pertanian, perternakan ataupun akuakultur.
Bagi tujuan akuakultur, proses bioakumulasi logam berat boleh terjadi yang akhirnya
berkemungkinkan memasuki rantai makanan ekosistem bekas lombong tersebut. Oleh itu, ikan yang
merupakan sebahagian daripada komponen rantai makanan dalam ekosistem tasik ini mungkin akan
turut tercemar dengan sisa logam berat tersebut. Keadaan yang sama mungkin juga berlaku kepada
radionuklid tabii seperti U-238, Th-232 dan K-40. Kemasukan logam berat dan radionuklid tabii
13
dalam rantai makanan ini akan akhirnya sampai kepada manusia yang memungkinkan pemakannya
menghadapi resiko kesihatan.
Oleh yang demikian objektif utama kajian ini adalah untuk menentukan aras kandungan logam berat
dan radionuklid tabii dalam ikan di tasik bekas lombong serta hubungkaitnya dengan kandungan pada
sediment, air dan tumbuhan.
Bahan dan kaedah.
a. Kawasan kajian
Tiga buah tasik bekas lombong (L1, L2 dan L3) yang dijadikan kawasan kajian terletak di kawasan
Puchong (Rajah 1). Ketiga-tiga tasik ini telah ditebusguna dan dijadikan kawasan ternakan
akuakultur.
b. Sampel kajian.
i. Ikan
Sampel ikan yang diguna adalah Ikan Keli Kayu (Clarias batrachus), Ikan Patin (Pangasius sutchi),
Ikan Tilapia (Oreochromiis mussambics), ikan dari famili Lidae seperti Ikan Toman (Ophiocehalus
micropeltes) dan Ikan Haruan (Ophiocehalus striatus), Ikan Tilapia (Oreochromiis Mussambics) atau
Ikan Belida (Notopterus notopterus)). Ikan ini ditangkap menggunakan jarring yang dipasang
semalaman.
ii. Air
Sama sebagaimana sampel ikan, sampel air (4L) juga diambil daripada tiga lokasi pada ketiga-tiga
tasik. Asid nitrik dititiskan kepada setiap sampel air yang diambil mengelak pertumbuhan mikrob dan
menghalang unsur daripada sampel air melekat pada dinding botol plastik yang diguna. Parameter air
turut diambil yang meliputi parameter fizikal (suhu, pH, konduktiviti, saliniti dan oksigen terlarut).
iii. Tumbuhan
Tumbuhan yang diambil bergantung kepada kehadiran pada setiap lokasi. Ianya terdiri dari
Eichhornia crassipes, Eragrostis atrovirens, Pennisetum puerperiu, Panicum repen, Pennisetum
polystachion, Melampodium divaricatu dan Brachiaria mufica.
iv. Sedimen
Sedimen diambil menggunakan penceduk plastik dan dimasukkan ke dalam beg plastik yang telah
dilabel.
c. Perawatan dan analisis sampel
i. Ikan.
Sampel ikan yang diambil dibasuh menggunakan air suling dan disiang untuk mengasingkan kepala,
isi perut, insang dan tisu daging ikan. Setelah dicuci menggunakan air suling, tisu daging ikan yang
diketahui beratnya dikeringkan menggunakan ketuhar pada suhu 80°C sehingga beratnya menjadi
malar. Sampel kering ini kemudiannya dikisar sekali lagi untuk mendapatkannya butiran halus sekitar
500 µm. Sebanyak 400 mg sampel kemudiannya dicampurkan dengan 3.0 ml asid nitrik, 3.0 ml asid
hidroklorik dan 0.5 ml asid peroksida. Campuran dihazam menggunakan kemudahan multigelombang
selama dua jam sehingga membentuk larutan jernih. Larutan sampel yang terhasil kemudiannya
dituras dan ditambah air suling sehingga isipadu larutan sampel mencecah 100 ml sebelum ditentukan
kandungan logamnya berat menggunakan peralatan ICP-MS.
14
Taman Putra
L1 Prima L2
L3
L1, L2 dan L3 Tasik bekas lombong.
RAJAH 3.1 Lokasi tasik kajian
ii. Air.
Sampel air yang telah diambil itu dibersihkan daripada kotoran dengan cara menapisnya
menggunakan penapis membran plasma. Sampel air yang digunakan untuk penentuan keradioaktifan
tabii dan logam berat di dalam kajian ini disediakan di dalam dua bentuk iaitu sampel air yang
dipekatkan dan yang tidak dipekatkan.
Penyediaan sampel air yang tidak melalui proses pemekatan ialah dengan cara menuras sampel air
yang telah dibersihkan menggunakan membran plasma menggunakan kertas turas sehingga
isipadunya mencapai 100 ml.
iii. Tumbuhan
Sampel tumbuhan itu dibasuh menggunakan air suling untuk menghilangkan kotoran yang melekat.
Tumbuhan kemudiannya dibilas menggunakan air suling sekali lagi. Berat basah tumbuhan yang
telah bersihkan dicatatkan. Keseluruhan tumbuhan (akar, daun dan batang) kemudiannya dikeringkan
menggunakan ketuhar pada suhu 105°C sehingga beratnya menjadi malar. Sampel kering ini
kemudiannya dikisar menggunakan pengisar untuk mendapatkan butiran halus sekitar 500 µm.
Sebanyak 400 mg butiran halus seterusnya dicampur dengan asid nitrik (3.0 ml), asid hidroklorik (0.5
ml) dan asid peroksida (9.0 ml). Campuran dihazamkan menggunakan kemudahan multigelombang
15
selama dua jam sehingga membentuk larutan jernih. Larutan sampel yang terhasil dari proses
penghadaman itu kemudiannya dituras dan air suling ditambah sehingga isipadu larutan sampel
mencecah 250 ml.
iv. Sedimen
Sampel sedimen itu dibersihkan daripada kotoran dengan membuang semua akar tumbuhan dan
bendasing lain. Berat basah sedimen yang telah dibersihkan dicatatkan. Sampel sedimen kemudiannya
dikeringkan menggunakan ketuhar pada suhu 105ºC sehingga berat menjadi malar. Sampel kering ini
seterusnya dikisar untuk mendapatkannya butiran halus sekitar 500 µm.
Sebanyak 300 mg butiran halus sedimen dicampurkan dengan asid nitrik (5.0 ml), asid hidroklorik
(2.0 ml) dan asid peroksida (0.5 ml) dan asid fluorida (3.0 ml). Campuran seterusnya dihazamkan
menggunakan kemudahan multigelombang selama dua jam sehingga membventuk larutan jernih.
Larutan sampel yang terhasil dari proses penghadaman itu kemudiannya dituras dan air suling
ditambah sehingga isipadu larutan sampel mencecah 100 ml.
d. Analisis sampel
Sampel yang telah disediakan dalam bentuk larutan dianalisis menggunakan Spektrometer Jisim
Gandingan Plasma Teraruh (ICP-MS) yang telah dikalibrasi menggunakan piawai (SRM MA-A-2
(TM) Fish Flesh Homogenate), piawai tumbuhan (SRM 1572 Citrus Leave) dan larutan Perkin Elmer
Pure Atomic Spectroscopy Standard.
e. Analisis data
Bagi penentuan kepekatan logam berat, setiap data yang diperolehi dikira semula kepada berat asal
sampel dan dinyatakan dalam unit mg/kg atau mg/L.
Manakala aktiviti radionuklid tabii (Bq/kg) pula boleh ditentukan menggunakan rumus:
dN = λӨw x NA
dt WM
Di mana;
Ө = Kelimpahan tabii radionuklid tabii di dalam semesta
w = Berat radionuklid tabii di dalam sampel
NA = Nombor Avogadro
M = Berat molekul radionuklid tabii
W = Berat sampel
Hasil dan perbincangan
a. Parameter fizikal
Jadual 1 menunjukkan bacaan pelbagai parameter fizikal lombong kajian. Oksigen terlarut yang
diukur in situ menunjukkan nilainya berada di antara 7.0 -7.6 mg/l. Kandungan oksigen ini
dipengaruhi oleh pelbagai faktor seperti proses pernafasan haiwan akuatik, forosintesis tumbuhan
akuatik serta pengoksidaan yang berlaku di bawah permukaan air (Elbering et al. [2]). Air tasik bekas
lombong biasanya bersifat asid (Muhamad Samudi Yasir et al. [3]). Sungguhpun begitu hasil kajian
ini menunjukkan sebaliknya, iaitu sedikit beralkali. Ini mungkin disebabkan oleh faktor masa, iaitu
telah melebihi 15 tahun berhenti beroperasi. Kajian yang dilakukan di Sepanyol juga menunjukkan
sifat kealkalian pada air tasik bekas lombong (Riba et al. [4]).
16
Jadual 1. Pelbagai parameter air dan sedimen tasik
parameter Tasik 1 (L1) Tasik 2 (L2) Tasik 3 (L3)

Oksigen terlarut (mg/L) 7.0 7.4 7.6
Nilai pH
a.air 7.21 7.47 7.92
b.sedimen 7.34 6.80 6.97
Suhu (oC) 30.6 30.0 32,3
Konduktiviti (µS/cm) 386 385 278
Nilai konduktiviti menggambarkan kandungan kation dan anion yang terlarut dalam air tasik. Hasil
kajian mendapati nilainya berada pada julat 278 – 386 µS/cm bagi ketiga-tiga tasik.
b. Logam berat
Logam berat yang ditentukan dalam setiap sampel ialah mangan, kuprum, zink, arsenik, kadmium,
stanum, antimoni, barium, merkuri, plumbum, zirkonium dan hafnium. Hasil kajian mendapati Zn
dan Cu merupakan logam berat yang paling banyak terkandung dalam tisu daging ikan dengan
masing-masing dalam julat (0.50 - 3.81 mg/kg) dan (0.50 - 2.27 mg/kg). Kepekatan tinggi kedua-dua
logam ini berbanding logam lain mungkin disebabkan kedua-duanya merupakan sebahagian daripada
nutrien utama yang diambil oleh ikan untuk hidup. Selain daripada itu, kajian yang dijalankan oleh
Papagiannis et al. [5] mendapati hubungan yang rapat di antara kandungan Zn dan Cu di dalam ikan
dan zooplankton di dalam ekosistem tasik. Oleh yang demikian, Zn dan Cu yang terkandung di dalam
ikan ini mungkin di ambil secara tidak langsung melalui makanan yang di ambil oleh ikan tersebut.
Nilai kepekatan logam berat yang diperolehi ini tidak melebihi tahap maksimum yang dibenarkan
dalam Akta Makanan 1983 (Akta 281)[6] iaitu 30 mg/kg dan 100 mg/kg untuk masing-masing Cu dan
Zn. Walau bagaimanapun kandungan Hg pada L2 (0.53 mg/kg) adalah 0.05 mg/kg melebihi takat
maksimum yang dibenarkan. Oleh itu bolehlah disimpulkan bahawa tasik yang tercemar dengan
logam berat akibat aktiviti perlombongan tidak semestinya akan menyebabkan peningkatan kepekatan
kesemua logam berat di dalam tasik terhadap yang hidup akuatik di dalamnya.
Secara amnya kandungan logam berat dalam air adalah lebih rendah berbanding dengan sedimen dan
tumbuhan kecuali barium yang mana kandungannya berada dalam julat 7.78 ± 1.80 hingga 27.67 ±
2.25 µg/l (Jadual 2). Barium terdiri daripada logam alkali yang mudah ditemui di dalam perut bumi.
Proses perlombongan menyebabkan sebahagian besar logam yang terkandung dalam perut bumi
dikeluarkan yang akhirnya menyebabkan kandungannya tinggi pada tasik bekas lombong.
Logam berat tertinggi yang dikesan pada tumbuhan adalah mangan iaitu 19.05 ± 8.10 mg/kg di tasik
L2, diikuti oleh barium (7.66 ± 0.66 mg/kg) juga di L2. Kajian yang dilakukan di kawasan tasik
bekas lombong arang batu di Poland oleh Samecka dan Kempers [7] mendapati kandungan Mn amat
tinggi (1142 – 2116 mg/kg) diikuti oleh Zn, Ba dan Pb. Hasil kajian ini didapati tidak mengikut corak
tersebut. Ini disebabkan oleh perbezaan jenis mineral yang pernah dilombong.
Kandungan Ba juga didapati tertinggi berbanding logam berat lain dalam sedimen, diikuti oleh Pb dan
Cu. Terdapat persamaan antara kepekatan Cu dalam sedimen dengan kepekatan Cu dalam tumbuhan
di mana kepekatannya adalah paling tinggi di L1, di ikuti oleh L3 dan L2. Corak ini sama dengan
nilai pH sedimen (Jadual 1). Ini mungkin disebabkan pada pH tinggi, Cu dalam sedimen dapat
diambil oleh tumbuhan yang berada berdekatan berbanding pada keadaan pH rendah. Kandungan Cu
di dalam sedimen mungkin meresap melalui air bawah tanah dan akhirnya diambil oleh akar
tumbuhan. Kepekatan Hf dan Hg pula adalah paling tinggi di L3 dan diikuti oleh L2 dan L1.
Kepekatan Hf dan Hg yang tinggi turut dikesan pada ikan dari tasik berkaitan. Oleh yang demikian
pencemaran logam Hg dan Hf boleh menyebabkan pemekatan dan akhirnya memasuki rantai
pemakanan akuatik.
17
JADUAL 2 Kepekatan logam berat di dalam ikan, sedimen, air dan tumbuhan
Kandungan logam berat dalam sampel (mg/kg) basah atau ug/L (air)
Unsur
L1 L2 L3
Ikan Sedimen Air tumbuhan Ikan Sedimen Air tumbuhan Ikan Sedimen Air tumbuhan
0.08 2.2 3.92 0.07 2.63 19.05 0.06 8.63
Mn ± ± 0.00 ± ± ± 0.00 ± ± 0.00 0.00 ±
0.03 0.85 2.58 0.01 0.02 8.10 0.01 5.56
0.50 2.47 1.00 5.97 2.27 1.0 1.00 1.79 0.13 2.28 1.00 2.49
Cu ± ± ± ± ± ± ± ± ± ± ± ±
0.02 1.29 0.01 1.33 0.48 0.51 0.01 0.03 0.01 2.22 0.01 1.16
0.50 0.40 0.88 4.09 3.81 0.38 13.5 0.75 0.65 0.51 7.73 1.38
Zn ± ± ± ± ± ± ± ± ± ± ± ±
0.10 0.21 0.02 0.67 2.91 0.01 2.65 0.24 0.21 0.30 1.03 0.54
0.11 0.34 2.83 0.07 0.32 2.25 0.06 0.35
As ± ± ± 0.00 ± ± ± 0.00 ± ± 0.00 0.00
0.01 0.20 0.29 0.01 0.11 0.29 0.01 0.14
0.07
Cd 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 ± 0.00 0.00 0.00
0.01
Sn 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
Sb 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
14.78 27.7 4.52 0.11 45.10 25.0 7.66 0.13 13.08 7.78 3.32
Ba 0.00 ± ± ± ± ± ± ± ± ± ± ±
5.30 2.25 2.50 0.03 9.26 0.29 0.66 0.03 4.20 1.80 1.19
0.17 0.20 0.53 0.33 0.11 0.60
Hg ± ± 0.00 0.00 ± ± 0.00 0.00 ± ± 0.00 0.00
0.01 0.10 0.20 0.10 0.02 0.13
6.09 1.00 2.26 13.55 1.05 3.57
Pb 0.00 ± ± ± 0.00 ± ± 0.00 0.00 ± 0.00 0.68
0.73 0.01 0.25 1.67 0.01 0.56
0.33 0.53 0.12 0.52
Zr 0.00 ± 0.00 0.00 0.00 ± 0.00 0.00 ± ± 0.00 0.00
0.12 0.20 0.01 0.16
0.72 2.83 1.40 3.00 0.14 1.65
Hf 0.00 ± ± 0.00 0.00 ± ± 0.00 ± ± 0.00 0.00
0.06 0.29 0.31 0.01 0.02 0.56
c. Radionuklid tabii.
Kandungan radionuklid tabii U-238, Th-232 dan K-40 ditunjukkan dalam Jadual 3. Hanya K-40 yang
dapat dikesan pada sampel ikan, air, tumbuhan dan sedimen dari ketiga-tiga tasik. Radionuklid U-238
dan Th-232 hanya terdapat pada sampel sedimen.
JADUAL 3. Kandungan radionuklid tabii dalam ikan, air, tumbuhan dan sedimen.
Kandungan radionuklid tabii (Bq/kg atau Bq/L)
Tasik 1 Tasik 2 Tasik 3
Ikan
K-40 7.78E-02 ± 0.01 7.34E-02 ± 0.01 6.52E-02 ± .01
Th-232 0 0 0
U-238 0 0 0
Air
K-40 1.11E-03 ± 0.00 1.07E-03 ± 0.00 1.23E-03 ± 0.00
Th-232 0 0 0
U-238 36.72 36.72 0
Tumbuhan
K-40 0.09 ± 0.05 1.12E-03 ± 0.00 0.10 ± 0.04
Th-232 0 0 0
U-238 0 0 0
Sedimen
K-40 1.57E-04 ± 4.48E-05 4.24E-04 ± 1.47E-04 9.44E-05 ± 5.40E-06
Th-232 30.76 ± 2.71 35.34 ± 0.27 34.06 ± 11.85
U-238 18.86 ± 2.60 26.32 ± 3.01 9.37 ± 2.30
18
Terdapat dua laluan utama yang membolehkan ikan di tasik bekas lombong dicemari oleh K-40, iaitu
melalui deposisi permukaan ataupun melalui rantai pemakanan ikan tersebut. Deposisi permukaan
boleh berlaku apabila terdapat pemindahan unsur kalium daripada komponen ekosistem tasik seperti
sedimen dan tumbuhan. Aktiviti perlombongan dijangka meningkatkan kandungan pelbagai
radionuklid tabii di kawasan sekitarnya. Kajian ini menunjukkan walaupun ketiga-tiga radionuklid
tersebut terdapat pada sediment, hanya K-40 yang dapat dikesan pada tumbuhan dan ikan.
Penyebaran unsur K-40, Th-232 dan U-238 dalam ekosistem akuatik boleh berlaku melalui ampaian
pepejal yang seterusnya membabitkan komponen akuatik lain seperti plankton, herbivor, karnivor
serta omnivor yang terdapat dalam sistem akuatik tersebut. Ketiadaan U-238 dan Th-232 dikesan
pada tumbuhan adalah disebabkan unsur tersebut diikat kuat oleh komposisi tanah dan sukar di ambil
oleh tumbuhan. Selain daripada itu, tumbuhan juga tidak mengambil uranium dan thorium sebagai
unsur nutrien. Kajian sebelum ini mendapati kandungan radionuklid tabii U-238 dan Th-232 dalam
sampel tanah di kawasan lombong aktif adalah masing-masing 3.75 Bq/kg dan 10.51 Bq/kg
(Muhamad Samudi Yasir et al.[8]), manakala kandungan dalam sistem air kawasan lombong aktif
pula adalah antara 26.3 – 36.2 Bq/L (Th-232) dan 26.8 – 35.4 Bq/L (U-238) (Ismail et al. [9]. Oleh
yang demikian kandungan radionuklid tabii dalam semua sampel yang di ambil adalah lebih rendah
dari hasil kajian sebelumnya.
Aktiviti U-238 dan Th-232 bagi sedimen di L1, L2 dan L3 adalah relatif lebih tinggi berbanding
aktivitinya di dalam tisu daging ikan, air dan tumbuhan. Tetapi aktiviti K-40 bagi sedimen di L1, L2
dan L3 adalah relatif lebih rendah berbanding aktivitinya di dalam tisu daging ikan. Fenomena ini
mungkin terjadi kerana faktor kedudukan tropik ikan yang tinggi menyebabkan berlaku pemekatan K-
40 dalam jumlah yang banyak.
Kesimpulan
Hasil kajian ini mendapati walaupun ekosistem tasik bekas lombong mengandungi pelbagai logam
berat dan radionuklid tabii, kandungannya kebanyakan logam berat dalam ikan adalah rendah dan
berada di bawah aras maksimum yang dibenarkan dalam Akta Makanan (1983). Oleh yang demikian
dari sudut pencemaran logam berat dan radionuklid tabii kawasan tasik bekas lombong ini berpotensi
untuk dimajukan sebagai kawasan ternakan akuakultur.
Penghargaan
Penulis merakamkan ucapan terima kasih kepada Universiti Kebangsaan Malaysia di atas
peruntukan kewangan yang disediakan.
Rujukan
1. Amran Ab Majid, Muhamad Samudi Yasir, Redzuwan Yahaya. 2004. “Taboran Radionuklid
Tabii (NORM) dan Kaitannya dengan Aktiviti Pembangunan di Negeri Selangor”. Dalam:
Indicator of Sustainable Development: Assessing Changes in Environmental Conditions. A.
Latiff et al. (ed), Institute For Environment and Development (LESTARI), Universiti Kebangsaan
Malaysia, ISBN 983-9444-58-1, 213 – 225.
2. Elbering B., Knudsen K.L, Kristensen P.H dan Asmund G., 2004. “Appliying foramineral
stratigraphy contaminationand mining impact in a fiord in West Greenland”. Marine
Enviromental Research 55 3: 235-256.
3. Muhamad Samudi Yasir, Ismail Bahari, Sahibin A.R., Dahlia Suriati Abd Rahim and Halim Abd
Rahman, 2001. “The Concentration of Natural Radiobuclides and Heavy Metals in Soils From A
Tin-Mining and Its Surrounding Area”. J. Sains Nuklear Malaysia, 19: 50 – 56.
4. Riba I., Conradi M., Forja J.M. dan Delvalls T.A. 2004. “Sediment Quality in the Guadalquivir
Estury: Lethal Effect Associated with the Azhalcollar Mining Spil”. Marine Pollution Bulletin. 48
1:144-152.
19
5. Papagiannis I, Kagalov I., Leonardos J., Petridis D. dan Kalfakou V. 2004. “Copper and Zink in
Four Freshwater Fish Species from Lake Pamvotis (Greece)”. Journal of Enviromental
International. 30 (3) :257-362.
6. Akta Makanan 1983. MDC, Kuala Lumpur.
7. Samecka C. dan Kempers. 2004. “Consentrations of heavy metal and plant nutrients in water,
sediments and aquatic macrophytes of anthropogenic lakes (former open cut brown coal mines)
differing in stage of acidification”. Environment International. 281 :87-98
8. Muhamad Samudi Yasir, Amran Ab Majid dan Redzuwan Yahaya, 2005. “Kandungan
Radionuklid Tabii Dalam Sampel Amang, Tanah dan Air di Sekitar Kawasan Industri
Perlombongan Di Dengkil Selangor”. Proceeding of The 6th ITB-UKM Joint Seminar on
Chemistry, 17 – 18 May, Bali Indonesia. ISBN: 983-29766-38-7653 - 658.
9. B. Ismail, M.S. Yasir, Y. Redzuwan and A.M. Amran, 2003. “Radiological Environmental Risk
Associated with Different Water Management System in Amang Processing in Malaysia”.
Pakistan Journal of Biological Science. 6: 1544 – 1547.
20
CHARACTERIZATION OF NOVEL ORGANOTIN(IV) COMPLEXES WITH O, N, O –

DONOR LIGAND DERIVED FROM CARBOHYDRAZIDE: X-RAY CRYSTAL
STRUCTURE OF [Ph2Sn(H2CBS)]
Y. Z. Liew 1*, M. A. Affan 1, Fasihuddin B. Ahmad 1, Mustaffa B. Shamsuddin 2 and Bohari M.

Yamin 3
1
Resource Chemistry, Faculty of Resource Science and Technology
Universiti Malaysia Sarawak, 94300 Kota Samarahan,
Sarawak, Malaysia
2
Department of Chemistry, Faculty of Science,
Universiti Teknologi Malaysia, 81310 UTM Skudai
Johor Darul Takzim, Malaysia
3
School of Chemical Sciences and Food Technology
Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia
Abstract
An interesting series of new organotin(IV) complexes has been synthesized by the reaction of
RnSnCl4-n with potentially O, N, O – tridentate carbohydrazone ligand derived from carbohydrazide.
Five new organotin(IV) complexes of carbohydrazide-bis(salicylaldehyde) ligand [H4CBS, (1)] with
RnSnCl4-n (n = 1, 2) have been synthesized in the presence of base and refluxing in 1:2:1 molar ratio
(metal:base:ligand). All complexes (2-6) have been characterized by different physico-chemical
techniques like molar conductivity measurements, elemental analysis, UV-Visible, IR and 1H NMR
spectral studies. All complexes (2-6) are non electrolytic in nature. Among them, diphenyltin(IV)
complex (4) is also characterized by X-ray crystallography diffraction analyses. In the solid state, the
carbohydrazone ligand (1) exist as the keto tautomer but in solution in the presence of base and
organotin(IV) chloride(s), it converted to the enol tautomer and coordinated to the tin atom in its
deprotonated enolate form. X-ray crystallographic analysis shows that the diphenyltin(IV) complex,
[Ph2Sn(H2CBS)] (4), is five-coordinate and has a distorted trigonal-bipyramidal geometry with the
ligand coordinated to the tin(IV) as a tridentate dinegative fashion through its phenolic-O, enolic-O
and imine-N atoms. The general bond length order is: oxo < phenolato < enolato. The Sn-O (enolato)
bond is longer than Sn-O (phenolato) bond by ~0.095 Å and is identical with Sn-O (carboxylate)
bond. The crystal of [Ph2Sn(H2CBS)] (4) is triclinic with space group P-1 with a=8.514(2)Å,
b=12.505(3)Å, c=12.794(4)Å, α=105.169(4)°, β=107.639(4)°, γ=96.232(4)°, V=1226.5(6) Å3, Z=1
and Dcalc=1.541 Mg/m3. The IR, UV and 1H NMR data are consistent with all the organotin(IV)
derivatives having similar geometry.
Keywords: Carbohydrazone ligand; Organotin(IV) compounds; Crystal structure.
Corresponding author: Tel.: 6082-679235; fax: 6082-672275

E-mail address: maaffan@frst.unimas.my
Introduction
Hydrazides and hydrazones have interesting ligational properties due to the presence of several
potential coordination sites. The chemistry of carbohydrazide compounds has been studied by Swamy
and Siddalingaiah [1]. Variety of metal complexes of symmetrical dihydrazones derived from
thiocarbohydrazides have been synthesized and their stereochemistry have been reported in the
literatures [2-3] and potentially useful biological activities [3]. However, the coordination chemistry
of the corresponding carbohydrazide derivatives is less explored [4]. Complexes of the
carbohydrazide with non-transition metal ions such as organotin(IV) have not received as much
21
attention. In view of the importance of tin compounds in medicinal chemistry and biotechnology [5]
and as part of our on going work on tin-hydrazones [6-7], here we report about the synthesis and
characterization of the carbohydrazide-bis(salicylaldehyde) ligand (Scheme 1) and of its
organotin(IV) complexes (Scheme 2) and together with the X-ray crystal structure of diphenyltin(IV)
complexes [Ph2Sn(H2CBS)] (4).
O H
H C H
2 O C
N N +
H2N NH2 HO
Carbohydrazide Salicylaldehyde
abs. ethanol
Keto form
+ 2H2O
Enol form
Scheme 1
Experimental
Synthesis
Chemicals were obtained from Fluka and Aldrich and were used without further purification. Solvents
were analytical grade and purified by standard methods [8]. The C, H, N elemental analyses were
performed on a Carlo Erba model EA 1108 analyser. Infrared spectra were recorded as KBr disc using
Shimadzu 8201 PC Fourier-Transform Spectrometer. 1H NMR spectra were recorded in DMSO-d6
solution on a Bruker 300 FT-NMR spectrophotometer. Electronic spectra were recorded on a
Shimadzu 2401 PC UV-Visible spectrophotometer.
22
Carbohydrazide-bis(salicylaldehyde) ligand (H4CBS) [C15H14N4O3] (1)
A mixture of carbohydrazide (0.005 mole, 0.450 g) and salicylaldehyde (0.0010 mole, 1.221 g) in
absolute ethanol (30 mL) were heated under reflux for 3-4 hours. The reaction mixture was allowed to
cool down to room temperature for half an hour. Then the white precipitate was filtered off and
washed several times using absolute ethanol. The crystalline white solid obtained was purified by
recrystallization from hot absolute ethanol and dried in vacuo over P2O5 overnight. Yield = 70.05%.
M. p. = 178-180 °C. Found: C, 59.20; H, 4.59; N, 18.40%. Calc. for C15H14N4O3: C, 59.21; H, 4.61;
N, 18.42%. λmax (nm) (DMF): 262, 292, 328. IR (νmaxcm-1 (KBr): 1681(-C=O), 1616 (C=N)+(C=C),
1272 (C–O, phenolic), 946 (N–N). 1H NMR (300 MHz, DMSO-d6): δ 10.86 (br, OH), δ 8.44 (s,
N=CH), δ 7.69 (br, CONH), δ 6.62–7.27 (m, aromatic-H).
Me2Sn(H2CBS) [Me2Sn(C15H12N4O3)] (2)
Carbohydrazide-bis(salicylaldehyde) ligand (1) (0.002 mole, 0.596 g) was dissolved in hot absolute
methanol (20 mL) under nitrogen atmosphere with potassium hydroxide (0.0042 mole, 0.236 g)
previously dissolved in methanol (10 mL). The colour of the solution changed from off-white to
yellow. The resulting mixture was refluxed for one hour and a solution of Me2SnCl2 (0.002 mole,
0.439 g) in methanol (10 mL) was added dropwise to the potassium salt of ligand solution, the color
of the solution became darker. The resulting solution was refluxed for four hours and allowed to cool.
The precipitated potassium chloride (KCl) was removed by filtration and the filtrate was evaporated
to obtain the yellow solid. The yellow micro-crystals were filtered off and washed with hexane and
dried in vacuo over P2O5 overnight. Yield = 69.51%. M. p. = 218-220 °C. Found: C, 45.91; H, 4.03;
N, 12.56%. Calc. for C15H12N4O3Me2Sn: C, 45.88; H, 4.05; N, 12.59%. λmax (nm) (DMF): 266, 343,
398. IR (νmaxcm-1 (KBr): 1604 (C=N)+(C=C), 1318 (C–O, phenolic), 960 (N–N), 608 (Sn–C), 566
(Sn–O), 470 (Sn–N). 1H NMR (300 MHz, DMSO-d6): δ 11.41 (br, OH), δ 10.89 (s, NH) δ 8.14 &
8.43 (s, N=CH)), δ 6.63–7.93 (m, aromatic-H). δ 0.68 (s, Sn-CH3).
Bu2Sn(H2CBS) [Bu2Sn(C15H12N4O3)] (3)
Complex (3) was synthesized in a similar way as in (2), using dibutyltin(IV) dichloride (0.002 mole,
0.688 g) instead of dimethyltin(IV) dichloride. Yield = 68.51%. M. p. = 225-227 °C. Found: C, 52.00;
H, 6.03; N, 10.55%. Calc. for C15H12N4O3Bu2Sn: C, 52.02; H, 6.02; N, 10.55%. λmax (nm) (DMF):
267, 340, 396. IR (νmaxcm-1 (KBr): 1604 (C=N)+(C=C), 1323 (C–O, phenolic), 952 (N–N), 590 (Sn–
C), 560 (Sn–O), 440 (Sn–N). 1H NMR (300 MHz, DMSO-d6): δ 11.45 (br, OH), δ 11.05 (s, NH) δ
8.15 & 8.41 (s, N=CH)), δ 6.65–7.89 (m, aromatic-H), δ 0.79 (t, 6H, (-CH3) of butyltin), δ 1.18 (m,
4H, (-CH2) of butyltin), δ 1.26-1.45 (m, 4H, (-CH2) of butyltin) δ 1.55-1.64 (m, 4H, -CH2-Sn).
Ph2Sn(H2CBS) [Ph2Sn(C15H12N4O3)] (4)
Complex (4) was prepared similarly to complex (2), using diphenyltin(IV) dichloride (0.002 mole,
0.688 g) instead of dimethyltin(IV) dichloride. Single crystals suitable for X-ray diffraction studies
were obtained by slow evaporation of dichloromethane-petroleum ether (40-60 °C) solution (1:1).
Yield = 64.33%. M. p. = 208-210 °C. Found: C, 56.65; H, 3.88; N, 9.87%. Calc. for
C15H12N4O3Ph2Sn: C, 56.69; H, 3.87; N, 9.85%. λmax (nm) (DMF): 266, 342, 398. IR (νmaxcm-1 (KBr):
1600 (C=N)+(C=C), 1325 (C–O, phenolic), 960 (N–N), 600 (Sn–C), 520 (Sn–O), 460 (Sn–N). 1H
NMR (300 MHz, DMSO-d6): δ 11.48 (br, OH), δ 11.15 (s, NH) δ 8.17 & 8.38 (s, N=CH)), δ 6.70–
7.62 (m, aromatic and Sn-C6H5 protons).
MeSnCl(H2CBS) [MeSnCl(C15H12N4O3)] (5)
The procedure described above for (2) was followed for the preparation of (5), with methyltin(IV)
trichloride being used instead of dimethyltin(IV) dichloride and the refluxing time for this preparation
23
was 2-3 hours. Yield = 65.46%. M. p. = 205-206 °C. Found: C, 41.19; H, 3.25; N, 12.05 %. Calc. for
C15H12N4O3MeSnCl: C, 41.23; H, 3.22; N, 12.02%. λmax (nm) (DMF): 267, 341, 398. IR (νmaxcm-1
(KBr): 1610 (C=N)+(C=C), 1340 (C–O, phenolic), 952 (N–N), 590 (Sn–C), 560 (Sn–O), 472 (Sn–N).
1
H NMR (300 MHz, DMSO-d6): δ 11.42 (br, OH), δ 10.96 (s, NH) δ 8.14 & 8.44 (s, N=CH)), δ 6.64–
7.90 (m, aromatic-H). δ 1.14 (s, Sn-CH3).
PhSnCl(H2CBS) [PhSnCl(C15H12N4O3)] (6)
The procedure described above for (5) was followed for the preparation of (6), with phenyltin(IV)
trichloride being used instead of methyltin(IV) trichloride. Yield = 64.23%. M. p. = 213-215 °C.
Found: C, 49.10; H, 3.63; N, 10.22 %. Calc. for C15H12N4O3PhSnCl: C, 49.15; H, 3.66; N,
10.25%.λmax (nm) (DMF): 268, 340, 399. IR (νmaxcm-1 (KBr): 1608 (C=N)+(C=C), 1330 (C–O,
phenolic), 956 (N–N), 615 (Sn–C), 520 (Sn–O), 451 (Sn–N). 1H NMR (300 MHz, DMSO-d6): δ 11.44
(br, OH), δ 11.10 (s, NH) δ 8.18 & 8.39 (s, N=CH)), δ 6.72–7.62 (m, aromatic and Sn-C6H5 protons).
X-ray Crystallography
Yellow single-crystal of (4) (size 0.46 x 0.17 x 0.09 mm) was grown from dichloromethane-petroleum
ether (40-60 °C) mixture at the room temperature. The measurements were performed at 273 (2) K on
Siemen SMART CCD diffraction using graphite-monochromated Mo-Kα radiation (λ = 0.71073 Å).
Orientation matrix and unit cell parameters were obtained from the setting angles of 25-centered
reflection. The crystals are monoclinic, space group P2(1)/c with a = 8.514(2), b = 12.505(3), c =
12.794(4) Å, α = 105.169(4)°, β = 107.639(4)°, γ = 96.232(4)°, V = 1226.5(6) A3, Z = 1, Dcalc = 1.541
Mg/m3, µ = 1.078 mm-1. The diffraction intensities were collected by ω scans (1.7 to 27.0°). A total of
13623 / 5308 reflections were collected (-10< =h< =10, -15< =k< =15, -16< =l< =16). The structure
was solved using direct methods and refined using the full-matrix least-square method on F obs2 using
the SHELXTL [9] software package. All non-H atoms were anisotropically refined. The hydrogen
atoms were located in a difference Fourier map and then were fixed geometrically and treated as
riding atom on the parent C atoms, with C-H distances = 0.97 Å.
Results and discussion
The carbohydrazone ligand [H4CBS, (1)] was synthesized by the condensation reaction of
carbohydrazide with salicylaldehyde in a 1: 2 ratio in absolute ethanol (Scheme 1). Five new
organotin(IV) complexes (2-6) of carbohydrazide-bis(salicylaldehyde) ligand (1) have been
synthesized by direct reaction of the appropriate organotin(IV) halide(s) with the ligand in the
presence of a base (Scheme 2). In all the cases, a base is added to the reaction mixture in order to
force the deprotonation of the ligand. The low molar conductance values (10-40 ohm-1cm2 mol-1) (see
Table 1) indicating non-electrolytic nature for all the organotin(IV) complexes [10].
Table 1. Molar conductance values for organotin(IV) complexes of ligand (1)
Compound Molar conductance, Λm (ohm-1 cm 2 mol -1)

[Me2Sn(H2CBS)] (2) 10
[Bu2Sn(H2CBS)] (3) 23
[Ph2Sn(H2CBS)] (4) 29
[MeSnCl(H2CBS)] (5) 40
[PhSnCl(H2CBS)] (6) 35
24
RnSnCl4-n + Ligand [H4CBS, (1)] + 2KOH
H
H N H
N C
C N N C
O
OH Sn
O
abs. MeOH
R
R
R = Me, Bu and Ph
OR
H
H N C N H
C N N C
O
OH Sn
O
R
Cl
R = Me and Ph
Scheme 2
Electronic absoption spectra
The UV-Vis electronic spectra of ligand (1) and its organotin(IV) complexes (2-6) measured at room
temperature in DMF (10-4 M) solutions over 200 – 800 nm range are given in Table 2.
Carbohydrazone ligand (1) exhibited three main bands at 262, 292 and 328 nm. The first and second
bands were attributed to benzene π – π* and imino (C=N) π – π* transition, which were not affected by
the chelation. The third band is assigned to n – π* transition. The appearance of two new bands at 266-
343 nm and at 396-399 nm regions showed the metal-ligand coordination in all the complexes (2-6).
The high wavelength bands in the region 396-399 nm can be attributed to a charge transfer transition
involving the tin atom [11].
Table 2. The λmax (nm) peaks of ligand (1) and its organotin(IV) complexes (2-6).
Compounds λmax (nm)

(1) H4CBS 328, 292, 262
(2) [Me2Sn(H2CBS)] 398, 343, 266
(3) [Bu2Sn(H2CBS)] 398, 342, 266
(4) [Ph2Sn(H2CBS)] 396, 340, 267
(5) [MeSnCl(H2CBS)] 398, 341, 267
(6) [PhSnCl(H2CBS)] 399, 340, 268
25
Infrared spectra
The IR data for the ligand (1) and its all organotin(IV) complexes (2-6) are described in the synthesis
section. The IR spectra of the complexes (2-6), the disappearance of the ν(C=O) band indicating
enolization of the amide group into C-OH enolic form through the amide-imidol tautomerism process
and also indicating deprotonation of the functional group during coordination to tin atom [12]. The
ν(C=N) band in the complexes (2-6) appeared as a strong band in the region 1600–1610 cm-1, is
considerably shifted towards lower frequencies with respect to that of the free ligand (1620 cm-1),
confirming the coordination of the azomethine nitrogen to organotin(IV) moiety [12]. This is also
apparent from the ν(Sn–N) band at 440–472 cm-1 in the IR spectra of all the complexes (2-6). The
hydrazinic stretching ν(N–N) band observed at 950 cm-1 for the ligand (1) is shifted in the higher
region at 952–962 cm-1 in the complexes (2-6) further supporting that azomethine nitrogen is
coordinated to Sn(IV) ion [11]. The high intensity band observed at 1272 cm-1 in the ligand (1)
attributed due to phenolic ν(C–O), appears as a medium band at 1319-1340 cm-1 in the IR spectra of
all the complexes (2-6). These observations favour the formation of Sn–O bond via deprotonation.
The medium and weak bands observed at 590-651, 520–560 and 440–472 cm-1 in the IR spectra of all
the complexes (2-6) are attributable to the ν(Sn–C), ν(Sn–O) and ν(Sn–N) vibration bands
respectively, indicating coordination of the free ligand to the central Sn(IV) atom via deprotonated
enolic oxygen and azomethine nitrogen in the complexes (2-6).
1
H NMR spectra
The 1H NMR data for the ligand (1) and its all complexes (2-6) are presented in the experimental
section. The 1H NMR spectrum of ligand (1) is characterized by four signals at 10.86, 8.44, 7.69 and
6.62–7.27 ppm, which are assigned to the protons associated with –OH, –N=CH, –CONH and
aromatic ring protons, respectively. In 1H NMR spectra of organotin(IV) complexes (2-6), azomethine
N=CH signal is splited into two signals at 8.14-8.17 and 8.38-8.43 ppm. This indicates that only one
of the azomethine nitrogen in ligand (1) could be bonded to the Sn(IV) ion in the complexes (2-6),
another one HC=N group could be free in the complexes (2-6). This evidence is confirmed from X-
ray crystallographic analyses. The disappearance of the signal due to –CONH proton in the complexes
(2-6) indicated the enolization of the form of the ligand (1) in the complexes (2-6) [6]. The new signal
at 10.89-11.15 ppm in the complexes (2-6) is due to the free NH group of ligand (1) which is not
involved in the coordination to tin(IV) in the complexes (2-6). A signal appeared at 11.41-11.48 ppm,
which is indicated that phenolic proton present in the complexes (2-6). The sharp signal attributed to
methyl group attached to tin atom is appeared as a singlet at 0.68 ppm in the dimethyltin(IV) complex
(2). Another sharp resonance signal attributed to methyl group attached to tin(IV) atom is appeared as
a singlet at 1.14 ppm in methyltin(IV) complex (5). In the complex (3), a multiplet in the region 0.79-
1.64 ppm is assigned to the butyl group attached to the tin(IV) atom. Complexes (4 and 6) showed a
multiplet in the region 6.70-7.62 ppm, which may be assigned to aromatic ring protons and Sn-Ph
protons, respectively. The signals could not properly assigned due to overlap of corresponding signals
of Ph-Sn and aromatic ring protons.
Crystal structure of [Ph2Sn(H2CBS)] (4)
The X-ray structural investigation of [Ph2Sn(H2CBS)] (4) (Figure 1) revealed that the carbohydrazide-
bis(salicylaldehyde) ligand [H4CBS, 1] is O,N,O-coordinated in complex (4). The crystal data and
structure refinement for compound [Ph2Sn(H2CBS)] (4) are summarized in Table 3. The selected bond
distances and angles for the molecule (4) are given in Table 4. The crystal structure of
[Ph2Sn(H2CBS)] (4) reveals the tin(IV) atom has a five coordination geometry in a distorted trigonal-
bipyramidal arrangement. The Sn(1) atom lies in the ligand plane and form five membered and six
membered chelate ring with the ligand. Thus the two phenyl groups and an imine nitrogen take the
equatorial position, while the oxygen atoms, O(1) and O(2), take up the axial sites around the Sn(1)
atom.
26
Table 3. Crystal data and the structure refinement for the complex [Ph2Sn(H2CBS)] (4)
Formula C27H22N4O3Sn
Formula weight 569.18
Crystal system Triclinic
Space group P-1
Z 1
a (Å) 8.514(2)
b (Å) 12.505(3)
c (Å) 12.794(4)
α (°) 105.169(4)
β (°) 107.639(4)
γ (°) 96.232(4)
V (Å3) 1226.5(6)
Dcalc (mg m-3) 1.541
Absorption coefficient (mm-1) 1.078
Temperature (K) 273(2)
Wavelength (Å) 0.71073
Final R indices [I>2sigma(I)] R1 = 0.0337, wR2 = 0.0805
R indices (all data) R1 = 0.0380, wR2 = 0.0829
Goodness-of-fit on F^2 1.135
The distorted trigonal-bipyramidal arrangement is a result of the strain imposed by the

tridentate ligand, and from the constraints imposed by the six membered ring, Sn(1)-/N(1)-C(7)-C(6)-
C(5)-/O(1) and five membered ring Sn(1)-/N(1)-N(2)-C(8)-O(2). The trigonal-biypramidal geometry
in complex (4) is distorted as indicated by the bond angles of 157.08(8)° for O(1)-Sn(1)-O(2) and the
deviation from 90° of the angles O(1)-Sn(1)-N(1) (84.96(8)°) and O(2)-Sn(1)-N(1) (72.89(8)°). The
sum of the angles O(1)–Sn(1)–N(1), 84.96(8)° and O(2)–Sn(1)–N(1), 72.89(8)° is 157.85°, and it is
almost the same with the angle O(1)–Sn(1)–O(2), 157.08(8)°, so that the atoms Sn(1), N(1), O(1) and
O(3) are co-planer. The sum of the angles C(21)-Sn(1)-N(1), C(22)-Sn(1)-N(1) and C(21)– Sn(1)–
C(22) is 358.62°, thus the atoms Sn(1), N(1), C(19) and C(23) are almost in the same plane.
Figure 1. Molecular structure of [Ph2Sn(H2CBS)] (4).

Thermal ellipsoids at the 50% level.
27
Table 4. Selected bond lengths (Å) and angles (°) of diphenyltin(IV) complex [Ph2Sn(H2CBS)] (4)
Bond lengths
Sn(1)-O(1) 2.0482(19) Sn(1)-N(1) 2.162(2)
Sn(1)-O(2) 2.1430(19) Sn(1)-C(21) 2.112(3)
Sn(1)-C(22) 2.117(3) O(1)-C(5) 1.340(3)
O(2)-C(8) 1.282(3) N(1)-C(7) 1.301(3)
N(1)-N(2) 1.376(3) N(2)-C(8) 1.310(4)
N(3)-N(4) 1.362(3) N(3)-C(8) 1.361(3)
C(6)-C(7) 1.424(4) N(4)-C(9) 1.272(4)
C(9)-C(10) 1.456(4)
Bond angles
O(1)-Sn(1)-C(21) 97.52(9) O(1)-Sn(1)-C(22) 99.19(10)
C(21)-Sn(1)-C(22) 120.45(9) O(1)-Sn(1)-O(2) 157.08(8)
C(21)-Sn(1)-O(2) 91.16(9) C(22)-Sn(1)-O(2) 94.52(9)
O(1)-Sn(1)-N(1) 84.96(8) C(21)-Sn(1)-N(1) 126.43(9)
C(22)-Sn(1)-N(1) 111.74(9) O(2)-Sn(1)-N(1) 72.89(8)
C(5)-O(1)-Sn(1) 127.47(18) C(8)-O(2)-Sn(1) 112.67(17)
C(7)-N(1)-Sn(1) 125.69(19) N(2)-N(1)-Sn(1) 116.73(16)
C(7)-N(1)-N(2) 117.4(2) C(9)-N(4)-N(3) 119.4(2)
O(1)-C(5)-C(6) 122.7(2 C(5)-C(6)-C(7) 125.2(2)
N(1)-C(7)-C(6) 117.0(3) O(2)-C(8)-N(2) 126.4(2)
The asymmetry of H4CBS ligand is strongly reflected in the Sn-O distances. In the complex
(4), the Sn(1)-O(2) bond, 2.1430(19)Å, is longer than the Sn(1)-O(1) bond, 2.0482(19)Å. This is a
consequence of O(2) being a carbonyl and O(1) being bound to a benzene ring. In Ph2SnSalAp (where
SalAp/salicylideneamino-o-hydroxybenzene), the Sn-C bonds are 2.118(5) Å and 2.111(5) Å [13], is
almost similar with the Sn-C bonds found in complex (4), 2.112(3) Å and 2.117(3) Å. The Sn–N,
2.162(2) Å bond of the compound (4) is a little longer than that of the compound {[Ph2Sn(2-
OC10H6CHNCH2COO)]SnPh2Cl2} 2.136 Å [14] and Ph2Sn[Ph(O)C=CH-C(Me)=N-N=C(O)Ph]
2.145(3) [15], but shorter than that of [SnMe2(Salop)] 2.221(3) Å [16] and {Ph2Sn[4-NC5H4–
(O)N2C(CH3)CO2](H2O)}2 .CH2Cl2 .H2O, 2.288(7) Å and 2.282(7) Å [17], and it is considerably
less than the sum of the van der Waals radii of tin and nitrogen, 3.75 Å [18]. Due to the involvement
of N(1) atom in tin binding, the bond length of N(1)-C(7) is significantly increased to 1.301(3) Å as
compared with the imine function N(4)-C(9) (1.272(4) Å) which is having a double bond character.
Conclusions
The synthesis and physical properties of a new series of organotin(IV) compounds with
carbohydrazone ligand (1) have been described. The ligand behaved as a tridentate dinegative fashion
towards to tin(IV). The complexes (2-6) are monometallic. The coordination around the tin(IV) ion is
established by means of single crystal X-ray diffraction analysis on [Ph2Sn(H2CBS)] (4).
Acknowledgement
The authors are very grateful to Universiti Malaysia Sarawak (UNIMAS), for the financial support
(Grant # -01(101)/453/2004(190). The authors also would like to thank the School of Chemistry
Sciences and Food Technology, Universiti Kebangsaan Malaysia (UKM), for the CHN analyses and
also X-ray single crystal determination. We would also like to thank the Ibnu Sina Institute, UTM, for
the help in obtaining the 1H NMR spectra.
28
References
[1] Swamy, H. M. V., Siddalingaiah, A. H. M. (2000) “Studies on some lanthanides(III)

complexes of o-, m- and p- chloro substituted diphenylcarbazones” Ind. J. Chem. 39A. 1150-
1156.
[2] Wang, C. X., Du, S. X., Li, Y. H., Wu, Y. J. (2005) “A novel 3D cadmium coordination
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pyridylcarboxyl)-2,6-pyridine dicarbohydrazide) ” Inorg. Chem. Comm. 8. 379-381.
[3] Niasari, M. S., Mostafa, R. A. (2004) “Template condensation reactions of formaldehyde with
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Zn(II), V(II), Co(II), Ni(II) and Cu(II) complexes derived from carbohydrazide Schiff base
ligands” Ind. J. Chem. 39A. 415-420.
[5] Tergioglu, N., Gurso, N. (2003) “Synthesis and anticancer evaluation of some new hydrazone
derivatives of 2,6-dimethylimidazo[2,1-b][1,3,4]thiadiazole-5-carbohydrazide” Euro. J. Med.
Chem. 38. 781-786.
[6] Affan, M. A., Fasihuddin,B. A., Ramli, B. H. Mustaffa, B. S., Bohari, M. Y. (2003) “In:
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Murtedza, Published by UNIMAS & ANALIS, Sarawak, Malaysia. 214-221.
[7] Affan, M. A. (1999) “Novel complexes of molybdenum carbonyls, organotin(IV) and
lanthanide(III) ions with hydrazones and azines” PhD Thesis, Universiti Teknologi Malaysia.
[8] Armarego, W. L. F., Perrin, D. D. (1998) “Purification of Laboratory Chemicals” 4th edition,
Butterworth Heinemann, Oxford, USA.
[9] Sheldrick, G. M. (1997) “SHELXTL V5.1. Software Reference Manual. Bruker AXS” Inc.,
Madison, WI, USA.
[10] Geary, W. G. (1971) “The use of conductivity measurement in organic solvent for the
characterization of coordination compound” Coord. Chem. Rev. 7. 81-122.
[11] Khalil, T. E., Labib, L., Iskandar, M. F. (1994) “Organotin(IV) complex with tridentate
ligand” Polyhedron 13. 2569-2578.
[12] Casas, J. S., Sanchez, A., Sordo, J., Lopez, V. A., Castellano, E. E. (1994) “Diorganotin(IV)
derivatives of salicylaldehyde thiosemicarbazone” Inorg. Chim. Acta 216. 169-172.
[13] Yearwood, B., Parkin, S., Atwood, D. A. (2002) “Synthesis and characterization of organotin
Schiff base chelates” Inorg. Chim. Acta 333. 124-131.
[14] Khoo, L. E., Xu, Y., Goh, N. K., Chia, L. S., Koh, L. L. (1997) “Molecular adducts of
diorganotin dichloride with N- (2-oxidoarylideneaminoacidato) diorganotin(IV) complexes.
Crystal structure of [Ph2Sn(2-OC10H6Ch=NCH2COO)]SnPh2Cl2” Polyhedron 16. 573-576.
[15] Dey, K. D., Lycka, A., Mitra, S., Rosair, G. M. (2004) “Simplified synthesis, 1H, 13C, 15N,
119
Sn NMR spectra and X-ray structures of diorganotin(IV) complexes containing the 4-
phenyl-2,4 butanedionebenzoylhydrazone(2−) ligand” J. Organomet. Chem. 689. 88-95.
[16] Hill, H. A. O., Morallee, K. G., Mestroni, G., Costa, G. (1968) “Electronic transmission in
some organometallic derivatives of cobalt(III) BAE and SALEN complexes” J. Organomet.
Chem. 11. 167-173.
[17] Yin, H. D., Hong, M., Wang, Q. B., Xue, S. C., Wang, D. Q. (2005) “Synthesis and structural
characterization of diorganotin(IV) esters with pyruvic acid isonicotinyl hydrazone and
pyruvic acid salicylhydrazone Schiff bases” J. Organomet. Chem. 690. 1669–1676.
[18] Ma. C., Jiang, Q. and Zhang, R. (2004) “Synthesis, characterization and crystal structures of
new organotin complexes with 2-mercapto-6-nitrobenzothiazole” Polyhedron 23. 779-786.
29
STUDY ON BIMETALLIC NI/CO CATALYST DOPED WITH PRASEODYMIUM

USING VARIOUS SUPPORT FOR CO2 / H2 METHANATION TOWARDS
PURIFICATION OF NATURAL GAS
Faridah Mohd Marsin, Nor Aziah Buang*, Wan Azelee Wan Abu Bakar, Mohd Hasmizam Razali
Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia,

81310, Skudai, Johor, Malaysia.
Abtract:In this study, a group of Ni bimetallic catalyst added with other elements to form Ni/Co/Pr catalyst with the atomic
ratio of 60:35:5 has been washcoated onto several selected support, namely cordierite (2MgO-2Al2O3-5SiO2), alumina
(Al2O3), and molecular sieve (Na12[(AlO2)12].x H2O), to determine the best support for the CO2 elimination catalyst. During
the investigations of its catalytic performance, amount of nickel loading, the reaction temperature, the durability, and the
phase formed during reaction were also examined. Based on the results obtained, cordierite was found to be the best support
for eliminating CO2 by converting to CH4 at approximate 100% at 350°C during catalytic testing. Further characterization
using X-Ray Diffraction and Scanning Electron Microscopy showed that the catalyst on cordierite support formed Ni2+ and
spinel compound Co3O4 was evenly distributed onto the support, which later will form Ni3+ and CoO after the first catalytic
testing that enhance the catalytic performance of the catalyst. The latter testing also showed no carbon decomposition of the
catalyst, which proves the durability of the catalyst against sintering.
Keywords: Ni based catalyst, CO2 elimination, cobalt, support
Abstrak: Di dalam penyelidikan ini, mangkin campuran nikel telah ditambah dengan beberapa elemen lain untuk
menyediakan mangkin Ni/Co/Pr dengan nisbah 60:35:5. Mangkin telah di isitepu ke atas beberapa penyokong terpilih iaitu,
cordierite (2MgO-2Al2O3-5SiO2), alumina (Al2O3), dan molecular sieve (Na12[(AlO2)12].x H2O). Mangkin yang disediakan
telah diuji untuk mendapatkan jenis penyokong terbaik bagi mangkin penyingkiran CO2. kajian ini memfokuskan kepada
keupayaan mangkin, suhu tindakbalas, fasa yang terbentuk, saiz zarah, dan ketahanan mangkin. Hasil diperolehi mendapati
kordierite merupakan penyokong yang terbaik bagi penyingkiran CO2. Ini kerana ia dapat menyingkirkan hampir 100% CO2
daripada sistem dengan menukarkan CO2 kepada CH4 pada suhu 350°C. Seterusnya, kajian pencirian menggunakan
pembelauan sinar X (XRD) dan Mikroskopi Pengimbas Elektron (SEM) mendapati bahawa Ni2+ dan sebatian spinel Co3O4
terbentuk di atas permukaan mangkin secara sekata dan kemudiannya berubah menjadi Ni3+ dan CoO selepas ujian
pemangkinan dijalankan.ujian pemangkinan seterusnya mendapati tiada karbon terbentuk diatas permukaan mangkin yang
menunjukkan ketahanan mangkin.
Kata kunci: mangkin berasaskan nikel, penyingkiran CO2, cobalt, penyokong
1.0 Introduction
Supported bimetallic catalysts have received much attention as often these systems exhibit
higher activity, selectivity and/or stability making them superior to the monometallic catalysts for a
given reaction [1,2]. This type of catalysts is largely employed in process in the petroleum industry
(hydrodesulfurization, hydrocarbon reforming, etc.), fine chemical synthesis (hydrogenation and
isomerization reactions), partial oxidations or automobile emission control catalysts.
Nickel oxide exhibited high activity and selectivity of methane due to the ability of NiO to
undergo reduction process owing to the presence of defect sites of the surface [3] Despite of the fast
catalyst deactivation and carbon deposition, NiO catalyst was still favored for its high thermal
stability [4], and its low price [5]. The addition of selected metal into nickel oxide base catalyst was
found to enhance the capability of the catalyst. The usage of catalyst will depend on the dopant used
in order to form a durable, sulfur tolerant, high catalytic activity catalyst.
Cobalt was mainly used for Fisher-Troposh Synthesis (FTS) and was chosen due to its
reducibility of CO. Jacobs et al. [6] found that addition of Ru and Pt exhibited a similar catalytic
effect on decreasing both the reduction temperatures of cobalt oxides. The hydrogenation of CO and
CO2 were found to catalyze by the larger cobalt clusters formed by three incipient wetness
impregnations [7]. Other research by Guczi et al. [8] found that Co–Pd samples are fully reducible
and form bimetallic particles that can be reversibly oxidized/reduced. CO hydrogenation takes place
in the range of 200–300oC producing mainly alkenes on pure cobalt catalyst with short chains.
30
Synergism on the addition of small amount of palladium to cobalt is observed and the rate of the CO
hydrogenation significantly increases.
In this research, bimetallic Ni was added with cobalt and praseodymium, in the optimized
atomic ratio of 60:35:5, on different supports; namely cordierite, alumina, and molecular sieve, and it
was used to eliminate CO2 with the presence of H2 with the hopes for CO2 hydrogenation to occur.
The reducibility and characteristics of the Ni based catalysts were tested by means of FTIR for
catalytic activity of CO2 eliminated and methane produced, X-Ray Diffraction for phase or structural
changes, Scanning Electron Microscopy for the determination of particle size and distribution.
2.0 Experimental
2.1 Catalyst preparation
Ni/Co/Pr solution in atomic ratio of 60:35:5 was prepared via optimized sol gel method. This
method includes the addition of its specific metal salts with its base metal, Ni(NO3)2.6H2O, and
second dopant, Pr(NO3)3.6H2O. Using incipient wetness method, the selected support was then
impregnated with the catalyst solution. It was then aged at 75°C for 48 hours before it was calcined at
400°C for 17 hours.
2.2 Catalytic activity measurement
The prepared samples were then tested for its capability to eliminate CO2 by methanation
reaction. This was done using a flow bed reactor of 10 mm inner diameter under atmospheric
pressure. The reaction gas mixture of CO2 and H2 was passed continuously through the catalyst that
was filled inside the sample tube that flows through the FTIR whereby the CO2 peaks from the
composition of the gas will be detected. The temperature of the sample ranged from 25-500°C. The
tabulated data gave percentages of CO2 eliminated and CH4 converted within the temperature range.
2.3 Characterization
1. X-ray Diffraction Analysis : The catalyst samples were characterized by using a Philips D5000
X-Ray Diffractometer (Cu-Kα radiation) with a degree ranging from 10-80°,
2. Scanning Electron Microscopy Analysis: Best performance catalysts were scanned using a
Philip XL 40 SEM
3.0 Results and disscussion
3.1 Catalytic activity measurements
Figure 1 showed that cordierite supported catalyst showed superiority than alumina and
molecular sieve supported catalyst. It was figured by the high percentage of CO2 elimination at 350°C
(90%). Whilst the others, Al2O3 manage to reach 90% at 500°C, and molecular sieve was at 78%, also
at 500°C. From the bar chart it is clearly seen that cordierite supported catalyst manage to yield the
highest percentage of CH4. The CH4 was detected as low as 300°C and increased steadily throughout
the testing. Meanwhile CH4 in Al2O3 supported catalyst was detected at a high temperature of 450°C,
hence molecular sieve did not detect any CH4.
As we varied the amount of Ni and Co in the catalyst, we found out that only a certain amount
of Ni and Co must be present in order to have an efficient methanation catalyst. This can be seen in
Figure 2, whereby to have a high catalytic activity the optimized ratio is Ni/Co/Pr with the ratio of
60:35:5, as it showed superiority in both CO2 elimination and CH4 conversion. With the addition of
Co in the catalyst, the amount of CO2 eliminated and CH4 detected constantly decreased. We can
conclude that the optimized ratio for the supported catalyst is 60:35:5.
31
80 100
90
70
80
60
CO2 elimination (% )
CH4 conversi on (%)
70
50
60
40 50
40
30
30
20
20
10
10
0 0
25 50 100 150 200 250 300 350 400 450 500
temperature
kd ab ms
kd ab ms
Figure 1: catalytic activity of supported catalyst
80 100
90
70
80
60
70
CH4 conversion (%)
CO2 elimination (%)

50
60
40 50
40
30
30
20
20
10
10
0 0
25 50 100 150 200 250 300 350 400 450 500
temperature
nicopr 60:35:5 nicopr 50:45:5 nicopr 47.5:47.5:5 nicopr 45:50:5

nicopr 60:35:5 nicopr 50:45:5 nicopr 47.5:47.5:5 nicopr 45:50:5
Figure 2: catalytic activity of cordierite supported Ni/Co/Pr with various ratios (a) 60:35:5, (b) 50:45:5, (c) 47.5: 47.5: 5,
and (d) 45:50:5.
percentage CH4 conversion/ CO2
100
90
80
70
elimination
60
50
40
30
20
10
0
1 2 3 4 5
experiments (TOF)
CH4 CO2
Figure 3: percentage of CO2 elimination and CH4 converted at 350°C using the same catalyst during several experiments
The best-supported catalyst was then put into test to perceive the life span of the catalyst.
Figure 3 showed percentage of CO2 elimination and the CH4 yielded at 350°C using the same catalyst
during several experiments. It was found out that after the first experiment (70% CO2; 40% CH4), the
catalytic activity of the catalyst enhances to 90% CO2 and 80% CH4. However as the experiments
continue, the catalytic activity of the catalyst started to decrease until at fifth experiment only 30% of
32
CO2 eliminated, while 20% CH4 yielded. It was predicted that the presence of Co3O4 spinel compound
enhances the catalytic activity, and as the spinel compound decreases, so does the catalytic activity. It
was assumed that the spinel compound bonded with the support to form ternary compound MgCoO.
Further study on the characterization of the catalyst was done in order to proof the assumption.
3.2 Characterization
3.2.1 X-Ray Diffraction analysis
(c)
(b)
(a)
6 1 2 3 4 5 6 7
24-0712 (*) - Magnesium Nickel Oxide -
02-1073 (N) - Magnesium Cobalt Oxide 2-Theta - Scale
42-1467 (*) - Cobalt Oxide - Co3O4
84-1221 (C) - Cordierite - Mg2Al4Si5O18
Figure 4: diffractogram of cordierite supported Ni/Co/Pr with the ratio of 60:35:5 (a) as prepared, (b) after first exposure of
CO2 and H2, (c) after fifth exposure of CO2 and H2.
Figure 4 showed the diffractogram of cordierite supported Ni/Co/Pr with the ratio of 60:35:5
(a) as prepared, (b) after first exposure of CO2 and H2, (c) after fifth exposure of CO2 and H2. As
prepared, the catalyst exhibits NiO, Co3O4 spinel compound, and also ternary compound, which are
MgNiO and MgCoO. As it was exposed to CO2 and H2, the spinel compound started to decrease and
we can see the rising of MgCoO peak, which means the addition to the ternary compound. At the fifth
exposure, the Co3O4 spinel has gone, while the MgCoO increased. This corresponds with the
catalytic activity of the catalyst, which has decreased throughout exposure.
3.2.2 Scanning Electron Microscopy
As we compare SEM micrograph of the three different supported catalysts, we can see a big
difference in its structure. Molecular sieve has the smallest particle size of 1.011 µm, followed by
alumina 1.375 µm, and the biggest particle size is by the best-supported catalyst, cordierite, with
4.375 µm. Though at this stage the particle size do not influence its catalytic activity. Though alumina
has the smallest particle size, it has the lowest catalytic activity. Both Al2O3 and molecular sieve is
homogenous, evenly distributed and covered with pores. However for cordierite, which has low
surface area, seems to form crystallites and agglomerated on the surface of the support. Thus pores
formed in between the sintering. The interaction of the catalyst and cordierite was presumed to be
strong, as it formed ternary compound in the former XRD diffractogram.
33
Figure 5: SEM alumina bead supported catalyst after calcined at 400°C for 17 hours (a) 1000 x and (b) 5000 x
magnification
Figure 6: SEM molecular sieve supported catalyst after calcined at 400°C for 17 hours (a) 1000 x and (b) 5000 x
magnification
Figure 7: SEM cordierite supported catalyst after calcined at 400°C for 17 hours (a) 1000 x and (b) 5000 x magnification
4.0 Conclusion and suggestions
From the study it was clear that cordierite supported catalyst showed superiority from the
other supports. Results showed that only optimized ratio of the catalyst on the support will show the
highest catalytic activity. The optimized ratio depicted was Ni/Co/Pr with the ratio of 60:35:5.
However the turnover frequency showed that after the second testing, the catalytic ability declined.
This was due to the transformation of Co3O4 spinel compound to ternary compound MgCoO, which
forms strong bonds between the catalyst and the support. The SEM micrographs showed
agglomeration and crystallization on the cordierite supported catalyst, compared to alumina and
molecular sieve, which have small particle size and evenly distributed particles. Further study should
be done in order to maintain the presence of Co3O4 spinel compound in the catalyst for a long period
of time to add the life span of the catalyst.
5.0 Acknowledgements
The writers would like to thank the Ministry Of Science Technology Education for a research grant.
The financial support of MOSTE under project (vot: 74178) is recognized.
6.0 References
1. Guczi, L., and Sarkany, A., in: J.J. Spivey, S.K. Agarwall (Eds.), “Catalysis” Specialist Periodical Report,
vol. 11, Royal Society of Chemistry, London, 1994, p. 318.
2. Ponec, V., Appl. Catal. A: Gen. 222 (2001) 31.
34
3. Jose, A., R., Jonathan, C., H., Anatoly, I., F., Jae, Y., K. and Manuel, P. (2001). Experimental and
Theoretical Studies on the Reaction of H2 with NiO. Role of O Vacan cies and Mechanism for Oixde
Reduction. J. Am. Chem.Soc. 124, 346-354
4. Wang, L., Murata, K., and Inaba, M. (2004). Control of the Product Ratio of CO2/ (CO+CO2) and
Inhibition of Catalyst Deactivation for Steam Reforming of Gasoline to Produce Hydrogen. Applied
Catalysis B: Environmental. 48: 243-248
5. Hou, Z., Yokota,O., Tanaka, T., and Yashima, T. (2004). Surface Properties of a Coke-free Sn Doped
Nickel Catalyst for the CO2 Reforming of Methane. Applied Surface Science. 233: 58-68.
6. Jacobs, G., Das, T.K., Zhang, Y., Li, J., Racoillet, G., and Davis, B.H. (2002). Fisher-Troposh synthesis:
support, loading, and promoter effects on the reducibility of cobalt catalyst. Applied Catalysis A: General
233: 263-281.
7. Zhang, Y., Jacobs, G., Sparks, D.E., Dry, M.E., and Davis, B.H. (2002). CO and CO2 hydrogenation
study on supported cobalt Fisher-Troposh synthesis catalysts. Catalysis Today 71: 411-418
8. Guczi, L., Schay, Z., Stefler, G., and Mizukami, F. (1999). Bimetallic catalysis: CO hydrogenation over
palladium–cobalt catalysts prepared by sol gel method. Journal of Molecular Catalysis A: Chemical. 141:
177–185.
35
KAJIAN KRISTALOGRAFI SINAR-X KOMPLEKS [µ-1(2-PIRIDIL-κN)ETANON 4-

FENILTIOUREASEMIKARBAZONATO-κ2N1,S]BIS[IODOMERKURI(II)] DAN
BIS(N-4-METOKSIBENZOIL)-N’-O-TOLILTIOUREA-κS)DIIODOMERKURI(II)
Mohd Sukeri Mohd Yusof1* & Bohari M. Yamin2.

1
Jabatan Sains Kimia, Fakulti Sains dan Teknologi Malaysia, Kolej Universiti Sains dan Teknologi Malaysia, Mengabang
Telipot, 21030 K. Terengganu. 2Pusat Pengajian Sains Kimia dan Teknologi Makanan, Fakulti Sains dan Teknologi,
Universiti Kebangsaan Malaysia, 43650 Bangi, Selangor.
Abstrak
Kompleks [µ-1(2-piridil-κN)etanon 4-feniltioureasemikarbazonato-κ2N1,S]bis[iodomerkuri(II)], dan

Bis(N-4-metoksibenzoil)-N’-o-toliltiourea-κS)diiodomerkuri(II), mempunyai formula molekul
masing-masing Hg2(L)2I2 dan Hg(L)2I2. Kedua-dua kompleks, mempunyai sistem hablur monoklinik
dengan kumpulan ruang masing-masing adalah C2/c, a=18.466(3)Å, b=16.745(2)Å, c=13.9140(19)Å,
β=129.174(2)º dan P21/c, a=10.848(3)Å, b=24.474(7)Å, c=14.038(4)Å, β=92.206(4)º. Dalam
kompleks merkuri-bes Schiff, atom Hg mempunyai geometri segi empat pyramid di mana ligan
terikat secara tridentat melalui atom NNS. Atom Sulfur kedua-dua ligan membentuk jejambat
menghubungkan dua atom Hg menjadikan bentuk keseluruhan molekul seperti kotak. Sebaliknya,
dalam kompleks merkuri-tiourea atom merkuri berkoordinat dengan ligan melalui atom S kumpulan
tion secara monodentat dan bersifat dimerik ,menjadikan geometri Hg tetrahedron terherot.
Abstract
Both [µ-1(2-pyridyl-κN)ethanone 4-phenylthioureasemicarbazonato-κ2N1,S]bis[iodomercury(II)], and

Bis(N-4-methoxybenzoyl)-N’-o-tolylthiourea-κS)diiodomercury(II), complexes have the molecular
formula of Hg2(L)2I2 and Hg(L)2I2 where L is the ligand, respectively. They also crystallized in
monoclinic system, but having space group is C2/c, a=18.466(3)Å, b=16.745(2)Å, c=13.9140(19)Å,
β=129.174(2)º and P21/c, a=10.848(3)Å, b=24.474(7)Å, c=14.038(4)Å, β=92.206(4)º, respectively. In
the first complex, the Schiff base ligands are coordinated to the mercury atom via their (NNS) donor
atoms in a tridentate manner. The sulfur atoms act as a bridge connecting the Hg atoms, forming a
rectangular base that almost perpendicular to the ligand planes, resulting in an open box-like structure.
The geometry of the mercury atom is close to a square pyramid. However, the mercury atom in the
mercury-thiourea complex is coordinated to the ligands via the thiono sulfur atoms in a monodentate
manner and form a distorted tetrahedral geometry.
Kata kunci: Merkuri kompleks, tiourea, benzoiltiourea
Pengenalan
Masalah pencemaran oleh logam berat terutama dalam sistem akueus seperti air buangan yang
disalurkan ke sungai masih merupakan ancaman hingga ke hari ini. Kilang-kilang yang bertanggung
jawab sudah pasti sepatutnya menerima baik sebarang kaedah atau teknik yang boleh memisahkan
logam berat dan toksik di loji masing-masing daripada menyalurkan ke dalam sungai. Untuk itu
penyelidikan mencari bahan dan teknik yang murah, mudah dan efisien untuk memisahkan logam-
logam toksik terus berkembang. Satu daripada kaedah pemisahan logam tersebut adalah melalui
pengkompleksan dan seterusnya terpisah daripada larutan. Kajian awal kami mendapati terbitan
tiourea mampu membentuk kompleks dengan beberapa logam kini mendapat tumpuan. Baru-baru ini
benzoiltiourea yang diubahsuai di atas silica mesoporos MCM-48 didapati menjerap raksa daripada
larutan akueus (Olkhovyk et al. 2004). N-benzoiltiourea yang diubahsuai bersama polimer PAMAM
dapat memisah secara terpilih dan mengumpul semula ion logam berat daripada buangan akueus
(Rether & Schunster, 2003). Beberapa sebatian terbitan tiourea telah menunjukkan kemampuan yang
36
tinggi untuk membentuk kompleks dengan logam berat terutamanya raksa dan kadmium (Yamin &
Yusof, 2004). Kajian struktur tentang kompleks merkuri-tiourea telah banyak dilaporkan, tetapi
kebanyakannya adalah kompleks merkuri-siklotiourea seperti Hg(C8H16N4S2)Cl2, Hg(C8H16N4S2)Br2,
Hg(C8H16N4S2)I2, Hg(C8H16N4S2)(SCN)2 dan Hg(C8H16N4S2)(CN)2 (Popovic et al., 2000). Hanya
terdapat beberapa laporan tentang kompleks merkuri-tiourea (gelang terbuka) seperti Bis(o-
klorofenilbenzoiltiourea-κS)-diiodomerkuri(II) (Yusof et.al.,2004) dan bis{µ-kloro-kloro[(N-
dietilaminotiokarbonil)benzimido-O-metil ester-S]merkuri(II) (Leβmann et al., 2000). Kertas ini
membincangkan struktur kompleks merkuri-bes Schiff dan merkuri-tiourea.
Eksperimen
Tindak balas pengkompleksan dilakukan dengan merefluks ligan N-fenil-2-(1-piridin-2-

iletil)hidrazinakarbotioamida dan N-4-metoksibenzoil)-N’-o-toliltiourea masing-masing dengan
merkuri iodida (HgI2) dalam pelarut etanol selama 3 jam. Hasil refluks dituras dan dibiarkan menyejat
pada suhu bilik dan menghasilkan hablur yang kekuningan. Hablur yang diperolehi dianalisis untuk
menentukan struktur hablur dengan menggunakan alat pembelauan sinar-X hablur tunggal.
Keputusan & Perbincangan
Kompleks Bis[µ-1(2-piridil-κN)etanone 4-feniltioureasemikarbazonato-κ2N1,S]bis[iodomerkuri(II)],

Hg2(C14H13N4S)2I2], (I) dan Bis(N-4-metoksibenzoil)-N’-o-tolyltiourea-κS)diiodomerkuri(II),
Hg[C16H16N2O2S]2I2, (II), masing-masing mempunyai sistem hablur monoklinik dengan kumpulan
ruang C2/c, sel unit a=18.466(3)Å, b=16.745(2)Å, c=13.9140(19)Å dan β=129.174(2)º dan P21/c,
a=10.848(3)Å, b=24.474(7)Å, c=14.038(4)Å dan β=92.206(4)º. Data hablur dan parameter
penghalusan kedua-dua kompleks ditunjukkan dalam Jadual 1.
Jadual 1: Data hablur dan penghalusan data hablur kompleks (I) dan (II)
Perkara Kompleks I Kompleks II

Formula empirik Hg2(C14H13N4S)2I2 Hg[C16H16N2O2S]2I2
Berat molekul 1193.67 1055.13
Sistem hablur, kumpulan ruang Monoklinik, C2/c Monoklinik, P21/c
Dimensi sel unit a = 18.466(3)Å a=10.848(3)Å,
b = 16.745(2)Å b=24.474(7)Å,
c = 13.9140(19)Å c=14.038(4)Å
β = 129.174(2)º β=92.206(4)º
Isipadu 3335.3(8) Å 3 3724.2(18)
Z, ketumpatan 4, 2.377 Mg/m3 4, 1.882 Mg/m3
Saiz hablur 0.38 x 0.18 x 0.13 mm 0.47 x 0.39 x 0.07 mm
Kawasan theta untuk pungutan data 1.8 hingga 27.6º 1.66 hingga 27.50
Set data -24<=h<=24, -21<=k<=21, -14<=h<=14, -31<=k<=21,
-18<=l<=18 -18<=l<=18
Pungutan pantulan/ unik 3875 / 3277 [R(int) = 0.033] 40517/8483[R(int) = 0.053]
Kesempurnaan kpd. theta=27.6/27.5 º 99.5% 99.1%
Pembetulan serapan Multi-scan Multi-scan
Mak. dan min. transmisi 0.9288 dan 0.8624 0.6811 dan 0.1667
Kaedah pemprosesan Kuasa dua terkecil matriks- Kuasa dua terkecil matriks-
lengkap lengkap
Data / kekangan / parameter 3175 / 0 / 193 8483/0/406
Nilai ketepatan struktur (GooF) 1.081 1.084
Indeks akhir R [I>2sigma(I)] R1 = 0.0423, wR2 = 0.1124 R1 = 0.0413, wR2 = 0.0911
Indeks R (semua data) R1 = 0.0506, wR2 = 0.1189 R1 = 0.0560, wR2 = 0.0975
Perbezaan puncak dan lubang 0.219 dan -0.237 e. Å-3 1.343 dan -0.842 e. Å-3
37
Bagi kompleks (I), ligan N-fenil-2-(1-piridin-2-iletil)hidrazinakarbotioamida merupakan ligan

tridentat iaitu berkoordinat dengan atom Hg1 melalui atom N3, N4 dan S1i ( Rajah 1). Ini adalah satu
contoh logam merkuri berkoordinat lebih daripada 2 yang jarang berlaku. Molekul kompleks adalah
dimerik dengan dua atom Hg dihubungkan dengan dua ikatan jejambat daripada dua atom S dan
molekul berbentuk seperti kotak yang terbuka. Panjang ikatan Hg-S [2.6840(16)Å] dalam kompleks I
adalah pendek sedikit berbanding dalam kompleks bis(o-klorofenilbenzoiltiourea-κS)-
diiodomerkuri(II) (Yusof et al., 2003). Panjang Ikatan C7-S1, 1.768(4) Å adalah lebih panjang
dibandingkan dengan kompleks bis(o-klorofenilbenzoiltiourea-κS)-diiodomerkuri(II) [1.690(6) Å].
Panjang ikatan dan sudut yang lain adalah dalam jarak yang normal (Allen et al., 1987; Orpen et al.,
1989) dan juga bersamaan dengan kompleks bes-Schiff yang lain seperti [Mn(C14H13N4S2)2] (Usman
et al., 2002). Sudut dihedral bagi kedua-dua ligan ialah 9.07(11)º.
Rajah 1: Gambarajah ORTEP molekul kompleks (I) pada kebarangkalian 30% tanpa atom H untuk
gambaran lebih jelas.
Geometri bagi kedua-dua atom Hg adalah antara trigonal bipiramid dan piramid planar tetapi
mendekati kepada piramid planar. Atom N3, N4, S1i dan I1 adalah sesatah dengan pesongan
maksimum sebanyak 1.491(1) Å. Sudut pusat atom Hg adalah antara 67.63(12) dan 107.74(2)º.
Jadual 2: Panjang ikatan dan sudut ikatan dalam kompleks (I)
Ikatan Panjang Ikatan, Å Ikatan Sudut Ikatan, º

Hg1-N4 2.387(4) N4-Hg1-N3 67.63(2)
Hg1-N3 2.398(3) N4-Hg1-S1i 140.71(9)
Hg1-S1i 2.6173(11) N3-Hg1-S1i 73.32(9)
Hg1-S1 2.6490(12) N4-Hg1-S1 95.73(9)
Hg1-I1 2.6635(4) N3-Hg1-S1 97.71(8)
N2-C7i 1.290(5) S1i-Hg1-S1 93.48(3)
Molekul kompleks distabilkan oleh ikatan hidrogen inter-molekul, N1-H1A…I1ii, [simetri kod; (ii)
3/2-x,1/2-y,1-z] membentuk rantaian zig-zag yang selari pada paksi-a (Rajah 2).
38
Rajah 2: Gambarajah padatan molekul dalam sistem hablur, dilihat ke dalam paksi c. Garisan putus-
putus menunjukkan ikatan hydrogen inter-molekul N-H…I.
Kompleks (II) adalah dimerik (Rajah 3) dengan dua molekul ligan berkoordinat dengan atom pusat,
Hg1 melalui atom sulfur, S. Atom pusat Hg1 mempunyai geometri tetrahedral terherot dengan sudut
antara 96.18(4)º dan 133.75(2)º. Panjang ikatan dan sudut ikatan adalah normal (Allen et al., 1989;
Orpen et al., 1980) dan berpadanan dengan kompleks bis(o-klorofenilbenzoiltiourea-κS)-
diiodomerkuri(II) (Jadual 4).
Rajah 3: Gambarajah ORTEP molekul kompleks dengan kebarangkalian 50% tanpa atom H untuk
gambaran lebih jelas.
Jadual 3: Panjang ikatan dan sudut ikatan dalam kompleks
Ikatan Panjang Ikatan, Å Ikatan Sudut Ikatan, º

Hg1-I1 2.6558(6) I1-Hg1-I2 133.75(2)
Hg1-I2 2.6700(6) I1-Hg1-S2 104.47(3)
Hg1-S1 2.6762(13) I2-Hg1-S2 110.47(3)
Hg1-S2 2.7192(14) I1-Hg1-S1 108.21(3)
S1-C8 1.698(5) I2-Hg1-S1 97.31(3)
S2-C24 1.709(4) S2-Hg1-S1 96.18(4)
39
Panjang ikatan Hg1-I1 dan Hg1-I2 [2.6558(6)Å dan 2.6700(6)Å], didapati lebih pendek jika
dibandingkan dalam kompleks Hg(C8H16N4S2)I2 [2.7123(5)Å] (Popovic et al., 2000) tetapi
bersesuaian dengan panjang ikatan seperti dalam kompleks bis(o-klorofenilbenzoiltiourea-κS)-
diidomerkuri(II) [2.6582(6)Å] (Yusof et al., 2004). Ikatan Hg1-S1 [2.7192(14)Å adalah lebih panjang
berbanding dalam kompleks bis(o-klorofenilbenzoiltiourea-κS)-diidomerkuri(II) [2.6840(16)Å].
Rajah 4: Gambarajah padatan molekul kompleks yang dilihat pada permukaan ab. Garisan putus-
putus menunjukkan ikatan hidrogen inter-molekul N-H…O dan C-H…I.
Terdapat enam ikatan hydrogen intra-molekul, N-H…S, N-H…O dan C-H…N [Jadual 3] yang
memberikan 4 gelang pseudo-enam ahli (Rajah 3). Dalam sistem hablur, molekul terikat melalui
ikatan hidrogen inter-molekul N-H…O dan C-H…I (Jadual 4) membentuk dimer yang dilihat melalui
permukaan ab (Rajah 4).
Jadual 4: Senarai ikatan hidrogen intra- dan intermolekul dalam kompleks (Å,°).
D-H…A D-H H…A D…A D-H…A

N1-H1A…S2 0.86 2.76 3.5135 147
N2-H2A…O1 0.86 1.98 2.6558 134
N3-H3A…S1 0.86 2.76 3.5762 159
N4-H4A…O3 0.86 1.98 2.6419 133
…
C16-H16A N2 0.96 2.38 2.8546 110
C32-H32A…N4 0.96 2.38 2.8660 111
N2-H2A…O3i 0.86 2.36 3.0661 139
N4-H4A…O1ii 0.86 2.32 3.0124 137
C16-H16B…I2i 0.96 2.97 3.9245 173
Simetri kod; (i) 1+x,y,z; (ii) -1+x,y,z
Penghargaan
Penghargaan diberikan kepada Kerajaan Malaysia, Universiti Kebangsaan Malaysia dan Kolej
Universiti Sains dan Teknologi Malaysia diatas kemudahan yang telah diberikan
40
Rujukan
Allen, F.H., Kennard, O., Watson, D.G., Orpen, A.G. & Taylor, R. 1987. J. Chem. Soc. Perkin Trans.
2:S1-19.
Leßmann, F., Beyer, L., & Sieler, J. 2000. Synthesis and X-ray structure of the first chloro-bridged
thiourea mercury(II) complex [C6H5C(OCH3)NC(S)N(C2H5)2HgCl2]2. Inorganic Chemistry
Communications. Vol 3: Issue 2:62-64.
Olkhovyk, O., Antochshuk, V., & Jaroniec, M. 2004. Benzoylthiourea-modified MCM-48
mesoporous silica for mercury(II) adsorption from aqueous solutions. Colloids and Surfaces A:
Physicochemical and Engineering Aspect. Vol. 236, 1-3:69-72.
Orpen, A.G., Brammer, L., Allen, F.H., Kennard, O., Watson, D.G., & Taylor, R. 1989. J. Chem. Soc.
Dalton Trans. S1-83.
Popovic, Z., Pavlovic, G., Matkovic, D., Soldin, Z., Rajic, M., Vikic, D., & Kovacek, D. 2000.
Mercury(II) complexes of heterocyclic thiones.: Part 1. Preparation of 1:2 complexes of mercury(II)
halides and pseudohalides with 3,4,5,6-tetrahydropyrimidine-2-thione. X-ray, thermal analysis and
NMR studies. Inorganica Chimica Acta, Vol. 306: Issue 2:142-152.
Rether, A. & Schuster, M. 2003. Selective separation and recovery of heavy metal ions using water-
soluble N-benzoylthiourea modified PAMAM polymers. Reactive and Functional Polymers, Vol. 57,
1:13-21.
Yamin, B. M. & Yusof, M. S. M. 2003. N,N'-Bis(benzamidothiocarbonyl)hydrazine
dimethylsulfoxidedisolvate. Acta Cryst. E59: o358-o359.
Yamin, B. M. & Yusof, M. S. M. 2003. N-[N-(Furan-2-carbonyl)hydrazine thiocarbonyl]benzamide.
Acta Cryst. E59:o124-o126.
Yamin, B. M. & Yusof, M. S. M. 2003. N'-Benzoyl-N-p-bromophenylthiourea. Acta Cryst.. E59:
o340-o341.
Yamin, B. M. & Yusof, M. S. M. 2003. N-Benzoyl-N'-phenylthiourea. Acta Cryst. E59: o151-o152.
Yamin, B. M., Kassim, M. B., Yusof, M. S. M. & Shah, N. M. 2004. N-Benzoyl-N'-ethylthiourea.
Acta Cryst. E60: o556-o557.
Yusof, M. S. M. & Yamin, B. M. 2004. N-Benzoyl-N’-(2-chlorophenyl)thiourea. Acta Cryst. E60:
o1403-o1404.
Yusof, M. S. M. & Yamin, B. M. 2003. 3-(3-Benzoylthioureido)propionic acid. Acta Cryst. E59:
o828-o829.
Yusof, M. S. M., Yamin, B. M. & Shamsuddin M. 2003. N-(N-Benzoylhydrazino
carbothioyl)benzamide. Acta Cryst. E59: o810-o811.
Yusof, M. S. M., Yamin, B. M. & Kassim, M. B. 2004. Bis(o-chlorophenylbenzoyl thiourea-
S)diiodomercury(II). Acta Cryst. E60: m98-m99.
41
SUBCRITICAL WATER EXTRACTION OF ESSENTIAL OIL FROM CORIANDER

(Coriandrum sativum L.) SEEDS
Norashikin Saim*, Rozita Osman, Wan Azriza Hirmi Md Yasin, Rossuriati Dol Hamid
Fakulti Sains Gunaan, Universiti Teknologi MARA

40450 Shah Alam, Selangor
e-mail: noras691@salam.uitm.edu.my
Abstract
Subcritical water extraction (SWE) is a technique based on the use of water as an extractant, at temperatures
between 100 and 374 ºC and at a pressure high enough to maintain the liquid state. As the temperature of liquid
water is raised under pressure, the polarity decreases and it can be used as an extraction solvent for a wide range
of compounds. The application of SWE in the extraction of essential oil from coriander (Coriandrum sativum
L.) seeds was studied. Ground coriander seeds (3-4 g) were subjected to SWE with water for an extraction time
of 15 min under several extraction conditions (pressures of 870 and 1000 psi and temperatures of 65, 100 and
150 ºC). The SWE method was compared with hydrodistillation performed by treating 10 g of ground coriander
seeds with 100 mL of water for 3 hours. Compounds were removed from the aqueous extract with hexane and
determined by gas chromatography mass spectrometry (GC-MSD). It was found that the efficiency (g oil/g of
coriander) of SWE was higher than that provided by hydrodistillation with reduced extraction time. The major
compounds found were linalool, isoborneol, citronellyl butyrate and geraniol. SWE method has the possibility
of manipulating the composition of the oil by varying the temperature and adjusting the pressure.
Keywords: Subcritical water extraction, Coriander, Coriandrum Sativum L.
Introduction
Before the essential oils of plants can be analyzed, they have to be extracted and concentrated, and a
number of different methods can be used for that purpose such as hydrodistillation, steam distillation
and solvent extraction. Air dried leaves are usually subjected to hydrodistillation or steam distillation
for about 3-8 hours and the essential oils are dried over anhydrous sodium sulfate. In solvent
extraction (cold or hot), extraction of the air-dried plant material is carried out using an appropriate
solvent depending on the nature of the essential oils. Several drawbacks such as low yield, losses of
volatile compounds, long extraction times, degradation of unsaturated compounds and toxic solvent
residue in the extract may be encountered with the current extraction methods [1]. These
shortcomings have led to the emergence of new techniques in the extraction of essential oils such as
supercritical CO2 extraction (SFE) [1,2] and solvent free microwave extraction [3].
In recent years extraction with superheated water or subcritical water extraction (SWE) has been used
widely for the extraction of polar and non polar analytes from environmental samples. SWE has also
been applied in the extraction of essential oil from plants [4,5]. In this work subcritical water
extraction of coriander seeds was compared to hydrodistillation in terms of extraction yields and
essential oil composition. The influence of extraction conditions (temperature and pressure) on the
composition of essential oil was analyzed.
Experimental
Sample preparation
Coriander seeds were purchased from local stores. The seeds were crushed before analysis.
Subcritical water extraction (SWE)

SWE was performed using an accelerated solvent extraction system, ASE 200, equipped with a
solvent controller unit from Dionex Corporation. Extractions were performed using water with a static
42
extraction time of 15 min at different extraction temperatures and pressures (SWE 1: 870 psi, 65 ˚C,
SWE 2: 870 psi, 100 ˚C, SWE 3: 870 psi, 150 ˚C, SWE 4: 1000 psi, 65 ˚C, SWE 5: 1000 psi, 100 ˚C,
SWE 6: 1000 psi, 150 ˚C) All extractions were performed in 22 mL extraction cells, containing 3 g
of sample. The extraction procedure was as follows: (i) sample is loaded into cell; (ii) cell is filled
with solvent up to a specific pressure and temperature (iii) a static extraction is performed for 15 min
(v) the cell is rinsed (with 60% cell volume using extraction solvent); and (vi) solvent is purged from
the cell with N2 gas. Following SWE, liquid-liquid extraction step was performed using hexane and
HCl was added to facilitate the breaking of emulsion. The obtained oil was separated and the yield
was determined in terms of dry basis yield. The oil was stored in a freezer at -4 °C until analyzed by
GC–MSD.
Hydrodistillation
Ten grams of crushed coriander seeds were subjected to hydrodistillation using 100 mL of distilled
water for 3 h after the mixture had reached boiling point (100 °C). Liquid-liquid extraction step was
performed using hexane. The obtained oil was separated, and the yield was determined. It was stored
in a freezer at -4 °C until analyzed by GC–MSD.
Gas chromatography-mass spectrometry (GC-MS) analyses

GC-MS analysis was performed on an Agilent gas chromatography model 6890N coupled to a mass
selective detector 5973 inert. Compounds were separated on a cross-linked fused silica capillary
column HP5-MS (30m x 250µm x 0.25µm). The head pressure of the carrier gas helium (high purity)
was 50 kPa. The temperature programmed was set at an initial 60 °C, followed by an increase by 10
°C min-1 to 200 °C and held for 15 minutes. The MS detector was operated in the full scan mode with
70 eV electrons ionization, by scanning a mass range of m/z 35-450 in 0.45 s. The system was
computer-controlled using Agilent GC-MSD ChemStation. Compounds were identified by matching
their mass spectra with the flavour spectral library with a resemblance percentage above 90 %.
Results and Discussion
Extraction yield and time

The yields (in terms of g of essential oil/1 g of coriander seeds) of SWE under these conditions (0.6 -
0.8%) were found to be higher than that obtained by hydrodistillation (0.06-0.1%). Lower yields (~
0.4%) of SWE were obtained at high temperature (150 °C) under both pressure conditions. The
decreased in yield at higher temperature was also reported by other researchers [4,6]. It was observed
that there were no significant difference in the amounts of extracted oil obtained under different
pressures but same temperatures. The reduced extraction time (3 hours for hydrodistillation against
15 min for SWE) is clearly advantageous for the SWE method.
Composition of essential oil

For SWE extraction of coriander seeds, experiments were performed at different combination of
extraction pressure and temperature. Identification of compounds extracted using these techniques
were performed by GC-MS. Figure 1 shows the total ion current (TIC) chromatogram of coriander
seed extract obtained using the two methods. Major compounds identified were linalool, isoborneol,
citronellyl butyrate and geraniol. Comparison in the composition of coriander extracts obtained using
various extraction techniques is shown in Table 1.
Table 1. Compounds (%)* extracted from coriander by HD and SWE under different conditions.
Compound HD SWE 1 SWE 2 SWE 3 SWE 4 SWE 5 SWE 6
(870 psi, (870 psi, (870 psi, (1000 psi, (1000 psi, (1000 psi,
65 ˚C) 100 ˚C) 150 ˚C) 65 ˚C) 100 ˚C) 150 ˚C)
Linalool 66.5 87.8 87.7 68.7 88.6 85.4 79.5
Isoborneol 6.8 6.1 3.8 0.8 6.1 2.9 4.3
Citronellyl butyrate 13.4 2.2 1.2 3.7 1.6 1.4 1.3
Geraniol 13.3 4.0 7.3 26.8 3.6 10.4 14.9
*Percent of component based on area normalization.
43
(a) (b)
Figure 1. Total ion current (TIC) chromatograms of compounds of coriander seeds extracted by
SWE (a) and HD (b).
In general, the relative amounts of the major compounds obtained using HD and SWE under different
conditions are comparable. Linalool is the most abundant composition of coriander isolated either by
SWE under all conditions (68.7-88.6%) and HD (66.5%). However, the relative amount of citronellyl
butyrate obtained using SWE is slightly lower than those obtained using hydrodistillation. For SWE,
a change in extraction temperature resulted in a slight change of composition of the extracted oil. The
percentage of geraniol increases with temperature while the percentage of linalool and isoborneol
decreases at high temperature. Therefore using SWE, specific composition of essential oils can be
obtained by optimizing the extraction temperature. Extraction pressure however did not seem to give
significant effect on the oil composition as shown by the similarity of the oil composition for the
extracts obtained under different pressures but same temperature ie. SWE 1 vs SWE 4 and SWE 2 vs
SWE 5. It is noteworthy that the coriander extracts obtained using SWE produced a pleasant odor
similar to the starting plant material.
Conclusions
The use of SWE has shown to be a good alternative for the extraction of essential oils. Essential oil of
coriander obtained using SWE (at temperature 65-150 ˚C and pressure 870-1000 psi) was comparable
to that obtained from hydrodistillation. SWE enables rapid extraction and has the advantage of being
selective because it is possible to manipulate the extract composition under a given working
conditions.
Acknowledgement
The authors would like to acknowledge financial support from IRDC, Universiti Teknologi Mara for
funding this project (IRDC Project number: 600-IRDC/ST 5/3/744).
References
[1] Khajeh, M., Yamimi, Y., Bahramifar, N., Sefidkon, F., Pirmoradei, M.R. (2005) “Comparison of
essential oils compositions of Ferula assa-foetida obtained by supercritical carbon dioxide extraction and
hydrodistillation methods” Food Chemistry. 91. 639-644.
44
[2] Diaz-Maroto, M.C., Pérez-Coello, M.S., Cabezudu, M.D., (2002) “Supercritical carbon dioxide extraction
of volatiles from spices. Comparison with simultaneous distillation-extraction” J. Chromatogr. A. 947.
23-29.
[3] Lucchesi, M.E., Chemat, F., Smadja, J., (2004) “Solvent-free microwave extraction of essential oil from
aromatic herbs: comparison with conventional hydro-distilation” J. Chromatogr. A. 1043. 323-327.
[4] Ozel, M.Z., Gogus, F., Lewis, A.C. (2003) “Subcritical water extraction of essential oils from Thymbra
spicata” Food Chemistry. 82. 381-386.
[5] Ozel, M.Z., Gogus, F., Kaymaz, H. (2004) “Superheated water extraction, steam distillation and Soxhlet
extraction of essential oils of Origanum onites” Anal Bioanal Chem. 379. 1127-1133.
[6] Soto Ayala, R., Luque de Castro, M.D. (2001) “Continuous subcritical water extraction as a useful tool
for isolation of edible essential oils” Food Chemistry. 75. 109-113.
45
ANALISIS TOLUENA RESIDU DALAM PEMBUNGKUS MAKANAN DENGAN KAEDAH

PENGEKSTRAKAN RUANG KEPALA KROMATOGRAFI GAS
Lim Ying Chin1 and Mohd Marsin Sanagi2

1
INTEC, Universiti Teknologi MARA Section 17 Campus, 40200 Shah Alam, Selangor.
2
Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 81310 Skudai, Johor.
Tel: 607-55227155, Fax: 607-55227109, e-mail: limyi613@salam.uitm.edu.my
Abstrak. Bahan polimer banyak digunakan sebagai bahan pembungkusan makanan. Toluena residu yang tertinggal di dalam
bahan pembungkusan boleh meresap masuk ke dalam makanan yang dibungkus dan seterusnya menjejaskan kualiti
makanan. Dalam kajian ini, penganalisisan ruang kepala yang manual telah berjaya direkabentuk dan dibangunkan.
Penentuan kepekatan toluena residu dilakukan dengan kaedah penambahan piawai dan kaedah pengekstrakan ruang kepala
berbilang (MHE) menggunakan kromatografi gas-pengesan pengionan nyala (GC-FID). Pengenalpastian toluena dilakukan
dengan perbandingan masa penahanan toluena piawai dan GC-MS. Kajian mendapati suhu pemanasan yang paling sesuai
ialah 180 oC dengan masa optimum ialah 10 minit. Kajian juga menunjukkan sampel berbilang warna mempunyai kepekatan
toluena residu yang lebih tinggi berbanding sampel satu warna. Sampel yang dianalisis dengan kaedah penambahan piawai
didapati mempunyai kepekatan toluena residu yang lebih tinggi berbanding dengan kaedah MHE. Tetapi, perbandingan
dengan kepekatan toluena residu yang diperoleh daripada makmal De'Paris, Perancis mendapati bahawa sampel
berkepekatan rendah yang dianalisis dengan kaedah MHE mempunyai peratus ketepatan yang tinggi. Sementara itu, sampel
berkepekatan tinggi pula mempunyai peratus ketepatan yang rendah disebabkan oleh ralat bersistem. Perbandingan kaedah
penentuan toluena mendapati kaedah MHE adalah lebih presis berbanding dengan kaedah penambahan piawai.
Abstract. Polymeric materials are used in many food contact applications as packaging material. The presence of residual
toluene in this food packaging material can migrate into food and thus affect the quality of food. In this study, a manual
headspace analysis was successfully designed and developed. The determination of residual toluene was carried out with
standard addition method and multiple headspace extraction (MHE) method using gas chromatography-flame ionization
detector (GC-FID). Identification of toluene was performed by comparison of its retention time with standard toluene and
GC-MS. It was found that the suitable heating temperature was 180 oC with an optimum heating time of 10 minutes. The
study also found that the concentration of residual toluene in multicolored sample was higher compared to monocolored
sample whereas residual toluene in sample analyzed using standard addition method was higher compared to MHE method.
But, comparison with the results obtained from De'Paris laboratory, France found that MHE method gave higher accuracy
for sample with low analyte concentration. On the other hand, lower accuracy was obtained for sample with high
concentration of residual toluene due to the systematic error. Lastly, comparison between determination methods showed
that MHE method is more precise compared to standard addition method.
Kata kunci: pengekstrakan ruang kepala, bahan pembungkusan makanan, kromatografi gas
Pengenalan
Pembungkusan merupakan suatu proses yang tersusun untuk menyediakan barangan bagi tujuan
pengangkutan, pengedaran, penyimpanan, pemasaran dan penggunaan akhir. Kini, bahan
pembungkusan bertindak sebagai alat komunikasi untuk membolehkan pengguna mengenalpasti
produk atau jenama yang baru, mengetahui kandungan nutrient yang hadir dan dalam sesetengah kes,
bahan pembungkus memberi maklumat tentang cara penyediaan makanan tersebut. Pembungkus
fleksibel seperti lamina mengandungi beberapa lapisan filem dan pelarut toluena biasanya digunakan
dalam percetakan grafik pada filem ini.
Perhatian perlu diberikan kepada bahan pembungkus apabila terdapat kemungkinan
berlakunya keracunan makanan yang disebabkan oleh resapan bahan pencamar ke dalam makanan
semasa pemyimpanan makanan. Peresapan bahan ini mempunyai kesan dari segi kualiti dan
ketoksikan makanan. Ia mungkin mempunyai bau dan boleh memberi kesan kepada rasa makanan.
Keadaan sebaliknya seperti penyerapan rasa makanan oleh pembungkus juga boleh mengurangkan
intensiti rasa makanan dan aroma makanan. Bahan pencemar dalam pembungkus makanan bukan
sahaja menjejaskan kualiti makanan malahan ia mungkin membawa kesan negatif ke atas kesihatan
pengguna sekiranya kepetan bahan pencemar melebihi had kepekatan yang dibenarkan.
Kajian ini melibatkan perekabentukan dan pengubahsuaian penganalisisan ruang kepala yang
manual. Dalam kajian ini, suhu dan masa pemanasan dioptimumkan dan seterusnya kuantiti toluena
residu akan ditentukan menggunakan kaedah penambahan piawai dan kaedah pengekstrakan ruang
46
kepala berbilang (MHE) menggunakan kromatografi gas-pengesan pengionan nyala (GC-FID).

Perbandingan kepekatan toluena turut dilakukan di antara sampel satu warna dan berbilang warna
dalam 6 jenis pembungkus makanan.
EKSPERIMEN
Reagen
Larutan piawai toluena (99 %) daripada keluaran J. T. Baker, Phillipsburg, USA. Heksana
(95 %) gred GC diperoleh daripada Fisher Scientific, New Jersey, USA. Hidrokarbon piawai,
nonana (C9H20) adalah gred analar daripada jenama BDH, England. Gas helium pula
dibekalkan oleh Syarikat MOX, Pasir Gudang, Johor.
Alat radas
Vial jenis ruang kepala dengan bukaan luas bersaiz 10 mL (22.5 mm × 46 mm) dan 2 mL (12 mm ×
32 mm) daripada Supelco, Bellefonte, PA, USA. Penutup pembebasan tekanan jenis aluminium yang
mempunyai septum dengan pelapis jenis silikon / PTFE yang bersaiz 20 mm dan penutup yang
mempunyai septum teflon (11 mm) juga daripada Supelco, Bellefonte, PA, USA. Penyuntik jenis
kedap udara dengan saiz 100 µL keluaran Hamilton, Reno, Nevada, USA. Penyuntik kedap udara
yang bersaiz 1 mL pula daripada SGE, Australia. Crimper tangan yang dapat dilaraskan bukaan
saiznya jenis 20 mm dan decapper tangan jenis 20 mm yang digunakan untuk membuka penutup vial
ruang kepala pula adalah keluaran Kimble Glass Inc, Vineland, NJ, USA.
Instrumentasi
Analisis toluena residu dijalankan dengan menggunakan kromatografi gas Hewlett Packard model
6890 GC dengan pengesan pengionan nyala (FID). Turus pemisahan yang digunakan ialah turus
rerambut Ultra 1 (dimetilpolisiloksana) keluaran Hewlett Packard, USA. Turus yang berukuran 30 m
ini berdiameter 0.25 mm dengan ketebalan filem 0.17 µm. Dalam kajian ini, helium digunakan
sebagai gas pembawa pada kadar alir 2 mL/min. Suhu liang suntikan pada 150oC dan suhu pengesan
ialah 250oC. Suhu GC isoterma digunakan pada 150 oC dan masa analisis yang diperlukan adalah 5
minit. Gas ruang kepala (1 mL) disuntik dengan menggunakan mod suntikan berpecah 1 : 10.
Pensampelan
Enam jenis pembungkus yang digunakan dalam kajian ini iaitu sampel TE, CCB, MCP, PM, TC dan
PDC disumbangkan oleh Kilang BBM Sdn. Bhd., Tampoi, Johor. Sampel diletakkan bersama untuk
membentuk blok yang padat. Blok sampel yang padat ini dibungkus ketat dengan kerajang aluminium
yang mempunyai ketebalan 30 - 40 µm. Nisbah di antara luas spesimen (dalam cm2) terhadap isipadu
vial (dalam mL) ditetapkan pada 5. Sebanyak 15-20 kepingan sampel dikeluarkan daripada "blok"
sampel tanpa memisahkannya. Setelah mengeluarkan sekurang-kurangnya satu lapisan kepingan
sampel daripada bahagian atas blok sampel, kepingan sampel yang seterusnya diambil dan spesimen
dipotong menggunakan templat. Spesimen itu digulung dan dimasukkan ke dalam vial dengan segera.
Selepas itu, vial ditutup dengan cepat menggunakan crimper tangan.
Prosedur
Dalam kajian ini, dua kaedah digunakan dalam penentuan kuantitatif toluena residu iaitu kaedah
penambahn piawai dan kaedah MHE. Kedua-dua kaedah ini merupakan kaedah piawai seperti yang
tercatat dalam EN 261 WI 190. Dalam pengoptimuman parameter dalam pengekstrakan ruang kepala,
suhu penyuntik kedap udara ditetapkan pada 100 oC dan suhu pemanasan vial yang mengandungi
sampel ditetapkan pada 180 oC. Sampel dipotong, dimasukkan ke dalam vial dan ditutup dengan
segera. Botol sampel bersama dengan sampel dipanaskan pada 180 oC selama 2, 5, 10, 15 dan 20
minit ± 10 saat. Dalam pengoptimuman suhu pemanasan, botol sampel bersama dengan sampel
dipanaskan pada 100,120, 140, 160 dan 180 oC berdasarkan masa pengoptimuman yang diperoleh.
Gas ruang kepala disuntik dan dianalisis dengan GC-FID.
47
Kaedah penambahan piawai

Dalam kaedah penambahan piawai, sebanyak 5 vial disediakan. Spesimen yang disediakan (10 cm × 5
cm) digulung dan dimasukkan ke dalam vial 10 mL. Vial ditutup dengan segera. Jisim vial bersama
sampel ditimbang. Seterusnya, larutan piawai dengan isipadu yang meningkat (0-4 µL) disuntik ke
dalam vial yang telah tertutup melalui septum.Setiap vial ditimbang untuk mendapatkan jisim larutan
piawai yang telah ditambah. Semua vial dipanaskan pada suhu 180 oC selama 10 minit secara
berterusan. Analit dalam fasa gas itu (1 mL) kemudian disuntik ke dalam GC-FID menggunakan
penyuntik yang telah dipanaskan pada suhu 100 oC.
Kaedah pengekstrakan ruang kepala berbilang (MHE)

Kaedah MHE yang digunakan adalah secara piawai dalaman. Kaedah ini melibatkan 2 bahagian iaitu
penentuan faktor gerak balas dan seterusnya penentuan toluena residu dalam sampel. Dalam
penentuan faktor gerak balas, vial 10 mL yang tertutup ditimbang. Sebanyak 1 µL larutan piawai
toluena disuntik ke dalam vial yang tertutup dan kemudian vial ditimbang sekali lagi untuk
mendapatkan jisim 1 µL larutan toluena. Kemudian, 1 µL larutan nonana (piawai dalaman) disuntik
ke dalam vial menggunakan penyuntik yang berbeza. Sekali lagi, vial ditimbang untuk mendapatkan
jisim nonana. Vial dipanaskan pada suhu 180 oC selama 10 minit. Analit dalam fasa gas disuntik ke
dalam GC-FID dengan menggunakan penyuntik kedap udara 1 mL yang telah dipanaskan pada 100 oC
terlebih dahulu. Dalam penentuan toluena residu dalam sample, vial yang mengandungi spesimen
ujian ditimbang. Kemudian, 1 µL larutan nonana disuntik ke dalam vial menerusi septum
menggunakan penyuntik dan vial ditimbang semula untuk mendapatkan jisim larutan nonana yang
disuntik. Vial dipanaskan pada suhu 180oC selama 10 minit. Analit (1 mL) dalam fasa gas disuntik ke
dalam GC-FID dengan menggunakan penyuntik kedap udara 1 mL yang telah dipanaskan pada 100 oC
terlebih dahulu. Vial yang masih panas kemudian melalui proses pembebasan tekanan. Dalam proses
ini, satu jarum yang telah disambung ke sumber gas helium dimasukkan ke vial melalui septum vial
dan satu lagi jarum terus dicucuk ke dalam vial. Gas helium dialirkan pada kadar alir 15 mL/minit
selama 1 minit ± 10 saat. Selepas itu, kedua-dua jarum dikeluarkan daripada vial dan vial dimasukkan
semula ke dalam ketuhar. Untuk pengekstrakan kali kedua dan seterusnya, langkah pemanasan vial,
analisis GC dan proses pembebasan tekanan diulang untuk vial yang sama sebanyak 4 kali.
Keputusan dan perbincangan
Pengoptimuma masa pengekstrakan

Daripada Rajah 1 yang ditunjukkan, didapati luas puncak toluena tidak memberikan perbezaan yang
ketara pada masa pemanasan 2 minit dan 5 minit. Ini disebabkan kedua-dua tempoh masa ini terlalu
singkat untuk toluena mencapai keseimbangan antara fasa gas dan fasa pepejal dalam vial tertutup.
60
Luas punc ak toluena
50
40
(pA)
30
20
10
0
0 5 10 15 20 25
M a s a ( m in it )
Rajah 1: Luas puncak toluena melawan masa pemanasan (minit)
Sebaliknya, masa pemanasan selama 10 minit merupakan masa pengestrakan yang paling
optimum. Ini menunjukkan keseimbangan toluena antara fasa gas dan fasa pepejal telah tercapai.
Daripada Rajah 1 juga, didapati masa pemanasan yang lebih lama iaitu selama 15 minit dan 20 minit
48
memberikan luas puncak toluena yang semakin menurun. Dalam erti kata yang lain, masa pemanasan
yang lama akan mengganggu keseimbangan fasa toluena dalam vial. Dengan ini, masa pemanasan
selama 10 minit dipilih sebagai masa pemanasan yang paling optimum.
Pengoptimuma suhu pengestrakan

Daripada Rajah 2, ia jelas menunjukkan luas puncak toluena adalah berkadar langsung dengan suhu
pemanasan iaitu semakin tinggi suhu pemanasan yang digunakan, semakin tinggi luas puncak toluena
yang diperoleh. Ini menunjukkan suhu 180oC merupakan suhu yang paling sesuai.
30
Luas puncak (pA)
25
20
15
10
5
0
80 100 120 140 160 180
o
Suhu ( C)
Rajah 2: Luas puncak toluena melawan suhu pemanasan
Suhu pengekstrakan melebihi 180 oC tidak digunakan disebabkan kekangan penganalisian

ruang kepala. Septum pada tudung aluminium mungkin akan lebur pada suhu yang terlalu tinggi.
Selain itu, pemegangan vial yang telah dipanaskan pada suhu yang tinggi, contohnya 200 oC adalah
sukar walaupun sarung tangan digunakan. Dengan ini, suhu 180 oC dipilih sebagai suhu
pengesktrakan yang paling sesuai dan digunakan dalam analisis ruang kepala dalam kajian seterusnya.
Pengenalpastian sebatian toluene secara masa penahanan dan GC-MS

Dalam kajian ini, masa penahanan toluena adalah 2.653 minit dan ia hampir sama dengan masa
penahanan toluena piawai (Rajah 3).
FID Kelimpahan
Toluen
minit Masa
Rajah 3: Pemisahan GC bagi sebatian Rajah 4: Kromatogafi ion

toluena dalam sample
Analisis GC-MS telah dilakukan untuk mengenalpasti puncak toluena dan puncak sampingan yang
terhasil apabila penyuntik dipanaskan. Kromatografi ion jumlah seperti yang ditunjukkan dalam Rajah
4 menunjukkan kehadiran 2 puncak dengan tertib pengelusian yang sama. Puncak 2 dapat dikenalpasti
49
sebagai puncak toluena. Perbandingan spektrum jisim toluena dengan spektum jisim toluena daripada
pangkalan data Wiley memberika kebarangkalian yang melebihi 90 %.
Perbandingan kepekatan toluena residu antara kaedah penambahan piawai dan kaedah MHE
Perbandingan kepekatan toleuan residu antara kaedah penambahan piawai dan kaedah MHE hanya
dapat dilakukan ke atas 3 jenis sampel sahaja iaitu sampel TC, PM dan MCM. Perbandingan ke atas
sampel CCB, TE dan PDC tidak dapat dilakukan disebabkan kehabisan sampel dari kelompok sampel
yang sama untuk dianalisis dengan kaedah MHE. Jadual 1 jelas menunjukkan kepekatan toluena
residu yang ditentukan melalui kaedah penambahan piawai adalah lebih tinggi berbanding dengan
kaedah MHE. Ini mungkin disebabkan anggaran kuantiti toluena residu dalam sampel perlu dibuat
sebelum analisis sampel sebenar dilakukan menggunakan kaedah penambahan piawai. Dengan erti
kata lain, anggaran toluena residu mempengaruhi keputusan analisis.
Jadual 1: Perbandingan kepekatan toluena residu antara kaedah penambahan piawai dan kaedah MHE
Kepekatan toluena (mg/m2)
Sampel Kaedah penambahan piawai Kaedah MHE
satu warna berbilang warna Satu warna berbilang warna
TC 15.55 ± 2.16 18.33 ± 1.14 5.09 ± 0.27 7.21 ± 0.18
PM 10.78 ± 1.57 16.82 ± 1.02 2.81 ± 0.17 4.53 ± 0.13
MCM 8.32 ± 1.12 12.09 ± 1.09 4.19 ± 0.15 5.89 ± 0.15
Ujian awal mendapati anggaran kepekatan toluena residu adalah sekitar 8 mg/m2. Oleh itu,
kepekatan toluena residu yang ditentukan melalui kaedah penambahan piawai adalah lebih tinggi
nilainya. Sebaliknya, anggaran sebegini tidak perlu dilakukan dalam kaedah MHE. Analisis toluena
residu dilakukan secara terus secara pengekstrakan berterusan dalam kaedah MHE.
Perbandingan kepekatan toluena residu antara makmal

Dalam kajian ini, perbandingan kepekatan toluena residu telah dilakukan di antara makmal
disebabkan kesukaran untuk mendapatkan bahan rujukan piawai bagi sampel bahan pembungkusan.
Perbandingan keputusan analisis antara makmal dilakukan bertujuan untuk mengatasi ketakpastian
kaedah dan seterusnya meningkatkan kebolehpercayaan hasil analisis. Bagi sampel TC, PM dan
MCM, perbandingan kepekatan toluena residu telah dilakukan di antara makmal UTM dan makmal
Dannone, Paris seperti yang ditunjukkan dalam Jadual 2.
Jadual 2: Perbandingan kepekatan toluena residu antara makmal

Sampel Makmal UTM Makmal Peratus ketepatan (%)
Kaedah MHE Kaedah Dannone, Paris Kaedah MHE Kaedah
penambahan penambahan
piawai piawai
TC 1 7.2114 18.3280 6.2 116.31 295.61
TC 2 5.0876 15.5480 6.2 82.06 250.77
PM 1 4.5293 16.8200 10.7 42.33 157.20
PM 2 2.8100 10.7766 10.7 26.26 100.72
MCM 1 5.8890 12.0946 11.1 53.05 108.96
MCM 2 4.1859 8.3186 11.1 37.71 74.94
Daripada Jadual 2 yang ditunjukkan, dapat diperhatikan bahawa kepekatan toluena residu
dalam sampel TC yang dianalisis dengan kaedah MHE mempunyai peratus ketepatan yang baik iaitu
sekitar 82.06 % hingga 116.31 % berbanding dengan kaedah penambahan piawai. Sebaliknya, untuk
sampel PM dan sampel MCM pula, kaedah penambahan piawai didapati mempunyai peratus
ketepatan yang lebih baik iaitu di sekitar 74.94 % hingga 157.20 % berbanding dengan kaedah MHE.
Corak peratus ketepatan yang dapat diperhatikan ialah sampel dengan kepekatan toluena residu yang
tinggi iaitu melebihi 10 mg/m2 apabila dianalisis dengan kaedah penambahan piawai memberikan
peratus ketepatan yang tinggi berbanding dengan kaedah MHE. Keadaan sebaliknya diperhatikan
untuk sampel dengan kepekatan toluena residu yang rendah iaitu kurang daripada 10 mg/m2.
50
disebabkan nilai anggaran kepekatan toluena residu dalam sampel yang dianalisis dengan kaedah
penambahan piawai ditetapkan pada nilai kepekatan yang agak tinggi iaitu 8 mg/m2.
Perbandingan kaedah penentuan toluena residu

Perbandingan kaedah dilakukan bertujuan untuk menguji sama ada kaedah MHE dan kaedah
penambahan piawai berbeza dari segi kejituan iaitu sama ada perbezaan dua sisihan piawai bererti.
Jadual 3 : Fkiraan bagi sampel TC, PM dan MCM

Kaedah Penambahan piawai
Sampel
MHE
TC 1 41.1555
TC 2 62.0135
PM 1 58.4552
PM 2 84.0046
MCM 1 50.3499
MCM 2 59.6905
*Daripada jadual F, F2,2 = 39.00
Daripada Jadual 3, dapat diperhatikan bahawa Fkiraan bagi semua sampel adalah lebih besar
daripada Fjadual. Ini menunjukkan terdapat perbezaan bererti antara sisihan piawai kaedah MHE dan
kaedah penambahan piawai pada aras 5 %. Oleh kerana varians kaedah penambahan piawai lebih
besar daripada varians kaedah MHE pada 5 % aras kebarangkalian, maka dapat disimpulkan bahawa
kaedah MHE adalah lebih presis dalam penentuan toluena residu dalam sampel.
KESIMPULAN
Dalam kajian ini, kaedah pengekstrakan ruang kepala menggunakan analisis GC-FID dapat
diaplikasikan dalam analisis toluena residu. Toluena terelusi dalam masa kurang daripada 3 minit.
Didapati masa pemanasan yang paling optimum adalah 10 minit sementara suhu pemanasan yang
paling sesuai pula ialah 180 oC. Kajian yang dilakukan menunjukkan bahagian berbilang warna dalam
sampel pembungkus yang dianalisis dengan kaedah penambahan piawai mahupun kaedah MHE
memberikan kepekatan toluena yang lebih tinggi berbanding dengan bahagian satu warna. Bagi
sampel dengan kepekatan toluena residu yang rendah (kurang daripada 10 mg/m2), analisis toluena
residu menggunakan kaedah MHE mempunyai peratus ketepatan yang baik iaitu sekitar 82.1% hingga
116.3 %. Sebaliknya, untuk sampel sampel dengan kepekatan toluena residu yang tinggi (melebihi 10
mg/m2), peratus ketepatan yang rendah diperoleh iaitu di antara 26.3 % hingga 53.1 %. Ini mungkin
disebabkan terdapatnya ralat sistematik dalam kerja analisis yang menyebabkan perbezaan kepekatan
toluena residu dalam sampel yang sama.Ujian F yang digunakan dalam perbandingan kaedah
penentuan toluena residu menunjukkan kaedah MHE adalah lebih presis dalam penentuan toluena
residu dalam sampel.
Rujukan
1. Paine, F. A. (1990). "The Packaging User's Handbook." London: Blackie and Son Ltd. 4-7.
2. Crosby, N. T. (1981). "Food Packaging Materials." London: Applied Science Publishers. 9-11, 43-47.
3. Marsihi, R. (1993)."Testing Packaging Material : Why and How". Weeks Publishing Company.
4. Oswin, C. R. (1975). "Plastic Films and Packaging" London: Applied Science Publishers Ltd. 20-21.
5. Hotchkiss, J. H. (1988). "Food and Packaging Interactions." Washington, D. C.: American Chemical Society. 1-9.
6. Vermeiren, L., Devlieghere, F., Beest, M. V., Kruijf, N. and Debevere, J. (1999). “Developments in the Active
Packaging of Foods.”Trends in Food Sci & Technol. 10. 77-86.
7. Aurela, B., Kulmala, H and Soderhjelm, L. (1999). "Phthalates in Paper and Board Packaging and Their Migration into
Tenax and Sugar." Food Addit. Contam. 16(12). 571-577.
8. Aurela, B, Ohra-aho, T and Soderhjelm, L. (2001). "Migration of alkilbenzenes from packaging into Food and Tenax."
Packag. Technol. Sci. 14. 71-77.
9. Korbo, L., Ladefoged, O., and Lam, H. R. (1996). "Neuronal Loss in Hippocampus in Rats Exposed to Toluene."
Neurotoxicology .17(2). 359-66.
51
10. Ono, A., Sekita, K., and Ogawa, Y. (1996). "Reproductive and Developmental Toxicity Studies of Toluene : Effects of
Inhalation Exposure on Fertility in Rats." J Environ. Pathol. Toxicol. Oncol. 15(1). 9-20.
11. Gudat, A. E. and Brillante, S. M. (1996). "Mutiple Headspace Extraction Capillary Gas Chromatography for the
Quantitative Determination of Volatiles in Solid Matrices." U.S.A: Agilent Technologies.
12. Kolb, B and Pospisil, P. (1981). "Quantitative Analysis of Residual Solvents in Food Packaging Printed Film by
Capillary Gas Chromatography With Multiple Headspace Extraction." J. Chromatogr. A. 204. 371-376.
52
SYNTHESIS OF 3-O-SUCCINYL-BETULINIC ACID AS ANTI –CANCER AGENTS

AGAINST HUMAN MYELOID LEUKAEMIA AND HUMAN T-4 LYMPHOBLASTOID
CELLS LINE
Mohd Tajudin Mohd Ali a,Faujan Ahmad b,, Abdul Manaf Ali c, and Nurhana Faujan d
a
Faculty of Applied Sciences, Universiti Teknologi MARA,
40450 Shah Alam, Selangor, Malaysia.
b
Faculty of Science and Environmental Studies, Universiti Putra Malaysia,
43400 UPM Serdang, Selangor, Malaysia.
c
Faculty of Biotechnology and Biomolecular Sciences
d
Institute of Bioscience, Universiti Putra Malaysia
*
Corresponding author : Tel: +603-55435735 Fax : +603-55444566
email : tajudinali@salam.uitm.edu.my
Abstract: Betulinic acid was esterified with succinic anhydride in the presence of anhydrous
pyridine. Optimization studies on the esterification reaction were carried out. The effect of varying
parameters such as the mole ratio, temperature and time course was investigated. The optimal
conditions for synthesizing 3-O-succinyl-betulinic acid were obtained by refluxing betulinic acid and
succinic anhydride in 1:8 ratio with the presence of 5 ml anhydrous pyridine for 24 hours at a
temperature of 115oC. The percentage conversion as detected by gas chromatography analysis was
53.5%. The cytotoxic study also revealed that 3-O-succinyl-betulinic acid was more potent than
betulinic acid in inhibiting human myeloid leukemia (HL 60) and human T-4-lmphoblastoid
(CEMSS) cells line with IC50 values were 1.3 µg/ml and 1.3 µg/ml, respectively.
Keywords: Esterification, betulinic acid, betulinic acid ester
1 Introduction
The lupane type pentacyclic triterpene betulinic acid (1), 3-β-hydroxy-lup-20(29)-ene-28-oic, was
isolated from higher plants. Betulinic acid has been reported to possess anti-tumor activity towards
cultured human melanoma cell in vitro and vivo model1 . Other biological activities reported for
betulinic acid include anti-inflammatory activity2 and inhibition of phorbol ester-induced epidermal
ornithine decarboxylase accumulation in the mouse-ear with subsequent inhibition of the
carcinogenic response in the two- stage mouse –skin model3.
Betulinic acid has three active sites at C-3, C-20 and C-28 , where modifications can be
performed to yield derivatives. Modification of betulinic acid involving the esterification reaction
was reported. Note that 3-O-succinyl-betulinic acid was reported in literature; obtained by allowing
betulinic acid and succinic anhydride to react in the presence hydrochloric acid as a acid catalyst
(Fujioka et al., 1994). The role of the acid catalyst is expected to promote attack of a nucleophilic
oxygen of the betulinic acid on the carbonyl carbon of the anhydride by protonation of the carbonyl
oxygen.
In this work, betulinic acid ester was synthesized using betulinic acid and succinic
anhydride in Scheme 1. The effect of mole ratio, time and temperature of the reaction was
investigated.
2 Experimental
2.1 Materials
Betulinic acid was extracted from Malaysian higher plants. Succinic anhydride was purchased from
Sigma Chemical Co. Thin layer chromatography (TLC) was performed on Kieselgel 60 F254 plates
(0.2 mm thick. Merck). Column chromatography was carried out with silica gel (Merck, code number:
1.07734.1). Other chemicals were of analytical grade.
53
C O O H
H O
O
H 2C C
P y rid in e ,
H 2C C
R e flu x e d
O
C O O H
H 2 C C O
H 2 C C O H
Scheme 1 : Reaction of betulinic acid and succinic anhydride
2.2 Esterification of betulinic acid ester.

Betulinic acid (100.0 mg, 0.22 mmole) and succinic anhydride (175.4 mg, 1.32 mmole) in
anhydrous pyridine (5 ml) were refluxed for 24 hours. The reaction mixture was then acidified with
0.75 M hydrochloric acid (10 ml) The product was extracted with chloroform (2 x 30 ml) and dried
over moleculer seive. The solvent was then removed using Rotary Evaporator.
The compounds were detected by TLC analysis using chloroform as a solvent and detected
under iodine vapour. Detection of the ester product was made by comparing with the Rf values of
betulinic acid and succinic anhydride spots on TLC plate which were developed by the same
solvent system.
The crude product was chromatographed on silica gel ( code number: 1.07734.1000) column
chromatography (3 cm, i.d. and 30 cm length) using chloroform as the eluting solvent to afford the
desired product.
2.3 Analysis of the product

The ester product was analyzed by thin layer chromatography (TLC) and developed in the
mobile phase of chloroform. The purified product was also analyzed using a gas chromatograph (GC-
17A, Schimadzu) equipped with BPX5 column (0.25mm, ID and 30m length). Both of non-split
/splitless injector and detector were set at temperatures of 280oC and 320oC. Helium was used as the
carrier gas with a pressure of 37Kpa. The initial temperature of the column was 60oC which was run
for 3 minute. The temperature was increased with a flow rate of 20 deg./min. up to 200oC for 3
minutes, then up to 320oC for 3 minutes.
The percentage conversion was determined by using mixtures containing betulinic acid,
succinic anhydride and betulinic acid ester. Infrared analysis was carried out using Perkin- Elmer
Fourier transform infrared model 1650 while mass spectrum of the product was studied using
Shimadzu gas chromatography –mass spectroscopy (GC-MS, model 7A). The structure of the product
was studied using JOEL nuclear magnetic resonance model ECA-400.
2.4 Effect of mole ratio

The effect of various molar ratio of betulinic acid/ succinic anhydride (1:2, 1:4, 1:6, 1:8, 1:10) was
investigated. The percentage conversion of ester was calculated based on the number of mole of ester
produced by GC analysis.
54
2.5 Effect of time

The reaction mixture was refluxed in anhydrous pyridine (5 ml) at different reaction time i.e.
8, 16, 24, 32 and 48 hours respectively. The percentage conversion of ester was calculated based on
the number of mole of ester produced by GC analysis.
2.6 Effect of temperature

The reaction mixture was heated in anhydrous pyridine (5 ml) at different temperatures i.e.
37, 85 and 115oC, for 24 hours using oil bath. The percentage conversion of ester was calculated
based on the number of mole of ester produced by GC analysis.
3. RESULT AND DISCUSSIONS
Analysis of the product

The purified product was analyzed using spectroscopic method and was identified as 3-O-
succinyl-betulinic acid as white crystal (melting point: 278-280oC, 49.2% yield). It had IR (cm-1) had
νmax 3402(OH), 2948 (CH2), 1812 (C=O :ester), 1734 (C=O : acid), 1248 (C-O), 886 (C=C) cm-1; 1H
NMR (CDCl3) ; δ 0.86 (s,3H), 0.87 (s,3H), 0.88 (s,3H), 0.99 (s,3H), 1.00 (s,3H), 1.70 (s,3H), 3.52 (q,
H-19), 4.50 (dd,H-3), 4.65 (s,1H,H-29), 4.75 (s,1H,H-29) , 2.06 (s,2H,C-32), 2.26 (s,2H,C-33); 13C
(CDCl3) ; δ 38.7 (C-1), 26.0 (C-2), 81.2 (C-3), 38.2 (C-4), 55.7 (C-5), 18.4 (C-6), 34.5 (C-7), 41.0 (C-
8), 50.9 (C-9), 37.4 (C-10), 21.6 (C-11), 23.9 (C-12), 38.1 (C-13), 42.7 (C-14), 30.0 (C-15), 31.8 (C-
16), 57.9 (C-17), 49.5 (C-18), 46.7 (C-19), 150.1 (C-20), 30.5 (C-21), 36.5 (C-22), 28.1 (C-23),16.7
(C-24), 16.5 (C-25),16.3 (C-26), 16.2 (C-27), 181.2 (C-28), 110.3 (C-29), 19.6 (C-30), 167.4 (C-31),
30.7 (C-32), 32.4 (C-33), 171.3 (C-34). MS; m/z 556 (M+), 511, 483, 438, 423, 395, 248, 220 and
189.
It’s 1H-NMR spectrum in d-chloroform, indicated the presence of six tertiary methyl groups
resonated as singlets at δ 0.86 (s, 3H), 0.87 (s, 3H), 0.88 (s, 3H), 0.99 (s, 3H), 1.00 (s, 3H) and 1.70
(s, 3H). The signals at δ 3.00 (m, H) was due to the proton at C-19 of the betulinic acid skeleton and
δ 4.50 (dd, H) was due to the proton at C-3; while signals at δ 4.65 (s, H) and δ 4.77 (s, H) were due
to methyelene proton of isopropenyl group at C-29 position. The signals at δ 2.06 and δ 2.26 were due
to -CH2 from succinyl group. This data is summarized in Table 6. It is in agreement with the spectra
data reported by Fujioka et al.,(1994) and is compared with the expected values.
The 13C-NMR spectrum of 3-O-succinyl-betulinic acid showed the down field shifts of C-20
and C-29 which is the site of terminal double bond resonatiy at δ 150.1 and 110.3, respectively. The
signal due to the carbonyl group of C-28 resonated at δ 181.2. The additional 13C-NMR signals
resonated at δ 167.4 and δ 171.3 were due to carbonyl group of ester at C-31 and C-34. The
methylene proton of succinyl group at C-32 and C-33 resonated at δ 30.7 and 32.4 respectively.
Other chemical shifts of 3-O-succinyl-betulinic acid are summarized in Table 1; and are in agreement
with the literature data Fujioaka T., et al (1994) . To the best of our knowledge this is the first 13C-
NMR data reported for 3- O -succinyl- betulinic acid . The comparison of this 13NMR data with the
ChemNMR programme are also included on the table.
The Rf values for betulinic acid, 3- O -succinyl-betulinic acid and succinic anhydride were
measured at 0.09, 0.33 and 0.83, respectively. The desired ester was expected to have greater
polarity compared to betulinic acid.
Effect of mole ratio

In all cases, the conversion percentage increased along with the mole ratio up to a certain
value in which after that point, the conversion percentage started to decrease (Figure 1). In the
reaction of betulinic acid with succinic anhydride, the optimum mole ratio was at 1:8, with
conversion percentage at 32.9%.
It is noted that, after the optimum mole ratio has been achieved , the conversion percentages
of the products started to decrease. These results were due to the fact that large quantity of the acid
used was blocked the attacks of the nucleophilic oxygen of betulinic acid at C-3 position on the
55
carbonyl carbon of the acid . At high mole of the substrates, the medium of reaction mixtures were
more viscous, thus achieving lower reaction rates and consequently with lower conversion
percentages.
Table 1 : The 13C-NMR data of 3-O-Succinyl-betulinic acid and comparision to the assignment of
betulinic acid
Carbon Betulinic acid1 Experiment2

C-1 38.8 38.7
C-2 27.4 27.3
C-3 79.0 79.0
C-4 38.7 38.3
C-5 55.3 55.3
C-6 18.3 18.3
C-7 34.3 34.3
C-8 40.7 40.6
C-9 50.5 50.5
C-10 37.2 37.0
C-11 20.8 22.6
C-12 25.4 25.4
C-13 38.4 37.1
C-14 42.4 42.4
C-15 29.7 29.7
C-16 32.1 31.9
C-17 56.3 56.3
C-18 49.2 49.2
C-19 46.7 46.8
C-20 (C=C) 150.4 150.4
C-21 30.5 30.5
C-22 37.0 35.9
C-23 28.0 28.0
C-24 15.3 16.0
C-25 16.1 15.3
C-26 16.0 14.7
C-27 14.7 14.1
C-28 (C=O) 180.8 180.0
C-29 (C=C) 108.3 109.7
C-30 19.4 20.8
C-31 (C=O) 167.5
C-32 38.8
C-33 44.0
C-34 (C=O) 177.5
C-35 23.0
C-36 23.8
Note
1
Betulinic Acid
2
Experiment Value
56
Percentage of Conversion ( %
45
35
25
15
-5 0 1/2 1/4 1/6 1/8 1/10

Mole Ratio ( mol betulinic acid / mol
succinyl anhydride)
Figure 1:The effect mole ratio of substrates (betulinic acid / succinic anhydride ) on the
esterification reaction of betulinic acid (Reaction time : 24 hours, Temperature : 115oC and
0.062mole Pyridine)
Effect of time
Figure 2, shows the time course for the esterification reaction of betulinic acid with succinyl
anhydride. The mole ratio used in the reaction of betulinic acid with succinic anhydride was 1:8
which conversion percentage at 47.3% at the 8 hour reaction time. The conversion percentage
increased up to the optimum value at 24 hour reaction before starting to decreased and its optimum
conversion was at 53.5 %. After achieving the optimum point, the conversion percentages seemed to
decrease. This may be due to the water excessive production during the esterification reaction and
caused the reverse hydrolysis reaction to occur.
Percentage of Conversion ( %)
50
40
30
20
10
0
0 8 16 24 32 40 48
Time (Hours)
Figure 2: The effect of time on the esterification of betulinic acid and succinic anhydride.
The reaction mixture consisted of betulinic acid (100mg), succinyl anhydride (192.1 mg)
and anhydrous pyridine (5ml) and temperature at 115oC.
57
Effect of temperature
The effect of temperature on the esterification reaction of betulinic acid with succinic anhydride
shown in Figure 3. The percentage of conversion increased with increasing temperature and reached
the maximum conversion percentage at 115oC (i.e. 29.8% yield).
50
Percentage of Conversion ( %)
45
40
35
30
25
20
15
10
5
0
0 37 85 115
Temperature o( C)
Figure 3: The effect of temperature on the esterification of betulinic acid and succinic
anhydride. The reaction mixture consisted of betulinic acid (100mg), succinic
anhydride (192.1 mg) and anhydrous pyridine (5ml).
Cytotoxic Effects
Betulinic acid inhibited human myeloid leukemia cell (HL60) with an IC50 values were 2.4 µg/ml
and inhibited human T4-lymphoblastoid cell (CEMSS) growth with an IC50 value of 1.8 µg/ml. On
the other hand, 3-O-succinyl-betulinic acid showed an inhibitory effects on human myeloid leukemia
cell with an IC50 values of 1.3 µg/ml, while its IC50 for inhibition of human T4-lymphoblastoid cell
growth was 1.3 µg/ml. These data indicated that 3-O-succinyl-betulinic acid is more potent than
betulinic acid in inhibiting the human myeloid leukemia and of human T4-lymphoblastoid cells. The
IC50 values against cancer cell growths are summarized in Table 2.
Table 2 : Cytotoxic effects (µg/ml) for betulinic acid and 3-O-succinyl-betulinic acid.
Compound IC50a IC50b
Betulinic acid 2.4 1.8
3-O-succinyl-betulinic acid 1.3 1.3
a
Concentration which inhibits human myeloid leukemia cell by 50%.
b
Concentration which inhibits human T4-lymphoblastoid cell by 50%.
58
Conclusion
3-O-succinyl-betulinic acid ester can be synthesized from the reaction of betulinic acid with succinic
anhydride and its cyototoxic effects on human myeloid leukemia cell and human T4-lymphoblastoid
cell is first examined.
ACKNOWLEDGEMENT
This project was financed by Universiti Teknologi Mara.
REFERENCES
1. Kim, D. S. H. L., Pezzuto, J. M. and Pisha, E.(1998) “ Synthesis of Betulinic Acid Derivatives
with Activity Against Human Melanoma” Bioorg. & Medic.l Chem. Lett. 8:1707-1712.
2. Recio, M. D. C., Giner, R, M., Manez, S., Gueho, J., Julien, H. R., Hostettmann, K. and Rios, J.
L. (1995) “ Investigations on the Steroidal Anti-Inflammatory Activity of Triterpenoids from
Diospyros leucom” Planta Med. 61:9-12.
3. Yasukawa, K., Takido, M., Matsumoto, T., Takeuchi, M . and Nakagawa, S. (1991) “Sterol and
Triterpene Derivatives from Plants Inhibit the Effect of a Tumor Promoter and Sitosterol and
Betulinic Acid Inhibit Tumor formation in Mouse Skin Twi-Stage Carcingenesis”Oncology 48:
72-76.
4. Hashimoto, F., Kashiwada, Y., Cosentino, L. M., Chen, C. H., Garrett, P. E. and Lee, K.H.
(1997) “Anti-Aids Agents. Synthesis and Anti-HIV Activity of Betulinic Acid and
Dihidrobetulinic Acid Derivatives” Bioorg. & Medic.l Chem. Lett. 5:2133-2143.
5. Kashiwada, Y., Hashimoto, F., Cosentino, L. M., Chen, C. H., Garrett, P. E. and Lee, K. H.
(1996) “ Betulinic Acid and Dihydrobetulinic Acid Derivatives as Potent Anti-HIV Agents” J.
Medicinal Chem. 39:1016-1017.
6. Ahmad, F. H., Omar, J. H. and Ali, A. M. (1999) “ Chemical Examination of Local Plant :
triterpenes from Leave of Malaysian Callistemon Speciosus D.E.” Ultra Science 11:357-360.
59
KEKABU SEED OIL EXTRACTION AND PUFA ISOLATION
Khairul Asmak Abd. Kadir, Jumat Salimon*
Oleochemistry Programme, School of Chemistry and Food Technology, Faculty of Science

and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia.
e-mail*: jumat@pkrisc.cc.ukm.my
Abstract:
Oil from kekabu seed was extracted using n-hexane in orbital shaker for 3 hours at 60 oC at 150 rpm. The seed oil was
obtained by solvent evaporation using rotary evaporator. Fatty acids compositions were analysed using gas chromatography
(GC). The major fatty acids found in kekabu seed oil (KSO) are linoleic (C18:2n6), palmitic (C16:0) and oleic (C18:1n9)
which accounts 95 %. The other remaining acids include gamma linolenate (C18:3n6), linolenate (C18:3n3) and lignoceric
(C24:0). After the hydrolysis process, the free fatty acids were separated using urea complexation method. The saturated
(SFA) and monoenoic fatty acids (MUFA) were separated from polyunsaturated fatty acid (PUFA). The ratio of fatty acid to
urea at 1:4, (w/w) was found to be suitable for linoleic acid enrichment in the non-urea fraction, yielding up to 82.9 %
linoleic acid.
Abstrak:
Minyak biji kekabu diekstrak dalam heksana dengan 150 putaran per minit selama 3 jam pada suhu 60 oC. Minyak
diperolehi dengan menyuling keluar pelarut menggunakan alat penyulingan berputar. Analisa menggunakan gas
kromatografi (GC) menunjukkan asid lemak major (95 %) dalam minyak biji kekabu terdiri daripada asid linoleik
(C18:2n6), palmitik (C16:0) and oleik (C18:1n9) manakala selebihnya terdiri daripada asid lemak gamma linolenat
(C18:3n6), linolenat (C18:3n3) dan lignoserik (C24:0). Selepas proses hidrolisis minyak kekabu, asid-asid lemak dipisahkan
menggunakan kaedah pengkompleksian urea. Asid lemak tepu dan monotaktepu dapat dipisahkan daripada asid lemak
politaktepu. Nisbah asid lemak kepada urea yang paling sesuai bagi pengkayaan asid linoleik dalam fraksi bukan-urea
adalah pada nisbah 1:4, (b/b) yang menghasilkan 82.9 % asid linoleik.
Katakunci: Asid lemak politaktepu (PUFA), asid linoleik, pengkompleksian urea, Ceiba pentandra
Introduction
Structured lipid (SL) possessing long-chain polyunsaturated fatty acids (PUFA) located at the mid
position with medium-chain fatty acids (MCFA) at the end positions have gained considerable
attention recently as nutritional and health supplements [1]. The beneficial health effects of the PUFA
are well established. Therefore, synthesizing SL containing PUFA located at the mid position with
MCFA at the end positions has been the interest of many researchers.
Linoleic acid (Figure 1) is one of the two parent essential fatty acids (EFA). EFA are long chain
polyunsaturated fatty acid (PUFA) that vital for normal cellular function. The term ‘essential’ implies
that they must be supplied in the diet because they are required by the human body but cannot be
endogenously synthesized [2]. The functions of linoleic acid (LN) include the role in cell growth,
skin and hair condition, reproduction and wound healing. These effects occur largely through the
actions of LN in cell membranes [3]. LN also reduces plasma cholesterol levels and reduces platelet
aggregation. In the presence ∆6-desaturase enzyme, γ-linolenic acid (GLA) is biosynthesized from
LN. GLA serves as a precursor of cell membrane arachidonic acid (AA) which in turn is a substrate
for the hormone-like substances namely eicosanoid (prostaglandin, thromboxanes and leukotrienes).
These substances play important roles in physiological functions of modulating immune and
inflammatory responses and are effective for treating atopic eczema and rheumatoid arthritis [4].
COOH
Figure 1: Linoleic Acid (C18:2n-6)
60
Kekabu tree or Ceiba pentandra belong to the family of Bombacaceae. The tree is known by
many local vernacular name. For instance, kapok or kabu kabu (Java), Nun (Thai), silk-cotton (West
Africa) and ceyba or ceiba (Latin Amerika) [5-8]. It fruits from its fifth year, producing cocoa-like
shaped fruits. They are green at first, ripening to a dull, blackish-brown which split at maturity,
releasing seeds covered with silky hairs/lint (Figure 2). Kekabu seed has traditionally been consumed
in Asian countries such as Indonesia, Myanmar, Thailand and Malaysia. The unripe seeds were eaten
whilst the matured one were fried first. In most West African countries, the seed were crushed and
roasted or added in a soup.
a
d
c
b
Figure 2: Ripened fruit (a); unripe fruit (b); seeds embedded in masses of fibers (c) and kekabu seed
(d)
It had been documented that kekabu seed oil (KSO) was comparable to a well-known oil crop,
cottonseed oil [9]. In particular, kekabu seed contains up to 25 % oil yield. Preliminary study
conducted reveals that kekabu seed oil consist of linoleic acid as the most dominant fatty acid (40-50
%). Therefore, KSO can be seen as a potential local inexpensive source of linoleic acid. Hence,
separation and the isolation and purification process of linoleic acid from the kekabu seed oil is
important due to it can be easily obtained from natural sources rather than by chemical synthesis.
Urea complexation is one of the most appropriate methods for PUFA enrichment. It allows the
handling of large quantities of material in simple equipment, inexpensive solvents, milder condition,
cost effective and versatile process. Its fractional characteristics can be altered simply by changing
the amounts of either solvent or urea ratio [10]. This method is also favored by many researchers
because complexation depends upon the configuration of the fatty acid moieties due to the presence of
multiple double bonds rather than pure physical properties such as melting point or solubility [11].
This work is a part of ongoing research project on synthesizing structured lipid possessing long-
chain polyunsaturated fatty acids (PUFA) located at the mid position with medium-chain fatty acids
(MCFA) at the end positions whilst this particular paper reports linoleic acid (LN) enrichment from
kekabu seed oil by urea inclusion as the sources of PUFA for the production of structured lipid.
Experimental Procedures
Materials: Fatty acid standards were obtained from Sigma. Chemicals and solvents were obtained
from Merck and Fisher. The solvents were used without any further purification process.
Oil extraction: The oil content in kekabu seeds was determined using the standard procedure
described in IUPAC Standard Method [12] with slight modifications. The crushed seeds and hexane
were put in an orbital shaker at 155 rpm for three hours at 60 oC. The solvent was then evaporated at
60 oC by rotary evaporator. The extracted oil was then placed in a vacuum oven and kept at 60 oC for
30 minutes. The oil percentage yield was calculated. The obtained oil was then placed in a dark
container and stored in a refrigerator at 0 oC.
Fatty acid analyses

The fatty acids compositions were determined using gas chromatography, Shimadzu GC-17A FID
equipped with capillary column BPX 70 (30m x 0.25mm x 0.25µm). The column temperature was
61
programmed at 120 oC with 3 oC increasing for 57 minutes. The injector and detector temperature
were set at 260 oC and 280 oC respectively. Nitrogen gas was used as the carrier gas with 0.3 ml/min
flow rate. The identification of the peaks was achieved by comparing the retention times with the
authentic standards that were analyzed under the same conditions.
Free fatty acid preparation: The procedures reported by Wanasundara et al. [13] was employed with
a few modifications. 25 g of the kekabu seed oil was treated with 200 ppm butylated hydroxytoluene
(BHT) and saponified by refluxing with potassium hydroxide solution [(5.75 g in 95% (v/v) ethanol
(66 ml) and water (11 ml)] for 1 hour in a nitrogen atmosphere. The saponified mixture was diluted
with cold water (50 ml) and the unsaponifiable matter was extracted thoroughly with 2 x 100 ml of
hexane and discarded. The aqueous layer was then acidified to pH=1.0 with hydrochloric acid (3N)
and the liberated fatty acids (FFA) were subsequently extracted with 2 x 25 ml hexane. The hexane
extract was dried over anhydrous sodium sulphate and the remaining solvent was evaporated to
dryness by using a rotary evaporator. The recovered free fatty acids were stored at -5 oC prior to
further analyses.
Urea complexation: The free fatty acids (FFA) obtained from the saponification process were
subsequently undergoing urea complexation according to method proposed by Hidarjat et al. [14] with
a slight modifications. The FFA was mixed with hexane and was heated with constant stirring until a
clear solution produced. The solution was then cooled and crystallized at 4 oC for 24 hours. After 24
hours, the urea complex fraction (UCF) was separated from the liquid fraction of non-urea complex
fraction (NUCF) using a Buchner funnel suction and pressed well to remove mother liquor before and
washed with cold solvent saturated with urea. The NUCF was diluted with equal amount of water and
subsequently acidified with HCI (6N) to pH 4-5. An equal volume of hexane was added and the
mixture was stirred for 1 hour. The hexane layer, containing the liberated fatty acids was separated
from the aqueous layer. The hexane layer was then dried over anhydrous sodium sulphate and the
solvent was evaporated. Fatty acids from the crystal or UCF were similarly recovered. The
percentage recovery from each fraction was calculated.
Esterification of free fatty acid (FFA): Acids (0.1 g) were treated with 1 ml 0.4 M methanolic
sulphuric acid (H2SO4) in 10 ml test tube with screw-capped. The test tube was then placed in a water
bath at 70 oC for 10 minutes. 0.5 ml of distilled water and 1.0 ml hexane was added. The mixture
was centrifuged at 500 rpm. The upper layer, which contained fatty acid methyl ester (FAME) was
directly injected to GC for fatty acids analyses.
Fatty acid composition

The fatty acid composition of kekabu seed oil (KSO) and their free fatty acids (FFA) after
saponification process are shown in Table 1. KSO consists mainly palmitic, stearic, oleic and linoleic
whilst the minor fatty acids include γ-linolenic, α-linolenic, myristic, arachidic, behenic and
lignoceric. From the ten fatty acids analyzed, 42.73 % is represented by PUFAs (linoleic acid, ALA
and GLA), 21.72 % of monounsaturated fatty acids, MUFA (oleic acid), and 35.22 % is saturated
fatty acids, SaFA (myristic, palmitic, stearic, arachidic, behenic and lignoceric acids). After the
saponification process, the content of free fatty acids was slightly increased. Meanwhile, a
considerably decreased in stearic acid content could be due to difficulty during the handling process.
This is because stearic acid is in a solid state at room temperature.
Urea complexation
Theoretically, urea (NH2.CO.NH2) normally crystallizes in tetragonal form with channel of 5.67 Å
diameter. However, in the presence of long straight-chain molecules it crystallizes in a hexagonal
structure with channels of 8-12 Å diameter within the hexagonal crystals [15]. The channels formed
are sufficiently large to accommodate aliphatic chains. Straight-chain saturated fatty acids with six
carbon atoms or more are readily adducted. The presence of double bonds in the carbon chain
increases the bulk of the molecule and reduces the probability of its complexation with urea.
62
Monoenes are more readily complexed as compared to dienes which, in turn, are more readily
complexed than trienes. The saturated fatty acids, monoenes and, to a lesser extent, dienes are
crystallized with urea (UCF) and non-urea complex fraction (NUCF) in the solution is separated by
filtration [11].
Table 1: Fatty acid composition of kekabu seed oil (KSO)
Fatty Composition* (%)

Acids A B
Major acids
Palmitic C16:0 22.58 ± 0.00 24.35 ± 0.25
Stearic C18:0 11.83 ± 0.34 3.643 ± 0.13
Oleic C18:1(ω-9) 21.72 ± 0.20 23.57 ± 0.11
Linoleic C18:2(ω-6) 39.99 ± 0.19 40.51 ± 0.06
Minor acids
γ-linolenic C18:3(ω-6) 2.159 ± 0.14 1.948 ± 0.11
α-linolenic C18:3(ω-3) 0.5690 ± 0.03 0.7170 ± 0.09
Myristic C 14:0 0.0800 ±0.01 0 ± 0.00
Arachidic C 20:0 0.1290 ± 0.02 0.5850 ± 0.09
Behenic C 22:0 0.4680 ± 0.02 0.5515 ± 0.21
Lignoceric C 24:0 0.1335 ± 0.01 3.015 ± 0.13
Others 0.0392 ± 0.01 1.139 ± 0.88
Σ SFA1 35.22 ± 0.04 32.14 ± 0.35

Σ UFA2 21.72 ± 0.20 23.57 ± 0.21
Σ PUFA3 42.72 ± 0.29 43.18 ± 0.20
* Values were means of duplicate samples ± standard deviation;

A - Kekabu seed oil (KSO);
B - Free fatty acid of KSO after saponification.
1
Saturated fatty acid (myristic, palmitic, stearic, arachidic, behenic & lignoceric)
2
Unsaturated fatty acid (oleic)
3
Polyunsaturated fatty acid (linoleic, α-linolenic & α -linolenic)
Table 2 showed the major fatty acid composition of kekabu seed oil free fatty acids (KSO-FFA)
after urea complexation. Due to the presence of two double bond, linoleic acid is crystallized with
urea in a lesser extent compared to palmitic (C16:0). stearic (C18:0) dan oleic (C18:1) acids. The
enrichment of linoleic acid in the non-urea complex fraction (NUCF) increased in parallel with the
increasing amount of urea until ratio of 1:4 (FFA:urea, w/w). At the ratio of 1:5, the percentage of
linoleic acid obtained was decreased. This could be as a result of the larger extent of linoleic acid
that complexed with urea as the amount of urea increased. The enrichment of linoleic acid performed
the best at the ratio of 1:4 (FFA:urea, w/w) with up to 82 % with a totally removal of palmitic and
oleic acid.
Conclusion
PUFA enrichment in kekabu seed oil was done with the most favorable result at the ratio free fatty
acids to urea 1:4. At this ratio, linoleic acid was concentrated up to two fold, from 40 % in the FFA to
82.9 % in the non-urea fraction.
63
Table 2: Fatty acid composition of saponified kekabu seed oil (KSO) after urea complexation process
Major Compositiona (%)

Fatty Acid The Ratio of Fatty acid to Urea
FFAb Fractionsc 1:1 1:2 1:3 1:4 1:5
Palmitic 24.35 ± 0.25 1 46.75 ± 0.15 47.93 ± 1.07 34.33 ± 0.78 30.18 ± 0.72 28.45 ± 0.73
(C16:0) 2 5.34 ± 0.08 0.41 ± 0.71 0 ± 0.00 0 ± 0.00 0 ± 0.00
Stearic 3.643 ± 0.13 1 5.79 ± 0.11 4.25 ± 0.08 3.08 ± 0.15 3.69 ± 0.59 1.12 ± 0.16
(C18:0)
2 0.54 ± 0.41 0.94 ± 0.04 1.66 ± 0.05 2.77 ± 0.29 0 ± 0.000
Oleic 23.57 ± 0.11 1 20.05 ± 0.01 32.34 ± 0.62 32.19 ± 0.78 28.51 ± 0.97 27.98 ± 0.77
(C18:1)
2 26.41 ± 0.41 11.66 ± 0.42 0 ± 0.00 0 ± 0.00 0.48 ± 0.07
Linoleic 40.51 ± 0.06 1 22.69 ± 0.15 13.05 ± 0.30 26.57 ± 1.13 33.04 ± 0.87 36.03 ± 0.06
(C18:2n-6)
2 56.52 ± 1.87 71.32 ± 1.54 80.53 ± 0.43 82.91 ± 0.09 77.05 ± 2.45
a
Values were means of duplicate samples ± standard deviation;
b
Free fatty acid of KSO after saponification.
c
1 = Crystal fraction / Urea complex fraction (UCF); 2 = Liquid fraction / Non-
urea complex fraction (NUCF)
Temperature = 4 oC; Time course = 24 hours
Acknowledgements
We would like to thank Universiti Kebangsaan Malaysia and the Ministry of Science and Technology
& Innovation (MOSTI) for research grant # 09-02-02-0115 EA 277 and the National Science
Fellowship fund for Miss Khairul Asmak Abdul Kadir
References
1. Halldorsson, A., Magnusson, C.D. & Haraldsson, G. (2001) “Chemoenzymatic Synthesis of Structured TAG”
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3. Smit, E.N., Muskiet, F.A.J. & Boersma, E.R. (2004) “The Possible Role of Essential Fatty Acids in the
Pathophysiology of Malnutrition: a review” Prostaglandins, Leukotrienes and Essential FattyAcids. 71: 241-250.
4. Berry, E.M. (2001) “Are Diets High in Omega-6 Polyunsaturated Fatty Acids Unhealthy? European Heart Journal
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5. H.G. Baker. (1965) “The Evolution of The Cultivated Kapok Tree” Res. Ser. Inst. Intnl. Studies, Uni.Calif. 9: 185-216
6. A.C. Zeven. (1979) “Kapok; Ceiba pentandra (Bombacaceae) in N. W. Simmonds (Ed.) Evolution of crop plants”,
USA: Longman Inc.
7. Allen, B.M. (1971) “Some Common Trees of Malaysia and Singapore”, Singapore: Singapore Offset Printing (Pte.)
Ltd
8. Fong, C.H. & Enoch., I.C. (1988) “Malaysian Trees in Colour”, Kuala Lumpur: Tropical Press Sdn. Bhd.
9. J.G.Vaughan. (1970) “The Stucture and Utilization of Oil Seeds”, Great Britain: The Chaucer Press.
10. Guil-Guerrero, J.L. & Belarbi, E.H. (2001) “Purification Process for Cod Liver Oil Polyunsaturated Fatty Acids” J.
Am. Oil. Chem. Soc. 78(54): 477-482
11. Shahidi, F & Wanasundara, U.N. (1998) “Omega-3 Fatty Acid Concentrates: Nutritional Aspect and Production
Technologies” Trends in Food Sciene and Technology. 9: 230-240.
12. International Union of Pure and Applied Chemistry. (1979) “Standard Methods for the Analysis of Oils, Fats and
Derivatives. (6th ed)”, Great Britain: Pergamon Press.
13. Wanasundara, U.N & Shahidi, F. (1999) “Concentration of Omega-3-Polyunsaturated Fatty Acids of Seal Slubber Oil
by Urea Complexations: Optimization of Reaction Conditions” Food Chemistry. 65: 41-49
14. Hidarjat, K., Ching, C.B., Rao, M.S. (1995) “Preparative-scale Liquid Chromatography of ω-3 Fatty Acids from Fish
Oil Sources. Journal of Chromatography A. 702: 215-221
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Press.
64
TOXICITY AND ANTITERMITE ACTIVITIES OF THE ESSENTIAL OILS FROM PIPER

SARMENTOSUM
Chieng T.C.*, Assim Z.B., and Fasihuddin B.A.

Faculty of Resource Science and Technology
Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak
* Phone: 082 679179; Fax: 082 672275; chieng@frst.unimas.my
Abstract. The leaves of Piper sarmentosum were hydrodistilled using the modified Clevenger-type
apparatus, and an average yield of essential oil of 1.10% (v/dry weight) was obtained. The leaf oils
were analyzed by GC and GC-MS. A total of 31 components were identified. Spathulenol (21.0%),
myristicin (18.8%), β-caryophyllene (18.2%) and (E,E)-farnesol (10.5%) were the major compounds
found in the leaf oil. The leaf oil showed inhibitory activity against the larvae of Artemia salina with
LC50 value of 35.2 µg/mL, and 100% mortality within two days at 1% concentration against the
subterranean termite (Coptotermes sp.). The crude extract was then subjected to bioassay-guided
isolation using silica gel column chromatography, and eluted with hexane containing increasing
volumes of ethyl acetate and yielded three pure compounds. Their toxicity and antitermite activities of
the three compounds were determined. Compound 2 showed the most potent activity against the
larvae of A. salina with LC50 value of 7.5 µg/mL, while the LC50 values for compound 3 and
compound 1 were 17.2 µg/mL and 22.5 µg/mL respectively. Compound 3 showed the strongest
inhibitory activity against the subterranean termite (Coptotermes sp.) with 100% mortality after 3
days at 0.1% concentration followed by compound 2 with the same mortality rate at 0.5%
concentration. Compound 1 showed the weakest inhibitory activity with 80% mortality after 3 days at
2% concentration.
Key words: Piper sarmentosum; essential oil; bioassay-guided isolation; toxicity; antitermite activity
Introduction
The genus Piper belongs to the family Piperaceae, comprising more than 700 species distributed
throughout the tropical and subtropical regions of the world [1]. Most of the species in this genus are
aromatic, woody perennial climbers and rarely shrub. The Piper species have high commercial,
economical and medicinal importance. Many species have been shown to possess antimicrobial,
antifungal, antioxidant, insecticidal, allelopathic and antitumour activities [1]. Various compounds,
including alkaloids/amides, propenylphenols, lignans, neolignans, terpenes, steroids, kawapyrones,
chalcones, flavones and flavanones have been isolated from different Piper species. The chemistry of
Piper species has been reviewed by several researchers [2, 3].
Piper sarmentosum Roxb., locally known as “kaduk”, is a creeping herb with erect, slender
branchlets about 30 cm tall. It is commonly found in damp open spaces, riverbanks cleared and
cultivated lands in Sarawak [4]. The leaves are used in folk medicine as counter-irritants in poultices
for headaches and pains in bones. A decoction of the boiled leaves may be utilized to treat coughs,
influenza, toothaches and rheumatism. The root is also a remedy for toothache and may be made into
a wash for fungoid dermititis on the feet [5]. A bioactive compound, isoasarone has been isolated
from the roots of P. sarmentosum and showed insecticidal property similar to DDT [6]. Four
phenylpropanoids isolated from the benzene-soluble fraction of the methanolic leaf extract showed
antimicrobial activity against Escherichia coli and Bacillus subtilis [7], and the methanol extract of
the leaves was found to possess a profound neuromuscular blocking activity in rat nerve-
hemidiaphragm preparation [8]. The chloroform extract of the plant showed considerable antimalarial
activity against Plasmodium falciparum and Plasmodium berghei parasites [9], while the water
extract of the whole plant had a hypoglycaemic effect in rats [10]. In addition, some amides isolated
from the hexane and methanol extracts of fruits exhibited antituberculosis and antiplasmodial
activities [11]. A resent study showed that this plant is a good source of natural antioxidants as the
65
methanol extracts were found to possess high antioxidant activity, which may be attributed to the high
contents of vitamin E and xanthophylls [12].
This paper reported three compounds isolated by means of bioassay-guided chromatographic
separation from the essential oil extracted from the leaves of P. sarmentosum. The bioactivity of these
compounds against brine shrimp larvae and subterranean termites were also studied.
Materials and Methods

Plant materials
The leave samples of Piper sarmentosum was collected around Kuching area. Sample was identified
and authenticated by a plant taxonomist. Voucher specimens was deposited at the Herbarium of
Universiti Malaysia Sarawak.
Extraction and analysis

The randomly picked leaves were air dried and ground. About 100 g of the ground samples were
hydrodistilled using the modified Clevenger-type apparatus for 6 hours, and the oily layers obtained
were separated and dried over anhydrous sodium sulphate and stored in vial at 4 – 5oC. The yield was
calculated based on dry weight of the plant material.
The oils were analyzed on a Shimadzu GC-17A chromatograph equipped with a flame ionization
detector using fused silica capillary column DB-5 (25 m × 0.22 mm ID and film thickness 0.25µm).
The operation parameters were: N2 as carrier gas at flow rate of 20 cm3 min−1, splitless, injector
temperature 280oC, and detector temperature 320oC. The column was programmed initially at 50oC
for 2 minutes, and ramped to 300oC at a rate of 6.5oC min−1 and held for 7 minutes.
The oils were also analyzed using GC-MS (Shimadzu GC-17A) fixed with the same type of
capillary column as GC and under ionization energy of 70 eV. The operation parameters were: He as
carrier gas at flow rate of 20 cm3 min−1, splitless, injector temperature 250oC, and detector
temperature 280oC. The column was programmed initially at 50oC for 2 minutes, and then ramped to
300oC at a rate of 6.5oC min−1 and held for 7 minutes.
The components were identified tentatively by comparing their Kovat’s retention indices with
literature values and their mass spectral data with those from NIST mass spectral database. The
retention indices were calculated for all components using a homologous series of n-alkanes as
standards [13]. For mass spectral analysis, the components were identified by matching the MS with
those of authentic standards held in the NIST library. Only similarity indices of 85 or higher were
taken as proof of identity [14].
Bioassay-guided fractionation
The biological activity of the extract was performed against brine shrimp larvae (A. salina) and
termites (Coptotermes sp.) using the methods described below.
About 2.0 mL of the essential oil was applied to a silica gel column, and eluted with hexane and a
gradient mixture of hexane:EtOAc (19:1, 9:1, 4:1, 1:1). A total of 70 fractions (25 mL each) were
collected and combined into 6 groups (PS1 to PS6) based on the similarities in TLC profiles. The
activity against larvae of A. salina was found in fractions PS1, PS4 and PS6. These 3 fractions were
then submitted to further fractionation and purification. Fraction PS1 (352 mg) was subjected to CC
on silica gel and eluted with gradient mixture of hexane:EtOAc (9:1, 4:1) yielding 30 fractions, which
were pooled into 3 groups. Preparative TLC of group 1[hexane:EtOAc (9:1)] yielded 252 mg of
compound (1). Fraction PS4 (212 mg) was subjected to CC on silica gel and eluted with a mixture of
hexane:benzene:chloroform (3:1:2) yielding 15 fractions, which were pooled into 3 groups.
Preparative TLC of group 2 [hexane:benzene:chloroform (3:1:2)] yielded 157 mg of compound (2).
Fraction PS6 (272 mg) was subjected to CC on silica gel and eluted with gradient mixture of
hexane:EtOAc (4:1, 1:1) yielding 15 fractions, which were pooled into 3 groups. Preparative TLC of
group 2 [hexane:EtOAc (4:1)] yielded 136 mg of compound (3). The bioactivities of these three
compounds were determined against larvae of A.salina and termites (Coptotermes sp.)
66
Toxicity assay
The protocol established by Mclaughlin [15] was adopted for the toxicity assay against the larvae of
A. salina. The test samples were prepared by dissolving separately 2 mg of each extract in 2 mL of
methanol. From these solutions, 500, 50, and 5 µL were transferred to vials. The solvent was removed
under vacuum and 5 mL of artificial seawater was added to each vial, resulting in final concentrations
of 100, 10 and 1 µg mL−1 respectively. Then 2 mL of these diluted solutions were transferred to 24
well multidish and second instar larvae of A. salina (10 per well) were added. After 24 hours contact,
the number of dead larvae in each well was counted and the percentage of death was plotted against
the concentrations (on a log scale). The LC50 values were determined graphically. The LC50 is defined
as the lethal concentration of the sample at which 50% of the larvae do not show visible mobility.
Controls with and without thymol (10 µg mL−1) were run simultaneously. All experiments were run in
triplicate.
Antitermite assay
The modified method established by Sakasegawa et al [16] was used for the antitermite assay against
a Coptotermes sp. collected around Kuching area. The termites were cultured for 2 – 3 days in a
plaster container at room temperature. In the bioassay for termiticidal activity, cut filter paper
(diameter 35 mm) were placed in each well of a 6-well multidish (3 rows × 2 lines, hole diameter 35
mm). The samples were diluted to 10.0%, 1.0% and 0.1% with methanol. Exactly 80 µL of these
diluted samples were pipetted onto the filter paper in each well of one line. Exactly 80 µL of methanol
were placed on the filter paper in each well of the other line, which acted as control. The methanol
was allowed to evaporate from the filter paper for several hours. 6 termites (5 undifferentiated
workers and a soldier) were placed on each well. The absence of soldier causes the initiation of
physiological process in which a certain number of workers become soldiers [17]. The multidishes
were closed tightly and kept at 25oC in an incubator. The numbers of living termites were counted
each day. Each treatment was performed 6 times (3 wells/multidish × 2 times).
Hydrodistillation of the air dried leaves of P. sarmentosum yielded 1.10% (v/dry weight) of essential
oil. There were 31 components (greater than 0.1%) identified in the leaf oil of P. sarmentosum (Table
1). Sesquiterpenoids were the main constituents with sesquiterpene hydrocarbons representing 30.3%
and oxygenated sesquiterpenes 61.0% of the oil. The major component of the sesquiterpenes
hydrocarbons was β-caryophyllene (18.2%) while spathulenol (21.0%), myristicin (18.8%) and (E,E)-
farnesol (10.5%) were the main oxygenated sesquiterpenes. Monoterpenoids only made up of 5.1% of
the oils with α-phellandrene the only monoterpene hydrocarbon present.
This leaf oil showed inhibitory activity against the larvae of A. salina with LC50 value of 35.2
µg/mL, and 100% mortality within two days at 1% concentration against the subterranean termite
(Coptotermes sp.). After first fractionation, the fractions obtained were subjected to biological
activity. The biological activity against larvae of A. salina was found in fractions PS1, PS4 and PS6
with all showing LC50 value of less than 30 µg/mL after 24 hours of contact (Table 2). The result
showed that the three fractions might contain bioactive components.
Bioassay-guided chromatographic separation of the three active fractions afforded three
compounds. All the three compounds were subsequently tested against A. salina and subterranean
termite (Coptotermes sp.). Compound 2 and 3 showed stronger biological activity against the larvae
of A. salina and termites compared to compound 1. Compound 2 showed the most potent activity
against the larvae of A. salina with LC50 value of 7.5 µg/mL, while the LC50 values for compound 3
and compound 1 were 17.2 µg/mL and 22.5 µg/mL respectively. Compound 3 possess the strongest
inhibitory activity against the subterranean termite with 100% mortality after 3 days at 0.1%
concentration (Figure 1) followed by compound 2 with the similar mortality rate at 0.5%
67
concentration (Figure 2). Compound 1 showed the weakest inhibitory activity with 80% mortality
after 3 days at 2% concentration (Figure 3). However, detailed structure-activity relationship should
be further investigated.
As there is no previous report on the activity of these three volatile oil components on termites,
the data obtained will be used for further study to develop environmental friendly termiticital
compounds.
Table 1: Chemical composition of the essential oils from the leaves of P. sarmentosum
Compound Kovat’s Index % RA

1
KI KI2
α-Phellandrene 1005 1007 0.78
Piperitone 1245 1245 0.67
Cinnamyl alcohol 1314 1312 0.18
Eugenol 1366 1364 1.80
α-Copaene 1377 1377 0.29
Methyl eugenol 1407 1407 1.63
α-Ionone 1422 1422 2.96
γ-Elemene 1426 1425 2.48
β-Caryophyllene 1465 1467 18.19
α-Humulene 1469 1467 0.86
β-Guaiene 1481 1483 0.43
Germacrene D 1488 1487 1.26
Ethyl laurate 1494 1494 1.09
α-Farnesene 1499 1500 0.21
Elemicin 1512 1514 0.88
Bicylogermacrene 1516 1517 0.34
δ-Cadinene 1519 1519 0.72
Cadinadiene 1526 1527 0.96
Myristicin 1533 1532 18.77
γ-Cadinene 1543 1543 2.26
Germacrene B 1562 1562 2.26
Guaiol 1587 1589 0.69
Dehydrocarveol 1591 1593 0.85
Spathulenol 1617 1619 20.98
T-Muurolol 1636 1635 0.21
β-Eudesmol 1648 1648 0.58
β-Bisabolol 1668 1666 0.26
δ-Cadinol 1673 1674 0.29
α-Cadinol 1677 1676 0.70
E,Z-Farnesol 1696 1696 2.19
E,E-Farnesol 1720 1722 10.50
KI1 = Kovat’s retention indices obtained on a DB-5 column using the series of n-alkanes.
KI2 = Kovat’s retention indices from literature. [13]
RA = Relative area (peak area relative to total peak area).
68
Table 2: LC50 values against larvae of A.salina.
Fraction PS1 PS2 PS3 PS4 PS5 PS6

LC50 (µg/mL) 29.1 >100 nd* 21.6 82.0 23.5
nd* – not done due to inadequate amount of sample
90.0
80.0
70.0
60.0 2.00%
% Death
1.00%
50.0
0.50%
40.0
0.10%
30.0
Control
20.0
10.0
0.0
1 2 3 4
Day
Figure 1: Antitermite activity of Compound (1) at different concentrations
120
100
2.00%
80 1.00%
% Death
60 0.50%
40 0.10%
Control
20
0
1 2 3 4
Day
120
100
2.00%
80
1.00%
% Death
60 0.50%
0.10%
40
Control
20
0
1 2 3 4
Day
69
Conclusion
The finding of this study showed that essential oil from leaves of P. samentosum contains larvicidal
and termiticidal components. Further studies are required to increase the lethality of these components
due to the synergy achieved by the presence of other minor components of the oil.
Acknowledgement
This research was financially supported by a fundamental grant 01/19/295/2002(33) from Universiti
Malaysia Sarawak.
References
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constituents of some Piper species. Flavour Fragrance. J., 14, 279 - 282.
2. Sengupta, S., & Ray, A.B., (1987). The chemistry of Piper species: a review. Fitoterapia, 58,
147-165
3. Parmer, V.S., Jain, S.C., Bisht, K.S., Jain, R., Taneja, P., Jha, A., Tyagi, O.D., Prasad, A.K.,
Wengel, J., Olsen, C.E., & Boll, P.M., 1997. Phytochemistry of the Genus Piper.
Phytochemistry, 46 (4), 597 – 673.
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Bong, C.E.J. & Ipor, I.B. (Eds). The Pepper Industry: Problems and Prospect. Universiti
Pertanian Malaysia, Bintulu Campus, 72 – 78.
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Kuala Lumpur.
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Hadi, A.H., (1999). Chemical constituents and insecticidal activity of Piper sarmentosum. In:
Manaf Ali, A., Khozirah, S & Zuriati, Z. (Eds.), Phytochemical and Biopharmaceutins from the
Malaysian Rain Forest. FRIM, Kepong, 62-66.
7. Masuda, T., Inamuzi, A., Yamada, Y., Padolina, W.G., Kikuzaki, H. & Nakatani, N., (1991).
Antimicrobial phenylpropanoids from Piper sarmentosum. Phytochemistry 30, 3227–3228
8. Ridtitid, W., Rattanaprom, W., Thaina, P., Chittrakarn, S., & Sunbhanich, M. (1998).
Neuromuscular blocking activity of methanolic extract of Piper sarmentosum leaves in the rat
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Antimalarial activity of extracts of Malaysian medicinal plants. J. Ethnopharmacol. 64, 249–
254
10. Peungvicha, P., Thirawarapan, S., Temsiririrkkul, R., Watanabe, H., Prasain, J. K., & Kadota, S.
(1998). Hypoglycemic effect of the water extract of Piper sarmentosum in rats. J.
Ethnopharmacol., 60, 27–32.
11. Rukachaisirikul, T., Siriwattanakit, P., Sukcharoenphol, K., Wongvein, C., Ruttanaweang, P.,
Wongwattanavuch, P. & Suksamrarn, A. (2004). Chemical constituents and bioactivity of Piper
sarmentosum. J. Ethnopharmacol., 93, 173-176.
12. Chanwitheesuk, A., Teerawutgulrag, A. & Rakariyatham, N. (2005). Screening of antioxidant
activity and antioxidant compounds of some edible plants of Thailand. Food Chemistry, 92,
491-497.
13. Acree, T. and Arn, H. (2004). http://www.Flavornet.com. [online], retrieved on 7 March, 2005.
14. Adam, R.P., 1995. Identification of Essential Oil Components by Gas Chromatography/Mass
Spectrometry. Allured, Carol Stream, IL., USA.
70
15. McLaughlin, J.L., (1991). Grown gall tumours on potato disc and brine shrimp lethality: two
simple bioassay for higher plants screening and fractionation. In: Hostettmann, K. (Ed), Assays
for Bioactivity, Academic Press, San Diego, 2-32.
16. Sakasegawa, M., Hori, K. & Yatagai, M., (2003). Composition and antitermite activities of
essential oils from Melaleuca species. J. Wood Sci., 49, 181-187.
17. Tellez, M.R., Khan, I.A., Kobaisy, M., Schrader, K.K., Dayan, F.E. and Osbrink, W., (2002).
Composition of the essential oil of Lepidium meyenii Walp.. Phytochemistry, 61, 149-155.
71
MICROBIAL INFLUENCED CORROSION CAPABILITY OF

SULFATE-REDUCING BACTERIA ISOLATED FROM MALAYSIAN
OF THE SHIPYARD AND ENGINEERING HARBOURS, PASIR GUDANG
Fathul Karim Sahrani∗, Zaharah Ibrahim∗, Madzlan Aziz ∗∗ and Adibah Yahya∗
∗Department of Biology, Faculty of Science, Universiti Teknologi Malaysia,

81300 Skudai, Johor Darul Takzim
∗∗Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia,
81300 Skudai, Johor Darul Takzim
e-mail: fathulreem@yahoo.com
Abstract. Sulfate-reducing bacteria (SRB), implicated in microbiologically influenced corrosion,

were isolated from the deep subsurface at Malaysian Shipyard and Engineering (MSE) harbours, Pasir
Gudang. Alloy steel specimens were exposed to natural seawater to study the weight loss of
biocorrosion in both biotic and abiotic environments. Open circuit potentials (OCP) for the specimens
in presence and absence of SRB grown in VMNI media have been performed. Scanning electron
microscopy (SEM) images were obtained for the specimens, which were exposed to SRB cultures.
Energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD) were used to analyse the
corrosion products. Open circuit potential measured for 15 days showed great differences in the
presence and absence of SRB in the VMNI culture media. SEM observations resulted in good
correlation with open circuit potential data which show that the experimentally observed strong
acceleration in pitting corrosion process , induced by the SRB. EDS analysis of specimens indicated
high counts of sulphur and this confirms the presence of sulphur in the corrosion product was
confirmed by XRD analysis.
Abstrak. Bakteria penurun sulfat (SRB) yang mempengaruhi kakisan telah dipencilkan daripada
dasar persekitaran MSE, Pasir Gudang. Spesimen keluli aloi telah di dedahkan dalam air laut sebenar
untuk mendapatkan kehilangan berat oleh proses kakisan bio dalam persekitaran biotik dan abiotik.
Keupayaan litar terbuka (OCP) terhadap spesimen dalam medium VMNI dengan dan tanpa kehadiran
SRB turut dijalankan. Imej dari spektroskopi imbasan elektron juga diperolehi dari spesimen yang
didedahkan kepada kultur SRB. Spektroskopi tenaga penyebaran (EDS) dan penyebaran sinar-x
(XRD) digunakan untuk menganalisis produk kakisan. Pemonitoran selama 15 hari bagi keupayaan
litar terbuka menujukkan terdapat perbezaan yang besar dengan dan tanpa kehadiran SRB. Hasil
pemerhatian daripada SEM menunjukkan terdapat korelasi yang baik bagi menerangkan data
keupayaan litar terbuka dimana berlaku peningkatan yang ketara dalam proses kakisan berlubang.
Analisis EDS terhadap spesimen menunjukkan kiraan yang tinggi kepada unsur sulfur dan dibuktikan
bahawa unsur ini wujud dalam produk kakisan melalui analisis XRD.
Keywords: Sulfate-reducing bacteria, alloy steel, microbiologically influenced corrosion, corrosion

potential, localized corrosion. Scanning electron microscopy.
Introduction
Sulfate-reducing bacteria (SRB) are usually part of the indigenous community of microorganisms in
an ecosystem and are of great utilitarian and academic interest. They are of significant economic
sectors and of environmental concern because of their roles in contamination of petroleum products
and anaerobic corrosion of steel.
The role of SRB in anaerobic corrosion processes is indisputable [1,2,3,4]. Corrosion

processes involve biotic and abiotic factors. The corrosion of stainless steel in marine systems, for
72
example, is influenced by micro organisms and may be entirely biotic. Biocorrosion processes on
metal surfaces depend on the physiology of the microbial community developing under the influence
of the chemical and physical parameters of the environment. The process of corrosion is directly
linked to the growth and activities of these bacteria. Open circuit potentials (OCP) for steel that are
exposed to natural seawater are ennobled, which may result in localized corrosion on their surfaces
[5,6]. Ennoblement of passive metal does not occur when exposed to sterile seawater [7]. Natural
SRBs are a main cause of OCP ennoblement, which be related to the occurrence of localized
corrosion. Due to its simplicity, open-circuit corrosion potential measurements have been used in MIC
studies for many years [8]. A plot of potential as a function of time could be useful to detect the
initiation of an accelerated attack of SRB [9]. Rapid changes in the corrosion potential can be used to
indicate depolarization, or enhancement of the anodic reaction, or to the formation of a semi-
protective film [10]. On the other hand, the important mechanisms potentially involved in case of
MIC can be classified as follows: (i) Cathodic depolarization; (ii) Formation of occluded area on
metal surface; (iii) Fixing the anodic sites and (iv) Under deposit acid attack. Cathodic depolarization,
also known as the ‘classical theory’ is based on the idea that certain strains of SRB possess the
hydrogenase enzyme and therefore are able to catalyse both the reversible reactions involving
hydrogen and the reduction of sulfate by molecular hydrogen. This paper primarily focuses on the
impact of bacteria on cathodic corrosion processes.
Previous work reported the possibility of detecting a cathodic depolarization under the impact
of these bacteria [11,12,13]. The cathodic curve obtained on alloy steel in sterile seawater has a
typical form for the corrosion process with intense depolarization on reaction with hydrogen. It means
that the speed of corrosion of the sample was determined by the removal of hydrogen layer on
working electrode. The effect of hydrogen removal from the metal surface and ferrous sulphide (FeS)
formation in SRB-induced corrosion was recently stressed by Little and Wagner (1997) [14]. The
contribution of FeS to the cathodic depolarization was investigated by Tiller and Booth (1962) [15].
The rate of corrosion was proportional to the amount of FeS added and dependent on the contact of
FeS with metal surfaces. These results allowed reinterpretation of the way metal corrosion is induced
by sulfate reducers.
Electrochemical methods have been used to follow corrosion processes [16]. The Eoc (open
circuit potential) is defined as the potential between the metal coupon (working electrode) and the
platinum counter electrode relative to the reference saturated calomel electrode (SCE). Measurement
of Eoc changing with time is important for estimating the effect of depolarizers on corrosion reactions.
An increase of Eoc means depolarization of cathode and corrosion. A drop in potential is evidence for
decreased corrosion [17]. The present paper aims at investigating an ennoblement of open circuit
potential (OCP) of alloy steel, which is responsible for the occurrence of localized corrosion. Weight
loss measurement, scanning electron microscopy (SEM) examinations and electron diffraction
spectroscopy (EDS), have been performed on steel samples exposed to filter-sterilized seawater and
VMNI media with and without living cultures of the SRB.
Experimental
Media and Cultivation Conditions
A growth medium for marine SRB was proposed by Zinkevich et al. (1996) which is modification of
Posgate’s Marine medium C [18]: KH2PO4 (0.5g); NH4Cl (1.0g); CaCl2.6H2O (0.04g); MgSO4.7H2O
(0.06g); sodium lactate (6.0ml); yeast extract (1.0g); FeSO4.7H2O (0.5g); sodium citrate (0.3g);
Na2SO4 (4.5g); casamino acids (2.0g); thioglycillic acid (0.1g); trace elements (1.0ml); vitamins
(2.0ml); tryptone (2.0g); ascorbic acids (0.1g). All of the chemicals were weighed and then made into
a 1 litre solution with filter-sterilized seawater. The media was degassed with N2 for 30 minutes to
create anaerobic condition and pH was adjusted to 7.2 using 1.0M NaOH before autoclaving at 121oC
it was left to cool room temperature before being inoculated with the SRB.
73
The SRB were left to grow for 3 – 5 days in anaerobic jars at temperatures 35oC. The
indication of successful incubation was blackening of the medium within 2-3 days and generation of
H2S were recorded as a positive indication for presence of SRB. Colony morphological characteristics
including size, shape, colour, margin, configuration and elevation were examined using
stereomicroscope (Leica Zoom 2000 Z45V). The 16S rRNA analysis were carried out and work is
currently on going.
Weight loss study in both biotic and abiotic environment
The composition (% wt) of the low- carbon alloy steels was as follows: C, 0.35; Cr, 1.0; Si, 0.8; Mn,
0.8; Mo, 0.25; Ni, 0.25; Fe (remaining %). Test coupons were prepared according to the ASTM
standard for cylindrical rods (dimension: 9.5 mm diameter x 3 mm thickness). Coupons were finished
by using 120, 320, 600, 800, 1000, 1200 grid SiC paper, washed with tap water and acetone and
finally air-dried.
Before immersion into the natural seawater, coupons were left in alcohol for about 24h,
weighed, marked and then exposed to the biotic and abiotic conditions. Seawater was collected from
the Malaysian Shipyard and Engineering harbours, Pasir Gudang. In abiotic conditions, seawater was
sterilized by filtering through a 0.45µm membrane before use. During the 6 months period that the
coupons were immersed in seawater, coupons were taken out monthly, cleaned, weighed after drying
and the corrosion rates, vc, were determined.
(a) (b)
Cuprum Saturated
wire calomel
electrode
working
Glass electrode
tube Nitrogen gas
Epoxy
polyester
Alloy steel
Coupons
Fig. 1. (a) Schematic diagram of alloy steel concentric electrode (working electrode) and (b)
Electrochemical cell
Potential change studies
Open-circuit potential, Ecorr changes were measured against a standard saturated Calomel electrode
placed in the same compartment (Fig. 1). The exposed electrodes used under various conditions were
embedded in polyester had approximate diameter of 9.8 mm. A copper wire was soldered at the rear
of the electrode which was housed in a glass tube to protect it from the test medium. The potential of
the alloy steel electrode immersed in various solutions: (a) filter-sterilized seawater (control) ; (b)
VMNI (control); (c) VMNI+SRB1; (d) VMNI+SRB11 and (e) VMNI+SRB1+SRB11 were measured
using a multimeter (model Megger M8013) and was incubated under semi-anaerobic condition for 15
days. For all measurements, samples collected were VMNI filter-sterilized seawater taken from the
74
vicinity of tested sites and passed through a 0.45µm filter membrane. Prior to the experiment, all glass
wares were autoclaved and aseptic procedures were followed. To identify the corrosion products, the
electrode used in monitoring potential change was analysed using XRD, SEM observations and EDS.
Weight Loss Study
The weight loss and corrosion rate of alloy steel with time under biotic and abiotic environment is
shown in Fig. 4(a) and 4(b). The overall corrosion rate was calculated from the weight change of the
coupon according to ASTM method [24,25]:
Corrosion rate = (K x W) / (A x T x D)
Where:
K = constant (8.76 x 104),
T = time of exposure in hours,
A = area in cm2,
W = mass loss in grams, and
D = density in g/cm3
35
30
25
Weight loss (mg/cm 2)
20
15
Abiotic
10
biotic
0
0 1 2 3 4 5 6 7
Tim e (m onth)
Fig. 4a. Weight loss as a function of time for alloy steel in biotic (natural seawater) and abiotic (filter-
sterilized seawater) environment.
250
200
Corrosion rate (um/year)
Abiotic
biotic
150
100
50
0
0 1 2 3 4 5 6 7
Tim e (m onth)
Fig. 4b. Corrosion rates as function of time for alloy steel in biotic (natural seawater) and
abiotic (filter-sterilized seawater) environment.
75
Results of weight loss study of alloy steel showed that in abiotic environment (ASS) after one
month was 10.4 mg/cm2 and 24.2 mg/cm2 after 6 month of immersion respectively (Fig. 4(a).
However the weight loss of alloy steel in biotic environment (ASNS) was 13.2 mg/cm2 after one
month immersion and 30.2 mg/cm2 after six months immersion. The results showed that the weight
loss of alloy steel in biotic environments was relatively higher compared to that of abiotic
environment. Figure 4(b) represents the results of corrosion rates for 6 months immersion in biotic
and abiotic environment. The highest corrosion rates under both conditions were observed after one
month immersion (ASS = 160.0 µm/year and ASNS = 204.3 µm/year). The rates were then slowly
deceased (ASS = 62.7 µm/year and ASNS = 78.0 µm/year) after six months immersion. In general,
corrosion rates of biotic environments were higher compared to that of abiotic environment. Higher
corrosion rate under biotic environment implied that seawater was a favourable environment for
microbial growth, thus were susceptible to microbiologically influenced corrosion (MIC).
The corrosion rate was highest after one month’s immersion after it which slowly decreased
with time. The observation was to be expected for metal immersed in seawater. Chloride is known to
induce localised corrosion and this may explain why the corrosion of alloy steel remained relatively
high even after six months of exposure of the coupon in seawater (ASS = 160.0 µm/year). When the
coupons were immersed, the metal oxidised into solution as cations i.e., a negative charge on the
metal until equilibrium potential is established between metal and solution. Further possibilities were
the formation of hydroxides or oxides (corrosion products) of the metal, which formed a passive layer
on the surface. This condition affected corrosion causing corrosion process to decrease slowly.
Open Circuit Potential Measurements
Results of open-circuit corrosion potential, Ecorr were measured against a standard saturated calomel
electrode in the culture solution with and without SRB is shown in Fig. 5. The plot of Ecorr as a
function of immersion time, shows large variations in the corrosion potentials. When filter-sterilized
seawater (SW) was used as the medium, the Ecorr shifted initially to more positive values starting at –
582 mVSCE and decrease slowly to a constant value of – 703 mVSCE after 15 days of exposure. While,
the Ecorr of medium VMNI, the initial point value at – 644 mVSCE and then slowly dropped to a value
of – 824 mVSCE. Significantly higher corrosion potential was recorded in the filter-sterilized seawater
compared to VMNI medium. The presence of the yeast extract in VMNI media may explain the
decrease in the corrosion potential observed over long periods of exposure, that is after two weeks.
According to Feron and Dupont (1997), yeast extract may be adsorbed on the electrode surface
inhibiting the corrosion of alloy steel [19]. Medium VMNI also contained these anions but it also
contains the organic nutrients, increasing the complexity of the system.
However, when alloy steel is exposed in VMNI inoculated with bacteria, the trends in the
plots were very different and varied significantly with exposure time compared to VMNI not bacteria.
In VMNI+SRB11 medium, the Ecorr shifted to more positive values after 3 days exposure starting at –
630 mVSCE until it reached a maximum of – 513 mVSCE after 6 days. The potential then dropped
slowly to a value of – 658 mVSCE. The starting potential of VMNI+SRB1 was the lowest that is – 682
mVSCE and increased suddenly to a maximum value of – 548 mVSCE before decreasing slowly to a
value of – 682 mVSCE after 15 days exposure. In medium VMNI+SRB1+SRB2, the corrosion potential
increased after 4th days exposure to reach maximum values of – 530 mVSCE after 11 days exposure.
The potential then dropped slowly to a value of – 721 mVSCE. The Ecorr shifted drastically to more
positive values after 3 or 4 days exposure in the presence of the SRB compared to that without SRB.
These results showed that the composition of microbial consortia within a biofilm could play an
important role in biocorrosion and that SRB isolates vary in their ability to influence the process of
steel deterioration.
76
(a)
Incubation time (day)
-500
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
-550
Potential Ecorr (mVSCE)
Filter-sterilized seawater
-600
VMNI
-650
-700
-750
-800
-850
Fig. 5a. Plots of open circuit potential Ecorr as a function of immersion time in filter-sterilized
seawater and VMNI media sterile.
(b)
Incubation time (day)
-450
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
-500
-550
Potential Ecorr (mVSCE)
-600
-650
VMNI
-700 VMNI+SRBI
VMNI+SRBII
-750 VMNI+SRBI+SRBII
-800
-850
-900
Fig. 5b. Plots of open circuit potential Ecorr as a function of immersion time in VMNI media sterile
(control) and VMNI with some of varies bacterial inoculation of SRB.
According to the corrosion mechanism generally accepted an accumulation of sulphide as

well as an increase of protons is brought about by the growth of the SRB, and this stimulated the
initial dissolution of iron at quite a high rate. On the other hand, when a film of ferrous sulphide
formed on the electrode surface and is trapped by biofilm layer may cause a partial inhibition in the
anodic dissolution of iron. In fact, visual observation of the steel samples after being exposed to
medium containing SRB show a black film over the electrode surface, while the same medium free of
SRB produces a grey film and an orange yellowish precipitate. Medium VMNI also contained these
anions but it also contained the organic nutrients, increasing the complexity of the system. The
presence of the yeast extract may explain the decrease in the corrosion rate observed over long periods
of exposure, that it after two weeks of incubation. According to Feron and Dupont (1997), yeast
extract adsorbed on the electrode surface inhibits the corrosion of steel [19].
77
(a) (b)
Fig. 6. SEM micrographs of alloy steel surface after four days of exposure to the SRB culture in
VMNI: (a) SRBI and (b) SRBII.
(a) (b)
Fig. 7. SEM image of alloy steel surface after the removal of bacterial biofilm and
corrosion products after 15 days of exposure in VMNI media (a) without SRB culture (b) with
SRB culture, showing localized corrosion.
The corrosion potential, Ecorr of alloy steel in the presence of mixed bacterial cultures are
presented in Fig. 5(b). The corrosion potential of the mix-culture (SRBI and SRBII) was only slightly
different than that of the pure-cultures. It has been reported that corrosion appears to be worse when a
wide variety of microorganisms is present [20]. The purecultures usually induced higher corrosion
rates initially, but with time the corrosion rates decreased compared to that of the control (without
bacteria). In another study, it was shown that the degree of corrosion varied with the bacterial species,
carbon sources and the metal used [21].
SEM examination biofilm on the alloy steel’s surface showed the corrosion pitting caused of
bothing bacteria (SRBI and SRBII) of about 5-6 µm long (Fig. 6). The pit morphology of alloy steel is
shown in Fig. 7. When VMNI was used in the presence of SRB (Fig. 7b), small size single pit was
found compared to that of the VMNI medium without SRB. The EDS spectra of the corrosion
products underneath the biofilm in the medium inoculated with and without of SRB (control) are
given in Fig. 8. The presence of sulphur, most probably related with iron sulphides is obvious in the
spectra corresponding to the culture medium with SRB. Analysis of EDS spectra indicates high counts
of sulphur. The presence of iron sulphide in the corrosion product is confirmed by XRD analysis (Fig.
9)
78
(a) (b)
Fig. 8. EDS of corrosion product: after 15 days of exposure in VMNI media (a) without SRB
(control) and (b) with SRB.
According to Anderco and Shuler (1997) [22] mackinawite is a particular form of iron sulphide
that can occur frequently in immersion corrosion studies. It is known that mackinawite is produced
easily from iron and iron oxides by consortia of microorganism including SRB. Moreover, the
presence of mackinawite in corrosion products formed in seawater is indicative of SRB-induced
corrosion [23].
It should be noted that, the importance of microbial consortia on the corrosion behaviour of
steel is a complex phenomenon requiring not only a better understanding of the ecology and
physiology of biofilms but also that of electrochemical reactions at metal/biofilm interface. To date,
relatively few studies have attempted to characterize biological and physicochemical processes in
mix-cultural biofilms comprising of a mono-culture formed on steel. By combining field and
laboratory investigations, our research provides further evidence of the ability of naturally occurring
consortia to influence corrosion rates of steel and emphasizes the necessity of a long term monitoring
of the behaviour of steel when exposed to microorganisms, in order to evaluate the influence of
biological components in the deterioration of steel in marine environment.
21
FeO-SRB
20
19
18
17
16
15
14
13
Lin (Counts)
12
11
10
20 30 40 50 60 70 80 90
2-Theta - Scale
FeO-SRB - File: FeO-SRB.RAW - Type: 2Th/Th locked - Start: 20.000 ° - End: 90.000 ° - Step: 0.050 ° - Step time: 1. s - Temp.: 25 °C (Room) - Time Started: 2 s - 2-Theta: 20.000 ° - Theta: 10.
Operations: Import
DIF - FeO-SRB - FeO-SRB.dif - Y: 95.24 % - d x by: 1. - WL: 1.54056 - 0 -
06-0696 (*) - Iron, syn - Fe - Y: 50.00 % - d x by: 1. - WL: 1.54056 - Cubic -
86-0389 (C) - Mackinawite, syn - FeS - Y: 50.00 % - d x by: 1. - WL: 1.54056 - Tetragonal - I/Ic PDF 5. -
Fig. 9. XRD spectrum of corrosion product in Fig. 6(a) indicating mackinawite of iron sulphide
compound.
79
Conclusions
The following factors have been shown to be important in our study of microbially influenced
corrosion of alloy steel in VMNI media. Both of SRB strains tested demonstrated the ability to
corrode the alloy steel specimen at rates above that seen with an uninoculated control.
Electrochemical measurements and SEM examination show that the experimentally observed strong
acceleration in pitting corrosion process due to an increase in corrosion potential, induced by the both
SRB strains. Under similar cultural conditions, SRB isolates vary in their ability to influence
corrosion of alloy steel. Electrochemical tests and surface techniques act as a good method in analysis,
giving much greater information on the biocorrosion process.
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7. Scotto, V& Lai, M.E. Corrosion Science. 40 (6). 1007-1018.
8. Dexter, S.C., Duquette, D.J., Siebert, O.W. & Videla, H.A. (1991) “Use and Limitations of
Electrochemical Techniques for Investigating Microbiological Corrosion” Corrosion, 308.
9. Tuovinen, D.H. & Cragnolino, G. A. (1986). A Review of Microbiological and Electrochemical
Techniques in the Study of Corrosion Caused by Sulphate-reducing Bacteria. In Corrosion
Monitoring in Industrial Plants Using Non-destructive. Testing and Electrochemical Methods.
Ed. Moran Labine. ASTM STP 908.
10. Dexter, S.C. (1995) “Microbiological effects” In Corrosion Testing and Standards, Application
and Interpretation, ed. R. Baboian, ASTM manual Series. MNL 20.
11. Booth, G.H. & Tiller, A.R. (1960) “Polarization studies of mild steel in culture of sulphare
reducing bacteria” Trans. Faraday Soc. 56: 1689-1696.
12. Booth, G.H. & Tiller, A.R. (1967) “Cathodic characteristics of mild steel of suspension of
sulphate-reducing bacteria” Corros Sci. 8: 583-600.
13. Iverson, W.P. (1966) “Direct Evidence for the Cathodic Depolarization Theory by Bacterial
Corrosion. Science. 151:986-988.
14. Little, B., Wagner, P. & Mansfeld, F. (1997). Microbiologically influenced corrosion. NACE
International, Houston.
15. Tiller A.K. & Booth G.H. (1962) “Polarization Studies of Mild steel in Cultures of Sulfate-
reducing Bacteria II. Thermophilic Organisms” Trans Faraday Soc. 58:110-115.
16. Potekhina, J.S. (1984) “Inhibition of Corrosion on Mild steel Using Aerobic Microorganisms” J.
Prot. Met. 3:469-470.
17. Romanov, B.B. (1965) “The Methods of Investigation Corrosion of Metals” Metallurgy, 280.
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18. Zinkevich, V., Bogdarina, I., Kang, H., Hill, M.A.W., Tapper, R.C. and Beech, I.B. (1996).
Characterization of exopolimers produced by different isolates of marine sulphate-reducing
bacteria. Int. Biodet. Biodeg. 163-172.
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potentials of stainless steels in natural seawater, in: D. Thierry (Ed.), EFC Publications No. 22,
Aspect of Microbially Induced Corrosion, The Institute of Materials, London, pp. 103-112.
20. Franklin, M.J., Niven, D.E., Mittelman, M.W., Vass, A.A., Jack, R.F., Dowling, N.J.E.,
Mackowski, R.P., Duncan, S.L., Ringleberg, D.B. & White, D.C. (1989). An analogue MIC
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measures. Corrosion 89, NACE Proc. Conf., New Orleans, pp. 187-201.
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corrosion of metals. J. Microbiol. Meth., 8, 191-198.
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iron sulphide species using stability diagrams, Computers & Geosciences. 23 (6) 647-658.
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United States.Designation: G1-03.
81
KANDUNGAN TOLUENA DAN XILENA DALAM DEBU JALAN DI SEKITAR KUALA

LUMPUR
Mohd Rozali Othman dan Nor Faizawati Zulkifli
Pusat Pengajian Sains Kimia dan Teknologi Makanan, Fakulti Sains dan Teknologi
Universiti Kabangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia.
e-mail: rozali@pkrisc.cc.ukm.my
Abstrak: Kajian telah dilakukan untuk menentukan kehadiran toluena dan xilena dalam debu jalan yang bersaiz <38 µm dan
38 – 63 µm di sekitar Kuala Lumpur. Kedua-dua bahan ini dipercayai digunakan dalam minyak petrol sebagai bahan tambah
untuk meningkatkan nombor oktana petrol berkenaan. Setelah dilakukan pengekstrakan untuk mengeluarkan kedua-dua
parameter kajian dari fasa pepejal (debu) kepada fasa cecair, analisis seterusnya dijalankan menggunakan Kromatografi Gas
yang dilengkapi dengan Pengesan Pengionan Nyala. Hasil kajian mendapati debu yang dikutip di kawasan pusat bandar
memberikan kepekatan toluena dan xilena yang lebih tinggi berbanding dengan yang dikutip di kawasan pinggir bandar.
Ujian ANOVA yang dilakukan menunjukkan bahawa terdapat perbezaan yang bererti antara lokasi persampelan yang
berbeza dari segi struktur kawasan dan bilangan kenderaan yang melaluinya dan juga antara saiz debu yang berbeza. Ini
menunjukkan bahawa kedua-dua bahan pencemaran ini adalah lebih bersifat setempat dan lebih bergantung kepada
puncanya.
Abstract: The objective of this study is to determine the content of toluene and xylene in street dust (<38 µm and 38 – 63
µm in diameters) collected at various sites in the vicinity of Kuala Lumpur. Both pollutants were believed had been used as
an additive in petrol to increase an octane number. After extraction by either soxhlet or agitating method, gas
chromatography completed with flame ionization detector were used to quantify the amount of both parameters in street dust
samples. Result obtained showed that the samples collected in urban areas recorded a higher concentration of toluene and
xylene. ANOVA test showed that there are a significant differences between locations due to traffic flows and between two
different sizes of dust particles.
Keywords: street dust, toluene, xylene, analysis
Pendahuluan
Kemerosotan kualiti alam sekitar merupakan salah satu masalah yang semakin meruncing dari semasa
ke semasa. Dunia semakin maju dan pesat membangun dalam pelbagai sektor, di mana ini secara
tidak langsung akan memberikan impak yang begitu besar terhadap alam sekitar malah juga
mengganggu kesihatan manusia sejagat. Pencemaran udara adalah salah satu masalah kualiti alam
sekitar yang semakin parah pada masa kini. Terdapat banyak punca yang menyebabkan berlakunya
pencemaran udara. Antaranya pembakaran secara terbuka, pembebasan asap daripada kilang–kilang,
penggunaan kenderaan dan sebagainya. Dalam bahan pencemaran udara terkandung pelbagai jenis
bahan pencemar yang berbahaya terhadap kesihatan manusia sama ada bahan-bahan organik atau
bukan organik.
Debu terutamanya debu jalan raya terhasil daripada pelbagai punca. Debu jalan raya juga
dikenalpasti sebagai unsur bahan pencemaran yang memberikan kesan buruk kepada manusia, haiwan
dan tumbuhan. Debu boleh dikelaskan mengikut sumber dan kesannya kepada kesihatan manusia dan
alam sekitar. Debu dikenali sebagai pepejal aerosol dan kebanyakannya tidak toksik [1], tetapi debu
juga boleh dikatogerikan sebagai toksik jika bersaiz kurang dari 10µm [2, 3].
Debu adalah merupakan zarah–zarah kecil pepejal yang terkandung dalam udara di
persekitaran. Zarah–zarah kecil ini wujud bersama–sama dengan gas, wap dan asap [4]. Debu juga
boleh terhasil daripada pelbagai proses yang lain contohnya seperti daripada asap dari ekzos
kenderaan, emisi dari industri dan pembakaran hutan, di mana zarah-zarah kecil ini akan bergabung
dengan unsur-unsur lain di atmosfera dan membentuk zarahan yang lebih besar dan kemudian akan
termendap dan membentuk debu. Debu merupakan zarah yang cuba memendap dalam masa yang
agak singkat [5]. Kehadiran debu dalam udara juga adalah hasil daripada proses penyepaian
mekanikal dalam proses pemecahan, peleburan atau penggerudian yang kemudiannya terampai ke
udara hasil daripada percampuran dengan gas [6]. Walaupun debu jalan raya mengandungi kepekatan
bahan pencemar yang tinggi, namun ia hanya mewakili penumpukan pencemar semasa [7].
Dengan adanya sifat debu yang boleh terampai, mempunyai saiz yang sangat kecil dan
berkemampuan untuk termendap, maka debu boleh digunakan sebagai matriks yang sesuai untuk
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analisis tahap pencemaran udara di sesuatu kawasan. Di samping itu debu boleh terampai dalam
atmosfera atau dalam gas-gas lain serta tidak boleh meresap tetapi boleh mengenap dengan pengaruh
graviti [8]. Salah satu contoh analisis terhadap debu yang telah dijalankan dalam banyak kajian
terdahulu ialah analisis kandungan logam berat [3, 9, 10]. Di samping itu debu juga sesuai untuk
analisis bahan pencemar organik seperti toluena dan xilena kerana dipercayai terkandungan dalam
minyak petrol kenderaan.
Toluena dan xilena merupakan suatu bahan yang merbahaya kepada hidupan di mana
pendedahan yang maksimum kepada sebatian ini boleh memudaratkan kesihatan manusia [11].
Kedua-dua bahan ini dapat memberi kesan jangka panjang dan jangka pendek kepada manusia
bergantung kepada tahap atau paras pendedahannya. Antara kesan jangka pendek ialah simptom
seperti pening kepala, alergi dan muntah [12]. Manaka kesan jangka masa panjang ialah
menyebabkan barah, kerosakan jantung, hati dan buah pinggang [13].
Bahan–bahan ini dipercayai ditambah ke dalam minyak petrol untuk meningkatkan nombor
oktananya. Bahan ini merupakan antara bahan yang ditambah ke dalam petrol tanpa plumbum untuk
menggantikan plumbum yang bersifat anti-ketuk, di mana bahan-bahan ini akan terbebas ke udara
sama ada bersama asap sekiranya pembakaran tidak sempurna ke atas petrol berlaku atau akan meruap
bersama-sama petrol sekirangnya proses peruapan berlaku [14]. Bahan pencemar ini dilaporkan
terkandung dalam minyak petrol kenderaan dalam kuanttiti yang kecil, tetapi yang menjadi
masalahnya kepada persekitaran ialah penggunaannya adalah tidak terhad iaitu tiada piawai yang
ditetapkan untuk kedua-dua bahan berkenaan [14, 15, 16]. Menurut kajian yang telah dilakukan
amaun toluena dan xilena yang ditambahkan ke dalam minyak petrol adalah sedikit iaitu dalam
jumlah antara 0.1 – 0.6% sahaja, terutamanya apabila penggunaan petrol tanpa plumbum semakin
meningkat [16].
Sehubungan dengan itu kajian ini dilakukan untuk melihat kandungan toluena dan xilena
yang terdapat di permukaan debu serta mengaitkannya dengan kesesakan lalulintas sesuatu lokasi
persampelan. Kawasan persampelan yang terlibat ialah kawasan bandar dan kawasan pinggir bandar
Kuala Lumpur di mana kedua-dua kawasan ini mempunyai kadar kesesakan lalulintas atau bilangan
kenderaan yang berbeza.
Eksperimental
Lokasi Kajian
Lokasi kajian atau kawasan persampelan terbahagi kepada dua kawasan iaitu kawasan bandar dan
kawasan pinggir bandar. Pemilihan kawasan persampelan adalah berdasarkan kepada pemerhatian
tahap kesibukan lalulintas kawasan berkenaan. Kedudukan stesen-stesen persampelan adalah seperti
yang diringkaskan dalam Jadual 1. Persampelan telah dilakukan sebanyak tiga kali di setiap lokasi.
Jadual 1. Kedudukan kawasan persampelan.
Kategori Singkatan Stesen

Kawasan bandar PD Puduraya
DM Dataran Merdeka
BSK Perhentian bas Klang
Kawasan pinggir bandar DK Danau Kota
BU Bukit Unggul
Bahan kimia dan alat radas
Bahan-bahan kimia utama yang digunakan dalam kajian ini adalah seperti petroleum eter (60-80oC
takat didihnya) sebagai pelarut, toluena dan xilena (kesemuanya bergred Analar, BDH Chemical
Limited). Manakala alat-alat radas yang digunakan ialah seperti alat-alat kaca yang berkaitan,
pengayak (bersaiz 38 dan 63 µm), pengekstrak soxhlet yang dilengkapi dengan didal (thimble)
selulosa, penggoncang mekanikal (Grant instrument), Penyejat berputar (Bünchi Rotavapor) dan
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Kromatografi Gas (Hewlett-Packard Company) yang dilengkapi dengan pengesan pengionan nyala
sebagai pengesan untuk analisis toluena dan xilena.
Persampelan dan Pengolahan Sampel
Pengambilan debu dilakukan dengan cermat iaitu menggunakan berus dan penyodok khas yang bebas
xilena. Di mana permukaan jalan disapu secara perlahan-lahan untuk memastikan hanya debu di
permukaan sahaja yang diambil. Kepanjangan jalanraya lebih kurang 100 m diperlukan untuk
memastikan sampel yang cukup diperolehi. Sampel diambil di kawasan pusat bandar dan kawasan
pinggir bandar. Sampel debu yang dikutip ini dimasukkan ke dalam beg plastik untuk dibawa ke
makmal. Berat debu kira-kira dua kilogram diambil bagi setiap stesen. Di makmal sampel debu
seterusnya dipindahkan ke atas piring kertas untuk proses pengeringan di udara sekurang-kurangnya
selama 72 jam untuk memudahkan proses pengayakan. Saiz debu yang dikehendaki adalah berukuran
<38 µm dan 38-63 µm. Saiz butiran debu ini diayak menggunakan pengayak bersaiz tertentu yang
telah ditetapkan.
Kedua-dua kaedah pengekstarakan secara soxhlet dan penggoncangan (menggunakan
penggoncang mekanikal) digunakan dalam kajian ini, dimana kedua-dua kaedah ini adalah merupakan
kaedah klasik yang digunakan untuk mengekstrak bahan organik dari sampel pepejal [17]. Dalam
kedua-dua kaedah pengekstrakan ini petroleum eter dengan takat didih dalam julat antara 60-80ºC dan
debu seberat lebih kurang 1.0 gram (yang telah ditimbang dengan tepat) digunakan. Proses
pengekstrakan ini dilakukan selama dua jam untuk kedua-dua kaedah pengekstrakan.
Alikuot sampel yang terhasil daripada proses pengekstrakan di atas seterusnya dikeringkan
sehingga ke isipadu kira-kira 5 ml menggunakan alat pengeringan penyejat berputar sebelum
dipindahkan ke dalam botol sampel yang bebas daripada toluena dan xilena [18]. Pemekatan alikuot
sampel ke isipadu kira-kira 1.0 ml seterusnya dilakukan menggunakan aliran gas nitrogen sebelum
disuntik ke dalam turus kromatografi. Sampel seterusnya dianalisis secara kualitatif dan kuantitatif
menggunakan kromatografi gas.
Analisis Kromatografi Gas dan Kuantifikasi
Larutan ekstraksi dianalisis menggunakan kromatografi gas (model GC-HP5890 series II, Hewlett-
Packard Company, USA). Keadaan pengoperasian alat kromatografi gas adalah seperti yang
diringkaskan dalam Jadual 2.
Jadual 2. Pengoperasian kromatografi gas
Parameter Peralatan/Keadaan/nilai
Pengesan Pengesan Pengionan Nyala (FID)
Turus Silika
Suhu pengesan 200°C
Suhu penyuntik 150°C
Gas pembawa Nitrogen
Masa pemisahan 40 minit
Nota: Masa pemisahan yang panjang digunakan untuk memisahkan bahan
lain yang hadir bersama
Pengesanan kehadiran toluena dan xilena ditentukan berdasarkan kepada masa penahanan
bagi larutan piawainya dan juga kaedah penambahan piawai. Satu siri kepekatan larutan piawai
toluena dan xilena yang sesuai digunakan untuk menyediakan lengkuk kalibrasi. Selain daripada
untuk menentukan kehadiran toluena dan xilena, kaedah penambahan piawai juga digunakan untuk
menentukan kepekatan toluena dan xilena dalam larutan sampel.
Hasil dan Perbincangan
Contoh kromatogram yang diperolehi dari analisis kromatografi gas untuk salah satu larutan sampel
debu yang telah diekstrak adalah seperti yang ditunjukkan dalam Rajah 1. Manakala Jadual 3 dan 4
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pula meringkaskan kepekatan toluena dan xilena dalam debu jalan yang telah diperolehi dalam kajian
ini.
Rajah 1. Contoh kromatogram bagi sampel yang diekstrak dengan kaedah pengekstrakan soxhlet
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Jadual 3. Kepekatan purata bahan cemar dalam sampel menggunakan kaedah pengekstrakan soxhlet dan pengoncangan
untuk saiz debu jalan <63 µm
Kaedah pengekstrakan Kepekatan (µgg-1)

Toluena Xilena
Soxhlet 203 847
Penggoncangan 163 183
%(Penggoncangan/soxhlet) 80.3% 21.6%
Jadual 4. Purata kepekatan toluena dan xilena dalam sampel debu
Saiz Stesen Kepekatan (µgg-1)

debu persampelan Toluen Xilen
a a
<38 PD 163 100
µm DM 114 td
BSK 203 172
DK 407 572
BU td td
38-63 PD 132 183
µm DM 572 td
BSK 491 938
DK 953 td
BU td td
Nota:td=tidak dapat dikesan
Dalam kajian ini dua kaedah pengekstrakan telah digunakan untuk mengekstrak sampel debu.
Kaedah pengekstrakan tersebut ialah pengekstrakan soxhlet dan penggoncangan. Kedua-dua kaedah
ini adalah merupakan kaedah klasik yang digunakan untuk mengekstrak bahan organik dari sampel
pepejal [17]. Hasil analisis menunjukkan kepekatan paling tinggi bagi bahan cemar organik toluena
dan xilena dikesan pada sampel debu yang diekstrak menggunakan kaedah pengekstrakan soxhlet. Ini
adalah kerana kesan daripada penggunaan isipadu pelarut yang digunakan dan juga kesan dari
pemanasan. Jika pelarut yang digunakan adalah banyak maka lebih banyak bahan organik yang dapat
dijerap keluar daripada permukaan butiran debu begitu juga dengan pemanasan. Aspek kelarutan dan
pemanasan juga mungkin memainkan peranan dalam menentukan peratus kecekapan pengektrakan.
Hasil kajian juga mendapati peratus perbandingan antara kedua-dua kaedah ini adalah lebih tinggi
bagi toluena berbanding xilena (Jadual 3). Keadaan ini menunjukkan bahawa toluena lebih mudah
terekstrak berbanding xilena walaupun kaedah pengekstrakan yang mudah seperti penggoncangan
digunakan. Analisis secara kuantitatif dilakukan untuk menghitung kepekatan kedua-dua bahan
pencemar organik toluena dan xilena dalam sampel debu menggunakan kalibrasi yang telah
disediakan. Jadual 4 menunjukkan purata kepekatan toluena dan xilena dalam sampel debu yang telah
digunakan.
Toluena dan xilena adalah merupakan bahan organik yang sangat berbahaya. Bahan pencemar
organik ini adalah sangat bersifat sangat toksik dan memberikan kesan kepada kesihatan manusia
walaupun bahan-bahan ini hanya wujud dalam kuantiti yang sedikit. Antara gejala yang wujud jika
tahap dedahan tinggi kepada seseorang individu ialah sesak nafas, sakit kepala dan keletihan. Hasil
daripada analisis yang telah dijalankan didapati kehadiran bahan organik berkenaan dalam kajian di
dalam sampel debu jalan adalah kecil, namun ianya tidak boleh dipandang ringan kerana kedua-
duanya merupakan antara bahan yang boleh menyebabkan pembentukan asbut fotokimia di udara dan
ini hanya dikesan pada permukaan debu sahaja. Kandungan bahan pencemar organik berkenaan yang
diperolehi dalam kajian ini adalah bergantung kepada latar belakang kawasan persampelan. Kehadiran
bahan organik kajian tertinggi dapat dikesan dalam sampel dari kawasan bandar yang sibuk dengan
kenderaan. Ini membuktikan bahawa bahan organik ini adalah terkandung dalam minyak petrol
kenderaan.
Penggunaan kenderaan di kawasan pusat bandar adalah lebih banyak jika dibandingkan di
kawasan pinggir bandar. Maka tahap kepekatan pencemaran yang disebabkan oleh toluena dan xilena
86
yang diperolehi adalah lebih tinggi di kawasan pusat bandar. Pelepasan bahan pencemar ini ke
atmosfera adalah mungkin secara tumpahan minyak petrol, kebocoran enjin kenderaan atau melalui
peruapan petrol. Bahan pencemar ini mempunyai sifat meruap, maka bahan-bahan ini akan meruap di
atmosfera dan bergabung dengan zarah-zarah debu yang terampai di udara dan kemudiannya akan
termendap secara graviti [8].
Hasil daripada kajian mendapati terdapat perbezaan kepekatan bahan organik yang dikesan
bagi kawasan persampelan yang berlainan. Kandungan toluena dan xilena yang paling tinggi dapat
dikesan adalah sampel kawasan pusat bandar iaitu stesen Puduraya. Kepekatan toluena dan xilena
bagi stesen yang lain boleh dilihat pada Jadual 4. Bagi sampel debu dari kawasan pinggir bandar iaitu
stesen Bukit Unggul tiada kehadiran toluena dan xilena dapat dikesan. Ini menunjukkan
kebergantungan antara kadar penggunaan kenderaan dengan amaun bahan organik yang dikesan.
Namun pegecualian dapat dilihat di stesen Danau Kota (Jadual 4). Keadaan ini berkemungkinan
berlaku kerana debu di kawasan berkenaan adalah debu yang telah lama, kerana kawasan berkenaan
tidak dibersihkan sekerap di kawasan pusat bandar, oleh itu penumpukan toluena dan xilena berlaku.
Ujian ANOVA sehala tanpa replikasi pada paras keyakinan 95% menunjukkan bahawa kawasan
persampelan di pusat bandar dan kawasan pinggir bandar memberi perbezaan yang bererti.
Dalam kajian ini, sampel debu yang diperoleh dari kawasan persampelan yang telah dipilih
terbahagi kepada dua iaitu dengan saiz butiran bersaiz <38 µm dan 38-63 µm. Tujuan menggunakan
saiz butiran debu yang berlainan ialah untuk mengetahui kebergantungan kepekatan bahan pencemar
organik yang dapat diekstrak dengan saiz butiran debu.
Hasil daripada analisis yang dijalankan bagi saiz butiran debu mempengaruhi kepekatan
toluena dan xilena yang terdapat di permukaan debu berkenaan (Jadual 4). Di mana didapati saiz debu
yang lebih besar iaitu antara 38-63 µm memberikan kepekatan toluena dan xilena yang lebih tinggi
(kecuali ada lokasi terentu memberikan nilai yang sebaliknya) (Jadual 4). Ini mungkin disebabkan
oleh peratus fraksi yang bersaiz <38 µm adalah kecil dan berbeza antara lokasi. Contohnya kawasan
yang berpasir mempunyai saiz butiran debu bersaiz kecil yang lebih rendah dan begitu juga
sebaliknya. Ujian ANOVA satu hala tanpa replikasi pada paras keyakinan 95% menunjukkan bahawa
secara amnya terdapat perbezaan yang bererti antara saiz butiran debu, iaitu debu yang bersaiz 38-63
µm memberikan kepekatan toluena dan xilena yang lebih tinggi berbanding debu bersaiz <38 µm
bagi hampir kesemua lokasi kajian.
Hasil kajian juga mendapati bahawa tidak terdapat trend yang tetap antara kandungan toluena
dan xilena (Rajah 3). Namun berdasarkan kepada Rajah 3, dapat dilihat dengan lebih jelas bahawa
sekiranya kaedah pengekstrakan soxhlet digunakan kandungan xilena adalah lebih tinggi berbanding
dengan toluena. Keadaan ini menunjukkan bahawa terdapat kemungkinan penggunaan xilena yang
lebih tinggi berbanding toluena dalam petrol dan juga mungkin kerana xilena lebih sukar meruap di
udara, menjadikannya lebih mudah termendap pada zarah-zarah debu.
Kesimpulan
Secara keseluruhannya, hasil daripada kajian yang telah dijalankan mendapati bahan pencemar oganik
seperti toluena dan xilena terdapat dalam sampel debu jalan raya yang dianalisis. Hasil daripada
analisis terhadap sampel debu dilakukan didapati kepekatan bahan pencemar organik toluena dan
xilena yang paling tinggi dapat dikesan adalah di kawasan pusat bandar. Perbezaan kedua-dua
kawasan persampelan ini adalah dari segi kesibukan kenderaan, maka ini membuktikan bahawa
kehadiran bahan pencemar organik ini adalah berpunca daripada minyak petrol kenderaan.
Penggunaan dua kaedah pengekstrakan yang berbeza menunjukkan bahawa toluena lebih mudah
diekstrak berbanding dengan xilena. Saiz butiran juga mempengaruhi kandungan toluena dan xilena
dalam debu jalan. Secara amnya kandungan xilena juga adalah lebih tinggi berbanding dengan toluena
dalam debu jalan yang digunakan yang mungkin disebabkan oleh pengunaannya yang lebih meluas.
Penghargaan
Penulis ingin merakamkan ribuan terima kasih kepada semua yang terlibat secara langsung dalam
menjayakan penyelidikan ini. Terima kasih juga kepada pihak UKM kerana bantuan kewangan yang
diberikan.
87
Rujukan
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162-172.
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Sekitar Kuala Lumpur, Kajang, Bandar Baru Bangi, Bangi Lama, Nilai dan Seremban”. Malays. J. Anal. Sci. 6(1).
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Garden Soils and House Dust in an Old Lead Mining Area of North Wales, Great Britain” Environ. Pollut. 9 (Series
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Sciences”, New York: Cambridge University Press.
8. Zaini, H. (1997) “Pengenalan Pencemaran Udara”. Kuala Lumpur: Dewan Bahasa Dan Pustaka
9. Rozali, M. O., Mohd Kamal Md. Zin (1997) “Kandungan Logam Berat Dalam Debu Jalan Di Sekitar Bandar Kota
Kinabalu, Sabah”. Malays. J. Anal. Sci. 3(1). 101-112.
10. Massadeh, A.M. (2003) “ Distributions of Copper and Zinc in Different Fractions of Particle Sizes in Road Dust
Samples in Irbid City, Jordan using Atomic Absorption Spectrometry” Res. J. Chem. Environ. 7(4). 49–54.
11. Dictionary Organic Compaunds (1983) Ed. Ke-5. New York: Chapman & Hall.
12. Butler, J.D. (1979) “Air Pollution Chemistry”, London: Academic Press.
13. Kupchella, C.E., Hyland, M.C. (1993) “Environment Science: Living within the System of Nature”, Ed. Ke-3
14. Simons, C. (1995) “The Lies of Unleaded Petrol - Part 2. Highly Toxic Chemicals are Replacing the Lead in Our
Fuel, yet Government Authorities Continue to Underestimate the Serious Risks to Public Health” NEXUS Magazine..
2(26). http://www.nexusmagazine.com/articles/ulp2.html
15. Crabtree, J.M, Roziah Hj. Mohamed, Zani bin Assim (1997) “Aromatics (Benzene, Toluene and Xylene) and
Oxygenate (Methly tert-Butyl Ether) Contents in Unleaded Gasoline. Abstracts 2nd International Conference on
Environmental Chemistry and Geochemistry In The Tropic (Geotrop)”, hlm. 31. Kuala Lumpur: Malaysia.
16. Kokosa, J.M., Andrzej, P. (2002) “Headspace Microdrop Analysis- An Alternative Test Method for Gasoline Diluent
and Benzene, Toluene, Ethlybenzene and Xylenes in used Engine Oils” J. Chromatogr. 983. 205-214.
17. Dean, J.R., Guohua Xiong, (2000) “Extraction of Organic Pollutants from Environmental Matries: Selection of
Extraction Technique” Trends in Anal. Chem. 19(9). 553-558.
18. Buch, A., Sternberg, R., Meunier, D., Rodier, C., Laurent, C. (2003) “Solvent Extraction of Organic Molecules of
Exobiological Interest for in-situ Analysis of the Martin Soil” J. Chromatogr. A(999). 165-174.
88
210
PO/210PB DALAM AIR HUJAN, AIR SUNGAI DAN AIR LAUT
Che Abd Rahim Mohamed, Wong Fatt Yin dan *Zaharuddin Ahmad
Pusat Pengajian Sekitaran & Sumber Alam
Fakulti Sains & Teknologi
Universiti Kebangsaan Malaysia
43600 Bangi, Selangor
*Malaysian Institute Technology Nuclear (MINT)

Bangi, Selangor.
E-mail: carmohd@pkrisc.cc.ukm.my, mohamed6566@yahoo.com
Abstrak: Kajian ini bertujuan untuk menentukan aktiviti 210Po dan 210Pb dalam air hujan, air sungai dan air laut.
Kawasan persampelan air hujan dan air sungai terletak di sekitar Kajang-Bangi, Selangor manakala air laut di
Kapar, Selangor dan Pulau Tinggi, Johor. Aktiviti 210Po dan 210Pb dalam sampel air ditentukan dengan
menggunakan Spektrometer α dan pembilang αβ paras rendah. Julat aktiviti 210Po dalam air laut di Kapar dan
Pulau Tinggi masing-masing adalah 0.020-0.153 dpm/L dan 0.046-0.216 dpm/L. Hasil kajian menunjukkan
terdapat perbezaan aktiviti 210Po dalam kedua-dua kawasan yang dikaji berpunca daripada keadaan geografi dan
aktiviti yang berlaku di kawasan tersebut. Manakala aktiviti 210Po dalam air hujan dan air sungai adalah agak
tinggi dengan julat aktiviti masing-masing ialah 0.117-0.598 dpm/L dan 0.111-0.251 dpm/L. Pelepasan sisa
industri dan kenderaan bermotor ke atmosfera adalah faktor yang mempengaruhi aktiviti 210Po dalam air hujan
dan air sungai. Untuk aktiviti 210Pb pula, julat aktivitinya pada air laut di Pulau Tinggi adalah di antara 0.046-
0.126 dpm/L dengan purata 0.087 dpm/L. Aktiviti 210Pb di Pulau Tinggi adalah berpunca daripada input Pb dari
atmosfera ke permukaan lautan. Dalam kajian ini, corak bagi jumlah keseluruhan aktiviti 210Po yang ditunjukkan
oleh ketiga-tiga sampel ialah air hujan> air sungai> air laut.
Abstract: This study was carried out to determine the activity of 210Po dan 210Pb in rainwater, river and
seawater. The sampling area for rainwater and river water was collected from Kajang-Bangi, Selangor
meanwhile seawater from Kapar, Selangor and Pulau Tinggi, Johor. Activity of 210Po and 210Pb in water sample
were measured using spectrometer α and low background αβ counting system. The activity range of 210Po in
seawater from Kapar and Pulau Tinggi are found to be 0.020-0.153 dpm/L and 0.046-0.216 dpm/L respectively.
The result from this study showed a different activity of 210Po in both areas. The variation was due to geography
condition and the activities in that area. On the other hand, the activity of 210Po in rainwater and river water was
a bit high with a range of 0.117-0.598 dpm/L and 0.111-0.251 dpm/L respectively. Industrial and automobile
waste releases to atmosphere were the factor which influenced the activity of 210Po in rainwater and river water.
Meanwhile, the activity range of 210Pb in seawater collected from Pulau Tinggi was 0.046-0.126 dpm/L with
mean concentration of 0.087 dpm/L. The activity of 210Pb in Pulau Tinggi was caused by the input of Pb from
atmosphere to sea surface. In this research, the pattern of total activity of three samples is rainwater > river
water > seawater.
Pendahuluan
210
Po dan 210Pb merupakan ahli kepada siri reputan 238U yang mana pereputannya berlaku secara semulajadi.
Sebelum ini, 210Po dan 210Pb merupakan dua unsur yang paling banyak digunakan sebagai penyurih radioaktif
untuk mengkaji proses-proses biogeokimia. 210Po dalam persekitaran marin merupakan produk reputan 210Pb
yang kebanyakannya diserap oleh zooplankton dan melekat pada partikel organik. Maka sifat 210Po dalam
persekitaran marin adalah berkaitan dengan aktiviti metabolik zooplankton dan ketumpatan zooplankton di
permukaan air (Tateda et al. 2003). Akan tetapi 210Pb adalah terhasil di atmosfera dan melekat kepada partikel
organik yang halus. Maka sifat 210Pb dalam sistem akuatik lebih dipengaruhi oleh pemendakan aluminosilikat
seperti partikel tanah liat dan debu dalam turus air (Hung & Chung, 1998). Oleh itu, dua nuklid ini merupakan
penyurih yang sesuai untuk partikel organik dan inorganik yang halus di persekitaran marin.
Di laut terbuka, keseimbangan di antara 210Po dan 210Pb di turus air mencapai hampir tahap yang stabil
kerana (i) beban sedimen yang rendah di mana ia mengawal proses skaveng untuk kedua-dua nuklid, (ii)
produktiviti biologi yang rendah di mana ia menyekat fluks penurunan kedua-dua nuklid dengan ‘fecal pellet’
dan penenggelaman serpihan biogenic dan (iii) input 210Po dari atmosfera dalam kuantiti yang sangat rendah
(Tateda et al. 2003). Akan tetapi, keseimbangan antara kedua-dua nuklid ini di tepi pantai atau estuari adalah
berubah-ubah kerana input 210Po dengan beban sedimen di sungai, proses skaveng untuk 210Pb yang terhad oleh
gangguan lateral 210Pb di kawasan air terbuka, kehilangan 210Po secara peruapan biologi dari permukaan air
(Momoshima et al. 2001), dan pembentukan 210Po dari degradasi berterusan partikel biogenik dan proses
pengaliran atasan.
Justeru itu tujuan kajian ini adalah untuk melihat perubahan aktiviti dalam tiga fasa yang berbeza iaitu
air hujan, air sungai dan air laut. Di samping itu juga untuk membuat perbandingan di antara kawasan
persampelan tersebut
Persampelan dan penganalisaan

Tiga jenis sampel air telah diambil dalam kajian ini iaitu sampel air laut, air sungai dan air hujan. Ketiga-tiga
jenis sampel air dikumpul dari lokasi yang berlainan (Jadual 1). Sampel air laut telah diambil dari dua lokasi
iaitu di Kapar, Selangor dan Pulau Tinggi, Johor (Rajah 1). Kapar merupakan kawasan muara sungai di pantai
barat Semenanjung Malaysia manakala Pulau Tinggi merupakan sebuah pulau yang terletak di Laut China
Selatan. Sebanyak 8 stesen dipilih untuk melakukan persampelan di Kapar dan 6 stesen di Pulau Tinggi.
Sampel air sungai pula diambil di sepanjang Sungai Langat yang mengalir dari Kajang ke Bangi,
Selangor (Rajah 1). Sebanyak enam stesen sampel air sungai diambil di sepanjang Sungai Langat yang mengalir
dari Kajang ke Bangi, Selangor dan sampel air hujan pula dikumpul di satu lokasi Universiti Kebangsaan
Malaysia, Selangor.
Penganalisaan 210Po dan 210Pb berpandukan kepada kaedah Theng and Mohamed et al (2005). Aktiviti
210
Po dibilang dengan menggunakan alfa-spektrometer (EG & G, ORTEC) dengan masa pembilangan 24 jam
untuk setiap sampel. Baki larutan dalam bikar selepas pemendapan spontan digunakan untuk analisis 210Pb iaitu
melalui proses pemisahan resin kation. Kerja-kerja pembilangan zarah-β dilakukan dengan mesin pembilangan
spektrometer gross α/β, Model LB5100-W, TENNELEC dan masa pembilangan ialah 100 minit untuk setiap
sampel.
Keputusan dan perbincangan

Aktiviti 210Po dalam air laut
Pulau Tinggi, Johor
Julat aktiviti 210Po yang diperolehi daripada lapisan permukaan, pertengahan dan dasar masing-masing adalah
0.057-0.096 dpm/L, 0.059-0.122 dpm/L dan 0.046-0.216 dpm/L. Manakala purata aktiviti 210Po yang diperolehi
masing-masing adalah 0.074 dpm/L, 0.076 dpm/L dan 0.100 dpm/L. Secara keseluruhannya, aktiviti 210Po
tertinggi wujud pada lapisan dasar pada stesen 5 dengan nilai 0.216 dpm/L. Secara puratanya, aktiviti 210Po
terendah pula berlaku pada lapisan permukaan lautan.
Pulau Tinggi yang terletak di Laut China Selatan adalah sebuah pulau yang di kelilingi oleh lautan luas
dan terbuka. Pulau Tinggi juga merupakan sebuah pulau yang telah di iktirafkan sebagai kawasan Taman Laut
Malaysia. Oleh itu, kawasan in kurang menerima pencemaran dari luaran atau dalaman kawasan berkenaan.
Input 210Po dari atmosfera ke lautan terbuka adalah rendah (Nozaki et al. 1991). Maka ini akan mengurangkan
aktiviti 210Po pada permukaan lautan.
Aktiviti 210Po yang rendah pada lapisan permukaan juga berkemungkinan adalah kerana proses biologi
yang terlibat di kawasan berkenaan. Ini adalah kerana aktiviti 210Po adalah berhubung kait dengan aktiviti
metabolik zooplankton dan ketumpatan zooplankton di permukaan air (Jefree et al. 1997). Maka permindahan
210
Po dari permukaan dikawal oleh aktiviti biologi dan proses skaveng yang terlibat (Heussner et al. 1990;
Tateda et al. 2003). Aktiviti 210Po dan 210Pb yang tinggi pada fasa terlarut pada dasar lautan juga ditingkatkan
oleh proses pemendapan sedimen dari permukaan dan sumber Pb terlarut di dasar (Theng & Mohamed 2005).
Oleh itu, aktiviti 210Po yang lebih tinggi wujud di bahagian dasar berbanding permukaan lautan.
Kapar, Selangor
Julat aktiviti yang diperolehi pada lapisan permukaan dan lapisan dasar masing-masing adalah di antara 0.020-
0.153 dpm/L dan 0.029-0.081 dpm/L dengan nilai purata 0.064 dpm/L pada permukaan air dan 0.058 dpm/L
pada lapisan dasar. Secara puratanya, aktiviti 210Po adalah lebih tinggi di lapisan permukaan berbanding dengan
lapisan dasar. Keadaan ini adalah berbeza dengan aktiviti 210Po di Pulau Tinggi, Johor di mana aktiviti 210Po
adalah lebih tinggi di lapisan dasar berbanding lapisan permukaan air (Rajah 3).
Kapar terletak di estuari Sungai Kapar Kechil berhampiran dengan Selat Melaka. Ia merupakan sebuah
kawasan penjanaan kuasa elektrik yang menggunakan arang batu. Kawasan ini tercemar dengan sisa buangan
dari stesen penjanaan kuasa elektrik melalui asap yang disalur ke atmosfera dan air panas yang disalur terus ke
lautan. Sumber utama pembebasan radioaktif dari pembakaran arang bukan U dan Th sahaja, tetapi juga produk
yang dihasilkan dari reputan radioaktif seperti radium, radon, polonium, bismuth dan plumbum (Gabbard 2002).
Maka peningkatan aktiviti 210Po pada lapisan permukaan air adalah berpunca daripada input dari atmosfera yang
kebanyakan sumber poloniumnya datang dari proses pembakaran arang batu. Selain itu, sumber polonium di
atmosfera juga wujud dari proses pereputan 222Rn (Yang dan Lin 1992) yang memasuki perairan atau kembali
ke permukaan bumi melalui pemendapan kering dan air hujan (Peck & Smith 2000).
Di samping itu, aktiviti 210Po di persisiran juga dipengaruhi oleh tindakan ombak, faktor geologi dan
input dari aliran air tawar (Wildgust et al. 1998; Theng & Mohamed 2005). Perairan kawasan Kapar yang
merupakan kawasan estuari banyak menerima input dari Sungai Kapar Kechil dan Sungai Kapar Besar. Ini
dapat menjelaskan bahawa lapisan permukaan stesen 5 menerima aktiviti 210Po yang paling tinggi iaitu 0.153
dpm/L.
Aktiviti 210Po yang rendah wujud pada lapisan dasar lautan kecuali stesen 7 dengan nilai purata 0.058
dpm/L. Kapar merupakan perairan yang cetek dengan kedalaman kurang daripada 10 m. Pada perairan yang
cetek proses pencampuran yang berpunca dari arus air dan ombak sering berlaku yang menyebabkan
pencampuran antara sedimen dasar dengan air laut. Ini menyebabkan kandungan polonium tidak termendak ke
dasar tetapi mengalir kembali ke permukaan air.
Pada stesen 7, aktiviti 210Po adalah tinggi di dasar dengan kepekatan 0.081 dpm/L berbanding lapisan
permukaan 0.020 dpm/L. Stesen 7 merupakan lokasi persampelan yang berdekatan dengan jeti. Di sini, jeti
digunakan sebagai kawasan kapal berlabuh untuk menurunkan barangannya seperti arang batu untuk dibawa ke
daratan. Semasa angkutan, kemungkinan arang jatuh ke lautan dan mendak ke dasar menyebabkan aktiviti 210Po
tinggi di dasar lautan. Di samping itu, sisa-sisa buang kapal yang terjatuh ke laut dan mendak juga boleh
menyebabkan kepekatan polonium bertambah.
Aktiviti 210Po dalam air sungai

Kajang-Bangi, Selangor
Stesen persampelan ini terletak di sekitar Sungai Langat yang mengalir dari Bandar Kajang ke Bangi. Daripada
maklumat analisis yang diperolehi, didapati aktiviti 210Po dalam air sungai adalah tinggi dan bertabur secara
tidak seragam bagi semua stesen yang dikaji. Julat aktiviti 210Po yang diperolehi ialah antara 0.111 dpm/L
hingga 0.251 dpm/L dengan purata aktiviti 210Po 0.160 dpm/L (Rajah 4).
Dalam kajian ini, nilai aktiviti 210Po dalam air sungai yang diperolehi adalah agak tinggi. Di
persekitaran Bandar Kajang, diketahui bahawa bandar ini merupakan suatu tempat yang sentiasa dipenuhi oleh
kenderaan bermotor. Penggunaan plumbum tetraetil dalam enjin kenderaan, pembakaran minyak fossil serta
hasil buangan gipsum fosfatik akan meningkatkan kepekatan 210Po dan 210Pb dalam persekitaran akuatik melalui
atmosfera (Stepnowski & Skwarzec 2000). Di samping itu, kawasan Bangi yang merupakan kawasan
perindustrian juga menyumbang kepada kandungan polonium ke dalam sungai ini. Aktiviti 210Po meningkat
dalam sistem akuatik kerana perlepasan dari sisa industri (Carvalho 1997) yang memasuki sungai secara terus
atau melalui atmosfera.
Aktiviti 210Po di sepanjang sungai ini yang paling tinggi wujud pada stesen 4 dengan kepekatan 0.261
dpm/L. Ini berkemungkinan berpunca dari perlepasan asap yang banyak dari kilang papan yang dibina
berdekatan dengan stesen persampelan berkenaan. Ini akan menyebabkan peningkatan penyerapan partikel 210Po
pada kawasan berkenaan. Di samping itu, berkemungkinan juga kilang ini menyalurkan sisa buangannya terus
ke dalam sungai menyebabkan kepekatan yang tinggi wujud di stesen ini.
Aktiviti 210Po yang tinggi wujud di persekitaran Sungai Langat juga dapat dijelaskan dengan merujuk
kepada keputusan untuk aktiviti 210Po dalam air hujan yang diperolehi. Aktiviti 210Po dalam air hujan juga
menunjukkan nilai yang agak tinggi dan ini menyumbang kepada jumlah kepekatan 210Po pada air sungai.
Kawasan persampelan air hujan juga adalah berdekatan dengan tempat persampelan air sungai. Maka interaksi
atmosfera dan air hujan dapat mempengaruhi dan menjelaskan aktiviti 210Po yang wujud dalam air sungai.
Aktiviti 210Po dalam air hujan di UKM, Bangi

Hasil kajian untuk aktiviti 210Po dalam air hujan dari 6 tarikh persampelan telah diperolehi (Rajah 5). Daripada
keputusan ini didapati aktiviti 210Po dalam air hujan adalah agak tinggi dan taburannya adalah secara tidak
seragam mengikut tarikh persampelan. Julat aktiviti 210Po yang diperolehi adalah antara 0.117-0.598 dpm/L
dengan purata aktiviti 0.277 dpm/L.
Kawasan Kajang dan Bangi merupakan kawasan daratan yang luas dan pesat dengan pembangunan.
210
Po dan 210Pb wujud di atmosfera sebagai produk pereputan gas 222Rn yang mengalir keluar dari daratan atau
kerak bumi (Cochran 1998). Aktiviti 222Rn pada udara di kawasan daratan adalah lebih kurang 102 kali lebih
tinggi daripada aktiviti 222Rn pada udara di kawasan perairan (Tateda et al. 2003). Di samping itu, aerosol yang
lama terkandung di atmosfera memperlihatkan sumber yang signifikan untuk 210Po dan 210Pb di atmosfera
(Tokeida et al. 1996). Oleh itu, tempoh tidak hujan yang lebih lama akan menyebabkan penambahan kandungan
210
Po di atmosfera.
Sumber kemasukkan 210Po ke atmosfera telah dicadangkan melalui pembakaran biomass (Le Cloarec et
al. 1995), letupan volkanik (Nho et al. 1996), pelepasan industri (Carvalho 1995) dan proses peruapan 210Po
secara biologi dari perairan (Tateda et al. 2003). Di persekitaran kawasan persampelan, kehadiran 210Pb dalam
air hujan adalah kemungkinan besar datang dari proses pembakaran biomass dan pelepasan sisa industri. Ini
kerana kawasan persampelan air hujan berdekatan dengan kawasan bandar dan perindustrian. Di samping itu,
perlepasan 210Po ke atmosfera melalui pembakaran boleh mencapai 100 kali ganda berbanding pemeruapan
210
Po secara semula jadi (Lambert et al. 1985). Maka jelaslah bahawa sumber utama aktiviti 210Po dalam air
hujan adalah berpunca dari pembakaran petrol oleh kenderaan dan pelepasan dari kawasan industri (Stepnowski
& Skwarzec 2000).
Hasil kajian ini mempunyai nilai yang signifikan dengan keputusan aktiviti 210Po dalam air sungai
(Rajah 4.3 dan Rajah 4.4). Kedua-dua keputusan menunjukkan nilai aktiviti 210Po yang agak tinggi di mana
purata aktiviti 210Po untuk air hujan dan air sungai masing-masing adalah 0.277 dpm/L dan 0.160 dpm/L. Oleh
kerana tempat persampelan antara air hujan dan air sungai adalah sama, maka wujud kaitan interaksi antara air
hujan dengan air sungai yang mengalir di permukaan bumi.
Daripada kedua-dua keputusan, kita dapat andaikan secara puratanya hampir 58% daripada aktiviti
210
Po di atmosfera akan memasuki sistem sungai apabila hujan turun. Selainnya iaitu 42% daripada aktiviti 210Po
di atmosfera akan hilang ke persekitaran seperti meresap masuk ke dalam tanah, diserap oleh akar tumbuhan dan
sebagainya. Keadaan ini dihitung dengan andaian bahawa tiada input 210Po lain yang memasuki sistem sungai
melalui persamaan di bawah.
% input 210Po melalui air hujan ke aktiviti 210Po di air sungai

= x 100%
sistem sungai aktiviti 210Po di air hujan
Aktiviti 210Pb dalam air laut di Pulau Tinggi, Johor

Aktiviti 210Pb dalam air laut di Pulau Tinggi bagi 6 stesen ditunjukkan dalam Rajah 6. Keputusan yang
diperolehi aktiviti 210Pb dalam air laut juga tertabur secara tidak konsisten. Julat aktiviti 210Pb untuk lapisan
permukaan, pertengahan dan dasar air masing-masing adalah antara 0.026-0.126 dpm/L, 0.072-0.125 dpm/L dan
0.046-0.120 dpm/L dengan purata aktiviti 210Pb masing-masing adalah 0.093 dpm/L, 0.079 dpm/L dan 0.070
dpm/L. Secara puratanya, tiada perbezaan aktiviti 210Pb yang ketara diperlihatkan di antara ketiga-tiga lapisan
dan ini mungkin menunjukkan proses percampuran yang sempurna telah berlaku di lautan Pulau Tinggi.
Untuk lapisan permukaan, input 210Pb adalah berkemungkinan dari atmosfera. 210Pb terbentuk di
atmosfera melalui reputan radioaktif gas 222Rn (T1/2 = 3.8 hari) yang terbebas dari kerak bumi. 210Pb dengan
separuh hayat yang panjang, pengeluarannya dari atmosfera lebih dikawal oleh proses yang bertindak terhadap
aerosol yang membawanya seperti sistem angin dan kadar kejatuhan di atmosfera berbanding reputan
radioaktifnya (Paatero et al. 2003). Maka, 210Pb yang dibawa dari daratan oleh tiupan angin memasuki
permukaan lautan sebelum ia sempat mereput dan menyebabkan kandungan 210Pb yang tinggi di permukaan air.
Di samping itu, didapati ralat yang diperolehi bagi sesetengah lapisan adalah mencatatkan nilai yang
lebih tinggi daripada hasil. Ada juga sesetengah lapisan yang tidak dapat diketahui nilai sebenar aktiviti 210Pb
seperti lapisan permuaan stesen 2 dan 3, lapisan pertengahan stesen 4, lapisan dasar stesen 5 dan ketiga-tiga
lapisan untuk stesen 6. Keadaan ini adalah disebabkan oleh nilai latar belakang yang tinggi dan nilai sampel
yang agak rendah pada lapisan berkenaan.
210
Po/210Pb
Julat nisbah 210Po/210Pb untuk lapisan permukaan, pertengahan dan dasar air di Pulau Tinggi masing-masing
adalah antara 0.450-2.831 dpm/L, 0.558-7.556 dpm/L dan 0.603-1.705 dpm/L. Secara keseluruhannya aktiviti
210
Pb adalah jauh lebih rendah berbanding dengan aktiviti 210Po (Jadual 2).
Keputusan ini menunjukkan bahawa terdapat lebihan kemasukan polonium-210 berbanding plumbum-
210 untuk setiap lapisan yang telah ditafsirkan. Secara umumnya, nisbah 210Po/210Pb dalam kawasan pantai
adalah lebih tinggi daripada uniti iaitu satu. Ini adalah kerana terdapat satu pengayaan 210Po berkaitan 210Pb yang
tidak boleh dijelaskan oleh input asas mereka dari atmosfera kerana nisbah 210Po/210Pb di atmosfera adalah kira-
kira 0.1 (Gasco et al. 2002). Di samping itu, sifat kimia yang berbeza bagi kedua-dua radionuklid di turus air
adalah digambarkan oleh afiniti yang kuat bagi 210Po terhadap partikel berbanding 210Pb (Gasco et al. 2002).
Kesimpulan
Kajian mendapati jumlah aktiviti 210Po yang ditunjukkan oleh ketiga-tiga jenis sampel air ialah; air hujan > air
sungai > air laut. Andaian bahawa aktiviti 210Po yang berada di atmosfera akan turun ke bumi bersama air hujan
dan di permukaan bumi ia akan meresap ke dalam tanah atau memasuki aliran air permukaan seperti sungai dan
akhirnya 210Po akan memasuki lautan melalui aliran sungai. Terdapat kehilangan aktiviti 210Po ke persekitaran
semasa ia diangkut dari atmosfera ke lautan iaitu jumlah aktiviti adalah menurun dari atmosfera ke lautan.
Penghargaan
Setinggi-tinggi penghargaan dan ribuan terima kasih diucapkan kepada kakitangan makmal Universiti
Kebangsaan Malaysia dan Malaysian Institute for Nuclear Technology di atas bantuan yang diberi. Terima kasih
juga ke IPRA: 09-02-02-0045-EA141
Rujukan
Carvalho, F.P. 1995. Origins and concentrations of 222Rn, 210Pb, 210Bi and 210Po in the surface air at Lisbon,
Portugal, at the Atlantic edge of the European continental landmass. Atmospheric Environment. 29:
1809-1819.
Carvalho, F.P. 1997. Distribution, cycling and mean residence time of 226Ra, 210Pb and 210Po in the Tagus
estuary. Sci. Total Environ. 196: 151-161.
Cochran, J.K., Frignani, M., Salamanca, M., Belluucci, L.G. & Guerzoni, S. 1998. Lead-210 as a tracer at
atmospheric input of heavy metals in the northern Venice Lagon. Marine Chemistry 62: 15-29.
Gabbard, A. 2002. Coal Combustion: Nuclear Resources or Danger. (atas talian)
http://www.ornl.gov/info/ornlreview/rev26-34/text/colmain.html (25 Feb 2005)
Gasco, C., Anton, M.P., Delfanti, R., Gonzalez, A.M., Meral, J. and Papucci, C. 2002. Variation of the activity
concentrations and fluxes of natural (210Po, 210Pb) and anthropogenic (239,234Pu, 137Cs) radionuclides in
the Strait of Gibraltar (Spain). J. Environ. Radioactivity. 62: 241-262.
Heussner, S., Cherry, R.D. & Heyraud, M. 1990. 210Po, 210Pb in sediment trap particles on a Mediterranean
continental margin. Continental Shelf Research. 10: 989-1004.
Hung, G.W. & Chung, Y.C. 1998. Particulates fluxes, 210Pb and 210Po measured from sediment trap samples in a
canyon off northeastern Taiwan. Continental Shelf Research. 18: 1475-1491.
Jeffree, R.A., Carvalho, F., Fowler, S.W. & Farber-Lorda, J. 1997. Mechanism for enhanced uptake of
radionuclides by zooplankton in French Polynesian oligotrophic waters. Environmental Science and
Technology. 31: 2584-2588
Lambert, G.M., Le Cloarec, F., Ardouin, B. & Bonsng, B. 1985. Volcanic emission of radionuclides and magma
dynamics. Earth and Planetary Science Letters. 76: 185-192.
Le Cloarec, M.F., Ardouin, B., Cachier, H., Liousse, C., Neveu, S. & Nho, E.Y. 1995. 210Po in savanna burning
plumes. J. Atmospheric Chemistry. 22: 111-122.
Momoshima, N., Song, L.X., Osaki, S. & Maeda, Y. 2001. Formation and emission of volatile polonium
compound by microbial activity and polonium methlation with methyl-cobalamin. Environmental
Science and Technology. 35: 2956-2960.
Nho, E.Y., Ardouin, B., Le Cloarec, M.F., Ardouin, B. & Tjetjep, W.S. 1996. Source strength assessment of
volcanic trace elements emitted from Indonesian arc. Journal of Volcanology and Geothermal
Research. 74: 121-129.
Nozaki, Y., Tsubota, H., Kasemsupaya, V., Yashima, M. & Ikuta, N. 1991. Residence times of surface water
and particles-reactive 210Pb and 210Po in the East China and Yellow seas. Geochimica et Cosmochimica
Acta. 55: 1265-1272.
Paatero, J., Hatakka, J., Holmen, K., Eneroth, K. & Viisanen, Y. 2003. Lead-210 concentration in the air at Mt.
Zeppelin, Ny-Alesund, Svalbard, J. Physics and Chem. Of the Earth. 28: 1175-1180.
Peck, G.A. and Smith, J.D. 2000. Determination of 210Po and 210Pb in rainwater using measurement of 210Po and
210
Bi. Journal of Analytica Chimica Acta. 422: 113-120.
Stepnowski, P. & Skwarzec, B. 2000. A comparison of 210Po accumulation in mollusks from the southern Baltic,
the coast of Spitsbergen and Sasek Wielki Lake in Poland. J. Environ. Radioactivity. 49: 201-208
Tateda, Y., Carvalho, F. P., Fowler, S. W. & Miquel, J. C. 2003. Fractionation of 210Po and 210Pb in Coastal
Waters of the NW Mediterranean Continental Margin. Continental Shelf Research 23: 295-316.
Tokeida, T., Yamanaka, K., Harada. K. & Tsunogai, S. 1996. Seasonal variations of residence time under upper
atmospheric contribution of aerosols studied with Pb-210, Bi-210, Po-210 and Be-7. Tellus. 48B: 690-
701.
Wildgust, M.A., McDonald, P. & White, K.N. 1998. Temporal changes of 210Po in temperate coastal waters. Sci.
Total Environ. 214: 1-10.
Yang, C.H. & Lin, H.C. 1992. ead-210 and Polonium-210 across the frontal region between Kuroshio and East
China Sea, Northeast of Taiwan. TAO 3 (3): 379-394.
Jadual 1. Lokasi stesen-stesen persampelan yang dilakukan semasa kajian
Kedalaman Jisim Lokasi

Nama Tempat Stesen
Air, m Air, L (Latitud, Longitud)
Kapar 1 2.3 m 15 L 03°06'44"U 101°20'06"T
21 Julai 2004 2 - 15 L 03°06'39"U 101°19'52"T
3 0.8m 15 L 03°07'40"U 101°19'03"T
4 8.3m 15 L 03°05'35"U 101°20'19"T
5 9.5m 15 L 03°06'00"U 101°19'41"T
6 4.8m 15 L 03°06'25"U 101°19'08"T
7 4.2m 15 L 03°06'33"U 101°19'00"T
8 4.1m 15 L 03°07'37"U 101°18'41"T
Pulau Tinggi 1 30m 15L 02°15'56"U 104°05'59"T
14-22 Ogos 2 30m 15L 02°18'01"U 104°05'02"T
2004 3 21m 15L 02°16'15"U 104°08'43"T
4 25m 15L 02°18'43"U 104°08'35"T
5 23m 15L 02°17'25"U 104°09'40"T
6 27.9m 15L 02°19'58"U 104°07'54"T
Kajang* 1 15L 02°59'30"U 101°47'53"T
4 Nov 2004 2 15L 02°59'29"U 101°47'30"T
3 15L 02°59'37"U 101°47'06"T
4 15L 02°57'51"U 101°47'02"T
5 15L 02°55'53"U 101°46''33"T
6 15L 02°55'06"U 101°45'33"T
UKM** 02°56'02"U 101°46'51"T
(*) sampel air sungai dan (**) sampel air hujan.
Kajang-Bangi
Kapar
Pulau
Tinggi
Rajah 1. Peta menunjukkan lokasi persampelan

Jadual 2. Aktiviti 210Po, 210Pb dan nisbah 210Po/210Pb dalam air laut di Pulau Tinggi, Johor
Stesen Aktiviti Po-210, dpm/L Aktiviti Pb-210, dpm/L 210

Po/210Pb
(Lapisan) ralat ralat
1 (S) 0.0736 0.0954 0.0260 0.0007 2.8309

(M) 0.1220 0.1350 0.1253 0.0017 0.9737
(B) 0.0871 0.1140 0.0629 0.0013 1.3861
2 (S) 0.0670 0.0984 - - -

(M) 0.0589 0.0807 0.0723 0.0013 0.8149
(B) 0.0463 0.0921 0.0455 0.0011 1.0171
3 (S) 0.0738 0.1720 - - -

(M) 0.0605 0.0978 0.1085 0.0016 0.5576
(B) 0.0881 0.1252 0.0516 0.0010 1.7048
4 (S) 0.0566 0.0921 0.1259 0.0020 0.4498

(M) 0.0649 0.1839 - - -
(B) 0.0727 0.1045 0.1205 0.0018 0.6030
5 (S) 0.0776 0.1228 0.1276 0.0016 0.6079

(M) 0.0709 0.1142 0.0094 0.0005 7.5557
(B) 0.2162 0.3001 - - -
6 (S) 0.0957 0.1435 - - -

(M) 0.0816 0.1366 - - -
(B) 0.0886 0.1364 - - -
S, M dan B masing-masing mewakili lapisan permukaan, pertengahan dan dasar.
0.25
0.20
Po, dpm/L
0.15
210
Aktiviti
0.10
0.05
0.00
1 2 3 4 5 6
Stesen
Perm ukaan Pertengahan D asar
Rajah 2. Aktiviti 210Po dalam air laut di Pulau Tinggi, Johor

0.18
0.16
0.14
Po, dpm/L
0.12
0.10
210
0.08
Aktiviti
0.06
0.04
0.02
0.00
1 2 3 4 5 6 7 8
Stesen
Permukaan Dasar
Rajah 3. Aktiviti 210Po dalam air laut di Kapar, Selangor
0.30
0.25
Po, dpm/L
0.20
0.15
210
Aktiviti
0.10
0.05
0.00
1 2 3 4 5 6
Stesen
Rajah 4. Aktiviti 210Po dalam air sungai mengikut stesen di Kajang-Bangi Selangor
0.7
0.6
0.5
Po, dpm/L
0.4
210
Aktiviti
0.3
0.2
0.1
0.0
25-Okt 2-Nov 3-Nov 25-Nov 28-Nov 10-Dis
Tarikh
Rajah 5. Aktiviti 210Po dalam air hujan di UKM, Bangi
0 .1 4
0 .1 2
0 .1 0
Po, dpm/l
0 .0 8
210
0 .0 6
Aktiviti
0 .0 4
0 .0 2
0 .0 0
1 2 3 4 5 6
S te s e n
P erm u k aan P e r te n g a h a n D asar
Rajah 6. Aktiviti 210Pb dalam air laut di Pulau Tinggi, Johor

INFLUENCE OF PRECIPITATING AGENT ON COPPER(II) OXIDE SYNTHESISED VIA
PRECIPITATION METHOD
Irmawati Ramli*, Hooi Hong Lau Y.H. and Taufiq-Yap
Department of Chemistry, Universiti Putra Malaysia,

43400 UPM Serdang, Selangor D.E., Malaysia.
Email: irmawati@putra.upm.edu.my
Abstract. Copper(II) oxide powders were synthesized by the precipitation of a copper nitrate solution in 1.0 M with
precipitating agents such as ammonium hydroxide, ammonium carbonate and sodium carbonate, respectively, in 1.5 M. By
using sodium carbonate, the crystallites of copper(II) oxide formed were smaller than by using the others two. On the other
hand, its high surface area promotes the total amount of oxygen removed.
Abstrak. Serbuk kuprum(II) oksida telah disintesiskan melalui pemendakan daripada 1.0 M larutan kuprum nitrat dengan
1.5 M agen pemendakan seperti ammonium hidroksida, ammonium karbonat dan sodium karbonat, masing-masing. Dengan
menggunakan sodium karbonat, hablur kuprum(II) oksida yang terbentuk adalah lebih kecil daripada hablur yang terhasil
daripada agen pemendakan yang lain. Di samping itu, luas permukaan hablurnya menggalakkan jumlah pelepasan oksigen
yang tinggi.
Keywords: copper(II) oxide, pH, precipitation, precipitating agent
Introduction
Copper(II) oxide, CuO occurs in nature as the black minerals. It crystallizes in a monoclinic structure.
In mineralogy, copper(II) oxide is known as tenorite. Its molecular weight is 79.54 g mol-1 and melts
at 1603 K. Sometimes, copper(II) oxide is also known as cupric oxide [1]. One of the largest
commercial applications of copper(II) oxide is in the production of compounds for wood preservation.
Copper(II) oxide is also used extensively as a feed additive and as a pigment in glass, ceramic and
porcelain enamels. In many catalytic reactions, it is a main component in various catalysts [1].
Copper(II) oxide catalysts have been widely used in many chemical reactions that involve hydrogen
as a reactant or a product. For example methanol synthesis from carbon monoxide or carbon dioxide
[2], the water-gas shift reaction and methanol steam reforming [3]. Besides, copper(II) oxide is also
used as catalyst in carbon monoxide oxidation [4] and propene oxidation [5]. In this work, influence
of precipitating agent on the physicochemical properties of CuO was studied.
Experimental
50 mL of copper nitrate solution in concentration of 1.0 M was placed in a round bottom flask under
moderate stirring. The solution was then titrated with 1.5 M of ammonium hydroxide, (NH4)OH in
drop wise manner. The precipitation process was stopped at pH 3.0. The solid was filtered and was let
to dry in a desiccator containing silica gel. The solid phase obtained after drying was weighed and
ground. The solids obtained were labelled as AHpre, where AH = 1.5 M of ammonium hydroxide; pre
= precursor. The whole procedure was repeated. This time, 1.5 M of sodium carbonate, Na2CO3 and
ammonium carbonate, (NH4)2CO3 were used as the precipitating agent, respectively. The precursors
obtained from the process were labelled as SCpre, and ACpre where SC = 1.5 M of sodium carbonate
and AC = 1.5 M of ammonium carbonate. All of the precursors were calcined in air at 623 K for 5
hours. The calcined samples were labelled as AH, SC and AC. The calcined samples were
characterised by using Shimadzu Diffractometer model XRD-6000 for X-ray diffraction (XRD)
analyses, ThermoFinnigan Sorptomatic 1990 for B.E.T. surface area measurements, JEOL scanning
electron microscope, model JSM-6400 for morphological studies and ThermoFinnigan TPDRO 1100
for temperature-programmed reduction in hydrogen (H2-TPR).
Figure 1 shows the XRD patterns of the precursors precipitated from different precipitating agents.
The diffractograms are similar to each other and show the presence of well crystallised copper
hydroxyl nitrate phase (JCPDS File No. 450594). All the precursors have monoclinic structure and is
in agreement with Guillou et al. [6]. The peaks of Cu2(OH)3NO3 are present at 2θ = 12.6, 21.4, 25.6,
31.8, 33.5, 36.2 and 58.1˚. The overall reaction of the precipitation process was the mixture solution
of copper nitrate and precipitating agent yielded the copper hydroxyl nitrate solid and ammonium
nitrate. Figure 2 indicates the XRD patterns of calcined samples. Calcination at 623 K transformed the
copper hydroxyl nitrate into copper(II) oxide phase (JCPDS File No. 5-0661). The obtained CuO
powders presented the main peaks at 2θ = 35.5, 38.7 and 48.7˚. These three peaks correspond to
( 1 11), (111) and (202) planes. Carbonate anion from sodium carbonate promotes the formation of
smaller crystallites in granular morphology. Figure 3(a) shows the morphology of AH in tabular plate-
like structure with indented contours. Figure 3(b) illustrates the morphology of AC in platelet
morphology. Figure 3(c) shows the morphology of SC is in dispersive granular morphology. The
dispersive granular copper(II) oxide is suggested to be a factor that caused the surface area of SC (7.7
m2 g-1) higher than AC (3.6 m2 g-1) and AH (2.3 m2 g-1). Figure 4 shows the hydrogen TPR profiles of
the copper(II) oxide prepared from 1.0 M of copper nitrate solution with 1.5 M of different
precipitating agent solutions (NH4OH, (NH4)2CO3 and Na2CO3). A single peak at maximum
temperature of 585 K was observed in sample AH, corresponding to activation energy of 97.8 kJ mol-
1
. Three reduction peaks at maximum temperature of 474, 490 and 528 K were observed in sample
AC, with corresponding activation energies of 79.3, 81.9 and 88.3 kJ mol-1. For sample SC, only
single peak was found at 498 K with calculated reduction activation energy of 83.3 kJ mol-1. The total
amount of oxygen removed being determined by quantification of the H2 consumed and is shown by
integrating the area under the TPR profile. The amount of oxygen removed obtained by hydrogen
TPR are shown in Table 1. The total oxygen atom removed is 6.7 x 1021 atom g-1 for AH and 6.9 x
1021 atom g-1 for AC copper(II) oxide. For SC, the total amount of hydrogen consumed is 7.8 x 1021
atom g-1. A correlation is observed between the total amount of oxygen atom removed and the BET
surface area of copper(II) oxide produced. As shown in Figure 4.34, there is an increment of total
amount of oxygen atom removed when the surface area of copper(II) oxide increased. This happening
may be due to a wide exposure of sample surface to the hydrogen when surface area of sample
increased.
40000 15000
SCpre
30000 SC
10000
Intensity (a.u.)
ACpre
AC
Intensity (a.u.)
20000
AH
AHpre 5000
10000
JCPDS No. 450594

Cu2(OH)3NO3
JCPDS No. CuO
5-0661
0
10 20 30 40 50 60
0
10 20 30 40 50 60
2θ (degree)
2θ (degree)
Figure 1. XRD patterns of uncalcined samples. Figure 2. XRD patterns of calcined samples.
(a) (b) (c)
Figure 3. SEM micrograph of (a) AH (b) AC and (c) SC.

498
Rate of Hydrogen Consumption

SC
474 490
528
AC
585
AH
350 400 450 500 550 600 650 700
Temperature (K)
Figure 4. H2-TPR profiles of copper(II) oxide.
Table 1. H2-TPR data of copper(II) oxide.
Samples Peak Tmax (K) Reduction Total Total

activation amount of amount of
energy, Er oxygen oxygen
(kJ mol-1) removed removed
(mol g-1) (atom g-1)
AH 1 585 97.8 1.1 x 10-2 6.7 x 1021
AC 1 474 79.3 5.0 x 10-3 3.0 x 1021
2 490 81.9 4.6 x 10-3 2.7 x 1021
3 528 88.3 2.0 x 10-3 1.2 x 1021
Total 1.2 x 10-2 6.9 x 1021
SC 1 498 83.3 1.3 x 10-2 7.8 x 1021
Conclusions
The phase structure of copper(II) oxide and its precursor was not influenced by the variation of
precipitating agents. Nevertheless, the variation in precipitating agent in preparation of copper(II)
oxide which affected the surface area and morphology of copper(II) oxide have greatly affected the
total amount of oxygen removed from copper(II) oxide. Results showed that higher surface area value
of CuO increased the removable of oxygen species in reduction reaction by hydrogen.
Acknowledgement
Financial assistance from Malaysian Ministry of Science, Technology and Innovation is gratefully
acknowledged.
References
1. Richardson, H. W. (2003) "Ullmann's Encyclopedia of Industrial Chemistry", Germany: Wiley-VCH.
2. Kung, H. H. (1989) "Transition Metal Oxides: Surface Chemistry and Catalysis", New York: Elsevier.
3. Ridler, D. E. and Twigg, M. V. (1989) "Catalysis Handbook", ed. Twigg, M. V., Frome, England: Wolfe
Publishing Ltd.
4. Huang, T.-J. and Tsai, D.-H. (2003) "CO Oxidation Behavior of Copper and Copper Oxides" Catalysis Letters 87.
173-178.
5. Lin, M. M. (2001) "Selective oxidation of propane to acrylic acid with molecular oxygen" Applied Catalysis A:
General 207. 1-16.
6. Guillou, N., Louër, M. and Louër, D. (1994) "An X-ray and neutron powder diffraction study of a new
polymorphic phase of copper hydroxide nitrate" Journal of Solid State Chemistry 109. 307-304.
PHASE BOUNDARY HETEROGENEOUS TIO2/ZRO2 CATALYST FOR EPOXIDATION
Izan Izwan Misnon, Hadi Nur and Halimaton Hamdan
Ibnu Sina Institute for Fundamental Science studies, Universiti Teknologi Malaysia,
Skudai, Johor 81000 Malaysia.
Abstract - A heterogeneous oxidation catalyst was prepared by fluorination of TiO2 impregnated

ZrO2 particles with ammonium hexafluorosilicate [(NH4)2SiF6] followed by alkylsilylation of n-
octadecyltrichlorosilane (OTS). Enhanced catalytic activity of the system was observed in a liquid
phase epoxidation of 1-octene with aqueous hydrogen peroxide to produce 1,2-epoxyoctane. The
catalytic activity of the modified TiO2/ZrO2 is attributed to the modified local environment of titanium
active sites and increased hydrophobicity.
Keywords: Epoxidation catalyst, ammonium hexafluorosilicate, fluorination, alkylsilylation, titanium,

zirconia,hydrophobicity
Introduction
Heterogeneous catalyst such as titanium silicalite (TS-1) has been successfully applied in a
variety of oxidation reactions, namely the selective epoxidation of olefins and diolefins, the selective
oxidation of primary alcohol to aldehydes and secondary alcohols to ketones, the ammoximation of
ketones to oximes, the hydroxylation of aromatic and aliphatic compounds [1,2]. Its unique activity is
mostly due to the isolated tetrahedral Ti species in the hydrophobic framework [3].
However, in TS-1, the active sites are located in the internal framework [4] which limits its
application to substrates with relatively small molecular sizes [3]. Consequently, large molecules with
ring structures are not accessible, reducing its catalytic capabilities. Due to the importance of Ti active
sites, there is a need to design a system where Ti can be located in the external framework or at the
external surface which enable it to freely directly interact with every substrate. Studies have shown
that extraframework tetrahedral Ti is formed when fluorides are used [5]. Other modification shows
that epoxidation reaction will increase via alkylsilylation [6]. Alkylsilylation is to increase the
hydrophobicity and protect Ti active sites.
In this research, zirconia will be used as the catalyst support. The epoxidation catalyst will be
further modified with fluorine and enhanced with alkylsilylation, in order to create more tetrahedral Ti
and increase hydrophobicity respectively.
Experimental
Catalysts preparation
Catalyst was prepared by impregnation of raw material. 500 µmole of [Ti(PrO)4] in addition of 1g of
[Zr(OH)4] was mixed with stirring until the solvent is completely dry. The sample was calcined at 500
o
C for 2h at a rate of 2 oC/min in the furnace. Then, the sample was grind and labeled as TiO2/ZrO2.
Double distilled water (10 mL) was mixed with [(NH4)2SiF6] to make 1M clear solution. Next, it is
added to the container that contains TiO2/ZrO2. The mixture was shaken for five min before
undergoing ultrasonic treatment for 10 min at 30 oC. The suspension was then collected by
centrifugation at 3500 rpm for 20 min using centrifugation machine. Excess of solvent was discarded
and the wet-precipitate was heated at 100 oC overnight. Then, the sample was grind and labeled as F-
TiO2/ZrO2. Preparation of O-F-TiO2/ZrO2 was done by surface modification of F-TiO2/ZrO2 through
alkysilylation of OTS. Toluene (10 mL) containing OTS (250 µmole) was added to the container that
contain F-TiO2/ZrO2 powder (1g). The mixture was shaken for 20 minutes at room temperature. The
suspension was collected by centrifugation. The yield obtained was heated at 100 oC overnight and
then the prepared samples are labeled as O-F-TiO2/ZrO2.
Characterization
Comprehensive characterizations of the catalysts resulted from zirconia surface modifications were
done by X-Ray Diffraction (XRD), UV-VIS Diffuse Reflactance, SEM, EDAX and Surface Analyzer.
These characterizations are done in order to confirm the surface structure of catalysts and their active
sites that undergo modifications.
Catalytic Measurement
In this study, 1-octene (4 mL), H2O2 (1 mL), and TiO2/ZrO2 fine sample catalyst powder (50 mg) were
placed in a glass tube before it was tightly closed. The glass tube was covered using aluminum foil.
This was carried out in order to prevent decomposition of H2O2 before the reaction started. The
reaction was performed under stirring for 24h at room temperature. The resulting product was
characterized using GC. The same procedure was repeated with other catalysts.
Figure 1 shows UV-Vis Diffuse Reflectance spectra of TiO2/ZrO2, F-TiO2/ZrO2 and O-F-TiO2/ZrO2.
The band in the range of 200–240 nm is attributed to a charge-transfer of the tetrahedral Ti sites
between O2- and the central Ti(IV) atoms, while octahedral Ti was reported to appear at around 260–
330 nm. It shows that, for samples containing fluor (F-TiO2/ZrO2 and O-F-TiO2/ZrO2), only single
high intense band at around 208 nm can be observed.
F- TiO2/ ZrO2
1.2
O-F-TiO2/ ZrO2
1.0
0.8
0.6 Tetrahedral
0.4 Octahedral
0.2
190 250 300 350 400 450 500

nm
Figure 1: UV-VIS diffuse reflectance spectra of TiO2/ZrO2, F-TiO2/ZrO2 and O-F-TiO2/ZrO2.
This band is attributable to Ti in the tetrahedral structure. The decrease in intensity in the
range of 260-330 nm, which correspond to the octahedral Ti, indicates the occurrence of tetrahedral Ti
structure; transformed from the octahedral Ti framework during fluorination.
The surface area of fluorinated TiO2/ZrO2 (F-TiO2/ZrO2) (21.24 m2/g) is almost similar to that
of alkylsilylated O-F-TiO2/ZrO2 (19.63 m2/g) suggesting that alkylsilylation does not affect the
surface area of the catalysts (see Table 1).
Data in Table 1 agree with the relative crystallinity results obtained by XRD as shown in Fig.
2. The major peaks tend to appear at 2Ө range of 20o to 40o. The X-ray diffractograms demonstrate
that all the modified catalysts possess a similar crystalline structure with characteristic peaks at 28.2o
and 31.5o, corresponding to the monoclinic phase of ZrO2. It was observed that there is no change in
the crystallinity of the modified ZrO2 after impregnation of TiO2 and modification with (NH4)2SiF6
and OTS. The highly stable properties of ZrO2 surface contributes to the unaltered surface area.
Table 1: Summary of the total surface area obtained for all samples.
Modified Catalyst Specific Surface Area

(m2/g)
TiO2/ZrO2 N/A
F-TiO2/ZrO2 21.24
O-F-TiO2/ZrO2 19.63
O-F-TiO2/ZrO2
F-TiO2/ZrO2
TiO2/ZrO2
ZrO2
10 20 30 40 50 60 70
2-theta
Figure 2: X-ray diffractograms pattern of unmodified ZrO2 support and modified TiO2/ZrO2, F-
TiO2/ZrO2 and O-F-TiO2/ZrO2.
The EDAX analysis indicates that the amount of zirconium (Zr) and carbon (C) in O-F-
TiO2/ZrO2 are 55.4 wt% and 19.5 wt% respectively, in which carbon originates from alkylsilyl group
attached on the surface of the sample. The amount of titanium (Ti) and silicon (Si) show only 4.0 wt%
and 2.0 wt%, respectively on the surface of the catalyst. Fluorine (F) and chlorine (Cl) only takes part
ca. 1.0 wt% and 1.8 wt% respectively. This result shows that the amount of F in O-F-TiO2/ZrO2 (ca.
1.0 wt%) is very much lower compared to the amount of [(NH4)2SiF6] added during the fluorination
process (ca. 28%).
SEM micrographs of TiO2/ZrO2, F-TiO2/ZrO2 and O-F-TiO2/ZrO2 samples revealed that there
is no significant difference in morphology and the particle size among the samples. This suggests that
fluorination and alkylsilylation processes did not change the morphology and particle size of ZrO2.
This is supported by the fact that the structure of ZrO2 is still maintained after introduction of fluorine
and alkylsilyl groups (see Fig. 2). Furthermore, the samples also showed the irregularity in their
morphology in multi particle size distribution.
As shown in Fig. 3, fluorination and alkylsilylation onto the surface of TiO2/ZrO2 increases
their epoxidation activity. This phenomenon may be explained in terms of the local environment of Ti
active site. It is generally accepted that isolated Ti in tetrahedral form are considered the most active
species in epoxidation reaction. Based on these findings, the effect of fluorination on increasing the
epoxidation activity of F-TiO2/ZrO2 and O-F-TiO2/ZrO2 can be explained by the presence of isolated
Ti in tetrahedral form (see Fig. 1). As shown in Fig. 4, tetrahedral Ti reacted with 1-octene to give
1,2-epoxyoctane.
catalytic activity
200
155,35
150
yeild (µmole)
100 71,8
50
10,37
0
ZrO2/TiO2 F-ZrO2/TiO2 O-F-ZrO2/TiO2
sample
Figure 3: Column chart of epoxidation-catalyzed reaction.
H 2 O 2
F F
F F
F F
S i
O O
T i
A considerable increase in the epoxide yield was observed when the surface of F-
Figure 4: Proposed model of F-TiO2/ZrO2 as catalyst for transformation of 1-octene to
1,2-epoxyoctane.
TiO2/ZrO2 was alkylsilylated with OTS (O-F-TiO2/ZrO2) (see Fig. 3). This can be explained
on the basis of an increase in the hydrophobicity of F-TiO2/ZrO2 by alkylsilylation. When the
substrates interact with the active sites on the Ti active sites, the H2O2 releases water as a side product.
If this happens, Ti active sites will be poisoned by water molecules and it might reduce the catalytic
activity. The long hydrophobic carbon chains of alkylsilyl groups would prevent the water to interact
with Ti active sites.
Conclusion
The results from the study indicate that TiO2/ZrO2, F-TiO2/ZrO2, and O-F-TiO2/ZrO2 are active
catalyts in the epoxidation of 1-octene. Activity of the catalysts is due to the presence of Ti active
sites on their surface. The increase in catalytic performance is brought about by fluorination and
alkylsilylation processes of TiO2/ZrO2. We have demonstrated that fluorination can change the
coordination of Ti active sites from octahedral to tetrahedral and also increase the hydrophobicity of
the catalyst. Alkylsilylation on the other side has absolutely increased hydrophobicity of the catalyst
by preventing the Ti active sites from being poisoned by water molecules. The catalytic activity of O-
F-TiO2/ZrO2 increase significantly in the epoxidation of 1-octene with aqueous H2O2 due to two
properties. The first property is the presence of tetrahedral Ti active sites and second, the hydrophobic
character of the catalyst. Based on its properties, fluorination and alkylsilylation of impregnated Ti
active site on the surface of a support is a promising technique for the enhancement of the catalytic
activity in epoxidation of alkenes with aqueous H2O2.
References
1. Hamilton, D., (1973). “Technology, Man and the Environment”, London, Faber and Faber Ltd.,
69-72.
2. Capel-Sanchez, M.C., Campos-Martin, J.M. and Fierro, J.L.G., (2003). “Impregnation treatments
of TS-1 catalysts and their relevance in alkene epoxidation with hydrogen peroxide”, Applied
Catalysis A: General, 246(1), 69-77.
3. Liu, X., Wang, X., Guo, X. and Li, G., (2004). “Effect of solvent on the propylene epoxidation
over TS-1 catalyst”, Catalysis Today, 93-95, 505-509.
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in catalysis”, Applied Catalysis A: General, 221(1-2), 63-72.
5. Lopez, A. et al., (1993). “Titanium in MFI-Type Zeolites: A Characterization by XANES,
EXAFS, IR, and 74, 49Ti and 17O MAS NMR Spectroscopy and H2O Adsorption”, J. Solid State
Chem., 102(2), 480-491.
6. Hadi Nur, Ikeda, S. and Ohtani, B., (2001). “Phase Boundary Catalysis of Alkene Epoxidation with
Aqueous Hydrogen Peroxide Using Amphiphilic Zeolite Particles Loaded with Titanium Oxide”,
J. Catal., 204, 402-408.
PRODUCTION OF WATER SOLUBLE AND LOW MOLECULAR WEIGHT
CHITOOLIGOSACCHARIDES USING COMMERCIAL
MIXED-GLUCANASE FROM ASPERGILLUS ACULEATUS
Lee Lih Fen1, Mohd Yusof Maskat1, Rosli Md IIlias2 and Osman Hassan1*
1
School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti
Kebangsaan Malaysia, 43600 UKM-Bangi, Malaysia
2
Department of Bioprocess Engineering, Faculty of Chemical Engineering and Natural Resources
Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Malaysia
*Correspondence: osman@ukm.my
Abstract. Degradation of chitinous waste by microorganisms usually proceeded through a series of

low yielding but complementary enzymes in producing breakdown of chitinous material. Viscozyme®
L, a commercial carbohydrase from Aspergillus aculeatus, containing a mixture of arabanase,
cellulase, hemicellulase, xylanase and β-glucanase, was chosen as a model for low-yielding
chitosanase acting on a chitinous material from shrimp waste. The enzymatic hydrolysis was
optimised using a statistical modelling, whereby the application of 50% (w/w) of enzyme on 4% (w/v)
chitosan will optimally hydrolyse the chitinous material at pH 4.52 and temperature of 54.2°C. The
products obtained from the hydrolysis can be easily solubilised in cold-water [up to 10% (w/v)] to
give clear, brownish and low-viscosity solution. Results from HPLC analysis indicated that the
chitooligosaccharides consisted of DP 1-5. In conclusion, the mixed glucanases enzyme is a workable
enzyme to model the production of water soluble and low molecular weight chitooligosaccharides
from chitinous material.
Keywords: Chitosan, commercial glucanase, chitinase, chitooligosaccharide.
Introduction
Chitosan is a biopolymer of polysaccharide derived from chitin found in the shell of crustacean
animal (shrimp and crab) and cell wall of fungi (mushroom). In food industry, chitosan has been used
as dietary fiber, anti-microbial agent [1] and coating film of fruit and vegetable [2]. Water insoluble
chitosan can be degraded enzymically by chitosanase (E.C.3.2.1.132) or chitinase (E.C.3.2.1.14) to
produce water soluble of oligomer or monomer of chitosan that is more suitable for human
consumption and absorption. Chitosan oligomer has been developed as prebiotic [3], which enhances
the growth of useful probiotics in human colon, while chitosan monomer, glucosamine, has been used
as treatment for osteoarthritis [4]. However, the application of the purified and specific chitosanase in
producing chitosan hydrolysates has been prevented by the scarcity and high cost of chitosanase
enzymes.
Viscozyme® L is a commercial enzyme containing a mixture of arabanase, cellulase,
hemicellulase, xylanase and β–glucanase. Although Viscozyme® L is not the specific chitosanase, it
was found to be able to hydrolyse chitosan [5]. Therefore, chitooligosaccharide can be produced at
lower cost using Viscozyme® L and coupled with optimized reaction conditions.
Experimental
Materials
Shrimp chitosan (ChitoChem Sdn. Bhd., Malaysia) was ground and sieved to 115 Mesh. The crude
carbohydrase mixture of Aspergillus aculeatus, named Viscozyme® L was supplied from Novozyme
(Germany). The dyeing agent of Remazol Brilliant Blue ® (RBB) was purchased from Sigma (USA).
Acid acetic solution (99.8%) (Hamburg Chemicals, Germany), sodium acetate trihydrate and Sodium
hydroxide (Merck, Germany) were used in colloidal chitosan preparation and as pH buffer in chitosan
hydrolysis. Acetonitrile (Merck, Germany), ammonium hydroxide (R&M, UK) and deionised water
were utilised as mobile phase in HPLC for chitooligosaccharides determination.
Filter paper with 20/25 µm pore size (Advantec, No. 1, Japan), polyethersulfone (PES)
Ultrafiltration (UF) membranes (Vivascience, Vivaflow 50, Germany) with 10 kDa molecular weight
cut-off, and methanol (Hamburg, Germany) were used in purification of chitosan hydrolysis products
to be subjected for characterizations.
Procedures
Colloidal chitosan was prepared and dyed with RBB according to the method of Lee et al. [5] prior to
study the model of its hydrolysis by Viscozyme® L. The model study was conducted in micro-scale
for screening the soluble products of hydrolysis spectrophotometrically at 595 nm. The statistical
analysis and optimization of Viscozyme® L reaction was conducted with central composite design
(CCD) of response surface methodology (RSM), employing a statistical package Design-Expert
version 6.0.10 (Stat-Ease Inc., Minneapolis, USA). The recovery of chitooligosaccharides was done
according to the optimized parameters, but using the original non-dyed chitosan powder as substrate.
The chitosan hydrolysates were purified before characterization.
Preparation of Colloidal Chitosan.
Chitosan powder (10g) was dissolved in 0.3 M acetic asid [1:25 (v/v)] at room temperature
for 30 min before being neutralized to pH 9 by 0.3 M NaOH. After neutralization, the
colloidal chitosan was wash thoroughly by distilled water until the pH became 7. The washed
colloidal chitosan was filtered and dried to approximately 92% moisture content before
dyeing.
Dyeing of Colloidal Chitosan.
The colloidal chitosan was suspended at a ratio of 10 to 1 (w/v) in 1% (w/v) RBB solution. The
mixture was boiled for 60 min with gentle stirring. Then, the excess dye solution was drained, and the
bonding of dye to colloidal chitosan was fixed by boiling the coloured material in 1.5% sodium
dichromate and 1.5% potassium sodium tartarate tetrahydrate for 10 min. The dyed colloidal chitosan
was washed thoroughly with boiling distilled water until the washing solution became colourless. The
dyed chitosan is dried and finally ground to 115 mesh for subsequent study.
Optimization of Viscozyme® L Hydrolysis.
The RBB-dyed chitosan powder samples were suspended 4% (w/v) in 1 ml of 0.2 M acetate buffer in
1.5 ml micro-tubes. After equilibration at desired temperature for 10 min, 50% w/w Viscozyme® L
was added into the solution to initiate hydrolysis. The enzyme activity was terminated and
precipitated by adding 0.5 ml of 2 N NaOH. The mixtures were centrifuged (6400 rpm, 5 min) and the
supernatants were transferred to a 96-well microplate (375 ml well, 1 cm height). The absorbance of
supernatants was read spectrophotometrically at 595 nm, by using a 96-well microplate reader
(VersaMax, Molecular Devices Inc., USA). Blanks were prepared similarly (3 replicates) but without
the addition of enzyme. One unit of Viscozyme® L was defined as the amount of enzyme that
produced an increase of 0.01 in the absorbance.
Experimental Design and Statistical Analysis.
Based on the first-order model of 2-level full factorial design experiment carried out previously (data
not shown), pH and temperature were found to be the main factors in controlling the enzyme activity.
Therefore, the optimum regions observed were used for subsequent second-order model design of a
rotatable central composite design (CCD) of response surface methodology (RSM). The CCD
consisted of 4 fractions of factorial and axial points, respectively, and 5 centre points (Table 1). The
center point of pH was 4.50 and temperature was 55.0oC. Whereas substrate concentration was fixed
at 4% (w/v) as the value of Km determined by Lineweaver-Burk Method for Viscozyme® L acivity in
preliminary study and the initial viscosity of the original chitosan gel used was found maximum at 4%
(w/v). Besides, enzyme concentration was fixed at 50% (w/w), which was the maximum point in the
linear trend of enzyme activity and considering of its lower cost of application. The response
encountered in this experimental design is Viscozyme® L activity and was expressed as U/min.
Initially, the polynomial of the second-order model fitted to the response was giving an equation of
the form:
Y = β0 + β1X1 + β2X2 +β12X12 + β22X22 + β1β2X1X2 (1)
Where X1 and X2 represented the levels of factors according to Table 1, and β0, β1 and β2 represented
coefficient estimates with β0 having the role of a scaling constant.
Recovery of Chitooligosaccharides
Chitosan hydrolysis was performed again based on the optimized parameters, but using the non-dyed
chitosan powder as substate. The hydrolysis was carried out for 24 hours. The hydrolysate was filtered
through the 20/25 µm filter paper (Advantec, No. 1, Japan) after centrifugation at 10 000 rpm for 15
min, to remove the undegraded chitosan substrate. The supernatant was filtered again through an
ultrafiltration membrane, with 10 kDa molecular weight cut-off, to remove the enzyme protein and
high molecular weight chitosan.
The filtered supernatant was concentrated to 1/5 volume by using a vacuum rotary evaporator
at 60oC. Chitooligosaccharide products were obtained by alcohol extraction. The concentrated sample
was precipitated in methanol (97%) at a ratio of 1 to 6 (v/v) to give the final methanol concentration
of more than 80% (v/v). After storage at -20oC for 48 hours, the precipitated chitooligosaccharides
was centrifuged, washed 3-4 times again with methanol, ethanol and acetone to remove buffer salt and
vacuum-dried finally to obtain chitooligosaccharides fraction 1 (COS-1) [6]. The supernatant was
evaporated and reprecipitated by ethanol, centrifuged and washed by ethanol and acetone before
drying to obtain chitooligosaccharides fraction 2 (COS-2) [9].
Characterizations
Both chitosan and its degraded chitooligosaccharides (mixtures of COS-1 and COS-2) powder were
dried and pelletised into potassium bromide (KBr) disk to determine its degree of deacetylation (DD)
by Fourier Transform Infrared (FTIR) spectroscopic [7]. The spectra of chitosan samples were
obtained using an FTIR spectrometer (Perkin-Elmer, GX, USA).
The components of water soluble and low molecular weight chitooligosaccharides were
analyzed by high performance liquid chromatography (HPLC). An HPLC system consists of a Jasco
Intelligent HPLC Pump (PU 1580, Japan), Waters 717 Plus Autosampler (USA) and Alltech Prevail
Carbohydrate ES 5µ (250 mm x 4.6 mm) column were used. The mobile phase applied was a
combination of 75% acetonitrile and 25% deionized water containing 0.04% ammonium hydroxide.
The flow rate was maintained at 1.0 ml/min. Injection volume of sample was 20 µl. The eluent was
monitored by an evaporative light scattering detector (ELSD) (Eurosep Instruments, DDL 31, France).
The chitooligosaccharides concentration to be analyzed was approximately 10% (w/v). To determine
the degree of polymerization (DP), standard saccharides consisting of fructose, glucose (DP1),
sucrose (DP2), maltotriose (DP3), stachyose (DP4) and maltopentaose (DP4), were used. All data
obtained from HPLC were collected and analyzed using the Borwin Chromatography Software.
Optimization of Viscozyme® L Hydrolysis
In the model study of a wide-spectrum enzyme application on chitinous material, the dyed chitosan
was chosen as the substrate of hydrolysis. The soluble hydrolysates were determined
spectrphotometrically at 595 nm. The substrate and the analytical method used were shown to be
faster and more sensitive than the usual DNS determination [5].
Table 1 showed the experimental design, observed and predicted results obtained. From the
results of the optimization method, Viscozyme® L activity (Y) obtained was about similar and fitted to
the predicted results. The second-order model developed with the adjusted R2 of 0.9752 and stated
below was significantly fitted well (p<0.05) by each variables as there is no significant lack of fit
(p>0.05) in the model.
Y (U/min) = 0.2568 + 0.0104 X1 – 0.0123 X2 -0.0948 X12 – 0.0479 X22 + 0.0339 X1X2 (2)
where X1 and X2 are factors presented in Table 1.
Table 1. Experimental design and results of RBB-chitosan hydrolysis by Viscozyme® L based on

central composite rotatable design (n=3) [8]
Standard Variables *Response Y:
Order Coded Actual Viscozyme® L activity (U/min)
X1 X2 pH Temperature
Actual Predicted
(oC)
1 -1 -1 4.00 50.0 0.143 0.150
2 1 -1 5.00 50.0 0.105 0.103
3 -1 1 4.00 60.0 0.047 0.058
4 1 1 5.00 60.0 0.144 0.146
5 -1.414 0 3.79 55.0 0.063 0.053
6 1.414 0 5.21 55.0 0.080 0.082
7 0 -1.414 4.50 47.9 0.180 0.178
8 0 1.414 4.50 62.1 0.150 0.144
9 0 0 4.50 55.0 0.264 0.257
10 0 0 4.50 55.0 0.245 0.257
11 0 0 4.50 55.0 0.273 0.257
12 0 0 4.50 55.0 0.257 0.257
13 0 0 4.50 55.0 0.243 0.257
* Viscozyme® L activity = Increase of 0.01 in the absorbance (595 nm) per min
Table 2. Analysis of Varians for Viscozyme® L activity [8]

Prob >
Source Sum of Squares DF Mean Square F Value
F
Model 0.2292 5 0.0469 299.6 < 0.0001 significant
X1 0.0026 1 0.0026 16.42 0.0003
X2 0.0036 1 0.0036 23.24 < 0.0001
X12 0.1876 1 0.1876 1199 < 0.0001
X22 0.0479 1 0.0479 306.0 < 0.0001
X1X2 0.0138 1 0.0138 88.21 < 0.0001
Residual 0.0052 33 0.0002
Lack of Fit 0.0010 3 0.0003 2.330 0.0944 not
Pure Error 0.0042 30 0.0001 significant
Cor Total 0.2396 38
Std. Dev. 0.0125 R-Squared 0.9784

Mean 0.1690 Adj R-Squared 0.9752
C.V. 7.403 Pred R-Squared 0.9721
PRESS 0.0067 Adq Precision 41.63
Contour plot presented in Figure 1 showed that the optimum condition for Viscozyme® L activity
was at pH 4.52 and 54.2oC. Although Viscozyme® L is not a specific chitosanase, it was proven that
the enzyme, under optimal reaction condition, can effectively degrade chitosan into
chitooligosaccahrides. According to some published discussions, cellulase [9] and hemicellulase [10]
were able to cleave β-1,4-glycosidic bond in chitosan. Hence, the reactivity of Viscozyme® L toward
chitosan is not unexpected because of the presence of cellulase and hemicellulase that behave as low-
yielding chitosanase.
DESIGN-EXPERT Plot
Viscozyme activity
X = A: pH
Y = B: T emperature
0.258
0.208
Viscozyme activity
0.158
0.108
0.058
60.00
5.00
57.50
4.75
55.00
4.50
B: Temperature
52.50 4.25
A: pH
50.00 4.00
Figure 1. Response surface plot of the effect of pH and temperature on RBB-chitosan hydrolysis at
4%w/v substrate and 50%w/w enzyme for 6 hours.
Characterizations
From FTIR analysis, the degree of deacetylation (DD) of the chitosan substrate was 70%. The
chitosan hydrolysis was repeated using the optimized parameters with 4%w/v chitosan substrate,
50%w/w Viscozyme® L, pH 4.52 and 54.2oC for 24 hours. The chitooligosaccharides fraction which
was recovered by alcohol extraction, was cold-water soluble (up to 10% w/v) to form clear, brownish
and non-viscous solution.
According to Figure 2 (a) and (b), Viscozyme® L was capable to degrade chitosan and
produce chitooligosaccharides, in the range between monomer and pentamer. Fructose and glucose
with molecular weight (MW) 180.2, sucrose (342.3), maltotriose (504.4), stachyose (666.6) and
maltopentaose (828.7) were used as standards for determining the degree of polymerization of
chitoolisaccharides (approximately 76% DD) ranged from DP1 (179.2-221.2), DP2 (340.2-382.4),
DP3 (501.6-543.6), DP4 (662.8-704.8) and DP5 (824.0-908.0), because the molecular weight of
sugars are almost similar to chitomonosaccharide (glucosamine or N-acetyl-glucosamine) and
chitooligosaccharides, (oligomers consisted of glucosamine: N-acetyl-glucosamine at least 7:3), and
yet the cost is lower.
Each peak of the standard sugars represented 1/6% w/v [Figure 2(a)]. Fraction COS-1 was the
first extract from alcohol extraction, which was precipitated with final methanol concentration of
more than 80% v/v, whereas, fraction COS-2 was the subsequent fraction by ethanol extraction,
yielded in the form of brownish powder. Fraction COS-1 [Figure 2(b)] collected as yellowish white
powder, contained more chitooligosaccharides with DP of more than 4 compared to fraction COS-2
[Figure 1(c)]. The chitooligosaccharides with DP up to 4 were more soluble in methanol, but
oligomers with DP of more than 5 were less soluble [8]. Based on the studies by Cabrera & Cutsem
[11], chitooligomers with DP up to 11 were soluble in 70 and 80% methanol and up to DP 8 only in
90% methanol. The precipitated fractions by 90% methanol were oligomers of DPs up to 16.
Glucosamine is soluble in water, but slightly soluble in methanol, and insoluble in ethanol
and acetone [6]. However, the solubility of chitooligosaccharides in those organic solvents will
decrease as the DP value increase. After most of the higher chitooligosaccharides (DP>4) were
removed as COS-1, the supernatant was dried and reprecipitated by ethanol and washed by acetone.
Subsequently, acetate salt free chitosaccharides (COS-2) with DP up to 4 were obtained [Figure 2(c)].
However, both of the fractions COS-1 and 2 contained very low amount of detected oligomers (20
and 30% respectively) due to the higher molecular weight chitooligosaccharides were removed during
filtration prior to HPLC injection or reprecipitated by mobile phase and trapped inside the column,
which caused an increase in backpressure of column, and thus remained as undetected products. The
total chitooligosaccharide extracts obtained by alcohol extraction was only around 54% because of the
low yielding activity of Viscozyme®L.
(a)
DP1 DP2 DP3 DP4 DP5
(b)
DP1 DP2 DP3 DP4
(c)
Figure 2. HPLC profiles of (a) standard sugars, (b) COS-1 and (c) COS-2
Conclusions
Water soluble and low molecular weight chitooligosaccharides ranged

from DP 1 to 5 can be obtained, by using a commercial mixed-glucanase from Aspergillus aculeatus,
named Viscozyme® L (50%w/w) on 4%w/v chitosan hydrolysis at pH 4.52 and temperature of 54.2oC
in 24 hours, but with only low yield of products.
Acknowledgement
This study was carried out with the financial support of National Biotechnology Directorate, MOSTI
through Project 09-02-02-0006/BTK/ER/31 at Food Science Program, School of Chemical Sciences
and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia.
Refereces
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PENGHASILAN SURFAKTAN TAK BERION BERASASKAN LAURIL ALKOHOL
DARIPADA TERBITAN MINYAK KELAPA SAWIT
Norhafipah Mohamad*, Zarina Edris & Mohd Ambar Yarmo

Makmal Pemangkinan, Pusat Pengajian Sains Kimia dan Teknologi Makanan, Fakulti Sains dan
Teknologi, 43600 Universiti Kebangsaan Malaysia, Bangi.
Email: norhafipah@yahoo.com
Abstrak. Penghasilan surfaktan tidak berion jenis lauril alkohol teretoksilat (LAT) berasaskan produk
lauril alkohol daripada minyak kelapa sawit berjaya dilakukan. Sintesis ini melibatkan tindak balas
pengetoksilan antara gas etilena oksida (EO) dengan lauril alkohol (C12). Antara parameter yang dikaji
dalam kajian ini adalah kesan perbezaan bilangan mol EO, tekanan dan suhu tindak balas. Tindak
balas dilakukan menggunakan kalium hidroksida (KOH) sebagai mangkin homogen. Hasil terbentuk
dianalisis menggunakan teknik HPLC, FTIR, ujian pembuihan, nilai Keseimbangan Hidrofilik-
Lipofilik (HLB) dan Ujian Detergensi (peratus penyingkiran kotoran). Pembentukkan molekul LAT
dapat ditunjukkan melalui spektroskopi FTIR yang menandakan wujudnya puncak regangan C-O
(kumpulan eter) pada jarak gelombang 900 – 1350 cm-1. Manakala pencirian menggunakan teknik
HPLC menunjukkan masa penahanan produk ialah 6.201 min untuk LAT-3 EO, 6.261 min untuk
LAT-5 EO dan 6.260 min untuk LAT-7 EO. Pencirian surfaktan melalui ujian pembuihan
menunjukkan produk LAT yang diperolehi mempunyai kestabilan dan kekuatan buih yang baik.
Secara umum hasil menunjukkan produk yang diperolehi berpotensi untuk kegunaan detergen dan
industri.
Abstract. Ethoxy nonionic surfactant type of ethoxylated lauryl alcohol (LAT) based on product of
fatty alcohol from palm oil product was produced successfully. Ethoxylation reaction was carried out
between EO with lauryl alcohol (C12). Different moles ratio of EO, reaction temperature and pressure
were among various parameters investigate. The reaction was performance with KOH as a
homogenous catalyst. LAT products were analyzed using FTIR, HPLC, foaming test, value of
Hydrophilic-Lipophilic Balance (HLB) and detergency. Molecule of LAT was produced with look the
sign of stretching peak C-O (group of ether) at 900 – 1350 cm-1 of wave number from FTIR.
Characterization with HPLC showed the retention time of product at 6.201 minutes to LAT-3 EO,
6.261 minutes to LAT-5 EO and 6.260 minutes to LAT-7 EO. Generally, the ethoxylated product has
a good potential to be used as detergent and industrial application.
Kata kunci: Tindak balas pengetoksilan, lauril alkohol teretoksilat, etilena oksida, minyak sawit.
Pengenalan
Malaysia merupakan negara pengeluar dan pengeksport terbesar minyak sawit. Sebanyak 90%
minyak sawit yang dikeluarkan diguna dalam industri makanan dan industri oleokimia seperti
pembuatan kosmetik, detergen dan sebagainya [1]. Disebabkan pasaran turun naik bagi bahan mentah
surfaktan yang berasaskan minyak petroleum maka satu alternatif baru telah diberi perhatian iaitu
penggunaan bahan mentah yang berasaskan bahan semulajadi seperti kelapa sawit dan sayur-sayuran
sebagai menggantikan penggunaan produk berasaskan petrokimia. Ini berikutan daripada tren dunia
tentang penjimatan kos dan juga konsep yang lebih mesra alam.
Surfaktan diambil daripada singkatan perkataan ‘surface active agent’ yang bermaksud agen
aktif permukaan. Surfaktan meliputi kimia organik sintetik yang mana ia membantu dalam
pembuihan, pemelarutan, pengemulsian, penyerakkan dan pembasuhan. Walaupun demikian, tidak
kesemua surfaktan mempunyai ciri-ciri yang sama kerana ia sangat bergantung kepada struktur kimia
molekul tersebut [2]. Surfaktan mempunyai 2 bahagian iaitu hidrofilik (suka air) dan hidrofobik (suka
minyak).
Industri surfaktan kini semakin terus berkembang pesat di mana penggunaan surfaktan dunia
terus meningkat kepada 11 million ton setiap tahun. Di samping itu penggunaan bahan pengemulsi,
agen pembasah dan lain-lain lagi telah lama berkembang dalam sektor perindustrian. Surfaktan juga
banyak digunakan dalam industri pemprosesan seperti industri makanan, farmaseutikal dan penjagaan
diri. Surfaktan bukan sahaja penting sebagai juzuk yang aktif tetapi juga penting sebagai pengemulsi,
penstabil dan pelembut fabrik. Rajah 1 menunjukkan kegunaan lemak dan minyak dunia. Pada 1990,
pengeluaran surfaktan dunia mencapai sehingga 81 million ton yang mana 11.4 million ton
menggunakan proses teknikal dan kimia [3].
Rajah 1. Kegunaan lemak dan minyak dunia
Industri surfaktan telah meningkat pada akhir dekad dan penggunaan bahan mentah
semulajadi telah diberi perhatian bagi menggantikan produk yang berasaskan petrokimia seperti
penghasilan gula berasaskan surfaktan seperti alkil glukosida [4]. Sebanyak 20 million ton bahan
mentah kimia yang dikitar semula telah diguna seluruh dunia setiap tahun seperti karbohidrat 15 %
(55 % daripada minyak dan lemak), glukosa, sukrosa dan bahan sorbitol (daripada penghidrogenan
glukosa) merupakan sebatian polihidroksi daripada bahan semulajadi. Sebatian-sebatian ini boleh
diguna tanpa had, mempunyai ekologi yang bagus, berketulenan tinggi serta kurang toksik. Oleh
kerana itu, kajian secara menyeluruh tentang penggunaannya dilakukan [3].
Lemak alkohol teretoksilat merupakan salah satu daripada kelas surfaktan tidak berion yang
penting. Tindak balas pengetoksilan lemak alkohol ini dilakukan dengan kehadiran mangkin.
Mangkin aluminium alkoksida sulfat telah dikaji tentang keberkesanannya dalam tindak balas
pengetoksilan. Hasil yang diperolehi adalah lebih tinggi berbanding mangkin bes. Namun demikian
banyak produk sampingan yang terhasil [5]. Sehingga kini, mangkin bes seperti KOH dan NaOH
banyak digunakan dalam industri [6].
Kajian tindak balas pengetoksilan telah dilakukan terhadap nonifenol oleh Santecaseria.
Kajian ini mengenai kinetik nonilfenol teretoksilat dengan menggunakan mangkin KOH [6]. Selain
itu, kajian tindak balas pengetoksilan juga telah dilakukan dengan menggunakan 1-dodecanol dan 4-
nonilfenol sebagai reaktan. Kajian dilakukan untuk melihat keberkesanan tindak balas pada pelbagai
suhu tindak balas dilakukan [7]. Formula umum bagi poliglikol eter yang terhasil daripada tindak
balas pengetoksilan seperti kajian yang telah dilakukan oleh Alfonso dan rakan-rakan pada 2004 [1]
adalah seperti berikut:
CH3 (CH2)m-(OCH2CH2)nOH
Eksperimen
Reagen dan Kaedah
a) Penyediaan mangkin
Mangkin KOH dalam bentuk pallet dengan ketulenan 99.9 % daripada Aldrich Chemical digunakan.
Sebanyak 1.34 g KOH dilarutkan dalam campuran 0.56 ml air suling dan 0.804 ml metanol daripada
Fischer Scientific dengan ketulenan 99.99 %.
b) Bahan kimia dan reaktor
CaCO3 (kalsium karbonat) daripada Fluka sebagai peneutral dalam bentuk serbuk dilarutkan dengan
air suling, lauril alkohol berketulenan 99 % daripada Cognis Oleochemical sebagai bahan hidrofobik,
gas etilena oksida (EO) daripada Fluka sebagai bahan hidrofilik dengan ketulenan 99% dan cecair
nitrogen daripada Taylor Wharton berfungsi sebagai pemerangkap atau pemegun wap metanol.
c) Pelarut dan bahan untuk ujian pencirian surfaktan
Pelarut kalium bromida (KBr) digunakan dalam penyediaan sampel untuk analisis FTIR. Air suling
dan metanol berketulenan 99% (Gred HPLC) daripada Fischer Scientific sebagai pelarut untuk HPLC.
Cotton AS9 pigment/oil digunakan dalam ujian detergensi untuk menentukan peratus penyingkiran
kotoran.
Kaedah Tindak balas Pengetoksilan
Reaktor pengetoksilan yang digunakan adalah daripada Vinci Technology dari Perancis. Reaktor ini
mempunyai sistem keselamatan yang tinggi bagi mengendalikan gas EO. Secara umumnya reaktor
tersebut terdiri daripada tangki EO, sistem vakum, penimbang EO, pam gas EO, reaktor(sistem
pengetoksilan), pengawal suhu reaktor, sistem pembuangan gas EO dan sistem kawalan komputer
seperti dalam skema reaktor dalam rajah 2.
P-14 P-15
Gas Nitrogen
Tangki EO
P-12 P-13
V-11 V-8
P-4 P-5 Kebuk penyahtoksik

V-6 P-10
P-12 P-11
komputer
P-9
V-7
P-2P-2 P-3 P-3 P-3 P-7 P-8 P-16

Pam vakum
P-1 P-2
P-17
GC
V-1
Pam
P-3
V-2 V-3 V-5
Kebuk tindak balas
Skema tindak balas pengetoksilan
Rajah 2. Skema reaktor pengetoksilan.

Reaktor pengetoksilan dilengkapi dengan gas EO sebagai reaktan dan gas nitrogen sebagai gas pembawa. Gas EO akan
ditukar kepada bentuk cecair melalui pam EO. Kemudian cecair EO yang terhasil akan disuntik ke dalam reaktor
pengetoksilan. Rajah 3 menunjukkan reaktor pengetoksilan yang diguna dalam menghasilkan surfaktan tidak berion.
Kaedah sintesis LAT merupakan satu tindak balas pengetoksilatan iaitu antara molekul etilena
oksida (EO) terhadap lauril alkohol. Sebelum itu mangkin bes iaitu 1.34 g KOH dilarutkan dalam
campuran 0.56 ml air dan 0.804 ml metanol. Larutan ini dimasukkan ke dalam reaktor dan dipanaskan
di bawah vakum pada suhu 90°C selama 1 jam di dalam reaktor pengetoksilan. Selepas 1 jam, larutan
ini akan kelihatan seperti gel berwarna putih.
Seterusnya suhu reaktor terlebih dahulu dinaikkan sehingga 100 °C. Setelah itu lauril alkohol
dimasukkan ke dalam reaktor dan apabila suhu berada dalam keadaan stabil iaitu pada 70 0C, EO pula
disuntik masuk melalui sistem reaktor mengikut bilangan mol yang ditetapkan. Tindak balas
pengetoksilan berlaku sejurus EO masuk ke dalam reaktor. Pengacauan terus dilakukan sepanjang
proses tindak balas. Selepas tamat tindak balas, suhu reaktor diturunkan dan gas nitrogen dilalukan
dalam reaktor bertujuan untuk menyingkirkan lebihan EO yang mungkin tidak bertindak balas.
Rajah 3. Rajah reaktor pengetoksilan.
Pencirian
Produk yang diperolehi dianalisis menggunakan teknik FTIR jenis Perkin Elmer Model GX dan
HPLC jenis Agilent Technologies Siri 1100 dengan menggunakan pengesan ELSD jenis Altech
Model 2000. Selepas itu pencirian seterusnya dilakukan bagi menentukan tahap pembuihan dan
kestabilannya serta peratus penyingkiran kotoran dengan melakukan ujian pembuihan dan ujian
detergensi.
a) Ujian Pembuihan LAT
Ujian LAT dilakukan untuk ujian pembuihan telah dilakukan terhadap surfaktan yang
diperolehi iaitu keupayaan pembuihan (foaming power) dan kestabilan pembuihan (foaming
stability).
Kaedah yang digunakan ini diperolehi daripada AOTD in-house method. Kaedah ini
menggunakan silinder 500 ml yang diisi dengan larutan surfaktan (0.2 g dalam 200 ml air ternyahion).
Larutan dikocak menggunakan rod selama 30 kali pada kadar yang tetap. Apabila buih terbentuk,
bacaan diambil bagi mengira kadar keupayaan pembuihan iaitu kemampuan surfaktan menghasilkan
buih, berdasarkan persamaan :-
Keupayaan pembuihan = [(takat atas) – (takat bawah)/ Jumlah ketinggian]
Manakala kestabilan pembuihan pula diukur selepas 5 minit kocakan. Kestabilan pembuihan adalah
kemampuan surfaktan mengekalkan tahap pembuihan dalam masa tertentu dan ia diukur berdasarkan
persamaan:-
Kestabilan pembuihan = [(takat atas) – (takat bawah)/ Jumlah ketinggian]
b) Peratus penyingkiran kotoran
Dalam pencucian, % keupayaan detergen diuji dengan menggunakan spektrofotometer

CM36000. Dalam eksperimen ini, detergen diuji ke atas cotton AS9 pigment/oil. Surfaktan
terhasil dicairkan dengan menggunakna 100 ml air suling dan dicampur dengan 50 ppm air
liat (hardness water) selama 3 minit. Selepas proses pencampuran, cotton AS9 pigment/oil
dimasukkan ke dalam bekas untuk pencucian selama 10 minit. Akhir sekali, cotton AS9
pigment/oil dibilas sebanyak 2 kali dengan 1000 ml dan 500 ml air liat. Peratus penyingkiran
kotoran dikira berdasarkan:
% penyingkiran kotoran = [(AW-BW) / (BS-BW)] x 100
AW = intensiti kecerahan selepas cucian terhadap cotton AS9 pigment/oil

BW = sebelum cucian terhadap cotton AS9 pigment/oil
BS = sebelum cucian dalam standard clothes
Proses pengetoksilan
Proses pengetoksilan lauril alkohol berjaya dijalankan pada julat suhu 100 0C - 135 0C dengan masa
tindak balas selama 2 jam. Tindak balas yang berlaku adalah seperti berikut:
CH3 (CH2)11-OH + n (CH2CH2O) Æ CH3 (CH2)11-(OCH2CH2)nOH

(Lauril alkohol) (EO) (LAT)
Kenaikan suhu reaktor dapat dilihat secara mendadak semasa EO disuntik ke dalam
reaktor iaitu dari 60 0C sehingga 120 0C melebihi suhu yang diset pada monitor. Ini
menandakan tindak balas eksotermik telah berlaku pada tindak balas pengetoksilan tersebut.
Rajah 4 (i) menunjukkan profil perubahan suhu reaktor semasa tindak balas pengetoksilan
berlaku dan rajah 4 (ii) pula menunjukkan graf suhu yang telah diset pada monitor.
Suhu reaktor dikawal sepenuhnya oleh komputer. Pada awalnya suhu dinaikkan sehinnga 90
0
C dan pada ketika suhu mencapai 50 0C lauril alkohol dimasukkan ke dalam reaktor (a). reaktor
dibiarkan seketika sehingga suhu yang tercatat pada komputer adalah stabil iaitu pada 700C dan pada
ketika ini, EO disuntik (b). Peningkatan suhu secara mendadak dapat dilihat setelah EO habis disuntik
iaitu daripada suhu 60 0C dan meningkat sehingga 120 0C. Ini menandakan tindak balas eksotermik
berlaku. Ini adalah kerana kereaktifan molekul EO dan aktiviti mangkin yang digunakan.
Pencirian LAT
a) FTIR
Perubahan warna daripada warna jernih lauril alkohol kepada warna kuning keemasan setelah proses
pengetoksilan tamat menandakan wujudnya sebatian baru iaitu LAT. Produk yang terhasil dianalisis
dengan menggunakan teknik FTIR. Sampel disedia dengan menggunakan teknik lazim. Kumpulan
berfungsi yang wujud dalam sebatian yang dikaji dapat diketahui melalui maklumat yang diperolehi
daripada spectrum FTIR. Rajah 5 menunjukkan spektrum bagi lauril alkohol dan sebatian-sebatian
LAT yang diperolehi.
Rajah 4. Profil suhu tindak balas pengetoksilan LAT.
(a) = Kemasukkan lauril alkohol.

(b) = EO disuntik.
(c) = Masa tindak balas.
Rajah 5. Spektrum FTIR bagi produk LAT dan lauril alkohol.
Spektrum FTIR bagi semua siri LAT dan lauril alkohol yang diperolehi menunjukkan
wujudnya puncak ikatan O-H (i) pada jarak gelombang 2500 – 3450 cm-1 dan puncak regangan C-O
(ii) pada jarak gelombang 900 – 1350 cm-1. Regangan C-O menandakan bahawa hasil yang diperolehi
adalah sebatian yang mempunyai kumpulan eter.
b) HPLC
Setelah itu, LAT yang diperolehi dianlisis dengan menggunakan teknik HPLC. Turus jenis Zorbax
300 SB-C18 digunakan bagi mendapatkan spektrum LAT. Pelarut yang digunakan adalah metanol
dan air dengan nisbah 90:10. Sampel disuntik pada kadar 2 µl dengan kadar alir bagi teknik ini adalah
1ml/min. Rajah 6 menunjukkan kromatogram HPLC bagi LAT yang diperolehi.
(a) (b)
(c) (d)
Rajah 6. Kromatogram HPLC bagi sebatian lauril alkohol (a), LAT-3EO (b), LAT-5EO (c) dan LAT-
7EO (d).
Peratus penukaran produk diperolehi berdasarkan teknik ini. Daripada analisis kromatogram,
peratus penukaran produk bagi LAT hampir mencapai 100 % seperti yang tercatat dalam
jadual 1 di bawah.
Jadual 1. Hasil analisis berdasarkan kromatogram HPLC.
Produk Bilangan Masa penahanan Peratus penukaran

mol EO (min) produk
Lauril alkohol 0 1.846 0
teretoksilat (LAT) 3 6.201 70.6
5 6.261 97.6
7 6.260 79.8
c) Ujian Detergensi
Ujian pencucian dilakukan terhadap LAT yang terhasil bagi menentukan peratus penyingkiran
kotoran. Dalam proses ini, 3 elemen diambil kira iaitu substrat, jenis kotoran dan larutan pembersih
dalam menentukan keberkesanan pencucian. Struktur kimia bagi panjang rantai kumpulan hidrofobik
dan hidrofilik menentukan peratus pencucian kotoran. Panjang rantai karbon yang lurus adalah lebih
baik daripada rantai karbon bercabang bagi kumpulan hidrofobik dan peningkatan panjang rantai
kumpulan ini akan meningkatkan lagi keberkesanan pemindahan kotoran berminyak [8].
Manakala peningkatan kumpulan hidrofilik dalam rantai polioksietilena (POE) menunjukkan
penurunan keberkesanan jerapan surfaktan ke atas suatu bahan dan ia berkaitan dengan penurunan
dalam detergensi, contohnya dalam ujian detergensi kain kapas pada suhu 30 0C oleh POE nonilfenol
dalam air suling menurun dengan peningkatan kumpulan EO daripada 9 kepada 20 [9]. Rajah 7
menunjukkan peratus penyingkiran kotoran LAT (surfaktan tidak berion) yang diperolehi.
30 -
29 -
Penyingkiran kotoran (%)
28 -
27 -
26 -
25 -
24 -
23 -
22 -
0
3 EO 5 EO 7 EO
Surfaktan tidak berion
Rajah 7. Peratus penyingkiran kotoran bagi surfaktan tidak berion.
Peratus penyingkiran kotoran bagi LAT-5 EO adalah lebih tinggi berbanding LAT-3
EO dan LAT-7 EO. Ini menunjukkan panjang rantai EO iaitu kumpulan hidrofilik
mempengaruhi peratus penyingkiran kotoran, iaitu semakin panjang EO maka semakin
rendah peratus penyingkiran kotoran. Namun demikian, rantai EO terlalu pendek juga
mempengaruhi peratus pencucian. Oleh sebab itu, LAT dengan 5 mol EO adalah amat sesuai
digunakan dalam detergen berdasarkan ujian detergensi yang dilakukan.
d) Ujian Pembuihan
Ujian pembuihan bergantung kepada panjang rantai alkil, darjah pengetoksilan dan nisbah
surfaktan yang hadir. Peningkatan darjah pengetoksilan akan menghasilkan surfaktan yang
lebih larut air [10].
Semasa pembuihan, permukaan udara-air terbentuk. Secara amnya ia akan
menghasilkan luas permukaan yang tinggi bagi jerapan surfaktan yang tidak lengkap. Luas
permukaan akan menurun dengan masa dan ini merupakan peningkatan jerapan surfaktan.
Ujian pembuihan menentukan tahap keupayaan buih dan kestabilan buih yang terhasil. Rajah
8 menunjukkan keupayaan dan kestabilan pembuihan bagi LAT yang diperolehi.
0 minit 5 minit
120
100
Ketinggian buih (ml)
80
60
40
20
0
3 EO 5 EO 7 EO
Lauril alkohol teretoksilat
Rajah 8. Keupayaan dan kestabilan pembuihan bagi semua siri LAT.
Keupayaan pembuihan LAT-7 EO yang terhasil adalah lebih tinggi berbanding LAT-3 EO
dan LAT-5 EO. Namun, LAT-5 EO memberikan kestabilan pembuihan yang lebih baik berbanding
LAT-3 EO dan LAT-7 EO. Secara amnya peningkatan bilangan kumpulan EO dalam surfaktan tidak
mempengaruhi peningkatan dalam keupayaan pembuihan dan kestabilan pembuihan.
e) Nilai Keseimbangan Hidrofilik-Lipofilik (HLB)
Produk LAT yang diperolehi dihitung nilai HLBnya berdasarkan persamaan (i). Jadual 2 di
bawah menunjukkan peratus hasilan lauril alkohol teretoksilat (LAT) yang diperolehi.
Berdasarkan jadual 3, nilai HLB bagi produk LAT yang diperolehi menandakan ia
mempunyai ciri-ciri sebagai agen pengemulsi yang sesuai digunakan dalam penghasilan
detergen.
20 xMh
HLB = Persamaan (i)
Mh + Ml
Mh = berat bahagian hidrofilik

Ml = berat bahagian hidrofobik
Jadual 2. LAT yang terhasil dengan berat molekul, nilai HLB dan peratus hasilannya.
Produk Berat molekul Nilai HLB % Hasilan

LAT + 3 EO 318 8.30 91.52 %
LAT + 5 EO 406 10.84 88.96 %
LAT + 7 EO 494 12.47 99.63 %
Jadual 3. Ciri-ciri surfaktan berdasarkan nilai HLBnya.
Nilai HLB Ciri-ciri surfaktan / Potensi kegunaan
3–6 air dalam pengemulsi minyak
7–9 agen pembasah
8 – 15 minyak dalam pengemulsi air
12 – 15 bersifat detergen
15 – 18 bersifat keterlarutan
Kesimpulan
Bahan kimia berasaskan bahan semulajadi daripada minyak sawit digunakan iaitu lauril alkohol. Ini
berikutan banyak kelebihan yang boleh diperolehi daripada bahan semulajadi ini. Selain daripada
senang diperolehi dan murah, penggunaannya juga akan menghasilkan produk yang tidak toksik dan
mudah terbiodegradasi.
Hasil analisis menunjukkan bahawa LAT telah berjaya dihasilkan berdasarkan teknik FTIR
dan HPLC. Teknik FTIR menunjukkan wujudnya satu sebatian baru selepas tindak balas
pengetoksilan dilakukan terhadap lauril alkohol. Terdapat puncak ikatan O-H pada jarak gelombang
2500 – 3450 cm-1 dan puncak regangan C-O pada jarak gelombang 900 – 1350 cm-1. Regangan C-O
menandakan hasil yang diperolehi adalah sebatian yang mempunyai kumpulan eter. Sebagaimana
yang diketahui, melalui tindak balas pengetoksilan lemak alkohol akan menghasilkan sebatian
poliglikol eter.
Manakala melalui teknik HPLC menunjukkan peratus penukaran produk yang tinggi
iaitu menghampiri 100 % bagi produk-produk LAT ini. Setelah pelbagai pencirian surfaktan
dilakukan ke atas semua siri produk LAT, dapat disimpulkan bahawa produk LAT-5 EO
adalah lebih sesuai untuk kegunaan industri dan detergen. Ini adalah berdasarkan pada nilai HLBnya
iaitu 10.84 dan ujian detergensi yang dilakukan menunjukkan peratus pencucian bagi LAT-5 EO
adalah tinggi daripada LAT-3 EO dan LAT-7 EO dan mempunyai keupayaan pembuihan yang rendah
berbanding LAT yang lain. Sebagaimana yang diketahui pembuihan yang tinggi tidak menentukan
detergen yang baik. LAT-5 EO adalah lebih berpotensi dalam penggunaan detergen berbanding
produk LAT yang lain berdasarkan ujian-ujian yang dilakukan ke atasnya.
Penghargaan
Setinggi-tinggi ucapan terima kasih diucapkan kepada pihak MOSTE ‘Ministry of Science
Technology and Innovation of Malaysia’ di atas geran penyelidikkan IRPA 09-02-02-0033
SR0004/05-03. Selain itu, jutaan terima kasih juga diucapkan kepada Jabatan Kimia, Pusat Sains
Kimia dan Teknologi Makanan, Fakulti sains dan Teknologi, Universiti Kebangsaan Malaysia
(UKM).
Rujukan
1. Alfonso, E. S., Citterio. A., Ramis-Ramos, G., Righetti, P. G. & Sebastiano, R. (2004)
“Separation of Fatty Alcohol Polyethoxylates by Capillary Electrophoresis Through Easy
Electroosmotic Flow Control with a Queternary Diammonium Salt” Journal of
Chromatography A 1053. 235-239.
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THE OPTIMIZATION OF THE CHITIN HYDROLYSIS PARAMETERS BY
HYDROCHLORIC ACID BY RESPONSE SURFACE METHODOLOGY
Kah Leong, Chee & Osman, H.

School of Chemical Sciences & Food Technology, Faculty of Science & Technology,
National University of Malaysia, 43600 Bangi, Selangor, Malaysia.
Correspondence email: osman @ ukm.my
Abstract
Acid hydrolysis of chitin is a conventional method to produce N-acetyl-D-glucosamine. The

performance of 6 types of acid hydrolysis (6N HCl, 6N CH3COOH, 6N H2SO4, 6N HNO3, 85%
H3PO4, and 6N CCl3COOH have been studied through high performance liquid chromatography
analysis. Then, the response surface methodology was applied to optimize the acid hydrolysis
parameters (substrate concentration, acid concentration and temperature) for the N-acetyl-D-
glucosamine production. Throughout the screening, the 6N HCl hydrolysis produces the highest yield
of N-acetyl-D-glucosamine (0.496%), followed by 6N HNO3, 6N CH3COOH, 6N CCl3COOH, 85%
H3PO4 and 6N H2SO4, hydrolysis that produce 0.285%, 0.240%, 0.117%, 0.032% and 0.009% of N-
acetyl-D-glucosamine respectively. Therefore, HCl was chosen to optimize the production. The
concentration of substrate (CC-RBB), HCl concentration and temperature were significant factors
after analyzed by 23-1 fractional factorial design. The steepest ascent method was used to locate the
optimal domain and a central composite design was used to estimate the quadratic response surface
from which the factor levels for maximum absorbance were determined. The optimal conditions of
concentration of substrate (CC-RBB), HCl concentration and temperature at which maximum amount
of N-acetyl-D-glucosamine formed were found to be 140mg/ml of CC-RBB, 10.18M of HCl
concentration, 90oC, and 2.5 hours respectively. The higher yield of N-acetylglucosmine was
produced using colloidal chitin (11.96%) compare to chitin (10.40%) as substrate.
Keywords: Acid hydrolysis; Chitin; N-acetyl-D-glucosamine; Optimization; Response surface methodology.
1.0 Introduction
Chitin, a β-(1→4)-linked 2-acetamido-D-glucopyranan, is the second most abundant biopolymer on

earth (1). Enzymatic or acid hydrolysis of chitin produced oligosaccharides consisting of β-(1→4)
linked N-acetyl-D-glucosamine (2-acetamido-2-deoxy-D-glucose; GlcNAc) and monomer. Recently,
GlcNAc have been promoted as a treatment or as nutraceutical agents for patients with osteoarthritis
and inflammatory bowel disease [2, 3]. Since GlcNAc is also a component of proteoglycan and a
part of GlcN is transformed to GlcNAc by metabolism procedure, the therapeutic activity on
osteoarthritis for GlcNAc would be similar to that of GlcN. In contrast to GlcN hydrochloride
or sulfate, both of which have a bitter taste, GlcNAc has a sweet taste that facilitates its use in
daily consumption. However, GlcNAc has not been widely commercialized mainly due to the
lack of an economical process for its production that is acceptable for food and medicine.
Currently, N-acetyl-D-glucosamine is produced by the acetylation of glucosamine, a product
of the acid hydrolysis of chitin. That process is limited by poor yields and the availability of raw
materials such as crab shells [4]. Therefore, the optimization of the acid hydrolysis is important. Many
factors such as substrate concentration, acid concentration, temperature and time are important
variables affecting the production of N-acetyl-D-glucosamine. It is difficult to find the most important
factors and to optimize them via classic or empiric methods because they have several limitations
toward complete optimization. Actually, the traditional one-factor at a time approaches to
optimization is time-consuming and incapable of reaching the true optimum due especially to
interactions among the factors. Moreover, it assumes that the various hydrolysis parameters do not
interact and the process response is a direct function of the single varied parameter. In contrast, the
observed hydrolysis results arise from the interactive influences of the various variables. Unlike
conventional optimization, statistical optimization methods can take into account the interactions of
variables in generating the process response [5].
Factorial design of optimization experiments is especially suitable to account for the
interactions. A combination of factors generating a certain optimum response can be identified
through factorial design and the use of response surface methodology [6]. Response surface
methodology, an experimental strategy for seeking the optimum conditions for a multivariable
system, described first by Box and Wilson [7], is a much more efficient technique for optimization
[8]. This method had been successfully applied to the optimization of medium composition [9, 13],
conditions of enzymatic hydrolysis [10], parameters of food preservation [11] and fermentation
processes [12].
In this study, we applied the experimental study for response surface design to examine the
acid hydrolysis process of chitin. This will allow one to choose the best condition for produce the N-
acetyl-D-glucosamine. As a preliminary study, fractional factorial design (FFD) was used for
screening factors that influence GlcNAc production significantly and insignificant ones were
eliminated in order to obtain a smaller, more manageable set of factors. By using this method,
interactions between variables also can be identified and quantified. After that, the critical subset of
factors is focused and finally central composite design is used to examine and optimize this process.
Design Expert ® Software Version 6.0 was used as the statistical experimental methods in this study.
2.0 Experimental
2.1 Material
Shrimp chitin flake was purchased from Ohka Enterprise Inc. (Malaysia). Acids and alkalis (HCl,
CH3COOH, HNO3, H3PO4, CCl3COOH, H2SO4, NaOH) of analytical grade were obtained from E.
Merck (Darmstadt, Germany). Sigma Chemicals (Malaysia) supplied the standards of GlcNAc,
Remazol Brilliant Blue R (RBB) and chemicals for the dyeing reaction.
2.2 Colloidal chitin preparation
Ten grams of chitin were ground in a laboratory hammer mill (IKA-WERKE) and to particles of
0.25mm. The obtained powder was added in 300ml of 85% phosphoric acid and kept in a refrigerator
(5°C) for 24 h. Thereafter, 2 l of distilled water were added and the gelatinous white material formed
was separated by filtration through filter paper. The retained cake was washed with distilled water
until the filtrate achieves pH 6.5 [14].
2.3 Dyeing of chitinous material [15]
Two hundred grams of colloidal chitin were homogeneously mixed with 400 ml of an aqueous
solution of RBB at 5.0%. The resulting suspension was heated in a boiling water bath for 60 min with
gentle stirring. The colloidal dyed material was removed by filtration and, to fix the RBB-colour, the
obtained material as such was suspended in 25 ml of an aqueous solution of 1.5% sodium dichromate
and 1.5% sodium potassium tartrate, without any pH adjustment, followed by heating with gentle
stirring into a boiling water bath for 10 min. The dyed material was then separated by filtration and
washed with hot water until the filtrate was colorless. The gelatinous blue material obtained was
sterilized by autoclaving, and freeze dried.
2.4 Preparation of Acid Hydrolyzed Samples
For acid screening, 5 g of chitin powder was added into 100 mL of 6N (HCl, CH3COOH, HNO3,
CCl3COOH, H2SO4) and 85% of H3PO4. The sample solution was in a water bath maintained at a
constant temperature of 90ºC for 2 hours. Then, the residue was filtered by the Whatman No.1
filter paper. The 50-mL sample solution was quickly cooled in an ice bath, freeze-dried at -40ºC
under vacuum. The solid particles obtained after drying were redissolved in 50 mL deionized water.
The above drying and dissolution processes were repeated twice to remove as much residual acid as
possible. The 50 mL sample solution was then neutralized with 1N NaOH, followed by filtration
through Whatman no. 1 filter paper to remove impurities. The filtrate was concentrated by
evaporation under reduced pressure until the final concentration was around 1mL. Methanol (HPLC
grade, 19mL) was then added to precipitate oligosaccharides with DP’s ≥8 [16]. The methanolic
solution was filtered through a Whatman membrane filter (0.45 µm pore size, 13 mm diameter)
before HPLC injection. The preparation hydrolyzed sample of the optimum condition is the same with
the optimum parameters.
2.5 Quantification of GlcNAc
The identification and amount of GlcNAc was determined using HPLC analysis with a Waters liquid
chromatograph (Millipore Corp., Milford, MA, USA). The HPLC system was fitted with a Waters
1525 binary HPLC pumps, a Waters 717plus autosampler and a Waters 2414 RI detector. A prevail
carbohydrate column (5 µm, 4.6mm X 250 mm) from ALLTECH was used to elute GlcNAc
isocratically at 0.8 mL/min with 70% acetonitrile ratio to 30% deionized water (0.1% NH4OH).The
injection volume was 20 µL. For the quantification of hydrolyzed samples, standard solutions of
GlcNAc (containing 500 ppm of each GlcNAc) were used for calibrations before daily analysis. The
ratios between the peak areas of samples and standards were used to calculate the weight of GlcNAc
in the original hydrolysate solution.
2.6 Experimental design
A series of statistically designed studies were performed to investigate the effect of 3 factors
on producing GlcNAc. The optimization process first entails identifying the most important factors in
this process and then focusing on the critical subset of factors. The steepest ascent was used to
determine the direction toward predicting higher responses. Finally a central composite design was
performed to optimize the critical factors and optimize the process. A 23-1 full factorial design (FFD)
was used to pick factors that influence GlcNAc production significantly and insignificant ones were
eliminated in order to obtain a smaller, more manageable set of factors. In FFD, low and high factor
settings were coded as –1 and +1 , the midpoint was coded as 0. Factor settings were shown in the
form of both coded values and natural values. Xi = (xi-xi0) / δi, where Xi is the coded value, xi is the
corresponding natural value, xi0 is the natural value in the center of natural domain and δi is the
increment of xi corresponding to 1 unit of Xi. The factors and coded values of FFD are shown in
Table 1. The linear model obtained is expressed as follows:
k
y = βο + ∑ β ixi
i =1 Eq (1)
If the mean of the center points exceeds the mean of factorial points, the optimum would be near or
within the experimental design space. If the mean of the center point had been less than the mean of
factorial points, the optimum would be outside the experimental design space and the method of
steepest ascent should be applied. The direction of steepest ascent is parallel to the normal of contour
line of response curve of equation (1) and passes through the center point of FFD [17].
In this study, full factorial was chosen to determine the significant effect in producing
GlcNAc. By using 23-1 design, 16 runs are required for duplicate. 6 center points were added to
estimate the experimental error and check the adequacy of the first-order model. Table 1 shows the
factors and coded values of FFD.
Once critical factors were identified via screening and significant gross curvature has been
detected in the design space, the central composite design was proceeded to obtain a quadratic model,
consisting of trials plus a star configuration to estimate quadratic effects and central points to estimate
the variability and reassess gross curvature, with GlcNAc production as response. For two factors, the
model obtained was expressed as follows:
y = βο + β 1 x1 + β 2 x 2 + β 12 x1 x 2 + β 11 x112 + β 22 x 22 2 Eq (2)
where y was the measured response, βo was the intercept term, β1 bone and β2 were linear coefficients,
β11 and β22 were quadratic coefficients, and x1 and x2 were coded independent variables. Low and high
factor settings were coded as –1 and +1, the midpoint was coded as 0. The factor settings of trails that
ran along axes drawn from the middle of the cube through the centers of each face of the tube were
coded as +1.414 or –1.414. The experimental design and results is showed in Table 3. Design Expert
Version 6 was used to analyze the results.
Table 1: Factors and coded values of FFD
Factors Level of factors

-1 0 +1
Temperature (x1 , ºC) 60.0 80.0 100.0
Acid conc. (x2, M) 4.0 6.0 8.0
CC-RBB weight (x3, mg) 1.0 5.5 10.0
2.6 Assay with RBB [15]
One milliliter of HCl with different concentration (4, 6 & 8M) respectively was added into
every treatment respectively as shown in Table 3. After 2.5 hours, the tubes are put into the ice
water for 10 minutes. Then, 1ml NaOH with the suitable Molarity (4, 6 & 8M) was added to
neutralize the hydrolysate. The dilution of sample was with adding 10mL of distilled water.
The tubes were centrifuged at 7000 rpm and the absorbance of the supernatants was read with
Versamax microplate reader at 595 nm.
3.0 Results and discussions
3.1 Screening of best acid
Throughout the screening for six types of acid in the preliminary study, only the 6N HCl
hydrolysis produces the highest yield of N-acetyl-D-glucosamine (0.496%), followed by 6N HNO3,
6N CH3COOH, 6N CCl3COOH, 85% H3PO4 and 6N H2SO4, hydrolysis that produce 0.285%,
0.240%, 0.117%, 0.032% and 0.009% of N-acetyl-D-glucosamine respectively. The first reason is
relevant to the chitin adsorption of acid ability. According to Giles et al. 1958 [18], the acid binding
capacity value is 4.8 eq X kg-1 at pH 2.0. At lower pH value a sudden futher rise in absorption occurs
and proceeds without apparent limit. Once the acetamido and amino groups are saturated, no other
acid combining group will be available unless the ether groups form oxonium salts. Even if this were
so, the total quantity of acid could not be accounted for. Therefore, the general reaction would be:
R-NHCOCH3 + H+X- → R-N+HCOCH3X- Eq (3)
Actually, the acetamido and amino groups of chitin are the only operative groups toward
hydroxylic solutes. The hydroxyl and ether groups on the glucosidic rings are probably
protected by solvated water and thus rendered inactive. Table 2 shows the adsorption of acids
by chitin at 50°C and pH 3.0 for various types of acids where the value for HCl and H2SO4 are
the highest.
Table 2: Adsorption of acids by chitin.

Acid mmol x kg-1
Hydrochloric 3800
Formic 385
Acetic 255
Chloroacetic 320
Sulfuric 3870
Nitric 235
Benzesulfonic 200
On the other hand, the acid strength plays the vital role because the first step in the acid
hydrolysis of chitin is the amine group is protonated and the C(2)-NH3 group shields the glycosidic
oxygen from protonation [19]. The BrØnsted-Lowry theory defines acid strength in terms of proton-
donating tendency. The greater the tendency of a substrate to donate a proton, the greater is its acid
strength; and the higher value of ionization constant (Ka). During the acid hydrolysis, the chain
scission appears to occur in random. Therefore, the HCl, H2SO4, and HNO3 should have the higher
yield of hydrolysis [20].
Refer to the results, HCl hydrolysis produce the highest yield followed by HNO3 hydrolysis, but
the H2SO4 hydrolysis produce the lowest quantity. Why? Rupley (1964) reported that the monomer,
dimer and trimer were found to be the predominant species of the chitin hydrolysis by HCl [19]. This
phenomena does not fit the pattern expected for random splitting of the chain, but Rupley explained
that the chitin chain are aggregated in solution and that the rate-limiting step is a disaggregation step,
followed by rapid hydrolysis. Hydrolysis of chitin by H2SO4 is different with HCl. According to
Nagasawa et al. (1971), the most obvious being that chitin hydrolysis is accompanied by O-sulphation
of chitin. Therefore, the greatest hydrolysis occurs at the time of dissolution, the subsequent
hydrolysis in solution being slower. Secondly, the chain scission appears to occur at random; after 4 h
hydrolysis at 5°C in 96 wt-% H2SO4 the number average DP of the dialyzable fraction, which
amounted to 94 % of the product, was 16, while all the products for hydrolysis times of 1-6 h at -20°C
were non-dialysable. Thus, unlike hydrolysis by HCl, the predominant species are not the monomer,
dimer and trimer [21].
3.2 Optimization of N-acetylglucosamine production

3.2.1 Fractional factorial design
According to the results above and primary studies, HCl was selected as the hydrolyzing
agent for chitin hydrolysis. The concentration of substrate (CC-RBB), HCl concentration and
temperature were the main factors to optimize the medium composition using 23-1 full factorial design
(FFD). Real values of factors, design and results of experiment were shown in Table 3.
The factorial analysis of variance indicated that all factors and interactions are giving the
significant influences to the acid hydrolysis, especially CC-RBB weight give the most
significant impact to the acid hydrolysis due to the highest coefficient estimate value (0.17)
(Table 4). A linear regression equation could be obtained from the regression results of fractional
factorial experiment:
Y = 0.21 + 0.018 x1 + 0.087 x2 +0.170 x3+ 0.028x1 x2+0.019 x1 x3+0.059 x2 x3+0.029 x1 x2 x3 Eq (4)
The regression coefficients and determination coefficient (R2) for the linear regression model of N-
acetylglucosamine production are presented in Table 4. The model was highly significant (p < 0.0001)
and the lack of fit (p= 0.2060) is insignificant indicating that the first-order model is very adequate in
approximating the response surface of the experimental design. This statement is further supported by
the satisfactory value of R2 = 0.9907. The mean of center point (0.165) is lower than the mean of
factorial points (0.19) indicated that optimal point is outside the experimental design space and the
method of steepest ascent should be applied.
Table 3: Experimental design and results of FFD
________________________________________________________________________
Factor 1 Factor 2 Factor 3 Response 1
Run Block A:Temperature B:Acid conc. C:CC-RBB wt. Adsorbance
deg C M mg A
______________________________________________________________________________________
1 Block 1 100.00 4.00 10.00 0.226
2 Block 1 80.00 6.00 5.50 0.151
3 Block 1 60.00 8.00 10.00 0.456
4 Block 1 100.00 8.00 10.00 0.657
5 Block 1 80.00 6.00 5.50 0.181
6 Block 1 80.00 6.00 5.50 0.150
7 Block 1 100.00 4.00 1.00 0.006
8 Block 1 80.00 6.00 5.50 0.180
9 Block 1 80.00 6.00 5.50 0.151
10 Block 1 100.00 8.00 1.00 0.065
11 Block 1 60.00 8.00 1.00 0.069
12 Block 1 80.00 6.00 5.50 0.181
13 Block 1 60.00 4.00 1.00 0.005
14 Block 1 60.00 4.00 10.00 0.257
15 Block 2 100.00 8.00 10.00 0.578
16 Block 2 100.00 8.00 1.00 0.054
17 Block 2 80.00 6.00 5.50 0.150
18 Block 2 80.00 6.00 5.50 0.180
19 Block 2 60.00 4.00 1.00 0.006
20 Block 2 60.00 4.00 10.00 0.249
21 Block 2 100.00 4.00 1.00 0.005
22 Block 2 80.00 6.00 5.50 0.151
23 Block 2 80.00 6.00 5.50 0.181
24 Block 2 80.00 6.00 5.50 0.150
25 Block 2 100.00 4.00 10.00 0.197
26 Block 2 60.00 8.00 1.00 0.059
27 Block 2 60.00 8.00 10.00 0.404
28 Block 2 80.00 6.00 5.50 0.180
Table 4: Coefficient value and p-value for the factors

Factor Coefficient Estimate p-value
Intercept 0.21
Block 1 6.821E-003
Block 2 -6.821E-003
A-Temperature (X1) 0.018 0.0015
B-Acid conc. (X2) 0.087 < 0.0001
C-CC-RBB wt. (X3) 0.17 < 0.0001
AB 0.028 < 0.0001
AC 0.019 0.0009
BC 0.059 < 0.0001
ABC 0.029 < 0.0001
Center Point -0.040
3.2.2 Steepest ascent path
The direction of steepest ascent path can be determined by Eq. (4) and regression results. All
the factors were significant and their coefficients are positive. This means that increasing the
temperature, acid concentration and CC-RBB weight will have positive effects on the N-
acetylglucosamine production. CC-RBB was chosen as standard because its coefficient is the biggest.
Design of experiment of the steepest ascent and corresponding results are shown in Table 5. GlcNAc
production of OBS 5 was the highest (0.861A), then adsorbent decreased following the concentration
changing. The results mean that factors value of OBS 5 was near optimal, thus OBS 5 was chosen as
the center point to optimize the medium composition. This result is agreed with the statement of a
hydrolysis mechanism in which the physical arrangement of the chitin chains is important in
determining the kinetic behaviour is in agreement with the observed high reaction order with respect
to acid concentration [19].
Table 5: Results of the steepest ascent path experiments

OBS. Step A(Temp.) B(Acid conc.) C(CC-RBB) Response
°C N % A
1 Source (Median) 80 6 5.5 0.235
∆ 0.106 0.512 1 -
2 S+ 2∆ 80.212 7.024 7.5 0.266
3 S+ 4∆ 80.424 8.048 9.5 0.364
4 S+ 6∆ 80.636 9.072 11.5 0.645
5 S+ 8∆ 80.848 10.096 13.5 0.861
6 S+ 9∆ 80.954 10.608 14.5 0.853
7 S+ 10∆ 81.060 11.120 15.5 0.843
3.2.3 Central composite design

A highly significant regression model (p< 0.0001) for N-acetyl-glucosamine
production with a satisfied R2 = 0.9589 was achieved. According to the insignificant Lack of
Fit value (p= 0.1626), this model is suitable for the screening experimental design.
Experimental design and results are shown in Table 6. A full second-order polynomial model was
obtained from regression analysis of data of experiment of central composite design:
Y = 0.13 +1.474E-003 X1 ++3.531E-003 X2 +4.240E-003 X3 +1.282E-003X12-4.021E-003 X22

+4.287E-003 X32 -2.125E-003X1 X2 Eq (5)
The resulting response surface showed the effect of CC-RBB weight, acid concentration and
temperature on the N-acetylglucosamine production (Fig. 1). The maximum yield was obtained in the
optimized condition when the temperature, concentration of acid and CC-RBB weight were 90oC,
10.18M and 14mg respectively. The maximum response predicted from the model was 0.1414A.
Repeated experiments were performed to verify the predicted optimum. The result of five replications
(i.e., 0.140, 0.142, 0.141, 0.142, 0.140 A) was coincident with the predicted value and the model was
proven to be adequate. The final parameters optimized with RSM are: 90oC, 10.18M of HCl
concentration, 14mg of CC-RBB and 2.5 hours. The optimum time course of chitin hydrolysis is 2.5 h
because the chitin had been fully hydrolyzed.
D E S IGN -E X P E R T P l o t
D e s i ra b i l i ty
X = A : T e m p e ra tu re
Y = B : A c id c onc .
A c tu a l Fa c to r
C : C C -R B B w t. = 1 400.7
.0709
0 .6 9 8
0 .6 1 6
Desirability
0 .5 3 4
0 .4 5 3
1 1 .0 0
9 0 .0 0
1 0 .5 0
8 5 .0 0
1 0 .0 0
8 0 .0 0
B: Acid conc.9 .5 0 7 5 .0 0
A: T emper atur e
9 .0 0 7 0 .0 0
Fig. 1. Response surface plot for the effect of temperature (X1) and acid concentration (X2) on
GlcNAc production.
Table 6: Design and results of central composition design

Run Block A: Temp. B: Acid conc. C: CC- RBB wt. Response
1 Block 1 90.00 9.00 14.00 0.136
2 Block 1 96.82 10.00 13.00 0.138
3 Block 1 80.00 10.00 11.32 0.134
4 Block 1 90.00 9.00 12.00 0.127
5 Block 1 80.00 11.68 13.00 0.126
6 Block 1 80.00 10.00 13.00 0.132
7 Block 1 80.00 10.00 14.68 0.150
8 Block 1 80.00 10.00 13.00 0.128
9 Block 1 70.00 9.00 12.00 0.122
10 Block 1 70.00 9.00 14.00 0.130
11 Block 1 80.00 8.32 13.00 0.111
12 Block 1 70.00 11.00 14.00 0.138
13 Block 1 90.00 11.00 14.00 0.138
14 Block 1 80.00 10.00 13.00 0.130
15 Block 1 80.00 10.00 13.00 0.131
16 Block 1 63.18 10.00 13.00 0.129
17 Block 1 70.00 11.00 12.00 0.134
18 Block 1 80.00 10.00 13.00 0.130
19 Block 1 80.00 10.00 13.00 0.129
20 Block 1 90.00 11.00 12.00 0.128
According to Muzzarelli (1977) [18] and Smucker & Pfister (1978)[22], 4-6 N HCI hydrolysis at
100°C for 12-20 h is necessary to achieve complete chitin hydrolysis. Weaker acid hydrolysis such as
0.5-2.0 N HCI or 0.5 M H2SO4 is ineffectual in chitin hydrolysis, but is more than sufficient for most
non-amino sugar hydrolyses. Chen et al. also reported that the yield of NACOs became higher than
60% by hydrolyzing chitin with 10 N or 12 N HCl at 50ºC because some portion of chitin particles
could only be hydrolyzed under higher HCl concentrations [23]. The hydrolysis at higher
temperatures favored the formation of lower molecular weight oligosaccharides. The amount of
(GlcNAc)6 or (GlcNAc)5 obtained at 90ºC was much lower than that obtained at 70ºC (Chang et al.,
2000) [24]. Therefore, the optimum HCl concentration is 10.18M.
3.3 Quantification of GlcNAc by HPLC analysis
Figure 2 shows the HPLC chromatograms of the hydrolyzed samples obtained after treatment
of HCl on chitin and colloidal chitin. The higher yield of N-acetylglucosmine was produced using
colloidal chitin (11.96%) compare to chitin (10.40%) as substrate. The optimized parameters have
increased the production of N-acetylglucosamine almost 21 times compare to the screening
parameters of chitin hydrolysis by using HCl.
Major variable in chitin substrates is the degree of crystallinity or fiber-openness to the
hydrolysing agent attachment. In the current work, the swelling of the substrate through phosphoric
acid treatment may have substantially increased its intial accessibility to HCl. The relatively higher
amount of N-acetylglucosamine production for colloidal chitin in the present work probably results
from an opening up of the polymer and the exposure of centre phase of the colloid [25].
4. Conclusions
Chloric acid is a suitable hydrolysing agent to produce N-acetyl-D-glucosamine, especially by

using colloidal chitin as the substrate. This methodology as a whole proved to be quite adequate for
the design and optimization of a hydrolysis process. Using the method of experimental factorial
design and response surface analysis it was possible to determine optimal condition to obtain high N-
acetyl-D-glucosamine production. In addition to establishing optimal condition for operation, the
present methodology also makes it possible to predict both yield and productivity when the system is
disturbed in some way. This is useful not only for the additional knowledge supplied about the
process, but also for the potentials for process control.
A B C
Figure 2: HPLC elution profiles of (A) a mixture of sugar standard from monomer to tetramer, (B) a
typical acid hydrolyzed chitin sample; and (C) a acid hydrolysed colloidal chitin sample.
5.0 Acknowledgements
This work was supported by grant 09-02-02-0006/BTK/ER/31 from National Biotechnology
Directorate (BIOTEK), MOSTI, Malaysia.
6.0 References
1. Knorr, D. (1984) Use of chitinous polymers in food. Food Technol. 38: 85-97.
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10. Ma, A.Y.M. & Ooraikul, B. (1986) Optimization of enzymatic hydrolysis of canola meal with
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11. King, V.A. (1993) Studies on the control of the growth of Saccharomyces cerevisiae by using
response surface methodology to achieve effective preservation at high water activities. Int J
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12. Rosi, I., Costamagna, L. & Bertuccioli, M. (1987) Wine fermentation by immobilized yeast: an
optimization study. In: Flavor Science and Technology. NY: John Wiley and Sons.
13. Souza, M.C.de O. & Roberto, I.C. (1999) Solid-state fermentation for xylanase production by
Thermoascus aurantiacus using response surface methodology. Appl Microbiol Biotechnol
52:768–772.
14. Rojas Avelizapa, L.I., Cruz Camarillo, R., Guerrero, M.I., Rodrı´guez Va´zquez, R. & Ibarra,
J.E., (1999) Selection and characterization of a proteo-chitinolytic strain of Bacillus
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(2004) Colloidal chitin stained with Remazol Brilliant Blue RR, a useful substrate to select
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16. Muraki, E., Yaku, F. & Kojima, H. (1993) Preparation and crystallization of D-glucosamine
oligosaccharides with dp 6-8. Cabohydr. Res. 239: 227-237.
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chitin digestion. Comp. biochem. Physiol. 83A/ 3: 489-493.
LONG-TERM STUDIES ON THE SALINITY (TDS) AND LEACHING CHARACTERISTICS
OF COAL FLY ASH FROM TNB POWER GENERATION PLANT
Khoo Kok Siong1,*, Sukiman Sarmani2

1,*
Nuclear Science Program, School of Applied Physics, Faculty of Science and Technology,
National University of Malaysia, 43600 Bangi, Selangor, Malaysia,
2
Program of Chemistry and Food Technology, School of Applied Chemistry, Faculty of Science and
Technology, National University of Malaysia, 43600 Bangi, Selangor, Malaysia.
e-mail: khoo_koksiong@yahoo.com
Abstract. Use of coal in power generation plant has led to increasing environmental concern especially in
disposal of ash residues. The problem related to coal fly ash disposal is the constituents of the residue. In this
regard, three laboratory leaching test methods consisted of the pore water test (PW), the acid neutralization
capacity test (ANC) and the tank leaching test (TL) have been carried out. Subsequently, study on leaching
behaviour of salinity (TDS) from coal fly ashes was reported. In Pore water test (PW), it was found that the total
dissolved solids (TDS) decreased when the liquid/solid ratios (L/S) increased. In Acid neutralization capacity
test (ANC), TDS only increased rapidly in higher acid and base concentrations (molarities). In Tank leaching
test (TL), cumulative release (g/m2) was ranged from 151 to 448 g/m2 in 64 days respectively. The studies
indicated that the results are pH supported.
Abstrak. Penggunaan arang batu di janakuasa elektrik telah memberikan kerisauan terhadap alam sekitar
terutamanya pembuangan dan sisa abu arang batu. Masalah yang dikaitkan dengan abu terbang arang batu ialah
kandungan unsure-unsur dalam sisa tersebut. Sehubungan itu, tiga kaedah ujian larut resap iaitu ujian PW, ujian
ANC dan ujian TL telah dilakukan. Seterusnya, kajian terhadap kelakuan larut resap bagi paras kandungan
garam (TDS) pada abu terbang arang batu juga dilaporkan. Dalam ujian PW, didapati bahawa paras kandungan
garam (TDS) menurun apabila nisbah cecair/pepejal (L/S) meningkat. Dalam ujian ANC, TDS hanya meningkat
dengan cepat pada kepekatan asid dan bes yang tinggi. Dalam ujian TL pula, pembebasan kumulatif (g/m2)
adalah dalam julat 151 – 448 g/m2 dalam 64 hari masing-masing. Kajian ini telah menunjukkan bahawa
keputusan adalah bergantung kepada pH.
Keywords: Coal fly ash, salinity (TDS), pore water test, acid neutralization capacity test, tank leaching test
Introduction
Tenaga Nasional Berhad (TNB) is the largest power company in Southeast Asia, TNB generates two-
thirds of the electricity on the Malaysian Peninsula. It operates the national power transmission and
distribution grid, which serves more than 6 million residential, industrial, and commercial customers.
Kapar coal fired power generation plant is the first coal based power plant commissioned since 1988
with 2x300 MW units and additional 2x500 MW units in 2001. The rapid development in Malaysian
economy has resulted increase of coal consumption. One of the by-products produced from power
generation plant is coal fly ash (the non-combustible minerals found in coal). The coal fly ash is
subsequently removed, transported and deposited which may create problem of its disposal.
Characterization of the leaching behaviour of coal fly ashes is a crucial step in the
environmental assessment for reuse or disposal reasons. Recent awareness and empowerment of the
public on the environmental concern has resulted in the need for the assessment of its behaviour.
Leaching is the process by which constituents in a solid material, which in this case as coal fly
ashes, is released into environment via contact with water. The interest of knowing the leaching
process is to define the potential environmental impacts via water-borne mechanisms involving the
contamination of soil, groundwater and surface water; human health from advantageous use and
disposal of wastes; designs and acceptance criteria for waste management facilities and degradation of
structural performance of certain elements in the environment.
Coal fly ash is generated from the combustion of coal in power generation plant boilers and
collected from off-gas combustion in electrostatic precipitators. Typical coal fly ash particles ranged
in size up to 1-150 µm and their composition depends on the nature of the coal being burned [1]. The
coal fly ash is utilized in applications of the cement and concrete industry, such as structural filling
material and recovery of metals [2]. However, the rate of production is greater than the consumption.
The unused coal fly ash is disposed into holding ponds, lagoons, landfills and heaps. The bulk of the
coal fly ash normally consists of silica (SiO2), alumina (Al2O3), iron oxide (Fe2O3), calcium oxide
(CaO), magnesium oxide (MgO), potassium oxide (K2O), sodium oxide (Na2O), sulfur trioxide
(SO3), titanium dioxide (TiO2) and significant quantities of various trace elements such as Ba, Co, Cr,
Ga, Nb, Ni, Rb, Sr, V and Zr [3]. Other nutrients usually present in coal fly ash are P and B and
radioisotopes that are undesirable for animals and human [4].
The release of metals from coal fly ash is influenced by different physical, chemical and
biological factors. In particular, the leaching studies can be divided into three categories:
a) characterization tests for the characterization of materials;
b) compliance tests for verifying agreement with the characterized material;
c) in situ tests to verify the identity of a waste before its disposal or recovery.
In this framework, study has been carried out on three laboratory leaching characterization
test methods for the assessment of the leaching behaviour of salinity (dissolved solids) from coal fly
ashes. Salinity is simply a measure of the amount of salt dissolved in water. Salts such as sodium
chloride (NaCl), calcium carbonate (CaCO3) and others are picked up by water as they run over and
through the soils of catchments. Low concentrations of these salts are important to the growth of
plants and animals but high concentrations can be problem for aquatic life and human uses such as
crop irrigation and ground water. Salinity can be measured directly using total dissolved solids (TDS)
method. It is measured as mg/L and although not identical to the true salinity but it is close enough for
most purposes. These three experimental tests comprised two equilibriums and one dynamic test [5]:
a). The Pore Water Test (PW), which assessed the beginning equilibrium composition of the pore
solution and the soluble species Maximum Mobile Fraction (MMF) for leaching.
b). The Acid Neutralization Capacity Test (ANC), which is inspired from the European pre-standard
“Influence of pH on leaching with initial acid/base addition”. The purpose of the test was to study the
influence of pH on the leach ability of inorganic species from ash material by addition of
predetermined amounts of acid or base to desired end pH values in steady state condition.
c). The Tank Leaching Test (TL), which allowed to determine the leaching behaviour of granular
ashes under dynamic conditions. It was used to determine the dominant release mechanisms of
inorganic species from regularly shaped specimens of monolithic wastes [6].
Methodology
Five samples namely B, C, L, M and ASP based on different kinds of coal fly ashes are considered.
The details of the coal fly ashes are shown in Table 1, Table 2, Table 3 and Table 4.
Table 1. Sources and physical characteristics of coal fly ash.

Melting point
Sample Type of fly ash Type of coal Specific gravity (kg/m3)
(0C)
B Coarse Bitumen
C Fine Lignite
L Coarse Bitumen 2050 - 2300 > 1400
M Coarse Bitumen
ASP Fine Bitumen
Table 2. Liquid/Solid ratio of coal fly ash used in PW. Applied to other as well.
Label Liquid/Solid ratio (L/S)
B1 1
B5 5
B10 10
B50 50
B100 100
B200 200
Table 3. Liquid/Solid ratio and pH of coal fly ash used in ANC. Applied to other as well.
Label Liquid/Solid ratio (L/S) pH
B_1 1
B_2 3
B_3 5
B_4 10 ml/g dried fly ash 7
B_5 10
B_6 12
B_7 14
Table 4. Liquid/Surface ratio, area and pH of coal fly ash used in TL.
Liquid/Surface ratio Area
Sample pH
(L/S) (cm2)
B
C
L 80 x Area 5.31 4
M
ASP
a). Pore Water Test (PW)

This test allows the evaluation of the species solubilization at steady state conditions between ash and
demineralised water in closed vessels for different liquid/solid ratios (L/S) at room temperature during
7 days of continuous shaking [5]. The samples were put into contact with demineralised water
(Conductivity: 1.15±0.07 µS/cm; TDS: 0 mg/L; pH~7) for L/S ratios: 1, 5, 10, 50, 100 and 200 ml/g.
The closed vessels were shook for 7 days and then were filtered using 0.45 µm membrane filter. The
plotting of total dissolved solids (mg/L) to L/S ratio is showed in Figure 1.
b). Acid Neutralization Capacity Test (ANC)
The samples were put into contact with demineralised water for L/S ratio of 10ml/g in all the parallel
running samples. The closed vessels were shook for 7 days at room temperature. After filtration (filter
porosity: 0.45 µm) the solution was analyzed. The plotting of total dissolved solids (mg/L) to different
pH values of samples is showed in Figure 2.
c). Tank Leaching Test (TL)
This test was modified from the compact granular protocol of Tank Leach Test NEN 7345 [7]. The
test was carried out on 4 granular cylindrical samples. The fly ash was embedded to polyethylene
vessel and the leach was demineralised water which acidified to pH of 4. The Liquid/Surface ratio of
80/cm2 was remained constantly throughout the test. The polyethylene vessels were used and closed
to prevent the intake of CO2 and water evaporation during the leaching test. The leach was renewed
after 0.25, 1, 2, 5, 8, 21, 36 and 64 days. Then, 8 eluates were analyzed after filtration (filter porosity:
0.45 µm). The plotting of pH against duration, and total dissolved solids (TDS) against days are
showed in Figure 3 and 4.
The total dissolved solids (TDS) were measured using calibrated HACH C0150 Conductivity
Meter with HACH model Conductivity Probe 50161 k=1.0 and pH values were recorded using
calibrated microprocessor pH meter (Hanna Instrument pH 211).
Figure 1 shows the total dissolved solids (TDS) of different coal fly ashes in equilibrium state. The
decrease of TDS on the L/S ratios supports the hypothesis that the leaching available quantities
deduced from the test for the soluble salts. It also can be noticed that, in general, TDS of sample C, L
and M are quite similar with the magnitude of C>L>M. On the other hand, TDS of sample B is
approximately three times higher than sample C whereas ASP coal fly ash is the highest among the
samples.
The result showed that the pH levels of the solutions were still in the range of 6.5 – 8.0. As
expected, without any binder which normally gives alkaline solution, the pH levels have not changed
too much from neutral. According to Raask [8], the alkaline solution obtained when fly ash is
suspended in water would be attributable to leaching of alkali metals such as Na, K and Ca from
aluminosilicate fractions of fly ash particles. Aluminosilicate fractions of fly ash particles could
increase the OH- concentration in solution. Thus, the dissolution of Na, K and Ca increase the
concentration of Ca2+ and OH- ions, and subsequently, this situation will affect pH level in water.
It also can be noticed that, in general, TDS of C, L and M samples are quite similar with the
magnitude of C>L>M. On the other hand, TDS of B sample is approximately three times higher than
C sample whereas ASP coal fly ash is the highest among the samples. B, C, L and M samples
presented low saline content whereas concentration of total dissolved solids for ASP sample was high
especially for L/S of 1, 5 and 10. The high salts solubility in ASP sample can be assumed that the
initial pore water corresponding to the solid phase of the salt properties.
4000 1200
3500 1000
3000
800
TDS (mg/L)
2500
TDS (mg/L)
2000 600
1500
400
1000
200
500
0 0
B1 B5 B10 B50 B100 B200 C1 C5 C10 C50 C100 C200
1200 1200
1000 1000
800 800
TDS (mg/L)
TDS (mg/L)
600 600
400 400
200 200
0 0
L1 L5 L10 L50 L100 L200 M1 M5 M10 M50 M100 M200
12000 8
10000
7.5
8000
TDS (mg/L)
pH
7
6000
4000
6.5
2000
6
0 0 50 100 150 200
ASP1 ASP5 ASP10 ASP50 ASP100 ASP200 L/S ratio

B C L M ASP
Figure 1. Total dissolved solids in PW of the materials.
Figure 2 shows the results of total dissolved solids as a function of ANC of the materials.
Regarding to the curves of the samples, we can observe a significant steep decrease curve from each
graph and expected to increase rapidly in pH 14. The TDS value is not showed in pH 14 is due to the
detection limit of the HACH C0150 Conductivity Meter. Apparently, the actual value would be higher
than value in pH 1. From the observation, TDS only increased rapidly in higher acid and base
concentrations (molarities). ANC test showed that the change of pH value in solution was
insignificant from pH 3 – pH 10. However, increase of salt concentration in solution was obvious in
high molarities. From here, we can assume that with high Ca content of coal fly ash, which is high
alkaline property, the high pH values may cause the constituents of coal fly ash probably reach the
environment such as groundwater.
8
7
7
6
6
5
TDS (g/L)
TDS (g/L)
4 4
3 3
2 2
1 1
0 0
B_1 B_2 B_3 B_4 B_5 B_6 B_7 C_1 C_2 C_3 C_4 C_5 C_6 C_7
0.75 100000
0.7
0.65
TDS (mg/L)
10000
TDS (g/L)
0.6
0.55
1000
0.5
0.45
0.4 100
L_1 L_2 L_3 L_4 L_5 L_6 L_7 M_1 M_2 M_3 M_4 M_5 M_6 M_7
10
9
8
7
TDS (g/L)
6
5
4
3
2
1
0
1
7
P_
P_
P_
P_
P_
P_
P_
AS
AS
AS
AS
AS
AS
AS
Figure 2. Total dissolved solids as a function of ANC of the materials.
From Figure 3, the comparison of pH for Tank leaching test applied to coal fly ashes at room
temperature within 64 days has been plotted. The TL test gives the view of physico-chemical of the
eluates at the end of each leaching sequence such as released salts and pH values. Initial pH of
demineralised water was acidified to pH 4 and what we can see from the graph is the change of the pH
value during the duration. Obviously, the pH level of L sample changed instantly from pH 4 to pH 7
within 0.25 day. In the other hand, C sample also showing the change after 1 day following with the
ASP sample and B sample increasing slowly. However, M sample was ranged within pH 4.22 – pH
5.48 during the duration.
8
7.5
6.5
pH
5.5
4.5
4
0.25 1 2 5 8 21 36 64
t (Days)
L M C B ASP
Figure 3. Comparison of pH for TL applied to coal fly ashes at room temperature within 64 days.
500
450
400
Cumulative release (g/m2)
350
300
250
200
150
100
50
0
0.25 1 2 5 8 21 36 64
t (Days)
B C L M ASP
Figure 4. Comparison of cumulative release for TL applied to coal fly ashes at room temperature in
64 days.
Figure 4 shows the comparisons of cumulative release (g/m2) or also called as flux for TL
applied to coal fly ashes at room temperature within 64 days. A first look at the experimental results
shows a low cumulative release for B, C, L and M samples. However, ASP sample shows a rapid
increase after 0.25 day and continue increasing until day-5. From the graphs, we can predict the fluxes
for all samples will be saturated after 64 days.
Compare to PW and ANC tests, the eluate concentrations are lower orders of magnitude in TL
test. The pH-dependent solubility, renewal time and flow of eluate are playing important roles in
controlling the release processes.
Conclusion
A study has been carried out on three laboratory leaching characterization test methods for the
assessment of the leaching behaviour of salinity (dissolved solids) from coal fly ashes. Basically, B,
C, L and M samples were showing quite similarly results for the three tests. In the other hand, ASP
sample was significantly higher than other. pH has been playing an important role in effecting the
release of the metals from coal fly ash.
Generally, the environmental impact assessment of coal fly ash leaching scenario requires
data like cumulative release, fluxes and concentrations of eluate. Thus, a model for environmental
prediction based on laboratory tests will be useful.
Acknowledgement
The authors wish to thank the financial support from Ministry of Science, Technology and Innovation
(MOSTI), Malaysia.
References
1. Roy, D. M., Luke, K., Diamond, S. (1985) “Characterization of Fly Ash and Its Reactions in
Concrete”, Materials Research Society Symposium Proceedings. 43. 3-11.
2. Golden, D. M. (1986) “Epri Coal Ash Utilization Research”, Energy. 11. 1377-1387.
3. Ugurlu, A. (2004) “Leaching Characteristics of Fly Ash”, Environ. Geo. 46. 890-895.
4. Furr, A .K., Parkinson, T. F., Helforn, C. L., Reid, J. T., Haschek, Gutenmann, W. M., Bache,
C. A., St. John Jr., L. E., Lisk, D. J. (1978) “Elemental Content of Tissues and Excreta of
Lambs, Goats and Kids Fed White Sweet Clover Growing on Fly Ash”, J. Agric. Food Chem.
26. 847-851.
5. Barna, L., Imyim, A., Barna, R. (2004) “Long-term Prediction of the Leaching Behavior of
Pollutants from Solidified Wastes”, Adv. Environ. Res. 8. 697-711.
6. Mehu, J., Moskowicz, P., Barna, R., Sanchez, F. (1997) “Waste Stabilized/Solidified with
Hydraulic Binders”, in: Van der Sloot, H. A., Heasman, L., Quevauviller, Ph. (Eds), Studies in
Environmental Science. Harmonization of Leaching/Extraction Tests. Elsevier Science,
Amsterdam. 171-186.
7. NNI (1994) “NEN 7345: Leaching Characteristics of Building and Solid Waste Materials –
Leaching Tests: Determination of the leaching Behavior of Inorganic Components from Shaped
Building Materials, Monolithic and Stabilized Waste Materials”, The Netherlands.
8. Raask, E. (1982) “Pulverised Fuel Ash Constituents and Surface Characteristics in Concrete
Applications”, in: Cabrera, J. G., Cuseus, A. R. (Eds.), Proceedings of the International
Symposium on the Use of PFA in Cncrete, Leeds. 1. 5-16.
PENENTUAN KEPEKATAN UNSUR-UNSUR SURIH DALAM TANIH SAWAH DI PERLIS
SECARA ANALISIS PENDARFLOUR SINAR-X (XRF)
Pui Vui Ming1,* & Sukiman Sarmani2

1
Program Sains Nuklear, Pusat Pengajian Fizik Gunaan, Fakulti Sains dan Teknologi,
Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor.
2
Pusat Pengajian Sains Kimia dan Teknologi Makanan, Fakulti Sains dan Teknologi,
Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor.
e-mail: pvuiming@yahoo.com
Abstrak. Kajian ini dilakukan untuk menentukan profil dan pola taburan unsur dalam tanah sawah
bagi mengetahui status kesuburan tanah dalam usaha meningkatkan hasil pengeluaran. Kandungan
unsur-unsur dalam tanih dan baja ditentukan dengan menggunakan teknik Analisis Pendarfluor Sinar-
X (XRF). XRF merupakan teknik instrumentasi tidak memusnah yang berkebolehan menentukan
unsur-unsur dalam sampel (dari Z=5 hingga Z=92) pada kepekatan dari julat beberapa mgkg-1
sehingga 100%. Sampel tanih dalam kajian ini telah dikumpul dari kawasan penanaman padi Perlis
dalam kawasan MADA. Penyediaan sampel tanih dengan melalui proses pengeringan, pengisahan,
penghalusan dan akhirnya penekanan dalam bentuk pellet sebelum dianalisis dan dibilang dengan
menggunakan pengesan spektroskopi sinar-X. Sebanyak 16 unsur-unsur surihan iaitu As, Ba, Ce, Co,
Cr, Ga, Hf, La, Nb, Pb, Sr, Th, U, V, Y dan Zr berjaya dikesan secara kualitatif dan kuantitatif melalui
teknik ini. Taburan kepekatan unsur untuk kebanyakan sampel adalah sekata. Unsur Ba dengan
perbezaan kepekatan unsur yang ketara antara 49-1617ppm. Unsur-unsur lain adalah Ce (22-575ppm),
Co (1.4-36ppm), Cr (0.3-23ppm), Pb (17-321ppm) dan Th (24-196ppm). Penentuan unsur logam berat
dan logam toksik juga dikenalpastikan. Faktor perbezaan kepekatan telah dibincangkan dalam kajian.
Abstract. Determination of trace elements in agriculture soil are to understand and increase soil
quality and crop yields as the aim of this study is to determine the profile and distribution patterns of
trace elements in the both soil and fertilizer samples. X-ray fluorescence spectrometry (XRF) were
used to determine trace elements in these study. An X-ray fluorescence method is an alternative multi-
element analytical tool for the routine non-destructive analysis over several orders of magnitude
concentration from levels often below mgkg-1, up to 100%. Samples were prepared after following
way of drying, refinement, pressing into pellets form, counting and analysis. The concentration of 16
(As, Ba, Ce, Co, Cr, Ga, Hf, La, Nb, Pb, Sr, Th, U, V, Y and Zr) elements were determined
quantitatively and qualitatively, which were collected from agriculture area located in the Perlis.
Generally, the concentration give a even distribution patterns almost in every samples. Concentration
of Ba show large differentiation range from 49ppm to 1617ppm. The concentrations of Ce (22-
575ppm), Co (1.4-36ppm), Cr (0.3-23ppm), Pb (17-321ppm) and Th (24-196ppm) in samples were
determined. The heavy metal elements were identified. Factors of elemental pattern were also
discussed .
Kata kunci: tanih sawah, baja, Analisis Pendarfluor Sinar-X (XRF)
Pengenalan
Sejak manusia menjadi petani, dia mula menaruh perhatian terhadap tanah dan hidupnya juga mula
bergantung kepadanya. Sebagai petani, manusia terutamanya mementingkan cara penggunaan tanah
yang lebih luas dan lebih produktif, iaitu satu keadaan yang tercapai hanya dengan menambahkan
pengetahuannya mengenai bahantara dan cara pembentukannya. Konsep tanah sebagai bahantara yang
berguna dan produktif untuk tanaman telah dihuraikan sebagai ‘tanah sebagai suatu sumber’ [1].
Padi merupakan makanan penting bagi negara-negara Asia terutamanya di negara-negara Asia
Tenggara. Laporan menunjukkan China merupakan pengeluar 1/3 daripada jumlah pengeluaran padi
sedunia. Sewajar dengan ilham oleh Perdana Menteri Malaysia YAB Dato’ Seri Abdullah Ahmad
Badawi untuk memajukan negara dalam bidang pertanian, serta menampung keperluan pada masa
hadapan. Perusahaan yang banyak telah dijalankan dalam bidang pertanian ini. Dalam Bajet Malaysia
2005 yang baru-baru ini juga telah memberikan peruntukan RM 1.5 bilion bagi projek pertanian
terutamanya kepada projek pekebun kecil. Peruntukan sebanyak RM 300 juta pula sebagai modal
permulaan bagi mengkomersialkan produk pertanian. Hala tuju kerajaan ini adalah untuk menjadikan
Malaysia sebagai pengeluar global yang kompetitif bagi produk pertanian berkualiti tinggi, selamat
dan memenuhi piawaian antarabangsa.
Tanah mengandungi pelbagai jenis kandungan seperti unsur logam berat, unsur logam toksik
dan unsur nadir bumi yang lain. Namun, seperti sesetengah unsur logam berat dan logam toksik
adalah berpotensi berbahaya kepada kesihatan manusia jika melebihi julat tahapnya walaupun pada
jumlah yang sedikit [2,3,4,5,6]. Kebanyakan toksik dalam badan biosistem manusia berasal daripada
pertumbuhan dan perkembangan industri dan pencemaran [7,8]. Perhatian telah diberikan kepada padi
tanaman, tanih dan baja yang digunakan. Kesan unsur-unsur surih tersebut diambil berat serta kajian
ini dijalankan untuk menentukan kandungan unsur yang hadir. Tanah dan baja merupakan faktor
penting dalam pengagihan unsur-unsur surih tersebut. Tanih merupakan kunci komponen bagi
ekosistem terestrial samada semula jadi atau dalam pertanian. Tanih menjadi keperluan asas untuk
pertumbuhan dan tumbesaran tumbuhan, mereput dan kitar semula bahan jisim yang mati. Tanih
mengandungi pelbagai jenis kandungan unsur logam berat dan unsur nadir bumi yang lain. Tanih
yang dikaji ini merupakan tanih sawah padi di kawasan negeri Perlis. Kandungan logam berat pada
tanih kemungkinan diserap oleh tumbuhan padi, hasil penuaian padi dijadikan beras, makanan
manusia seharian. Kandungan pada baja dan tanih juga dikenalpasti dan banyak dikaji kerana prihatin
kepada persekitaran memandangkan daripada penambahan penggunaan baja jenis organik.
XRF, sinar-x pendarfluor juga banyak digunakan dalam penentuan unsur-unsur surih di mana
ia merupakan kaedah analisis unsur-unsur dengan mengenal unsur dan menentu kepekatan pelbagai
unsur dalam sampel dengan mengukur keamatan tenaga sesuatu sinar-x yang dapat dilepaskan kepada
pembilang. Keamatan tenaga sesuatu sinar-x adalah berkadar dengan kepekatan unsur dalam sampel
berkenaan [9,10].
Metodologi
Sampel-sampel tanih dan baja dari kawasan tanih sawah padi Perlis telah dipilih dan dikumpul untuk
kajian penganalisis unsur. Pengumpulan sampel-sampel tanih dan baja tersebut telah dibantu dan
kerjasama dari pihak MADA (Lembaga Kemajuan Pertanian MUDA) dan Pertubuhan Peladang
kawasan (PPK). Tiga sampel dikumpul dari satu lokasi kampung yang sama. Kesemua 24 sampel
tanih (dari lapan kampung yang berada dalam tiga daerah berlainan masing-masing) yang dikumpul
telah dikering dan dihalus. Semua sampel yang dianalisis dalam kajian ini telah disenaraikan dalam
JADUAL mengikut label, lokasi dan nilai pH. Pelabelan sampel adalah mengikut lokasi daerah seperti
Rajah 1 yang disertakan.
Sampel yang telah dihaluskan tersebut akan ditimbang dengan berat sampel ±1g dan berat serbuk asid
borik ±6g, kemudian acuan campuran ditekan dalam bentuk pellet setelah siap. Bentuk pellet tersebut
dilabel dan dianalisis oleh pengesan pendarflour sinar-x. Keputusan yang dibilang adalah dalam nilai
ppm.
Hasil keputusan daripada Jadual 3 dan 4 menunjukkan kepekatan unsur Ba berada pada tahap antara
500–1000 ppm, tetapi sampel A1KB2, A1KG2, E1KS2 dan E1KSBT1 pula memberikan nilai
kepekatan unsur Ba yang agak tinggi berbanding dengan yang lain. Semua kepekatan Ba mengikut
lokasi kampung pula bertabur dengan sekata kecuali pada lokasi A1KG dan E1KS. Merujuk kepada
Rajah 1, kepekatan Ba dalam lokasi tersebut besar bezanya yang ketara dalam tapak sawah dengan
tapak sawah walaupun berasal dalam satu kampung yang sama. Keadaan yang sama berlaku pada
A1KG, A1KT dan E1KPKB untuk kepekatan unsur Ce. Taburan kepekatan Ce secara keseluruhannya
berada dalam lingkungan 22 – 575 ppm dengan purata kepekatan pada 245 ppm. Keseluruhannya
penaburan kepekatan Ce adalah tidak sekata pada semua lokasi kampung kecuali yang paling ketara
pada kawasan E1KPKB, A1KG dan A1KT. Kepekatan unsur naik turun dengan ketara pada kawasan
lokasi yang sama.
Rajah 1. Lokasi kawasan pengumpulan sampel
Jadual 1. Pelabelan sampel bagi tanih

SAMPEL LOKASI (DAERAH) LOKASI (KAMPUNG) pH
A1KT1 Arau Kampung Teluk 4.52
A1KT2 5.74
A1KT3 5.97
A1KB1 Kampung Banat 6.57
A1KB2 5.72
A1KB3 5.64
A1KG1 Kampung Gobek 5.44
A1KG2 5.22
A1KG3 5.31
D1KTTT1 Tambun Tulang Kampung Tambun Tulang Timur 5.02
D1KTTT2 4.53
D1KTTT3 3.85
D1KLJ1 Kampung Lahar Jambu 5.00
D1KLJ2 4.90
D1KLJ3 5.54
E1KS1 Simpang Empat Kampung Kuala Sanglang 6.44
E1KS2 6.26
E1KS3 6.57
E1KPKB1 Kampung Pauh Kepala Batas 5.81
E1KPKB2 6.24
E1KPKB3 6.84
E1KSBT1 Kampung Sungai Baru Timur 6.00
E1KSBT2 5.49
E1KSBT3 5.87
Jadual 2. Pelabelan sampel bagi baja
Sampel Jenis baja
bc Baja Campuran
moc Baja Mineral Organic Compost
u UREA 46%N
bpt Baja Padi Tambahan lembing Sakti
Merujuk kepada Rajah 2 di bawah, didapati semua kepekatan adalah hampir sekata kecuali
pada D1KTTT3 dan E1KPKB3 di mana kepekatan unsur Th dan Pb masing-masing mempunyai nilai
yang lebih tinggi berbanding dengan yang lain. Perkara yang sama juga berlaku pada unsur Nb di
E1KS3 seperti pada Rajah 3(c). Lain-lain kepekatan unsur yang mempunyai pemerhatian penaburan
kepekatan mengikut lokasi yang tidak sekata adalah di lokasi kawasan A1KB untuk unsur La, Th dan
Pb manakala lokasi A1KT untuk unsur Ce, Co dan La, lokasi D1KLJ untuk unsur Pb dan lokasi
D1KTTT untuk unsur Th serta kawasan E1KPKB untuk unsur Th dan unsur Co. Rujukan untuk
pemerhatian yang tidak normal ini boleh didapati pada Jadual 3. kepekatan unsur lain memberikan
keadaan yang hampir sekata untuk semua sampel tanih dan baja.
1600
1400
1200
kepekatan (ppm)
1000
800
600
400
200
0
bpt
A1KB1
A1KB2
A1KB3
A1KG1
A1KG2
A1KG3
A1KT1
A1KT2
A1KT3
D1KLJ1
D1KLJ2
D1KLJ3
D1KTTT1
D1KTTT2
D1KTTT3
E1KPKB1
E1KPKB2
E1KPKB3
E1KS1
E1KS2
E1KS3
E1KSBT1
E1KSBT2
E1KSBT3
u
bc
moc
Ba
Ce
Rajah 1. Kepekatan unsur Ba dan Ce untuk semua sampel tanih dan baja
350
300
La
250
kepekatan (ppm)
Th
200 V
150 Y
Pb
100
Co
50
0
A1KB1
A1KB3
A1KG2
A1KT1
A1KT3
D1KLJ2
D1KTTT1
D1KTTT3
E1KPKB2
E1KS1
E1KS3
E1KSBT2
Rajah 2. Kepekatan unsur La, Th, V, Y, Pb dan Co untuk semua sampel tanih
40 35
35 U 30
30
ke p e ka ta n ( p p m )
Cr
25 25
k e p e k a ta n ( p p m )
U
Nb
20 20 Cr
Zr
15 Nb
Sr 15
10 Zr
5 As 10 Sr
0 Ga 5 As
Hf Ga
0
D1KLJ1 D1KLJ2 D1KLJ3 D1KTTT1 D1KTTT2 D1KTTT3 Hf
(a) (b)
70
60
U
ke p e ka ta n ( p p m)
50 Cr
40 Nb
30 Zr
20 Sr
As
10
Ga
0
Hf
E1 K PK B 1
E1 K PK B 2
E1 K PK B 3
E1 K S 1
E1 K S 2
E1 K S 3
E1 K S B T1
E1 K S B T2
E1 K S B T3
(c)
Rajah 3 (a)-(c). kepekatan unsur U, Cr, Nb, Zr, Sr, As, Ga dan Hf untuk semua sampel tanih
U
30 1500
Cr
25 V
kepekatan (ppm)
Nb
k e p e ka tan (p p m)
1000
20 Zr
Y
15
Sr 500
10 Pb
As
5 0
Ga
bc moc u bpt
0 Hf
bc moc u bpt Co Ba Ce La Th
(a) (b)
Rajah 4 (a) (b). Kepekatan unsur untuk sampel baja
Kepekatan unsur pada baja melihat juga terdapat ketidakserataan seperti tunjukkan pada
Rajah 4. Kepekatan unsur-unsur As, Ba, Ce, Th, V dan Zr telah menunjukkan penaburan unsur yang
tidak teratur. Graf menunjukkan kepekatan unsur-unsur tersebut turun-naik dengan mendadak dan
tidak seimbangan pembahagi pada baja. Baja UREA 46%N menunjukkan mempunyai kepekatan
unsur V dan Zr yang rendah berbanding dengan sampel baja lain, tetapi memberikan kepekatan yang
sebalik untuk unsur As, Ce dan Th. Baja Padi Tambahan lembing Sakti pula memberikan kepekatan
unsur Ba yang jauh lebih tinggi dalam antara sampel baja tersebut.
Pemerhatian penaburan yang tidak sekata serta sesetengah kepekatan unsur yang tinggi
kemungkinan disebabkan oleh perbezaan nilai pH yang telah mempengaruhi kandungan nutrient yang
terdapat dalam tanih. Mengikut Bear [11] analisis tanih untuk jumlah kuantiti bagi sesetengah unsur
yang jumpa dalam tanih memberikan variasi yang banyak. Ia bergantung bukan sahaja kepada
keadaan batuan semula jadi yang membentuk tanih tersebut, tetapi juga bergantung kepada umur tanih
tersebut dan keupayaan ia melarutkan air. Walaupun kawasan tersebut mempunyai jumlah hujan,
suhu, batuan dan mineral asal yang sama, tetapi tiada pun dua tanih yang mempunyai kandungan
kimia yang sama.
Faktor lain dalam pengaruhan sifat kandungan adalah kemungkinan faktor geologi kawasan
tanih tersebut. Pemilihan kawasan kajian adalah kawasan A1 di sebelah dalaman yang mendekati
dengan kawasan penggunungan dan sumber penghiliran air serta kawasan E1 pula merupakan
kawasan tepi dan berhampiran dengan laut, manakala kawasan D1 merupakan kawasan pertengahan
yang terletak di antara kawasan pedalaman dan tepi laut. Perbandingan menunjukkan taburan
kepekatan unsur pada kawasan A1 dan E1 sememangnya banyak yang tidak sekata berbanding
dengan kawasan D1 yang taburan kepekatan unsur lebih sekata. Kebarangkalian perbezaan kepekatan
unsur adalah tinggi. Perbezaan taburan kepekatan unsur yang ketara dapat dilihat daripada graf
taburan kepekatan unsur Ba dan Ce keseluruhan.
Penyerapan unsur-unsur oleh tumbuhan banyak dipengaruhi oleh faktor geokimia pada tanih
tersebut, terutama pada komponen dan kandungan mineral yang terdapat dalam tanih [12]. Sifat kimia
tanih dan kesan penggunaan baja juga boleh memberi kesan kepada kandungan kepekatan unsur-
unsur. Mengikut Coker [13] dan Kalpagé [14] pembajaan tanih biasa digunakan untuk mengekalkan
dan meningkatkan kualiti tanih tersebut serta meningkatkan penghasilan dan kualiti tanaman, tetapi
jika tidak digunakan dengan wajar sekali akan mengakibatkan kesan negatif untuk haiwan dan
keselamatan manusia.
Kesimpulan
Dalam penyelidikan ini, penentuan unsur-unsur surih dalam tanih dan baja secara analisis pendarflour
sinar-X (XRF) telah dapat menentukan dengan kesemuanya 16 unsur. Unsur-unsur tersebut telah
dikumpulkan kepada unsur NORM (U, TH) dan unsur REE (Ce dan La) serta unsur berat bertoksik
(As, Ba, Co, Cr, Ga, Hf, Nb, Pb, Sr, V, Y dan Zr). Data telah dianalisis dengan kepekatan yang
terkandung dan dapat memperbaiki dan membaikpulihkan sifat tanih tersebut serta data digunakan
untuk kajian selanjutnya. Nilai data ini akan digunakan membuat perbandingan dengan kaedah
Analisis Pengaktifan neutron Instrumentasi (APNI).
Penghargaan
Penulis ingin memberi penghargaan atas bantuan dan kerjasama daripada pihak MADA (Lembaga
Kemajuan Pertanian MUDA) dan Pertubuhan Peladang kawasan (PPK) yang mengumpul sampel-
sampel.
Rujukan
1. Cruickshank, J.G. (1983) “Geografi Tanah-Tanih”, Ismail B. Ahmad (pnjh). Kuala Lumpur:
Dewan Bahasa dan Pustaka.
2. Abdel-Haleem, A.S., Sroor, A., El-Bahi, S.M., Zohny, E. (2001) “Heavy Metal and Rare
Earth Elements in Phosphate Fertilizer Components Using Instrumental Neutron Activation
Analysis”, J. Applied Radiation and Isotopes 55. 569-573.
3. Alloway, B.J. (1990) “Heavy Metal in Soils”, Glasgow & London: Blackie & Sdn.Ltd.
4. Smith, D.G. (1986) “Heavy Metals in the New Zealand Aquatic Environment. A Review Water
& Soil Directorate”, New Zealand: Wonistry of Works & Development.
5. Ahmad, S., Chaudhary, M.S., Mannan, A., Qureshi, I.H. (1982) “Determination of Toxic
Elements in Tea Leaves by Instrumental Neutron Activation Analysis”, J. Radioanalytical
and Nuclear Chemistry 78. 375-383.
6. Frederick, W.O. (1978) “Toxicity of Heavy Metals in Environment”, New York: Marcel
Dekler Inc.
7. Nada, A., M. Abdel-Wahab, A. Sroor, A.S. Abdel-Haleem, M.F. Abdel-Sabour. (1999)
“Heavy Metals and Rare Earth Elements Source-Sink in Some Egyptian Cigarettes as
Determined by Neutron Activation Analysis”, J. Applied Radiation and Isotopes 51. 131-136.
8. Pollock, Eugene N. (1974) “Trace Impurities in Coal by Wet Chemical Methodsz”, dlm:
Babu, Suresh P. (pnyt). Trace Element in Fuel. (Advances in Chemistry Series 141)..
Washington, D.C.: American Chemical Society. 23-34.
9. Hamzah Mohamad. (2002) “Spektrometri Sinar-X”, nota kuliah. Bangi: Jabatan Geologi.
Universiti Kebangsaan Malaysia.
10. Wilkins, Carolyn. (1983) “X-Ray Fluorescence Analysis”, dlm: Smith, Keith A. (pnyt). Soil
Analysis- Instrumental Techniques and Related Procedures. New York: Marcel Dekker, Inc.
195-225.
11. Bear, Firman E. (1977) “Soils in Relation to Crop Growth”, New York: Robert E. Krieger
Publishing Company.
12. Wang, Y. Q., Sun, J. X., Chen, H. M., Guo, F. Q. (1997) “Determination of the Contents and
Distribution Characteristics of REE in Natural Plants by NAA”, J.Radioanalytical and
Nuclear Chemistry 1. 219. 99-103.
13. Coker, E.G. (1971) “Horticultural Science & Soils- Volume 2: Soils & Fertilisers”, London
Macdonald Technical & Scientific.
14. Kalpagé, F.S.C.P. (1979) “Soils and Fertilizers for Plantations in Malaysia”, Kuala Lumpur:
Kuala Lumpur Incorporated Society of Planters.
Jadual 3. Kepekatan Unsur-unsur dalam sampel tanih (dalam ppm)
Sampel Kepekatan Unsur (ppm)
Ba Ce La U Th Cr V Nb Zr Y Sr Pb As Ga Hf Co
A1KB1 885.478 355.496 69.446 21.640 33.985 4.085 99.427 20.360 33.662 58.006 99.698 1.132 20.273 10.090
A1KB2 1348.481 308.866 92.729 18.126 66.109 3.927 85.919 26.198 12.474 38.376 60.346 1.465 20.247 10.640 36.638
A1KB3 555.336 128.160 27.308 49.155 6.445 96.137 22.838 24.588 38.899 7.811 37.248 3.625 20.802 8.438
A1KG1 49.084 575.320 55.611 23.907 131.252 12.738 102.371 15.740 20.930 59.838 54.785 3.417 20.857 7.061 15.104
A1KG2 1124.447 95.704 105.552 29.859 80.387 0.308 120.381 20.528 28.674 44.788 33.826 4.747 20.795 8.576 19.998
A1KG3 825.735 162.916 19.032 163.377 0.938 104.449 17.336 15.182 59.707 23.988 8.029 20.700 6.924
A1KT1 631.572 22.430 91.379 29.632 94.665 1.252 101.505 20.150 19.125 46.882 105.259 3.708 20.754 6.098 9.231
A1KT2 512.087 81.931 18.182 102.696 4.085 94.405 20.780 21.025 53.949 75.317 6.617 20.674 8.300 1.401
A1KT3 765.992 375.481 50.886 26.004 97.342 9.592 104.276 13.808 15.467 28.953 73.178 3.999 20.864 7.612 19.020
D1KLJ1 377.667 95.704 65.397 21.810 100.019 6.917 172.505 20.360 25.016 50.023 41.098 2.296 20.692 10.503 2.380
D1KLJ2 556.893 42.414 68.096 20.903 67.894 99.427 26.871 19.315 46.097 17.999 7.489 20.915 8.713
D1KLJ3 377.665 82.382 76.532 25.551 114.297 3.613 93.019 23.846 34.184 59.576 97.560 20.003 6.678 16.083
D1KTTT1 138.621 21.243 133.929 23.752 90.941 24.308 30.384 34.319 62.057 8.195 20.689 5.685 12.168
D1KTTT2 467.279 135.673 86.993 23.454 81.280 13.840 91.634 24.812 15.895 32.356 80.022 7.904 20.707 9.126 6.295
D1KTTT3 571.829 322.189 109.264 28.045 196.394 5.815 93.366 20.024 13.329 46.228 3.127 20.627 9.677
E1KPKB1 328.851 105.552 26.741 62.540 6.287 98.561 26.619 22.925 49.892 48.797 6.035 20.722 8.576
E1KPKB2 945.219 95.704 107.576 22.773 24.169 1.410 95.617 26.955 23.353 39.946 69.756 3.459 20.962 6.235 15.104
E1KPKB3 751.057 548.674 82.269 30.482 86.634 2.669 109.644 18.428 28.436 34.973 321.259 2.462 20.809 7.474 23.914
E1KS1 302.987 183.163 26.174 109.835 11.637 91.980 26.030 15.990 34.450 1.866 54.785 1.340 20.831 5.685
E1KS2 1109.511 442.093 97.791 25.494 128.575 3.141 93.712 20.360 27.723 38.637 29.976 4.622 20.565 7.612
E1KS3 676.377 117.362 203.676 62.656 27.391 17.830 2.695 53.502 19.864 7.750 13.147
E1KSBT1 1184.190 62.398 145.370 17.049 79.495 97.695 23.216 14.137 51.724 1.720 22.704 3.750 20.922 4.171 6.295
E1KSBT2 188.963 88.680 25.211 100.019 3.770 95.617 27.165 14.700 46.751 73.178 5.079 21.126 8.438 16.083
E1KSBT3 721.184 49.075 112.301 20.280 105.373 6.130 97.522 28.887 24.018 56.304 4.041 20.718 5.823 30.765
Jadual 4. Kepekatan unsur-unsur dalam sampel baja (dalam ppm)

Sampel Kepekatan Unsur (ppm)
Ba Ce La U Th Cr V Nb Zr Y Sr Pb As Ga Hf Co
bc 706.250 55.736 158.530 26.846 13.053 23.578 12.338 24.303 7.229 11.807 24.415 3.584 19.989 6.373
moc 437.408 122.350 101.503 2.765 164.269 24.097 0.956 23.020 17.044 10.065 20.222 5.410
u 736.121 455.416 91.379 580.109 4.714 3.143 15.752 14.884 21.010 7.337
bpt 1288.738 215.608 97.116 10.474 99.127 17.931 14.226 7.508 16.655 22.541 7.988 19.999 5.547 20.977
PENENTUAN KUALITI AIR TASIK KEJURUTERAAN UKM
KAMPUS BANGI: KE ARAH SISTEM PENGURUSAN SUMBER AIR BERSEPADU
Mazlin Bin Mokhtar1, Othman A. Karim2 & Irene Lee Pei Ngo3
1
Institut Alam Sekitar dan Pembangunan (LESTARI)
2
Jabatan Kejuruteraan Awam & Struktur, Fakulti Kejuruteraan
3
Pusat Pengajian Sains Sekitaran dan Alam Sekitar, Fakulti Sains & Teknologi
43600 UKM BANGI, Selangor Darul Ehsan, Malaysia
Kata kunci: Kualiti air, tasik, UKM, pengurusan, bersepadu

Key words: Water quality, lake, UKM, management, integrated
Abstrak. Sistem pengurusan sumber air bersepadu (IWRM) merupakan suatu proses yang mempromosikan
pembangunan penyelarasan dan pengurusan air, tanah dan sumber lain yang berkaitan untuk memaksimumkan
manfaat ekonomi dan sosial secara seimbang tanpa menjejaskan kelestarian ekosistem. Kajian mengenai kualiti
air Tasik Kejuruteraan, UKM Kampus Bangi dijalankan untuk menentukan kualiti air tasik tersebut,
membandingkannya dengan nilai Interim Kualiti Air Kebangsaan (INWQS) (JAS, 2001), dan menganggarkan
nilai Indeks Kualiti Air berdasarkan enam parameter terpilih. Kajian ini juga bertujuan mengenalpasti sumber
dan tahap pencemaran air tasik berkenaan. Kesan hari hujan dan hari kering ke atas kualiti air tersebut juga telah
dinilai. Parameter yang diukur adalah pH, suhu, oksigen terlarut (DO), konduktiviti, kekeruhan, jumlah pepejal
terampai (TSS), permintaan oksigen biokimia (BOD), permintaan oksigen kimia (COD), nitrogen ammonia
(NH3-N), plumbum dan kadmium. Parameter suhu, pH, konduktiviti, oksigen terlarut dan kekeruhan diukur
secara in situ dengan menggunakan meter yang telah dikalibrasi. Kandungan logam berat ditentukan dengan
menggunakan spektrofotometer serapan atom (AAS). Kaedah pensampelan dan analisis dilakukan mengikut
garis panduan yang dicadangkan oleh American Public Health Association (APHA, 1998). Pada keadaan biasa,
kadar aliran masuk bagi air tasik dianggarkan sebanyak 0.057 ± 0.024 m3/s manakala, kadar aliran keluar adalah
0.052 ± 0.018 m3/s. Secara teori, masa mastautin air tasik dengan kedalaman purata tasik (1.5 m) dan keluasan
(18,000 m2) adalah 62.5 ± 37.6 hari. Jumlah anggaran bagi bahan yang diukur yang berada dalam tasik adalah
DO (200.88 ± 28.25 kg), TSS (163.78 ± 18.19 kg), NH3-N (12.65 ± 13.90 kg), BOD (41.90 ± 23.95 kg), COD
(1605 ± 75 kg), Pb (9.50 ± 0.90 kg) dan Cd (2.81 ± 0.24 kg). Anggaran jumlah berat bahan yang mengalir ke
Sungai Langat setiap hari dari tasik ini adalah TSS (27.81 ± 9.29 kg), NH3-N (2.12 ± 0.71 kg), BOD (7.01 ±
2.34 kg), COD (268.9 ± 89.78 kg), Pb (1.59 ± 0.53 kg) dan Cd (0.47 ± 0.15 kg). Hasil ujian ANOVA dua hala
menunjukkan perbezaan yang sangat bererti (p<0.001) antara kepekatan parameter semasa hari hujan dan kering
bagi Pb, p < 0.05 antara hari hujan dan kering bagi TSS. Perbezaan bererti p < 0.05 dicerap bagi data antara
stesen bagi parameter suhu, DO dan BOD. Semua stesen pensampelan kajian ini didapati mempunyai kualiti air
Kelas II kecuali Stesen S2 yang berada pada Kelas III (sedikit tercemar). Secara keseluruhannya, kualiti air
Tasik Kejuruteraan UKM boleh dikategorikan sebagai Kelas II-III. Beberapa langkah yang perlu diambil ke
arah sistem pengurusan sumber air bersepadu dalam kampus UKM Bangi bagi mewujudkan kualiti air yang
baik, sihat dan harmoni yang selaras dengan tema UKM ‘Universiti dalam Taman’ telah juga dicadangkan.
Abstract. Integrated Water Resources Management (IWRM) is a process, which promotes the coordinated
development and management of water, land and related resources, in order to maximise the resultant economic
and social welfare in an equitable manner without compromising the sustainability of vital ecosystems. A study
on the water quality of the “Engineering Lake”, UKM Bangi Campus was carried out to determine the its water
quality, and compare it with the Interim National Water Quality Standard (INWQS) (DOE, 2001), followed by
estimation of its Water Quality Index (WQI) based on six selected parameters. The purpose of this study was to
identify the possible causes of the water pollution and level of this pollution at the lake. The comparisons of
concentration values measured during dry days with those on rainy were performed using suitable statistical
methods. Water quality parameters that were measured are pH, temperature, dissolved oxygen (DO),
conductivity, turbidity, total suspended solids (TSS), biochemical oxygen demand (BOD), chemical oxygen
demand (COD), ammoniacal-nitrogen, lead and cadmium. Temperature, pH, conductivity, dissolved oxygen and
turbidity were measured in situ by using calibrated meters, whilst metal concentrations were determined by
using Atomic Absorption Spectrophotometry (AAS). Methods of sampling and water analyses were performed
according to recommendations that were outlined by the American Public Health Association (APHA, 1998).
On normal days, the inflow and outfow of the lake were estimated to be 0.057 ± 0.024 m3/s inflow and 0.052 ±
0.018 m3/s outflow. The theoritical retention time of the lake water with a mean depth of 1.5 m and area of
18,000 m2 was 62.5 ± 37.6 days. On normal days, the estimated total amounts of materials that were present in
the lake were DO (200.88 ± 28.25 kg), TSS (163.78 ± 18.19 kg), NH3-N (12.65 ± 13.90 kg), BOD (41.90 ±
23.95 kg), COD (1605.58 ± 74.68 kg), Pb (9.50 ± 0.90 kg) and Cd (2.81 ± 0.24 kg). The estimated total amount
of polluted materials which flowed into the Langat River daily were TSS (27.81 ± 9.29 kg), NH3-N (2.12 ± 0.71
kg), BOD (7.01 ± 2.34 kg), COD (268.9 ± 89.78 kg), Pb (1.59 ± 0.53 kg) and Cd (0.47 ± 0.15 kg). The results
from two way ANOVA showed that there were significant differences (p<0.001) between rainy days and dry
days for Pb. There were also significant differences (p<0.05) between rainy days and dry days in term of TSS.
There were significant differences (p<0.05) between stations in terms of temperature, DO and BOD. All
sampling stations were categorized as having Class II water, that means a reasonably good water quality except
S2 which was categorized as a Class III, which means it was slightly polluted. On the overall, the Engineering
Lake water of UKM Bangi Campus was categorized as Class II-III. Some measures of IWRM were suggested
for improvement of the lake’s water quality and its environment, in achieving a healthy lake ecosystem which is
in line with UKM’s theme of being a ‘University in A Garden’.
Pengenalan
Konsep pembangunan lestari telah dikemukakan oleh World Commission on Environment and Development
(Brundtland Commission) dalam laporan Our Common Future pada tahun 1987 (Bruce, 1997). Pembangunan
lestari menekankan pengurusan sumber alam yang terdiri daripada biodiversiti, air, tanah dan sumber lain. Di
Malaysia, konsep pembangunan lestari telah dimaktubkan dalam beberapa dasar dan dokumen rasmi kerajaan,
seperti Rancangan Jangka Panjang Malaysia Ketiga 2001 – 2010 (“Third Outline Perspective Plan 2001 –
2010), yang merangkumi Rancangan Malaysia ke-8 (2001-2005) dan Rancangan Malaysia ke-9 (2006-2010).
Malaysia menyertai Persidangan Pertubuhan Bangsa-bangsa Bersatu mengenai alam sekitar dan pembangunan
(UNCED) di Rio de Janeiro, Brazil pada Jun 1992, bila mana konsep pengurusan air secara bersepadu telah
dimajukan untuk memastikan bekalan air tawar dunia adalah mencukupi untuk semua penduduk di semua
negara bagi tujuan hidupan seharian dan pembangunan lestari. Dalam Millenium Summit of the United Nations
di New York (2000), the International Freshwater Conference di Bonn (2001) dan World Summit on
Sustainable Development di Johannesburg (2002), peserta dari pelbagai kerajaan, NGO dan organisasi telah
digalakkan untuk menyatakan iltizam dalam pengurusan air termasuk untuk menghebahkan dan melaksanakan
agenda pendekatan Pengurusan Sumber Air Bersepadu (“Integrated Water Resources Management”, IWRM)
(World Water Forum 2003). Global Water Partnership (GWP) atau Pakatan Air Sedunia mendefinisikan IWRM
sebagai suatu proses yang mempromosikan pembangunan dan pengurusan sumber air, tanah dan sumber lain
yang berkaitan untuk memaksimumkan manfaat ekonomi dan sosial secara seimbang tanpa menjejaskan
ekosistem lestari (GWP, 2000b).
IWRM boleh dijalankan dalam beberapa skala atau peringkat iaitu kebangsaan, negara, negeri,
tempatan atau lembangan sungai. Pengurusan Bersepadu Lembangan Sungai (Integrated River Basin
Managemen/ IRBM) merupakan cara pengurusan bersepadu dalam mengatasi pelbagai masalah dan
isu sumber air seperti pencemaran air, kekurangan air dan banjir dalam konteks isu guna tanah dan
pembangunan di sesebuah lembangan sungai. Lembangan Langat di Semenanjung Malaysia adalah
terdiri daripada empat buah daerah, iaitu Hulu Langat, Kuala Langat, dan Sepang di Selangor; serta
empat mukim di Negeri Sembilan. Lembangan ini mempunyai keluasan kira-kira 2 940 km2 dan
merupakan suatu pusat perindustrian, pembangunan dan pentadbiran di Malaysia yang merangkumi
projek nasional seperti Putrajaya, KLIA dan Koridor Raya Multimedia (MSC) (Mazlin et al., 2004).
Sungai Langat ini juga mengalir bersebelahan dengan kawasan UKM Kampus Bangi dan sungai ini
juga menerima air luahan dari beberapa saluran dan jasad air tertentu dalam kampus tersebut. Satu
daripada jasad air tersebut adalah “Tasik Kejuruteraan” UKM yang dinamakan sedemikian kerana
kedudukannya yang hampir dengan fakulti tersebut. Tasik ini telah dipilih sebagai kawasan kajian ini
memandangkan kualiti airnya yang kelihatan tidak berapa menarik dan juga kerana ia
menyumbangkan air luahan kepada Sungai Langat.
Permasalahan Kajian
Pencemaran air telah mula kelihatan berlaku di Tasik Kejuruteraan dan kualiti airnya diandaikan tidak
baik berdasarkan pandangan estetik. Antara penyebab pencemaran yang disyaki adalah projek
pembinaan Kolej Kediaman Pelajar Kausar dan kompleks Fakulti Teknologi dan Sains Maklumat
(FTSM) yang melibatkan kerja pembersihan tanah; serta aktiviti perniagaan di kantin dalam bangunan
Fakulti Kejuruteraan serta blok-blok makmal di sepanjang saluran menuju ke tasik. Kajian kualiti air
perlu dijalankan untuk mengenalpasti sumber dan tahap pencemaran air tasik dengan menggunakan
parameter fizikal dan kimia. Seterusnya, Indeks Kualiti Air (IKA) bagi kualiti air tasik ini telah juga
ditentukan.
Objektif Kajian
Objektif kajian ini adalah: (1) mengkaji sistem saliran ke Tasik Kejuruteraan, kampus UKM Bangi;
(2) menentukan kualiti air pada hari kering dan hari hujan dan kemudian membandingkannya dengan
piawaian Interim Kualiti Air Kebangsaan (INWQS) dan piawaian lain yang sesuai untuk menentukan
status pencemaran; (3) memberi cadangan tentang langkah-langkah yang perlu diambil untuk
mengurangkan pencemaran air dan mengindahkan tasik bagi menghasilkan suasana sihat dan harmoni
selaras dengan tema UKM ‘Universiti dalam Taman’ dan juga sebagai inisiatif ke arah pengurusan
sumber air bersepadu (IWRM) dalam kampus.
Lokasi Kajian
Tasik Kejuruteraan ini sebenarnya telah diubahsuai dari sebuah kawasan paya ke tasik buatan manusia
pada tahun 1989. Airnya mengalir dari bukit yang berhutan di hulu ke dalam tasik dan air tasik
kemudiannya mengalir ke Sungai Langat. Kawasan sekelilingnya telah dikembangkan menjadi taman
rekreasi dengan lorong jalan kaki, pondok dan menawarkan aktiviti mengayak untuk pelajar dan
warga kampus. Anggaran keluasan Tasik Kejuruteraan ini adalah kira-kira 1.8 hektar. Kedalamannya
secara purata adalah 1.5 m. Jabatan Pengurusan Pembangunan (JPP) UKM bertanggungjawab ke atas
pengurusan tasik ini dan ia juga bertanggungjawab memantau dan menjaga kebersihan tasik. Kini,
pencemaran air telah mula kelihatan dan kualiti air disyaki menjadi semakin teruk. Nilai estetik di
tasik kelihatan menurun saban hari dan pemandangan di tasik tidak menyenangkan mata. Punca
pencemaran boleh dibahagi kepada pencemar tentu dan pencemar tidak tentu (Krenkel & Novotny
1980). Di Tasik Kejuruteraan, pencemar tentu yang dikenalpasti termasuklah hakisan permukaan
tanah akibat projek pembinaan Kolej Kausar dan longkang kompleks Fakulti Teknologi dan Sains
Maklumat, kantin dan bangunan Fakulti Kejuruteraan. Menurut JPP, semua air buangan samada air
dari makmal, tandas atau kantin disalirkan oleh paip pembentungan khas ke tangki kumbahan yang
kemudiannya disalirkan ke kolam pengoksidaan kumbahan UKM untuk proses perawatan sebelum ia
dialirkan ke Sungai Langat. Air larian hujan dan hakisan kelodak di sekeliling tasik merupakan
pencemar tidak tentu. Kajian kualti air ke atas Tasik Kejuruteraan ini dilakukan di lima lokasi yang
telah ditentukan sebagai stesen pensampelan (Jadual 1).
Bahan & Kaedah
Parameter-parameter kajian
Parameter-parameter yang diukur adalah pH, suhu, oksigen terlarut (DO), konduktiviti, kekeruhan,
jumlah pepejal terampai (TSS), permintaan oksigen biokimia (BOD), permintaan oksigen kimia
(COD), nitrogen ammonia (NH3-N), plumbum dan kadmium. Parameter suhu, pH, konduktiviti,
oksigen terlarut dan kekeruhan diukur secara in situ dengan menggunakan meter yang telah
dikalibrasikan. Kandungan logam ditentukan dengan menggunakan spektrofotometer serapan atom
(AAS). Kaedah pengawetan dan penyimpanan sampel sebelum analisis adalah seperti yang
disarankan oleh APHA (1998).
Secara ringkas, pengukuran suhu, pH, DO, konduktiviti diukur dengan meter oksigen terlarut model
YSI 556. Meter turbiditi 2020 berjenama Lamotte digunakan untuk mengukur kekeruhan. TSS
ditentukan dengan kaedah APHA 2540D melalui penurasan dan pengeringan pada 103-105 0C. BOD
adalah sepertimana saranan Kaedah APHA 5210B kaedah elektrod. COD ditentukan melaui kaedah
refluks terbuka APHA 5220B. Nitrogen ammonia diukur dengan kaedah HACH Quick Program 380
(Kaedah Nessler) menggunakan spektrometer HACH DR2000 pada jarak gelombang 425 nm. Halaju
aliran air diukur dengan meter aliran elektromagnet model Valeport 801 Ver 3.10.
Jadual 1: Stesen pensampelan dalam kajian ini
Stesen Koordinat Perihalan ringkas
1 U 02o55’12.1” Aliran air sungai dari bukit
T101 o 46’20.3” selepas melalui Fakulti Teknologi
dan Sains Maklumat.
2 U 02o55’12.4” Perangkap lumpur yang
T101 o 46’23.6” berdekatan dengan makmal dan
kantin Fakulti Kejuruteraan.
3 U 02o55’24.0” Kawasan yang menerima aliran
T101 o 46’24.5” air dari longkang
bangunan Fakulti Kejuruteraan.
4 U 02o55’28.4” Di tengah-tengah tasik.
T101 o 46’20.7”
5 U 02o55’28.2” Kawasan aliran keluar air tasik ke
T101 o 46’17.4” Sungai Langat.
Pengiraan Data Hidrologi

Hasil ukuran keluasan melintang bagi aliran masuk ialah 0.151 m2 dan aliran keluar ialah 0.092 m2.
Kadar aliran isipadu, Q (m3/s)= V × A……..Persamaan(1)
di mana V= halaju aliran (m/s),
A = keluasan melintang (m2)
Kadar aliran isipadu masuk, Qin = Vin × A

= 0.376 m/s × 0.151 m2
= 0.057 ± 0.024 m3/s
Kadar aliran isipadu keluar, Qout = Vout × A
= 0.567m/s×0.092 m2
= 0.052 ± 0.018 m3/s
Luahan aliran masuk & keluar, Q = Qin– Qout ……..Persamaan (2)
Q = 0.057 m3/s – 0.052 m3/s

= 5.0 × 10-3 ± 0.003 m3/s
Masa mastautin, th = I / Q…..Persamaan(3)

di mana I= isipadu tasik (m3),
Q= Luahan aliran masuk & keluar (m3/s)
Masa mastautin, th = I/ Q
= 27,000 m3
(5.0 x 10-3 m3/s)
= 5.4 x 106 s
Jadi, dijangka air tasik berada dalam tasik selama 62.5 ± 37.6 hari.
Bagi pengiraan sesuatu pencemar, purata kepekatan di lima stesen diambilkira:

Jumlah berat, M = C × I……Persamaan(4)
di mana C = kepekatan sebatian,
I = isipadu air
Kadar aliran berat, L = C × Qout .…..Persamaan (5)

Persamaan (5) boleh digunakan untuk menganggar jumlah pencemar yang mengalir dari Tasik
Kejuruteraan ke dalam Sungai Langat.
Parameter-parameter
Nilai purata dan sisihan piawai bagi setiap parameter kajian pada pensampelan hari kering dan hari
hujan diringkaskan dalam Jadual 2 dan Jadual 3.
Suhu
Analisis statistik ANOVA dua hala menunjukkan tiada perbezaan yang bererti (p>0.05) di antara
keadaan hari hujan-kering bagi lima stesen dengan P=0.717. Manakala, terdapat perbezaan yang
bererti di antara stesen dengan p=0.006. Suhu berada dalam julat 28.00 ± 0.01 0C hingga 30.84 ± 1.57
0
C.
pH
keadaan hari hujan-kering dengan P=0.148. Tiada perbezaan yang bererti (p>0.10) antara stesen
dengan P=0.913. Nilai pH stesen kajian berada dalam julat 6.06 ± 0.07 hingga 7.57 ± 1.61, iaitu
berada pada subkelas IIA. Berdasarkan INWQS (1998), nilai pH yang melebihi julat 6 hingga 9,
menunjukkan pencemaran air telah berlaku. Namun menurut APHA (1992), kebiasaannya nilai pH air
yang neutral ialah di antara julat 4 hingga 9. Nilai ini adalah merupakan nilai biasa yang menunjukkan
kehadiran ion bikarbonat dan karbonat dalam keadaan alkali dan logam alkali bumi. Nilai pH air
semulajadi dipengaruhi oleh bahan organik tanah seperti asid humik, asid tanik, asid uronik dan asid
mineral hasil aktiviti tanah (Mokhtar et al., 2003).
Konduktiviti
keadaan hari hujan-kering dengan P=0.339. Tiada perbezaan yang bererti (p> 0.10) antara stesen
dengan P=0.712. Julat konduktiviti berada dalam julat 168.33 ± 13.61 µS/cm hingga 207.67 ± 8.14
µS/cm. Menurut Chapman (1992), kondukivitit air permukaan biasanya berada dalam julat 10 µS/cm
hingga 1000 µS/cm, tetapi mungkin melebihi nilai julat ini terutamanya dalam air yang tercemar atau
sistem saliran yang menerima air larian permukaan. Konduktiviti mengukur bahan bukan organik
terlarut yang terion membentuk elektrolit. Konduktiviti dan jumlah pepejal terampai adalah
berkadaran terus antara satu sama lain (Mokhtar et al., 2003).
Kekeruhan
keadaan hari hujan-kering dengan P=0.163. Manakala, tiada perbezaan yang bererti (p>0.10) antara
stesen dengan P=0.399. Nilai kekeruhan berada dalam julat 12.19 ± 14.56 NTU hingga 192.27 ±
287.48 NTU. Semua stesen berada pada subkelas I kecuali S2 yang melampaui paras maksimum 50
NTU yang disarankan oleh subkelas IIB. Stesen S2 adalah paling keruh (192.27 ± 287.48 NTU) pada
hari hujan kerana merupakan perangkap lumpur yang memerangkap sedimen dari projek pembinaan
Kolej Kausar dan kompleks FTSM.
TSS
Analisis statistik ANOVA dua hala menunjukkan terdapat perbezaan yang bererti (P<0.05) di antara
keadaan hari hujan-kering dengan P=0.049. Manakala, tiada perbezaan yang bererti antara stesen
dengan P=0.527. TSS adalah lebih tinggi pada hari hujan kerana air larian permukaan sewaktu hujan
mempunyai daya hakisan yang kuat dan mengakibatkan kadar hakisan tanah yang kuat dan
menyumbangkan kepada peningkatan pepejal terampai di kawasan tanah rata terutamanya yang
terdedah akibat proses pembangunan. Nilai TSS berada dalam julat 5.33 ± 2.57 mg/L hingga 69.00 ±
92.15 mg/L. Pada hari kering, semua stesen terletak pada subkelas I. Manakala pada hari hujan,
semua stesen dikategori sebagai subkelas I kecuali S2 berada pada subkelas III. Stesen S2 mempunyai
TSS yang paling tinggi (69.00 ± 92.15 mg/L) pada hari hujan kerana merupakan perangkap lumpur
yang memerangkap sedimen dari projek pembinaan Kolej Kausar dan kompleks FTSM. Pada
keadaaan normal, jumlah berat TSS dalam tasik adalah 163.78 ± 18.19 kg. Anggaran jumlah pepejal
terampai yang mengalir masuk ke Sungai Langat adalah 27.81± 9.29 kg setiap hari.
Oksigen Terlarut
keadaan hari hujan-kering dengan p=0.068. Manakala, terdapat perbezaan yang bererti antara stesen
dengan P=0.014. DO berada dalam julat 5.47 ± 1.16 mg/L hingga 8.53 ± 0.80 mg/L. Pada hari kering,
semua stesen berada pada subkelas I kecuali S3 berada pada subkelas IIA. Pada hari hujan, S4 dan S5
berada pada subkelas I manakala S1, S2, S3 berada pada subkelas IIA. S3 mempunyai nilai DO yang
paling rendah (5.47 ± 1.16 kg) kerana kehadiran bahan organik yang tinggi di S3. Bakteria
menggunakan oksigen terlarut semasa proses penguraian bahan organik tersebut. Pada keadaan
normal, dianggarkan 200.88 ± 28.25 kg oksigen terlarut berada dalam air tasik.
Nitrogen Ammonia
keadaan hari hujan-kering dengan P= 0.545. Tiada perbezaan yang bererti (p>0.10) antara stesen
dengan p=0.476. Nilai nitrogen ammonia berada dalam julat 0.08 ± 0.03 mg/L hingga 1.54 ± 0.85
mg/L. Pada hari kering, Stesen S3 terletak dalam subkelas I; S2 dan S3 berada pada subkelas IIA,
manakala S5 terletak pada subkelas III dan S4 terletak pada subkelas IV. Pada hari hujan, Stesen S1,
S3, S4 dan S5 dikategori sebagai subkelas III dan S2 terletak pada subkelas IV. Ini mungkin
disebabkan air kumbahan domestik yang dilepaskan ke dalam Tasik Kejuruteraaan dan pembuangan
air sisa makanan dari kantin. Menurut Jabatan Pengurusan Pembangunan (JPP), semua air buangan
samada air dalam makmal dan tandas disalirkan oleh paip pembetungan dalam sistem pembetungan
ke kolam pengoksidaan kumbahan UKM untuk proses perawatan sebelum ia dialirkan ke Sungai
Langat, namun dijangkakan terdapat juga luahan air buangan yang disalirkan ke dalam longkang yang
akhirnya mengalir ke dalam tasik kajian. Pada keadaaan normal, dianggarkan 12.65 ± 13.90 kg
nitrogen ammonia berada dalam tasik. Anggaran jumlah nitrogen ammonia yang mengalir masuk ke
Sungai Langat adalah 2.12 ± 0.71 kg setiap hari.
Permintaan Oksigen Biokimia (BOD)

keadaan hari hujan-kering dengan p=0.354; manakala, terdapat perbezaan yang bererti antara stesen
dengan p=0.018. BOD berada dalam julat 0.49 ± 0.37 mg/L hingga 3.48 ± 2.03 mg/L. Pada hari
kering, S1 dan S5 terletak pada subkelas I manakala S2, S3 dan S4 berada pada subkelas IIA. Pada
hari hujan, S1 dan S5 terletak pada subkelas I, S3 dan S4 berada pada subkelas IIA, manakala S2
berada pada subkelas III. Stesen 2 mempunyai nilai BOD yang paling tinggi, iaitu 3.48 ± 2.03 mg/L
sewaktu hari hujan. Ini mungkin kerana stesen ini terletak berdekatan dengan bengkel fakulti
kejuruteraan. Kawasan bengkel ini mungkin menyebabkan saliran air buangan berjaya masuk ke
dalam longkang pada hari hujan yang mengandungi bahan berorganik tinggi yang boleh
meningkatkan paras kepekatan BOD dalam sistem air. Pada keadaaan normal, jumlah berat BOD
yang dianggarkan hadir dalam tasik adalah 41.90 ± 23.95 kg. Anggaran BOD yang mengalir masuk ke
Sungai Langat adalah 7.01 ± 2.34 kg setiap hari.
Permintaan Oksigen Kimia (COD)

Analisis statistik ANOVA dua hala menunjukkan tiada perbezaan yang bererti (p>0.10)) di antara
keadaan hari hujan-kering dengan p=0.729. Tiada perbezaan yang bererti (p>0.10) antara stesen
dengan P=0.955. Secara keseluruhan, nilai COD adalah tinggi, iaitu dalam kategori subkelas IV (51
mg/L hingga 100 mg/L) berdasarkan INWQS disebabkan kehadiran bahan organik dan bahan bukan
organik yang banyak dalam tasik mungkin disebabkan air larian sisa makanan dari kantin dan air sisa
buangan dari makmal dan bengkel fakulti. Menurut JPP, semua air buangan samada air dalam
makmal dan tandas disalirkan oleh paip pembetungan dalam sistem pembetungan ke kolam
pengoksidaan kumbahan UKM untuk proses perawatan sebelum ia dialirkan ke Sungai Langat, namun
terdapat kemungkinan air sisa ini juga mengalir dalam saluran longkang dan mengikut air larian
sewaktu hari hujan. Nilai COD yang tinggi menunjukkan pembuangan air sisa yang mungkin
dialirkan secara langsung atau tidak langsung dari kawasan bangunan fakulti. Pada keadaaan normal,
dianggarkan air tasik mengandungi jumlah berat COD sebanyak 1605.58 ± 74.68 kg. Anggaran COD
yang mengalir masuk ke Sungai Langat adalah 268.90 ± 89.78 kg setiap hari. Nilai ini adalah amat
tinggi dan memberikan beban terhadap kualiti air Sungai Langat yang semakin tercemar.
Plumbum
Analisis statistik ANOVA dua hala menunjukkan terdapat perbezaan yang sangat bererti (p<0.001) di
antara keadaan hari hujan-kering dengan p=0.001. Julat Pb ialah 0.32 ± 0.06 mg/L hingga 0.40 ± 0.08
mg/L sewaktu hari kering manakala julatnya pada hari hujan adalah 0.12 ± 0.10 mg/L hingga 0.19 ±
0.11 mg/L. Nilai Pb adalah rendah semasa hujan disebabkan proses pencairan air tasik. Tiada
perbezaan yang bererti antara stesen dengan p=0.345 hasil ujian ANOVA. Secara keseluruhan, nilai
Pb adalah amat tinggi dan melebihi had piawai JAS, iaitu 0.05 mg/L. Kebanyakan plumbum yang
dibebaskan ke atmosfera berpunca daripada bahan api kenderaan bermotor yang mengandungi bahan
kimia antiketuk, tetraetil plumbum, (C2H5)4Pb. Plumbum yang termendap di tempat letak kereta dan
jalanraya boleh dibawa oleh air larian permukaan dan mengalir ke dalam tasik. Menurut Camp dan
Meserve (1974), berdasarkan pencemaran yang diakibatkan daripada kawasan bandar, terdapat
peningkatan masalah kontaminasi logam berat semasa berlakunya air larian permukaan di kawasan
bandar. Menurut Spliethoff dan Hemond (1996), kewujudan plumbum bukan sahaja boleh dicerap di
kawasan berkepadatan dengan kenderaan, tetapi juga dalam atmosfera yang turun dengan air hujan
dan diangkut ke sedimen akuatik dengan berkesan berbanding dari permukaan kawasan pertanian.
Pada keadaaan normal, dianggarkan terdapat kandungan Pb sebanyak 9.50 ± 0.90 kg. Anggaran
jumlah Pb yang mengalir masuk ke Sungai Langat adalah 1.59 ± 0.53 kg setiap hari. Nilai ini adalah
amat tinggi kerana logam berat ini amat toksik kepada manusia walaupun dalam kuantiti kecil.
Kadmium
keadaan hari hujan-kering dengan p=0.404. Tiada perbezaan yang bererti (p>0.10) antara stesen
dengan P=0.931. Julat Cd ialah 0.09 ± 0.02 mg/L hingga 0.11 ± 0.04 mg/L. Nilai kepekatan Cd adalah
amat tinggi dan melebihi had piawai JAS, iaitu 0.01 mg/L. Ini mungkin disebabkan kehadiran Cd
dalam debu dan termendap di tasik. Kewujudan Cd dalam atmosfera biasanya berpunca dari
pembakaran bahan api dalam kenderaan bermotor, kegiatan perindustrian dan aktiviti pembinaan yang
amat pesat di sekitar lembangan. Pada keadaan normal, kandungan Cd dalam tasik dianggarkan 2.81 ±
0.24 kg. Anggaran jumlah Cd yang mengalir masuk ke Sungai Langat adalah 0.47 ± 0.15 kg setiap
hari. Nilai ini adalah tinggi dan membahayakan hidupan jika ia terkumpul dalam tisu badan.
Indeks Kualiti Air (IKA)

Jadual 4 menunjukkan nilai IKA secara purata bagi hari hujan dan hari kering di stesen kajian ini.
Jadual 4: Nilai IKA secara purata di stesen kajian ini
Stesen IKA Pengelasan Status
1 81.32±3.33 II Baik
2 72.85±2.45 III Sedikit tercemar
3 81.22±0.02 II Baik
4 81.99±3.24 II Baik
5 82.50±2.62 II Baik
Semua stesen pensampelan berada pada Kelas II, iaitu mempunyai kualiti air yang baik kecuali stesen
S2 dikategorisebagai Kelas III, iaitu sedikit tercemar dan memerlukan rawatan ekstensif. Secara
keseluruhannya, Tasik Kejuruteraan dikategorikan sebagai Kelas II-III, iaitu merupakan bekalan air
yang memerlukan rawatan konvensional dan membenarkan kegunaannya untuk aktiviti rekreasi yang
melibatkan sentuhan. Hujan mempengaruhi kualiti air Tasik Kejuruteraan, terutamanya pada musim
tengkujuh, yang menyebabkan nilai bagi parameter TSS meningkat dengan banyak dan menyebabkan
air tasik kelihatan keruh dan berwarna coklat.
Cadangan
Berikut merupakan cadangan tentang langkah-langkah yang perlu diambil untuk menuju ke arah
IWRM dalam kampus bagi menghasilkan suasana ekosistem sihat dan harmoni selaras dengan tema
UKM ‘Universiti dalam Taman’. Di peringkat kampus, Pembuatan Keputusan Kolaboratif atau
‘Collaborative Decision Making” atau “CDM” (Mazlin et al. 2004b) sangat digalakkan sebagai
inisiatif bersama pelbagai pihak berkepentingan termasuk pihak pengurusan tertinggi universiti yang
terdiri daripada Naib Canselor, pihak pengurusan profesional dan eksekutif seperti jurutera dan
peringkat lain merangkumi warga universiti yang terdiri daripada pengajar, pelajar dan kakitangan
sokongan serta pengguna lain seperti kontraktor projek bertujuan untuk memperbaiki sistem
pengurusan sumber air yang sedia ada. CDM menggalakkan proses perkongsian dan pertukaran
maklumat di antara pelbagai pihak, dan secara tak langsung turut memperbaiki alat sokongan
membuat keputusan. Orang ramai yang memasuki kampus seperti ibu bapa pelajar dan orang yang
menggunakan kemudahan universiti juga harus didedahkan tentang konsep CDM dan IWRM.
Pihak pengurusan universiti bertanggungjawab membuat dasar dan peraturan, menetapkan mesyuarat
bersama. eksekutif dan menggalakkan proses berkongsi data dan maklumat, serta cara menangani isu
bersama. Jurutera atau saintis berperanan dalam pemonitoran kualiti air. Pengajar perlu menerapkan
konsep IWRM kepada setiap pelajar. Kakitangan sokongan mesti juga terlibat membantu mengawal
pelepasan sisa secara tidak bertanggungjawab. Pelajar harus menjaga kebersihan tasik seperti tidak
membuang sampah ke dalam tasik manakala kontraktor bertanggngjawab dalam pengaliran air
buangan berkelodak dan mengandungi pelbagai bahan buangan lain serta perlu juga mereka
mengambil inisiatif untuk menjamin kebersihan air tasik.
Selain itu, dana kewangan dan sumber manusia yang berpatutan juga perlu disediakan oleh pihak
pengurusan universiti, bermula dengan menerapkan konsep IWRM dan CDM kepada golongan staf
sedia ada. Kakitangan di Jabatan Pengurusan Pembangunan khususnya perlu didedahkan dengan
konsep tersebut agar dapat mengurus tasik ini dengan baik. Daya tampung bagi bahan pencemar
dalam tasik harus dinilai oleh pihak jurutera. Jenis spesies hidupan yang ada di dalam tasik perlu
diketahui melalui kajian masa depan. Pangkalan data harus dibina dan dikongsi bersama oleh semua
pihak berkepentingan dalam universiti, dan masyarakat yang berminat.
Kesimpulan
Sistem saliran air ke Tasik Kejuruteraan telah dikaji dan menunjukkan bahawa parameter kekeruhan
di S2 melampaui paras maksimum 50 NTU yang disarankan oleh subkelas IIB bagi INWQS. Jumlah
TSS di S2 tergolong dalam subkelas III pada hari hujan. Parameter NH3-N di S5 dan S4 telah
melebihi paras maksimum subkelas IIB pada hari kering manakala semua stesen telah melebihi paras
maksimum subkelas IIB pada hari hujan. Kandungan BOD di S2 tergolong dalam subkelas III pada
hari hujan. Nilai COD adalah tinggi, iaitu dalam kategori subkelas IV. Nilai kepekatan Pb adalah amat
tinggi dan melebihi had piawai JAS, iaitu 0.05 mg/L. Nilai kepekatan Cd adalah amat tinggi dan
melebihi had piawai JAS, iaitu 0.01 mg/L. Tasik Kejuruteraan dikategori dalam Kelas II-III bagi
INWQS. Analisis ANOVA dan kolerasi telah dilakukan. Beberapa langkah untuk memperbaiki kualiti
air dan keadaan saliran air tasik serta sistem sumber air di kampus telah juga dicadangkan.
Rujukan
1. AHPA.AWWA.WEF. 1998. Standard Methods for the Examination of Water and Wastewater
Ed.20, American Public Health Association, Washington.
2. Bruce Mitcell, 1997. Resource and Environmental Management. England. Wesley Longman
Limited.
3. Camp, T.R. & Meserve, R.L. 1974. Water and its impurities. Ed. Ke 2. Stroudsburd: Dowden
Hutchinson & Ross, Inc.
4. DID. 2000. Urban Stormwater Management Manual for Malaysia (Manual Saliran Mesra Alam
Malaysia). Vol 5: Runoff Estimation. KL.
5. DID. 2000. Urban Stormwater Management Manual for Malaysia (Manual Saliran Mesra Alam
Malaysia). Vol 8: Retention. KL.
6. Global Water Partnership, 2000b. Integrated water resources management. TAC Background
papers, Nomer 4, Stockholm, Sweden.
7. JAS. 2001. INWQS (atas talian) http://www.jas.sains.my/jas/river/interim_2-3c.htm (2 Feb
2005).
8. Mazlin bin Mokhtar, Mohd. Talib Latif, Lee Yook Heng. 2003. Kimia Air. KL.Utusan
Publications & Distributors Sdn. Bhd.
9. Mazlin B. Mokhtar, Shaharudin Idrus, Sarah Aziz. 2004. Kesihatan Ekosistem Lembangan
Langat. Prosiding Simposium Penyelidikan Ekosistem Lembangan Langat 2003, Penerbit
LESTARI, UKM.
10. Mazlin B. Mokhtar, Rahmah Elfithri and Abdul Hadi Harman Shah. 2004b. “Collaborative
Decision Making as a Best Practice in Integrated Water Resources Management: A Case Study
on Langat Basin, Malaysia”. Proceedings (Technical Papers), First Southeast Asia Water
Forum, 17-21 November 2003, Chiang Mai, Thailand, Volume 2: pp. 333-354. ISBN 974-241-
776-8.
11. World Water Forum 2003 (atas talian) http://www.water-forum3.com (21 Oktober2004).
12. Spliethoff, H.M. & Hemond, H.F. 1996. History of toxic dischange to surface waters of the
Aberjona Watershed. Journal of Environmental Science & Technology 30(1): 121-127.
REMOVAL OF BASIC AND REACTIVE DYES: A COMPARISON OF SORPTION AND
PHOTODEGRADATION STUDY
S.T. Ong, C. K. Lee and Z. Zainal
Chemistry Department, Faculty of Science,

Universiti Putra Malaysia, 43400 UPM, Selangor, Malaysia.
e-mail: cklee@fsas.upm.edu.my
Abstract. A comparative study on the effectiveness of using ethylenediamine modified rice hull
(MRH) and titanium dioxide (TiO2) under ultraviolet irradiation to remove both basic and reactive
dyes from aqueous solutions was carried out. The sorption characteristics of Basic Blue 3 (BB3) and
Reactive Orange 16 (RO16) by MRH were studied under various experimental conditions. Studies on
the sorption of both dyes showed that sorption was pH and concentration dependent. Langmuir
equation was employed to model the sorption behavior of MRH. Maximum sorption capacities
calculated from the Langmuir model are 3.29 and 24.88 mg/g for BB3 and RO16, respectively. The
effect of initial concentrations as well as light source was carried out in the photodegradation of BB3
and RO16. BB3 with concentration of 50 mg/l was totally degraded after 6 hours of contact with
TiO2 under UV illumination whereas RO16 at the same concentration was completely decolorized at
illumination time of 5 hours. The decolorizing efficiency decreased with increasing dye concentration
and a higher efficiency was obtained under solar light illumination.
Abstrak. Satu kajian perbandingan terhadap keberkesanan menggunakan sekam padi terubahsuai
etilenadiamina (MRH) dan titanium dioksida (TiO2) di bawah penyinaran ultraungu bagi
menyingkirkan kedua-dua pewarna basik dan reaktif daripada larutan akues telah dijalankan. Sifat-
sifat erapan basik biru 3 (BB3) dan reaktif oren 16 (RO16) oleh MRH telah dikaji di bawah pelbagai
keadaan eksperimen. Kajian erapan terhadap kedua-dua pewarna menunjukkan erapan adalah
dipengaruhi oleh pH dan kepekatan. Persamaan Langmuir telah diaplikasikan untuk memodel sifat
erapan oleh MRH. Kapasiti erapan maksimum dikira daripada model Langmuir adalah masing–
masing 3.29 dan 24.88 mg/g bagi BB3 dan RO16. Kesan kepekatan dan juga sumber cahaya telah
dijalankan ke atas fotodegradasi BB3 dan RO16. BB3 dengan kepekatan 50 mg/l telah
didegradasikan sepenuhnya selepas 6 jam dengan kontak TiO2 di bawah penyinaran UV manakala
RO16 dengan kepekatan yang sama telah dilunturkan warna sepenuhnya selepas diiluminasikan
selama 5 jam. Keberkesanan pelunturan warna menurun dengan peningkatan kepekatan pewarna dan
effisiensi yang lebih tinggi diperolehi di bawah illuminasi solar.
Keywords : Reactive dyes; Basic dyes; Sorption; Photodegradation
Introduction
The widespread usage of dyes in many industries has led to a tremendous increase in the production
of colored wastes. It is well known that colored waste has a strong impact on the ecosystem and will
lead to disturbances to aquatic life. Hence, the environmental issues surrounding the removal of color
in effluents has become a great concern. Owing to the nature of synthetic dyes, conventional
biological treatment methods are ineffective for decolorizing such waste water [1].
There is thus a need to search for new and economical process that could remove dyes that are
commonly used in the industry. Adsorption and photocatalytic degradation are two noteworthy
treatment processes. Activated carbon is perhaps the most widely used adsorbent for the removal of
many organic contaminants which are biologically resistant [2]. However activated carbon is costly
and difficult to regenerate. Thus, there has been intensive research exploring the potential of
alternative low-cost materials as sorbents for dyes.
Biological waste materials such as peanut hull, sugarcane dust, saw dust, corn corb, barley
husk and rice hull have been studied as alternatives for activated carbon in the removal of dyes in
textile wastewater [3-7]. Most of the materials investigated are efficient in binding either basic or
reactive dyes but not both. As they do commonly exist together in wastewater it is of great interest to
have a material that can remove both types of dyes at the same time.
Photocatalytic oxidation of pollutants, especially organic pollutants has generated much
interest. In this regard, much attention has been focused on one of the most commonly used
photocatalyst, titanium dioxide (TiO2). Investigation of photocatalytic degradation using TiO2 has
gained considerable interest not only because it is comparatively cheap, but also due to its absence of
toxicity. As TiO2 is illuminated by light rays with wavelength below 380 nm, electron-hole pairs are
generated. The valence band holes will react with water molecules or hydroxide ions (OH-) to
produce hydroxyl radicals (·OH). Both hydroxyl radicals and valence band holes are powerful
oxidizing species that will oxidize the organic pollutants to carbon dioxide, water and some simple
acids [8]. Oxygen is usually supplied as electron acceptor to prolong the recombination of electron-
hole pairs during photocatalytic oxidation [9].
A series of studies have reported that TiO2 photocatalysis is an effective method for
decolorizing and oxidizing organic dyes in wastewater [10-14]. The present paper describes a
comparative study on the removal of both basic and reactive dyes using two methods: sorption using
ethylenediamine (EDA) modified rice hull (MRH) and photodegradation using TiO2.
Experimental
2.1 Materials
Rice hull was washed several times to ensure the removal of dust and ash. It was then rinsed several
times with distilled water and dried overnight in an oven at 50 °C. The dried rice hull was ground to
pass through a 1 mm sieve and labeled as natural rice hull (NRH). Modification was carried out by
treating NRH with EDA in a ratio of 1.00 g rice hull to 0.02 mole of EDA in a well-stirred water bath
at 80 oC for 2 hours.
The TiO2 powder P-25 (mainly in anatase form, mean particle size of 30 nm, BET surface
area of 50 m2/g) from Degussa (Frankfurt, Germany) was used as the photocatalyst.
Basic Blue 3 (BB3, Sigma, 40 % dye content) and Reactive Orange 16 (RO16, Aldrich, 50 % dye
content) were selected as the sorbates in this study. Standard dye solutions of 2000 mg/l were
prepared and subsequently diluted when necessary.
2.2 Sorption experiments

All the batch experiments were carried out in duplicate and the results given are the averages. A
control without sorbent was simultaneously carried out to ascertain that the sorption was by MRH and
not by the wall of the container. Sorption experiments were performed by agitating 0.05 g of sorbent
in 20 ml of 100 mg/l dye solution at their natural pH of 4.7 for BB3, 6.0 for RO16 and 5.5 for binary
solution in a centrifuge tube at 150 rpm at room temperature (25 + 2 oC). The sorbent-sorbate mixture
was then centrifuged at 3.0 x 103 rpm for phase separation and the dye concentration of the
supernatant was analyzed using Shimadzu 160B double beam UV-vis spectrophotometer. All
measurements were made at the wavelengths corresponding to maximum absorption; for BB3, λmax =
654 nm and for RO16, λmax = 494 nm.
To study the effect of pH, a series of 100 mg/l dye solutions of BB3 and RO16 were prepared.
The pH of the dye solutions was adjusted to the range of 2 to 10 by adding dilute HCI or NaOH. Each
dye solution was shaken with MRH for 4 hours.
The effect of initial concentration was studied by varying the dye concentration from 50 to
100 mg/l for both dyes. At predetermined intervals samples were withdrawn and analyzed for dye
concentration.
Sorption isotherms were obtained by agitating samples of sorbent using dye concentrations of
5 to 150 mg/l.
2.3 Photodegradation experiments

Irradiation experiments of dye solutions were carried out by stirring 500 ml of 100 mg/l dye solutions
at their natural pH in a 1000 ml beaker with 1.25 g of TiO2 at room temperature (25 + 2 oC) for 8
hours unless otherwise stated. Aeration was provided by bubbling air into the reaction solution by an
air pump to ensure a constant supply of oxygen. UV-irradiation was provided by a high pressure
mercury lamp. At given intervals of irradiation, approximately 5 ml of the solution was withdrawn
from the reservoir. The solution was then filtered through a 0.45 µm membrane filter (Milipore) to
remove trace of TiO2 particles.
To study the effect of initial concentration and contact time, all the conditions described
above apply except the initial concentrations. The initial concentrations were varied from 50 to 100
mg/l for both dye solutions. At predetermined time intervals the mixtures were removed, filtered and
analyzed for their dye concentrations.
The effect of different light source was studied by using four different light sources namely
white fluorescent light (18 watt), halogen lamp, UV lamp and sunlight between 11.00 to 3.00 p.m.
The experiments were conducted by removing 5 ml of dye solutions from the beaker at predetermined
time intervals for analysis.

3.1 Sorption studies
3.1.1 Effect of pH
The effect of pH on the uptake of BB3 and RO16 by MRH in both single and binary dye solution is
shown in Figure 1. For BB3 in single dye solution, the percentage uptake increased from 1.32 to
66.99 % with increase in pH from 2 to 10. It is suggested that, at low pH, the carboxyl groups on the
surface of rice hull that are responsible for binding with BB3 are predominantly protonated (-COOH),
hence incapable of binding BB3. With increasing pH, the number of positively charged sites
decreases and the number of negatively charged sites increases. This phenomenon favors the sorption
of positively charged dye due to electrostatic attraction [15]. The reverse trend was observed in the
removal of RO16 where the percentage of uptake was much higher at lower pH. This is because at
low pH, the amine groups on the surface of MRH are protonated (–NH4+), thereby increasing
electrostatic attractions between negatively charged dye anions and positively charged sorption sites
resulting in an increase in dye sorption [16,17]. Similar observations were obtained in binary dye
solutions where uptake was higher.
3.1.2 Effect of initial concentration and contact time

The rate of sorption of BB3 and RO16 in single dye solution by MRH as a function of initial
concentration is shown in Figure 2. The uptake follows the typical sorption pattern shown by most of
the biosorbent, where the initial uptake was rapid followed by a more gradual process, irrespective of
the initial dye concentration. In the binary system, there was a noticeable and sudden increase in
sorption of RO16 at around 240 min intervals, indicating that there might be more than one step
involved in the sorption process. In order to explore the kinetics involved in dye sorption, the
experimental data were examined using different equations [18,19]:
k 1t
log( q e − q t ) = log( q e ) − (pseudo first-order) (1)
2 . 303
and
t 1 t
= + (pseudo second-order) (2)
qt h qe
where qe = the amount of dyes sorbed at equilibrium (mg/g), qt = the amount of dyes sorbed at time, t
(mg/g), k1= the rate constant of pseudo first-order sorption (1/min), h = (k2qe2 ) = the initial sorption
rate (mg/g min) and k2= the rate constant of pseudo second- order kinetics (g/mg min). The rate
constants and correlation coefficients for these two sorption kinetics are listed in Table 1. In many
studies, the kinetics of sorption by biological materials are described by the first order Lagergren
equation. However, it has been shown that, pseudo second-order provides a better fit than the first-
order Lagergren model [20]. In this study, it was found that the equilibrium sorption capacities
calculated by using the second-order kinetic model gave closer values with those determined
experimentally (Table 1). The correlation coefficients for the second kinetic model obtained at
various concentrations are in general higher than those of the first-order equation. This indicates that
pseudo second-order kinetics could better explain the data obtained and implies that the rate limiting
step may be chemical sorption or chemisorption involving valency forces through sharing or exchange
of electron between sorbent and sorbate [19].
120
BB3- single
RO16- single
100
BB3- binary
RO16- binary
80
% Uptake
60
40
20
0
0 2 4 6 8 10 12
pH
Figure 1: Effect of pH on the sorption of BB3 and RO16 in single and binary dye solutions by MRH
100
BB3- 50 mg/l BB3- 75 mg/l BB3- 100 mg/l
80 RO16- 50 mg/l RO16- 75 mg/l RO16- 100mg/l
60
% Uptake
40
20
0
0 60 120 180 240 300 360 420 480
Time (min)
Figure 2: Effect of initial concentration and contact time on BB3 and RO16 sorption from single dye
solution by MRH
3.1.3 Sorption Isotherm

Several sorption isotherms have proven useful in understanding the process of sorption. The simplest
isotherm is attributed to a pioneer in the study of surface processes, Langmuir, and is called the
Langmuir isotherm. The linearised Langmuir equation can be written as
C 1 C
e
= *
+ e
*
(3)
N e N b N
where Ce = equilibrium concentration of the dye (mg/l), Ne = amount of dye sorbed at equilibrium
(mg/g), N* = maximum sorption capacity (mg/g), b = constant related to the energy of the sorbent
(l/mg). The linear plots of Ce/Ne versus Ce obtained from the sorption data for both single and binary
dyes by MRH indicate the applicability of Langmuir isotherm. The constants derived from the
equations are shown in Table 2. In the binary solution, the maximum sorption capacities, N* of BB3
and RO16 were enchanced by 4.5 and 2.4 times, respectively. The actual mechanism involved in this
synergistic effect is not clear.
However, fitting the model to the sorption process does not necessarily imply any physical
interpretation attached to them since the biosorbent’s surface is non-homogeneous and there could be
more than one type of sorption sites on the biosorbent’s surface [21]. Nevertheless, this isotherm
model provides some valuable insight on the maximum sorption capacities of the sorbent studied.
Table 1: Pseudo first and second order constants for the sorption of single and binary dye solutions
Dye Initial Pseudo first order Pseudo second order Experimental

concentration
sorption R2 sorption R2 sorption
capacities capacities capacities
(mg/l) (mg/g) (mg/g) (mg/g)
BB3 50 0.550 0.9654 2.233 0.9997 2.232

(single) 75 1.387 0.9510 3.683 0.9940 3.152
100 2.182 0.9576 3.162 0.9936 3.564
RO16 50 9.979 0.8786 12.626 0.9954 11.520

(single) 75 12.465 0.9857 15.244 0.9849 14.956
100 13.960 0.9663 16.835 0.9824 16.904
BB3 50 4.333 0.9288 6.650 0.9875 6.548

(binary) 75 6.434 0.9444 9.320 0.9911 8.956
100 8.855 0.8814 11.442 0.9765 10.924
RO16 50 12.882 0.9971 17.271 0.9946 15.272

(binary) 75 22.320 0.9759 28.249 0.9898 25.600
100 23.988 0.9762 31.153 0.9837 29.054
Table 2: Langmuir constants for the sorption of single and binary dye solutions
Langmuir constants
Dye N*(mg/g) b (l/mg) R2
BB3 3.286 0.102 0.956

RO16 24.876 0.099 0.991
BB3 (binary) 14.680 0.003 0.941
RO16 (binary) 60.241 0.028 0.995
3.2 Photodegradation studies

3.2.1 Effect of initial concentration and contact time
The photodegradation rate of both dyes in single and binary system decreased with increasing initial
concentration. Similar findings have been reported [9, 22]. BB3 with concentration of 50 mg/l was
totally degraded after 6 hours of contact with TiO2 under UV illumination. RO16 at the same
concentration was completely decolorized at illumination time of 5 hours (Figure 3). As for the
binary systems, the formation of residues was observed at the end of the photodegradation
experiment. The degradation of various organic contaminants over illuminated TiO2 has been
reported to conform to the Langmuir- Hinshelwood first order kinetics model (23). Re-plotting the
data using the equation ln (C/Co) versus Time (Figure 4), it is clear that photodegradation of BB3 and
RO16 also follows the pseudo first-order kinetics
Co
In = kt (4)
C
where Co/C is the normalized BB3 and RO16 concentration and k the apparent rate constant (1/min).
3.2.2 Effect of different light source

Figure 5 shows the comparison of color removal efficiency of single BB3 and RO16 with various
light sources. Different light sources have different wavelength ranges and if the provided light source
has a wavelength shorter than 400 nm or equivalent energy higher than 3.2 eV, it can photoexcite
TiO2 and produce electron-hole pairs. Thus it is expected that sunlight or UV light (wavelength range
< 380 nm) should be the most effective followed by fluorescent light (wavelength < 400 nm) and
other artificial light (wavelength range > 400 nm). The results obtained were in accordance with this
order. The decolorized efficiency of single BB3 and RO16 with solar light irradiation was more
efficient than that with artificial UV light irradiation. This phenomenon was partially attributed to a
higher temperature of dye solution found under solar light irradiation. Previous work [9] also
reported that the color removal rate of methylene blue with solar light irradiation was almost twice
that of artificial UV light irradiation.
160.00
BB3- 50 mg/l BB3- 75 mg/l BB3- 100mg/l
RO16- 50 mg/l RO16- 75 mg/l RO16- 100 mg/l

120.00
% Photodegradation
80.00
40.00
0.00
0 60 120 180 240 300 360 420 480 540
Time (min)
Figure 3: Effect of initial concentration and contact time on BB3 and RO16 for the photodegradation
of single BB3 and RO16
4.00
BB3- 50 mg/l BB3- 75 mg/l BB3- 100 mg/l
2.00 RO16- 50 mg/l RO16- 75 mg/l RO16- 100mg/l
0.00
ln C/Co
-2.00
-4.00
-6.00
0 60 120 180 240 300 360 420 480 540
Time (min)
Figure 4: Graph ln C/Co versus Time (min) for the effect of initial concentration and contact time for
the photodegradation of single BB3 and RO16
2.00
UV light (BB 3) UV light (RO 16)
Sunlight (BB 3) Sunlight (RO 16)
1.60
White fluorescent (BB 3) White fluorescent (RO 16)
Halogen lamp (BB 3) Halogen lamp (RO 16)
1.20
C/Co
0.80
0.40
0.00
0 60 120 180 240 300 360 420 480 540
Time (min)
Figure 5: Effect of different light sources for the photodegradation of BB3 and RO16 in single dye
solutions
In the binary dye solution, however, the experimental results of photodegradation showed that
the removal of both BB3 and RO16 was more effective using UV light than solar light. This may be
due to the fluctuation in sunlight intensity. Nevertheless, sunlight can serve as an effective alternative
and cheaper light source for the photodegradation of color pollutant.
4. COMPARISON OF SORPTION AND PHOTODEGRADATION
Kinetic modeling has shown that, in the sorption study, the sorption capacities estimated from pseudo
first-order model are generally lower than those obtained experimentally, whereas the pseudo second-
order kinetic model which is based on the assumption that the rate-limiting step may be chemisorption
yields good agreement between estimated and experimental sorption capacities. Similar phenomena
have been reported in the biosorption of dyes on biosorbent [17, 19, 24, 25]. On the other hand, the
kinetics of photodegradation using TiO2 catalyst fits the pseudo first-order model well.
The plots of Ct/Co versus Time (min) for single BB3 in both sorption and photodegradation
are shown in Figure 6 for comparison. For the range of dye concentrations studied, photodegradation
using TiO2 suspension is a more efficient method than sorption in the removal of BB3 in aqueous
solution. Similar trend was observed in the removal of RO16 and in binary solution. Though TiO2
suspension have been reported to be efficient due to the large surface area of the catalyst available for
reaction and the efficient mass transfer within such system [26], there are, however, problems
associated with using TiO2 in suspended form. First, it is difficult to separate the particle at the end of
the process. Secondly, for photodegradation it is essential to ascertain sufficient supply of oxygen and
efficient light source. Both of these would add to the capital and operating costs of the treatment
process. In the present study,the formation of residues was also observed in the binary solution at the
end of the experiments. The nature of the residue is currently order investigation.
1.60
Sorption- 50 mg/l Photodegradation- 50 mg/l
Sorption- 75 mg/l Photodegradation- 75 mg/l
1.20 Sorption- 100 mg/l Photodegradation- 100mg/l
Ct/Co
0.80
0.40
0.00
0 60 120 180 240 300 360 420 480 540
Time (min)
Figure 6: Graph Ct/Co versus Time (min) for the effect of initial concentration and contact time for
the sorption and photodegradation of single BB3
5. CONCLUSION
The present study has shown the effectiveness of MRH and TiO2 in the removal of BB3 and RO16
from synthetic solutions. Sorption was pH dependent and in the pH range of 4 to 7 appreciable
amount of both dyes could be sorbed by MRH, both in single and binary dye solutions. The
equilibrium data conform to Langmuir isotherms. A study of the kinetic models on sorption showed
that sorption fitted the pseudo second-order model whereas photodegradation was explained by the
pseudo first-order kinetics model. Both dyes can be separately degraded in an aqueous TiO2
dispersion under irradiation by ultraviolet lamp. However, due to the micrometric size of TiO2
particles, the use of aqueous suspension is not suited for practical applications.
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KAJIAN KECEKAPAN ANOD KORBANAN ALOI AL-5.5ZN-2.0MG-XSN
DI DALAM AIR LAUT TROPIKA
Abdul Razak Daud1, Mahdi Che Isa1, Muhamad Daud2,

Mohd Yazid Ahmad3, Nik Hasanudin Nik Yusof3
1
Pusat Pengajian Fizik Gunaan, Fakulti Sains & Teknologi,
Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor
2
Institut Penyelidikan Teknologi Nuklear Malaysia (MINT), 43000 Kajang, Selangor
3
Institut Penyelidikan Sains & Teknologi Pertahanan (STRIDE), 43000 Kajang, Selangor
email: ard@pkrisc.cc.ukm.my; mahdicheisa@yahoo.com
Abstrak. Suatu kajian telah dijalankan bagi menentukan kecekapan aloi aluminium dengan komposisi nominal
unsur aloian, Zn 5.5%bt., Mg 2.0%bt, x%bt. Sn dengan x = 0.1, 0.5, 1.0, 1.5 dan 2.0 bagi kegunaan sebagai
anod korbanan untuk kawalan kakisan dalam persekitaran marin. Aloi berkenaan dihasilkan dengan kaedah
tuangan. Penciriannya dilakukan menggunakan kaedah pengutuban elektrokimia dan kapasiti arus. Keputusan
kajian menunjukkan penambahan unsur Sn ke dalam aloi Al-5.5Zn-2.0Mg telah meningkatkan kecekapan
anodnya. Keputusan juga menunjukkan kehadiran 1.34 %bt. Sn telah mengaktifkan aloi Al-5.5Zn-2.0Mg
dengan berkesan menyebabkan kadar kakisan meningkat disamping menghasilkan kecekapan anod yang tinggi
iaitu masing-masing 9.80 x102 µm.tahun-1 dan 89.05%.
Abstract. A study on the anode efficiency of aluminium alloys with nominal composition of alloying elements,
5.5 wt.% Zn,, 2.0 wt.% Mg and x wt.% Sn where x = 0.1, 0.5, 1.0, 1.5 and 2.0 for use as sacrificial anode in
corrosion control in marine environment. The alloys were prepared by casting method. They were characterized
using electrochemical polarization method and current capacity. The results obtained showed that the addition of
Sn in the Al-5.5Zn-2.0Mg alloy has increased its anode efficiency. It also showed that the presence of 1.34 wt.%
Sn has effectively activated the Al-5.5Zn-2.0Mg alloy which increased corrosion rate and anode efficiency to
9.80 x102 µm.year-1 and 89.05% respectively.
Kata kunci: Aloi aluminium; kadar kakisan; kecekapan anod
Pengenalan
Keunikan aluminium seperti ringan dan tahan kakisan telah menjadikannya sangat berguna dalam
pelbagai industri seperti industri pembinaan, pengangkutan, pembungkusan makanan dan sebagainya.
Penggunaannya sebagai bahan bagi mengawal serangan kakisan bagi struktur-struktur keluli telah
mendapat perhatian ramai penyelidik[1-3]. Walaupun aloi aluminium secara relatifnya agak baru
sebagai bahan anod korbanan, namun ia mempunyai merit yang cukup baik kerana mempunyai
beberapa kelebihan seperti ringan, mudah direkabentuk, nilai teori bagi kapasiti arus yang tinggi
(2800 Aj/Kg) dan mudah diperolehi [1, 2].
Aluminium adalah logam yang reaktif, oleh itu mudah membentuk lapisan oksida apabila
terdedah kepada oksigen. Kehadiran lapisan oksida menyebabkan logam aluminium menjadi pasif dan
kurang berkesan sebagai bahan anod korbanan. Jesteru itu, campuran dengan logam pengubahsuai
seperti zink dan magnesium amat perlu bagi mengubah sifat elektrokimianya. Kajian terdahulu
melaporkan campuran Zn dan Mg ke dalam aloi aluminium berjaya mengubah keupayaan kakisannya
kearah lebih elektronegatif. Namun begitu, bagi memastikan aloi ini terus aktif di dalam persekitaran
operasinya, bahan pengaktif turut diperlukan. Walaupun unsur seperti Hg atau In dapat meningkatkan
keaktifan aloi aluminium tetapi penggunaan Hg khasnya bertentangan dengan sensitiviti perlindungan
alam sekitar sementara In pula sukar didapati dan mahal, maka unsur pengaktif alternatif adalah
diperlukan [3]. Sehubungan dengan itu, Sn menjadi pilihan sebagai bahan pengaktif aluminium
berdasarkan ciri-ciri seperti tidak membentuk sebatian antaralogam dengan matriks aluminium,
mudah di dapati dan harganya lebih murah daripada indium. Tujuan kajian ini adalah untuk
memfabrikasi aloi aluminium yang ditambah dengan Sn dan mengkaji kemampuannya sebagai anod
korbanan bagi perlidungan keluli.
Kaedah
Penyediaan Sampel
Sebanyak enam aloi aluminium berbeza komposisi telah difabrikasi melalui teknik peleburan
konvensional di dalam persekitaran gas argon. Sn ditambah kedalam campuran bahan aloi iaitu
aluminium dan unsur-unsur lain seperti Zn dan Mg apabila suhu pemanasan mencapai 600oC. Leburan
dipanaskan sehingga suhu 850 oC sebelum dituang kedalam acuan keluli. Komposisi nominal aloi
yang dibina ialah Zn dan Mg masing-masing sebanyak 5.5%bt. dan 2.0 %bt. sementara Sn sebanyak
0.1 %bt. hingga 2.0 %bt. dan selebihnya Al. Spesimen daripada aloi yang telah difabrikasi dianalisis
komposisinya menggunakan spektrometer serapan atom mengikut piawai ASTM E 34-85[4]. Selepas
komposisi sebenar aloi diperolehi, beratara aloi di kira menggunakan persamaan 1 berikut[5],
m
fiAi
Beratara = ∑ g/mol (1)
i =1 ni
dengan fi = Pecahan mol jisim bagi unsur ke-i

Ai = Berat jisim molekul relatif unsur ke-i
ni = Valensi bagi unsur ke-i
Penentuan Kadar Kakisan

Kadar kakisan ditentukan daripada plot Tafel yang diperolehi melalui kaedah pengutuban
elektrokimia dengan kadar imbasan keupayaan 0.5 mV/saat pada julat keupayaan ± 250 mV disekitar
keupayaan kakisan (Ekakis)[6]. Kadar kakisan dikira berdasarkan persamaan 2. Bagi kajian ini
potentiostat model PC4/750 buatan Gamry yang dikawal oleh perisian Gamry Framework V4.2 telah
digunakan. Data diproses oleh perisian EChem Analyst V1.2 yang dipasang pada sistem peralatan
berkenaan.
0.13 B Ikakis
Kadar Kakisan = mil per tahun (2)
d
dengan B = beratara sampel (g.mol-1)

d = ketumpatan sampel (g.cm-3)
1 mil per tahun = 25.4 mikrometer per tahun.
Penentuan Kecekapan Anod dan Kapasiti Arus
Kecekapan anod menunjukkan keberkesanan sesuatu anod melindungi logam lain daripada serangan
kakisan. Ia dikira menggunakan data kapasiti arus sebenar dan kapasiti arus teori melalui persamaan
3. Kapasiti arus ditentukan dengan mengukur pemindahan cas (Amper.jam) per kehilangan jisim
sampel. Bagi kajian ini keberkesanan aloi aluminium sebagai anod korbanan di dalam persekitaran
marin ditentukan. Nilai kapasiti arus sebenar iaitu nilai uji kaji dan kapasiti arus teori ditentukan
masing-masing melalui persamaan 4 dan 5 [7]. Sampel aloi dijadikan anod dan kepingan keluli
dijadikan katod. Medium ujian ialah air laut tropika yang diambil disekitar pantai Lumut, Perak.
Sampel aloi adalah berbentuk cakera berdiameter 1.80 cm dan berketebalan 0.30 cm manakala keluli
(katod) pula dipotong berbentuk segi empat dengan dimensi 0.8 cm x 4.1 cm x 16.0 cm dengan nisbah
luas permukaan dedahan bagi anod dan katod adalah 1:15 seperti yang dilakukan oleh penyelidik
lain[8]. Sebelum ujian dimulakan semua sampel anod korbanan dan kepingan keluli dicanai
menggunakan kertas SiC sehingga grit 1200, dibasuh dengan air suling dan dibilas dengan aseton.
Kapasiti arus sebenar

Kecekapan anod = Kapasiti arus teori x100 % (3)
Q
Kapasiti arus sebenar = x1000 (Aj/kg) (4)
(M1 – M2)
dengan Q = Jumlah cas yang dipindahkan (Aj)

M1 = berat aloi sebelum ujian (kg)
M2 = berat aloi selepas ujian (kg)
(96480 kulom /3600 saat

Kapasiti arus teori = (Aj/kg) (5)
Beratara Al aloi, Kg
Berat awal setiap sampel anod korbanan direkodkan. Anod dan katod dipasang pada tangki
ujian dengan jarak antara permukaan masing-masing adalah 10 cm. Ketumpatan arus yang
dibekalkan kepada sistem ialah 0.5 A/cm2 selama 72 jam. Jumlah pemindahan cas (Aj) direkodkan
menggunakan kulometer. Selepas ujian, sampel anod dan keluli dibasuh dengan air dan dibersihkan
dengan berus kemudian dikeringkan dalam aliran udara kering. Seterusnya sampel direndamkan pula
ke dalam asid kromik (50 g/liter) selama lebih kurang 30 saat [9]. Sampel kemudian dibasuh dengan
air suling dan dibilas dengan aseton dan seterusnya dikeringkan dalam aliran udara kering. Sampel
seterusnya ditimbang bagi menentukan kehilangan jisim.
Keputusan dan Perbincangan
Komposisi sebanar aloi yang difabrikasi setelah dianalisis dengan spektrometer serapan atom adalah
seperti pada Jadual 1. Secara umumnya, komposisi sebenar aloi aluminium yang difabrikasi didapati
menghampiri nilai komposisi nominal yang digunakan. Nilai beratara aloi yang dikira berasaskan
komposisi sebenar aloi menggunakan persamaan 1 disenaraikan dalam Jadual 2.
Di samping itu, kehadiran unsur Fe dan Cu juga turut dikesan yang mungkin berpunca
daripada bendasing yang terdapat di dalam logam-logam utama yang digunakan. Beberapa kajian
terdahulu turut melaporkan kehadiran bendasing boleh membawa kesan buruk kepada sesetengah aloi
bagi kegunaan dalam perlindungan kotod dimana ia turut mempengaruhi kecekapan atau prestasi anod
korbanan[10,11]. Namun begitu, beberapa unsur pengubahsuai seperti Zn dan Mg manakala unsur
pengaktif seperti Sn ditambah kepada aloi aluminium bertujuan meningkatkan prestasi aloi tersebut.
Jadual 1: Komposisi sebenar aloi aluminium yang dianalisis menggunakan

spektrometer serapan atom.
Sampel Komposisi (% bt.)

(Komposisi nominal) Zn Mg Sn Cu Fe Al
Al-5.5Zn-2Mg 5.51 1.82 TD 0.005 0.006 Baki

Al-5.5Zn-2Mg-0.1Sn 5.49 1.85 0.07 0.004 0.008 Baki
Al-5.5Zn-2Mg-0.5Sn 5.51 1.91 0.43 0.005 0.009 Baki
Al-5.5Zn-2Mg-1.0Sn 5.52 1.89 0.96 0.002 0.008 Baki
Al-5.5Zn-2Mg-1.5Sn 5.47 1.95 1.34 0.001 0.012 Baki
Al-5.5Zn-2Mg-2.0Sn 5.52 1.79 1.95 0.019 0.032 Baki
TD : Tidak dapat dikesan
Nilai kapasiti arus sebenar, kapasiti arus teori dan kecekapan anod bagi aloi aluminium
berkenaan telah dikira dan disenaraikan pada Jadual 2. Secara umumnya menunjukkan yang
kehadiran Sn meningkatkan kecekapan anod bagi aloi Al-5.5Zn-2Mg sebelum kecekapannya kembali
menurun apabila aloi berkenaan mengandungi Sn sebanyak 1.95 %bt. Aloi Al-5.5Zn-2Mg-1.5Sn yang
mengandungi 1.34 %bt. Sn menunjukkan kecekapan anod yang paling tinggi iaitu sekitar 89.05 %
berbanding dengan aloi aluminium yang lain yang dikaji dalam kajian ini. Bagi aloi Al-5.5Zn-2.0Mg
tanpa kehadiran Sn hanya menunjukkan kecekapan 74.98% sahaja. Di samping itu, beberapa faktor
lain seperti mikrostruktur, kewujudan fasa sekunder dan pengaruh bendasing juga boleh menyumbang
kepada penurunan kecekapan [10,11]. Selain itu, prestasi aloi juga turut dikaitkan dengan kaedah
fabrikasi dimana kecacatan yang ketara kepada aloi turut mempengaruhi kapasiti arus dan kecekapan
anod tersebut [12]. Namun begitu aloi yang difabrikasi bagi kajian ini tidak mempunyai kecacatan
fizikal yang ketara seperti retak atau liang.
Jadual 2: Beratara aloi dan kecekapan anod bagi aloi berkenaan sebagai anod korbanan di
dalam air laut tropika pada suhu 27°C dan pH 8.1
Sampel Beratara Kapasiti arus sebenar Kapasiti arus teori Kecekapan

(Komposisi nominal) (g/mol) (Aj/kg) (Aj/kg) (%)
Al-5.5Zn-2Mg 9.5379 2109.12 2812.91 74.98

Al-5.5Zn-2Mg-0.1Sn 9.5368 2279.46 2809.30 81.14
Al-5.5Zn-2Mg-0.5Sn 9.5826 2335.14 2794.90 83.55
Al-5.5Zn-2Mg-1.0Sn 9.6492 2470.96 2776.98 88.98
Al-5.5Zn-2Mg-1.5Sn 9.6906 2457.03 2759.15 89.05
Al-5.5Zn-2Mg-2.0Sn 9.7743 2263.03 2741.41 82.55
Rajah 1 dan Jadual 3 menunjukkan kehadiran unsur Sn menyebabkan keupayaan kakisan aloi Al-
5.5Zn-2Mg berubah ke arah lebih negatif, ini bermakna aloi yang mengandungi Sn bersifat lebih anod
berbanding dengan aloi tanpa Sn. Kehadiran Sn juga menunjukkan berlaku peningkatan kadar kakisan
Rajah 1 Plot Tafel bagi menentukan kadar kakisan aloi Al-5.5Zn-2.0Mg-xSn

di dalam medium air laut tropika pada suhu 27 °C, pH 8.1. (x%bt.
Sn adalah komposisi nominal)
yang ketara berbanding dengan aloi Al-5.5Zn-2.0Mg tanpa Sn (Jadual 3). Kadar kakisan aloi-aloi ini
jelas menyokong keputusan yang diperolehi bagi kecekapan anod dimana aloi yang mempunyai
kecekapan yang rendah memberikan nilai kapasiti arus sebenar yang rendah. Keputusan ini
mengesahkan yang Sn memain peranan sebagai pengaktif aloi Al-5.5Zn-2.0Mg yang mana aloi yang
mengandungi 1.34 %bt. Sn di dapati mempunyai kecekapan sehingga 89.05%.
Dipercayai kehadiran Sn dalam aloi Al-5.5Zn-2.0Mg telah menyebabkan lapisan oksida
pelindung yang wujud pada permukaan aloi mengalami kecacatan dan tidak homogen. Kecacatan
lapisan oksida pelindung akan menurunkan rintangan permukaan aloi apabila dikutubkan dan
menyebabkan aloi menjadi aktif dan kadar kakisannya meningkat. Peranan Sn sebagai unsur pengaktif
aloi aluminium menjadi lebih ketara pada kepekatan yang bersesuaian di dalam aloi tersebut.
Sekiranya kandungan Sn berlebihan, kadar kakisan dan kecekapan anod akan kembali menurun.
Dalam kajian ini penurunan kecekapan anod dan kadar kakisan berlaku apabila kandungan Sn dalam
aloi berkenaan ditingkatkan kepada 1.95 %bt. (Jadual 2 dan 3). Ini kerana berlaku penomena tindak
balas katod setempat memandangkan Sn lebih bersifat katod berbanding matriks aluminium. Ini akan
menyebabkan kecekapan arus berkurang kerana sebahagian daripada elektron digunakan untuk
melindungi katod setempat tanpa dialirkan kepada logam struktur atau katod yang hendak dilindungi.
Jadual 3: Keupayaan dan kadar kakisan bagi aloi Al di dalam air laut yang
diperolehi daripada plot Tafel.
Ekakis Ikakis Kadar Kakisan

Bil Aloi (µm.tahun-1)
V(EKT) (µA.cm-2)
1 Al-5.5 Zn-2Mg -1.370 8.660 1.00 x 102
2 Al-5.5Zn-2Mg-0.1Sn -1.420 36.90 4.22 x 102
3 Al-5.5Zn-2Mg-0.5Sn -1.430 33.50 3.84 x 102
4 Al-5.5Zn-2Mg-1.0Sn -1.540 21.90 2.51 x 102
5 Al-5.5Zn-2Mg-1.5Sn -1.580 81.80 9.80 x 102
6 Al-5.5Zn-2Mg-2.0Sn -1.650 61.50 7.05 x 102
EKT = elektrod kalomel tepu
Kesimpulan
Kajian yang telah dijalankan menunjukkan kandungan Sn dalam aloi Al-5.5 Zn-2Mg memberi kesan
yang ketara terhadap sifat elektrokimia aloi aluminium. Aloi Al-5.5Zn-2Mg yang mengandungi 1.34
%bt. Sn memberikan kecekapan tertinggi berbanding komposisi lain bagi aloi aluminium yang dikaji.
Berdasarkan kepada nilai kecekapan aloi, ternyata kehadiran unsur Sn yang bertindak sebagai
pengaktif berjaya menambahkan kapasiti arus dan juga meningkatkan kadar kakisan aloi. Aloi
aluminium dengan komposisi kimia unsur-unsur utama 5.47 %bt. Zn, 1.95 %bt. Mg dan 1.34 %bt. Sn
merupakan anod korbanan paling cekap berbanding dengan aloi yang dikaji iaitu mempunyai
kecekapan anod sebanyak 89.05%.
Penghargaan:
Para penulis merakamkan penghargaan kepada Kerajaan Malaysia di atas sokongan kewangan melalui
geran penyelidikan, IRPA 09-02-02-0118-EA295. Penghargaan juga kepada Institut Penyelidikan
Sains dan Teknologi Pertahanan (STRIDE), Kementerian Pertahanan Malaysia di atas kemudahan
peralatan penyelidikan yang dibekalkan.
Rujukan
1. Lin, J. C. and Shih, H. C. (1987). “Improvement of the current efficiency of an Al-Zn-In anode
by heat treatment”. J. Electrochem. Soc. 134(4). 817-823.
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SQUARE WAVE CATHODIC STRIPPING VOLTAMMETRIC TECHNIQUE FOR
DETERMINATION OF AFLATOXIN B1 IN GROUND NUT SAMPLE
Mohamad Hadzri Yaacob1, Abdull Rahim Hj. Mohd. Yusoff2 and Rahmalan Ahamad2
1
School of Health Sciences, USM, 16150 Kubang Krian, Kelantan, Malaysia
2
Chemistry Dept., Faculty of Sciences, UTM, 81310 Skudai, Johor, Malaysia
Abstract An electroanalytical method has been developed for the detection and determination of the
2,3,6a,9a-tetrahydro-4-methoxycyclo penta[c] furo[3’,2’:4,5] furo [2,3-h][l] benzopyran-1,11-dione
(aflatoxin B1, AFB1) by a square wave cathodic stripping voltammetric (SWSV) technique on a
hanging mercury drop electrode (HMDE) in aqueous solution with Britton-Robinson Buffer (BRB) at
pH 9.0 as the supporting electrolyte. Effect of instrumental parameters such as accumulation potential
(Eacc), accumulation time (tacc), scan rate (v), square wave frequency, step potential and pulse
amplitude were examined. The best condition were found to be Eacc of -0.8 V, tacc of 100 s, v of 3750
mV/s, frequency of 125 Hz, voltage step of 30 mV and pulse amplitude of 50 mV. Calibration curve
is linear in the range of 0.01 to 0.15 uM with a detection limit of 0.125 x 10-8 M. Relative standard
deviation for a replicate measurements of AFB1 (n = 5) with a concentration of 0.01 uM was 0.83%
with a peak potential of -1.30 V (against Ag/AgCl). The recovery values obtained in spiked ground
nut elute sample were 94.00 +/- 0.67 % for 3.0 ppb, 91.22 +/- 1.56 % for 9 ppb and 92.56 +/- 2.00 %
for 15.0 ppb of AFB1. The method was applied for determination of the AFB1 in ground nut samples
after extraction and clean-up steps. The results were compared with that obtained by high
performance liquid chromatography (HPLC) technique.
Abstrak Satu kaedah elektroanalisis telah dibangunkan untuk mengesan dan menentukan 2,3,6a,9a-
tetrahydro-4-methoxycyclo penta[c] furo[3’,2’:4,5] furo [2,3-h][l] benzopyran-1,11-dione (aflatoxin
B1, AFB1) menggunakan teknik voltammetri perlucutan katodik denyut pembeza di atas elektrod
titisan raksa tergantung (HMDE) di dalam larutan akuas dengan larutan penimbal Britton-Robinson
(BRB) pada pH 9.0 bertindak sebagai larutan penyokong. Kesan parameter peralatan seperti
keupayaan pengumpulan (Eacc), masa pengumpulan (tacc), kadar imbasan (v), frekuensi gelombang
bersegi, kenaikan keupayaan dan amplitud denyut telah dikaji. Keadaan terbaik yang diperolehi
adalah Eacc; -0.8 V, tacc; 100 s, v; 3750 mV/s, frekuensi; 125 Hz, kenaikan keupayaan; 30 mV dan
amplitud denyut; 50 mV. Keluk kalibrasi adalah linear pada julat di antara 0.01 ke 0.15 uM dengan
had pengesan pada 0.125 x 10-8 M. Sisihan piawai relatif untuk 5 kali pengukuran AFB1 dengan
kepekatan 0.01uM ialah 0.83 %. Nilai perolehan semula di dalam larutan elusi sampel kacang yang
disuntik dengan 3.0 ppb, 9 ppb dan 15.0 ppb AFB1 adalah 94.00 +/- 0.67 %, 91.22 +/- 1.56 % dan
92.56 +/- 2.00 % masing-masingnya. Kaedah ini telah digunakan untuk menentukan kandungan AFB1
di dalam sampel kacang tanah selepas proses pengekstraksian dan pembersihan dijalankan. Keputusan
yang diperolehi telah dibanding dengan keputusan dari kaedah kromatografi cecair berprestasi tinggi.
Keywords: square wave stripping voltammetry, HMDE, aflatoxin B1, ground nut
Introduction
Aflatoxins (AF), the mycotoxin produced mainly by Aspergillus flavus and parasiticus and
display strong carcinogenicity [1]. They are dangerous food and contaminants and represent a
worldwide threat to public health. AFB1, B2, G1 and G2 and their metabolites M1 and M2 are the
most common, and of these, AFB1 and AFG1 are observed most frequently in food [2]. Of these,
researches have shown AFB1 (Fig 1) exhibits most toxic [3] with the order of toxicity is AFB1 >
AFG1 > AFB2 > AFG2 indicates that the terminal furan moiety of AFB1 is a critical point for
1
Corresponding address:
Chemistry Dept, Faculty of Science, UTM,
81310 Skudai, Johor, Malaysia
tel: 07-553 4492
determining the degree of biological activity of this group of mycotoxins [4]. Many countries
including Malaysia have stringent regulatory demands on the level of aflatoxins permitted in imported
and traded commodities.
O
O
O O
Figure 1 Chemical structure of AFB1
One of the foodstuffs which is most occurrence of AFB1 is ground nut. In Malaysia, the
AFB1 level in peanut is regulated with maximum level that cannot be greater than 15 ppb [5]. Several
analytical techniques for quantitative determination of the AFB1 in ground nut have been proposed
such as thin layer chromatograhpy [6], high performance liquid chromatography[ 7,8,9 ]and an
enzyme-linked immunosorbent assay, ELISA [10], All these methods, however, require specialist
equipment operated by skilled personel and expensive instruments and high maintenance cost [11].
Due to all these reasons, a voltammetric technique which is fast, accurate and require low
cost equipment [12,13] is proposed. A square wave cathodic stripping which is presented in this paper
is one of the voltammetric technique that in particularly, has a few advantages compared to other
voltammetric technique such as high speed, increased analytical sensitivity and relative insensitivity
to the presence of dissolved oxygen [14]. Previous experiment using cyclic voltammetric technique
showed that AFB1 reduced at mercury electrode and the reaction is totally irreversible [15]. This
work aimed to study and develop a SWSV method for determination of AFB1 at trace levels and to
determine this aflatoxin in ground nut samples. No such report has been published regarding this
experiment until now.
Experiment
Apparatus
Square-wave voltammograms were obtained with Metrohm 693 VA Processor coupled with a
Metrohm 694 VA stand. Three electrode system consisted of a hanging mercury drop electrode
(HMDE) was used as the working electrode, Ag/AgCl/3 M KCl reference electrode and a platinum
wire auxiliary electrode. A 20 ml capacity measuring cell was used for placing supporting electrolyte
and sample analytes. All measurements were carried out at room temperature. All pH measurements
were made with Cyberscan pH meter, calibrated with standard buffers at room temperature.
Reagents
AFB1 standard (1mg per bottle) was purchased from Sigma Co. and was used without further
purification. Stock solution (10 ppm or 3.21 x 10-5 M) in benzene:acetonitrile (98:2) were prepared
and stored in the dark at 14 0 C. The diluted solution were prepared daily by using certain volume of
stock solution, degassed by nitrogen until dryness and redissolved in Britton-Robinson buffer (BRB)
solution at pH 9.0. Britton-Robinson buffer solutions (prepared from a stock solution 0.04 M
phosphoric (Merck), boric (Merck) and acetic (Merck) acids; and by adding sodium hydroxide
(Merck) 1.0 M up to pH of 9.0. All solutions were prepared in double distilled dionised water (~ 18M
Ω cm). All chemicals were of analytical grade reagents.
Procedure
For voltammetric experiments, 10 ml of Britton-Robinson buffer solution with pH 9.0 was

placed in a voltammetric cell, through which a nitrogen stream was passed for 600 s before recording
the voltammogram. The selected Eacc = - 800 mV was applied during the tacc = 100 s while the
solution was kept under stirring. After the accumulation time had elapsed, stirring was stopped and
the selected accumulation potential was kept on mercury drop for a rest time (tr = 10 s), after which a
potential scan was performed between -1.00 as initial potential (Ei) and finished at -1.400 V as final
potential (Ef ) by SWSV technique.
Procedure for the determination in ground nut samples
AFB1 was extracted according to the standard procedure developed by Chemistry

Department, Penang Branch, Ministry Of Science, Technology and Innovation, Malaysia [16]. 1ml
of the final solution from extraction and clean-up steps in chloroform was pipette into amber bottle,
degassed with nitrogen and redissolved in 1ml of BRB solution. 200 ul of this solution was spiked
into 10 ml supporting electrolyte in volumetric cell. After that, the general procedure was applied and
voltammogram of sample was recorded. This experiment was followed by a standard addition of 10
ppb of AFB1 standard addition and voltammogram of sample with AFB1 standard was recorded.
Result and discussion
Using previous differential pulse stripping voltammetry (DPCSV) optimum parameters [17],
SWSV was run to determine 0.1 uM AFB1 in BRB pH 9.0. It gave a single reduction signal with
peak height (Ip) of 250 nA at peak potential (Ep) of -1.26 V (against Ag/AgCl). The SWS
voltammograms shows improved Ip compared to that obtained bt DPCSV where the Ip was increased
almost 4 times as shown in Figure 2.
Cyclic voltammogram of AFB1 in BRB pH 9.0 is shown in Figure 3 which only produced a
cathodic peak that indicates the non-reversibility of the electrode process [18].
Figure 2 Voltammograms of 0.1 uM AFB1 obtained by

(a) DPCSV and
(b) SWSV techniques. Experimental condition; for DPCSV: Ei = -1.0 V, Ef = -1.4 V, Eacc
= -0.6 V, tacc = 80 s, υ = 50 mV/s and pulse amplitude = 80 mV and for SWSV: Ei = -
1.0 V, Ef = -1.4 V, Eacc = -0.6 V, tacc = 80 s, frequency = 50 Hz, voltage step = 0.02,
amplitude = 50 mV and υ = 1000 mV/s.
Figure 3 Cyclic voltammogram of AFB1 in BRB pH 9.0. Experimental conditions: Ei = 0, Elow = -
1.5 V, Ehigh = 0 and scan rate = 200 mV/s.
The effect of the pH of the BRB on the stripping was studied in a pH range of 6 – 13 (Figure
4). The Ip increased slowly with increasing pH up to 8.0 followed with sharply increased for pH 8.0
to 9.0, then decreased in pH 10.0 and continuously decreased in a range of 10 – 13. Thus pH 9.0 was
chosen for the analysis. This result is not contradicted with that found by Smyth et al. (1979) when
they performed polarographic study of AFB1 [19].
300
250
200
I p (nA)
150
100
50
0
5 6 7 8 9 10 11 12 13 14
pH of BRB
Figure 4 Influence of pH of BRB on the Ip of 0.10 uM AFB1 using SWSV technique. The
instrumental parameters are the same as in Figure 2.0.
Figure 5 shows a dependence of the Ep on pH. Shifting of the Ep towards the negative
direction at higher pH implies that the reduction process takes up hydrogen ions [20]. A double bond
in aromatic ring conjugates with ketone group, in general, undergoes a reduction at mercury electrode.
The suggested mechanism of this reaction in BRB pH 9.0 is illustrated in Figure 6 as reported by
Smyth et al. (1979) [21].
1.34
1.32
1.3
1.28
Ep (-V)
1.26
1.24
1.22
1.2
1.18
5 6 7 8 9 10 11
pH
Figure 5 Relationship between Ep of AFB1 with pH of BRB
O O
O O
O
O
2e-
2 2H + 2
CH3
CH3
2 H 2O
D im e r + 2 OH-
Figure 6 Mechanism for reduction of AFB1 at mercury electrode in BRB pH 9.0
Optimisation of condition for the stripping analysis
The voltammetric determination of analytes at trace levels normally involves very small
current response. For that reason it is important to optimise all those parameters which may have an
influence on the measured current. The effect of Ei , Eacc , tacc, frequency, voltage step and amplitude
were studied. Square-wave voltammetric technique was used with stirring. For this study, 1.0 x 10-7 M
of AFB1 was spiked into supporting electrolyte.
A study of the influence of Ei showing a peak height (Ip) was obtained for Ei = -1.0 V (Figure
7). The Ip was slowly decreased for Ei more negative than -1.0 V. This value was chosen for
subsequent studies further optimisation step. Influence of Eacc to Ip of AFB1 was investigated where
the Eacc was varied between 0 to -1.4 V. The maximum value of Ip obtained at -0.8 V (366 nA) as
shown in Figure 8. This value was selected for subsequent experiments.
The dependence of Ip on tacc was studied. The effect of tacc on the Ip was studied where tacc was
varied from 0 to 160 s. The result is shown in Figure 9 which reveals that there is linearly up to 100s
according to equation y = 3.8557x + 21.383 (n=6) with R2 = 0.9936, then it increased rather slowly
leveling off at about 140 s. At 160 s, Ip decreased which may be due to the electrode saturation [21].
Thus, 100 s was appeared to be an optimum tacc for the pre-concentration prior to stripping.
300
250
200
Ip (nA) 150
100
50
0
0.2 0.4 0.6 0.8 1 1.2 1.4
Ei (-V)
Figure 7 Effect of Ei on the Ip of 0.1 uM AFB1 in BRB pH 9.0
400
350
300
25 0
Ip (nA)
200
150
100
50
0
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6
Eacc (-V)
Figure 8 Effect of Eacc on the Ip of 0.1 uM AFB1 in BRB pH 9.0
450
400
350
300
Ip (nA)
250
200
150
100
50
0
0 40 80 120 160 200
T acc (s)
Figure 9 Effect of tacc on the Ip of 0.1 uM AFB1 in BRB pH 9.0
For other instrumental condition such as square wave frequency, step potential and pulse
amplitude were examined, varying one of them and maintaining constant others. The variable ranges
were: 25 to 125 Hz for the frequency, 0.01 to 0.04 for the voltage step and 25 – 100 mV for pulse
amplitude. Generally, the Ip increase by increasing all of these instrumental parameters [22]. At
higher potential values the peak width increase; at higher frequency values the current background
increases. Finally, the condition selected were: frequency = 125 Hz, voltage step = 0.03 V and pulse
amplitude = 50 mV (Figures 10 to 12). Under optimised parameters, Ip of AFB1 was 956 nA which is
16 times higher compared to that obtained by DPCSV. Figure 13 shows voltammograms of 0.1 uM
AFB1 obtained by SWSV and DPCSV techniques.
1000
800
600
Ip (nA)
400
200
0
0 25 50 75 100 125 150
Fre que ncy (Hz)
Figure 10 Dependence of the Ip of AFB1 on SWSV frequency
1200
1000
800
Ip (nA)
600
400
200
0
0 0.01 0.02 0.03 0.04 0.05
Voltage step (V)
Figure 11 Dependence of the Ip of AFB1 on SWSV voltage step
1200
1000
800
Ip (nA)
600
400
200
0
0 0.025 0.05 0.075 0.1 0.125
Amplitude (V)
Figure 12 Dependence of the Ip of AFB1 on SWSV pulse amplitude

Figure 13 Voltammograms of AFB1 obtained by (a) DPCSV and (b) SWSV techniques in BRB pH
9.0.
Voltammetric determination of AFB1 and analytical characteristics of the method
Using the selected conditions already mentioned, a study was made of the relationship
between Ip and concentration. There is a linear relationship in the concentration range 0.01 to 0.15
uM as shown in Figure 14. Limit of detection (LOD) was 0.389 ppb (0.125 x 10-8 M) which was
determined by standard addition of low concentration of AFB1 until obtaining the sample response
that is significantly difference from blank [23]. Relative standard deviation (RSD) of the analytical
signals at several measurement (n=5) of 0.10 uM was 0.83%.
1400
1200
1000
Ip(nA)
800
600
y = 62.373x + 246.15
400
R2 = 0.9956
200
0
0 5 10 15 20
[AFB1] / 10-8 M
Figure 14 Calibration plot of AFB1 in BRB pH 9.0 obtained by SWSV technique.

Determination of AFB1 in ground nut samples
The proposed method was applied to the analysis of the AFB1 in ground nut samples.
Recovery studies were performed by spiked with difference concentration levels of AFB1 standard
into eluate of ground nut sample. In this case the concentrations used were 3 ppb (0.963 x 10-8 M), 9
ppb (2.889 x 10-8 M) and 15 ppb (4.815 x 10-8 M). The results of these studies are shown in Table 1.
For analysis of AFB1 in ground nut samples, the standard addition method was used in order to
eliminate the matrix effects. Figure 15 show voltammograms of real sample together with spiked
AFB1 standard. Table 2 listed the content of AFB1 in 6 samples obtained by proposed technique
compared with that obtained by HPLC. The results show that there are no significant different of
AFB1 content obtained by both techniques.
Table 1 Percent recovery of AFB1 spiked in real samples (n=3)
Amount added (ppb) Amount found (ppb) Recovery (%) n=3
3.00 2.82 +/- 0.02 94.00 +/- 0.67
9.00 8.21 +/- 0.14 91.22 +/- 1.56
15.00 13.88 +/- 0.30 92.53 +/- 2.00
Figure 15 SWS voltammograms of real sample (b) and spiked AFB1 (c) obtained in BRB pH 9.0 as
the supporting electrolyte (a)
Table 2 AFB1 content in ground nut samples by proposed technique compared with those obtained
by HPLC.
No of sample AFB1 content in real sample
By SWSV By HPLC
1 ND ND
2 ND ND
3 ND ND
4 9.21 8.25
5 13.92 14.34
6 36.00 36.00
Conclusion
A SWSV technique was successfully developed for the determination of AFB1 in ground nut
as an alternative method for determination of AFB1 which is sensitive, accurate and fast technique.
The results are not significant different with that obtained by accepted technique used for routine
analysis of AFB1.
Acknowledgement
One of the author would like to thank University Science Malaysia for approving his study
leave and financial support to carry out his further study at Chemistry Dept., UTM. We also indebt to
UTM for short term grant (Vot No 75152/2004) and to Chemistry Department, Penang Branch,
Ministry of Science, Technology and Innovation (MOSTI) for their help in analysis of aflatoxin in
ground nut samples using HPLC technique.
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ASSESSMENT OF ELEMENTAL POLLUTION IN THE STRAIT OF MELAKA
Awad A. Alzahrany1, 3, Ab. Khalik. H. Wood2, Elias Saion1, Md Suhaimi Elias2, Zainal Abidin
Sulaiman1, Wee Boon Siong2
1
University Puta Malaysia (UPM), College of Sciences, Department of physics, 43400 Serdang,
Selangor, Malaysia
2
Malaysia Institute for Nuclear Technology Research (MINT) BANGI, 43000
Bangi, Malaysia
3
Atomic Energy Research Institute, King Abdul-Aziz City for Science and Technology (KACST), P.O.
BOX 6086, RIYADH 11442, Saudi Arabia
ABSTRACT
In this study, 35 marine sediment samples along Strait of Melaka, representing the west coast
of peninsular of Malaysia were analyzed by using instrumental neutron activation analysis (INAA).
This work is carried out to evaluate the level of elemental pollution in the water body. In addition,
inductively coupled plasma-mass spectroscopy (ICP-MS) technique was used for non-measurable
elements by INAA. The concentration of Ag, Al, As, Ba, Br, Ca, Cd, Ce, Co, Cr, Cs, Cu, Eu, Fe, Hf,
K, La, Lu, Mg, Mn, Mo, Na, Ni, Pb, Rb, Sb, Sc, Sm, Sr, Ta, Th, Ti, U, V, Yb, and Zn were
determined in grab sediment samples of the Strait of Melaka. The total concentration of the most
elements found lower than the concentrations estimated for average shales and mean crustal materials
except As, Ag, Cd, Sb, Cs, Hf, Rb, Br, Th, U, Yb and Pb. K0 Instrumentation Neutron Activation
Analysis standardization technique is valuables in analyzing various samples types including sediment
samples. Moreover, K0-INAA is simple, flexible, accurate, and precise.
1. Introduction
Nowadays environmental pollution represents a major concern worldwide [1, 2, 3, and 4].
Especially the marine environmental pollutions consider as a dangerous pollution because of the
easiest way of its wide extend over long distance within a short period of time. The Strait of Melaka
situated at west coast of peninsular Malaysia and surrounded by four countries, which are namely
Malaysia, Singapore, Indonesia and Thailand. The length of Strait of Melaka is about 1111 Km, with
the wider width of about 407 Km near the north-west entrance and the shortest width of about 15 Km
at the south-east entrance near the Riau Archipilago [5]. Nationally, approximately 70% of the total
land of fish in Malaysia produces from Strait of Melaka [6]. Today the Strait of Melaka is one of the
most important and busiest waterways in the world. One quarter of global trade pass through the strait
everyday. Moreover, approximately a half of the world’s oil passes through the strait everyday. This
will lead to increasing the pollution in the strait, in particular the heavy metals. So, the sediments may
serve as a mean to evaluate the extent of contamination in aquatic system. The heavy metals
contamination may extent its hazards effect to aquatic life, system and human health, especially to
human consumers of seafood’s through bioaccumulation in food chain. For example between 1975
and 1993 there were 75 shipping incidents in the Strait of Melaka reported by the Marine Department
of Malaysia. Where 34 of 75 resulted in oil spills [7].
The main objectives of this research are: a) to determine the horizontal and vertical distributions
of the heavy metals pollution in strait of Melaka, b) to establish base-line data for the heavy metals
pollution in strait of Melaka, and c) To determine the sediment pollution trends. This paper will cover
only the grab sediment samples that collected from strait of Melaka.
There have been limited studies of elemental pollution along the Strait of Melaka. Din
[8] has study some trace elements concentrations of Strait of Melaka using ICP-MS and
AAS, he presented the results as only log-log plots of metal contents vs. Al contents expect
Cu, Zn and Cr he presented in range values as 3.3-18.0 ppm for Cu, 29.0-119.6 ppm for Zn,
and 10.7-69.1 ppm for Cr.
1. Experimental
1.1 Sampling and sampling preparation
In this research, 35 grab marine sediment samples along strait of Melaka from 12 sampling
location were collected. The 35 sampling location and map of Strait of Melaka are shown in Figure 1.
The coordinates of sampling locations were determined by Global Positioning System (GPS) as listed
in Table 1. The sediment samples were collected between February-March 2004.
Figure 1. Map of Strait of Melaka showing the location of collection samples

The grab sediment samples were collected from the top of about 23 cm layer of bottom sediments
using standard ponar grab sampler which has a 8.2 liter volume. The wet sediment samples were
stored in polyethylene bottles and the samples refrigerated at -5 ºC immediately after the researchers
return back to the laboratory, to prevent any change in heavy metals distribution among different
phases [9]. Then, the sediment samples dried in an oven at 65 ºC for about 5-10 days until to constant
dry weight was achieved. After that, the dried sediment samples were grinded manually and grinded
by using electronic agate homogenizer up to obtain homogenous powders of about 2mm mesh size
and stored in polyethylene bottles for analysis.
In neutron activation analysis (NAA), each dried sediment sample was shaked enough manually
and a duplicate weight ranging from 0.05 to 0.1 g for short irradiation and 0.15-0.20 g for long
irradiation were taken into a polyethylene vial and heat-sealed. On the other hand, the multi-elements
standard solutions for short and long irradiation were prepared by evaporated on filter paper in the
same polyethylene vial and heat-sealed. Moreover, to ensure accuracy and precision of geen nerated data
a proper quality contro
controll and qua
quality assu
ssurrance has been incorporated in the analytical procedures.
Accuurracy and precision of the INAA tecch hnique analysis were assured by two methods. First, by
irradiating and analyzin
analyzing certiffiied ref
eference material
als (CRM) such as
as soill--7 along with each group of
samples. As shown in Table 2, tth he average results found for IAEA-SOIL-7 Certified Reference
Material (CRM) with certified and recommended values the recoveries of thhee elements in the range of
70 % to 115 %. Second, by irradiatinng g Aluminum wire consisting of 0.1% gold (Au, diameter
0.5mm, length 5.5 mm, IRMM) along with each group of samples and apply K0-INAA
standardization technique. Also, as shown in Table 2, the average results found for IAEA-SOIL-7
Certified
ified Reference Material (CRM) M) with certified and recommended values the recoveries of the
elements in the range of 80 % to 126 %.
2.3 Irradiation facility and counting systems
2.3.1 Irradiation facility
The irradiations were performed in The TRIGA Mark II at Malaysia Institute for Nuclear
Technology Research (MINT) reactor. Reactor TRIGA II was installed by General Atomic (San
Diego, U.S.A) in 1981 and 1982 [10]. The reactor, with a maximum steady state power of 1 MWth,
consists of a circular core containing a circular grid plate and a graphite assembly surrounding the
core. In this reactor there are 40 position annular racks that rotate slowly around the core which used
for long irradiation with neutron flux of 4-5× 10ˆ¹² n/cmˆ².sec for long-lived isotopes such as As, Ba,
Ce, Co, Cr, Fe, Hf, La, Lu, Sb, Sc, Sm, Tb, Th, U, and Zn. and the pneumatic transfer system (PTS)
used for short irradiation for immediate counting of short-lived isotopes such as Al, Ti, V, Mn, Mg,
Na, K, and Ca.
In case of the short irradiation, each sample, standard solutions and certified reference
material were irradiated for a period of 30-60 second on the same position and the cooling time varies
from 5-20 minutes for first counting and the second counting after 24 hours for determination of Na
and K. For K0-INAA method three gold wires were attached to the top surface of the samples, the first
gold wire attached to the first sample, second gold wire attached to middle sample and the third gold
wire attached to the last sample to ensure and evaluate any fluctuations in neutron flux.
In case of the long irradiation, each batch which contains the sediment samples, standard
solutions, certified reference material and enough gold wires which were distributed carefully among
the batch were irradiated for 6 hours. The cooling times for first counting varies 3-6 days depending
on the dead time of the samples and 3-4 weeks for second counting.
2.3.2 Counting systems
After the sediment samples, standard solutions and certified reference materials were
irradiated for short and long irradiation. Then, the samples were analyzed spectroscopically and
gamma-ray spectra emitted from sediment samples were measured using different coaxial high
resolution hyper-pure germanium spectrometers (HpGe) with energy resolution better than 1.90 KeV
for the 1332 KeV gamma-ray line of Co-60, and with relative efficiencies ranging from 10 to 30 %.
Energy and efficiency calibration of the gamma spectroscopy systems in the energy range from 60 to
2000 KeV have been carried out in the same geometrical configuration used for measuring sediment
samples using a set of Ba-133 , Co-60 , Co-57, Cs-137, Mn-54 and Am-241 standard sources and at
different distance from the detectors.
Gamma-ray spectra emitted from the sediments samples were collected in 8000 channels of the
PC for a proper time period. The different photo peaks of measured spectra were identified and
analyzed using Gamma Vision and Genie 2000 analysis software’s.
In case of determination of heavy metals that non-measurable by INAA such as Pb, Ni, and Cd, a
Perkin-Elmer SCIEX Elan 6000 ICP-MS used for analysis of those heavy metals. A duplicate sample
of approximately 200-230 mg dry weight was transferred to a 100 ml valve Teflon beaker, 3 ml of
conc. HF and 10 ml of conc. HNO3 added to the weighed sample. The valve Teflon beaker was
closed and heated at 440 Watts inside a commercial microwave oven for 15 minutes, and then the
heated samples cooled to room temperature and reheated at 600 Watts for 20 minutes. After that, the
samples were cooled to room temperature and the valve Teflon beakers were opened carefully in fume
hood. The clear solutions transferred to 100 ml polyethylene bottle and diluted to about 50 ml by
double distilled water. A blank was carried out with each group of samples. Moreover, enough
samples from IAEA-SOIL-7 certified reference material was digested with some digestion group to
assure the accuracy of the analytical procedures. A 100 ppm of 28 multi-elements calibration standard
solution in 5% nitric acid was used for preparations of 50, 100, 250 ppb standard solutions
respectively for determination of concentration calibration curve.
The applicability of the analytical technique employed and the quality control of ICP-MS was
performed by analyzing the CRM, i.e. IAEA’s SOIL-7 obtained from IAEA. In general as shown in
Table 3, the accuracy of our measurements as compared to certified values of the CRM are
acceptable.
2. Results And Discussion
Results of the analyzed sediment samples are given in Table 4, Table 5 and Table 6 for
anthropogenic elements group, non-anthropogenic elements group and rare earth elements group,
respectively. The tables shows the measured concentrations in (%) for Al, Ca, K, Na, Fe, Ti and Mg,
while all other elements in (ppm) dry-weight. The last three rows of Table 4, Table 5, and Table 6
represent the mean, maximum, and minimum concentration for each element. Table 7, shows
comparison of elemental concentration ranges in grab sediments of Strait of Melaka with those of
Strait of Johor and with those in average shale, mean crustal material and guideline of sediment
quality of Canadian and Netherlands.
The anthropogenic elements that identified in the grab sediment samples were Ag, As, Ba,
Cd, Co, Cr, Cu, Mo, Ni, Pb, Sb, Sr, V, and Zn. In the anthropogenic group, the total concentrations of
Cr, Cu, Zn, Ni, Co, and V are lower than the concentrations estimated for average shales and mean
crustal materials while the total concentrations of Sr, Ba, and V are lower than the concentration
estimated for mean crustal materials. On the other hands, the total concentrations of As, Ag, Cd, and
Sb are greater than the concentration estimated for mean crustal materials and lower than the
concentration for average shales except Sb. Finally, the concentration of Pb is approximately two
times of the total concentrations estimated for average shales and means crustal material. This high
concentration of Pb in grab sediment samples from Strait of Melaka may due to industrial activities
such as automotive emissions.
The non-anthropogenic elements that identified in the grab sediment samples were Al, Br, Ca,
Cs, Fe, Hf, K, Mg, Mn, Na, Rb, Ta, Ti, Th, and U. The total concentrations of Fe, Mn, Al, and Ti are
lower than the concentrations estimated for average shales and mean crustal materials, while the total
concentrations of Na, K, Mg, Ta, and Ca are lower than the concentration estimated for mean crustal
materials only. On the other hands, Cs, Hf, Rb, Br, Th, and U shows a total concentrations greater
than the concentrations estimated for mean crustal materials.
The rare earth elements identified that in the grab sediment samples of Strait of Melaka were
Ce, Eu, La, Lu, Sc, Sm, and Yb. The total concentrations of Eu, Lu, and Sc are lower than the
concentrations estimated for average shales and mean crustal materials. The total concentration of Cr,
Sm is lower than the concentrations estimated for mean crustal materials and slightly above the
concentration for average shales. Finally, the total concentration of Yb is greater than the
concentrations estimated for both average shales and mean crustal materials.
3. Conclusion
The K0-INAA, a newly developed NAA quantitative technique at MINT has been
successfully tested of its capability. The K0-INAA that does not require the use of standard solutions
is a very useful technique, apart from facilitating determination of elements without standards, the
technique can also be an additional quality control of the analytical procedure.
The concentration of Ag, Al, As, Ba, Br, Ca, Cd, Ce, Co, Cr, Cs, Cu, Eu, Fe, Hf, K, La, Lu,
Mg, Mn, Mo, Na, Ni, Pb, Rb, Sb, Sc, Sm, Sr, Ta, Th, Ti, U, V, Yb, and Zn were determined in grab
sediment samples of the Strait of Melaka. The total concentration of the most of elements found lower
than the concentrations estimated for average shales and mean crustal materials except As, Ag, Cd,
Sb, Cs, Hf, Rb, Br, Th, U, Yb and Pb.
Table 1: Geographic coordinates, distance from the offshore and water depth of marine sediment
samples
SAMPLE SAMPLE LATITUDE LONGITUDE WATER Distance from

No. CODE DEPTH offshore (Km)
(m)
1 WC07A 06o 20.343' N 099o 38.256' E 9.1 2.96
2 WC07B 06o 09.90' N 099o 51.40' E 9.8 4.07
3 WC07C 06o 15.64' N 099o 57.67' E 8.0 5.93
4 WC08A 06o 10.90' N 100o 10.50' E 9.5 8.70
5 WC08B 06o 04.94' N 100o 12.91' E 11.0 7.87
6 WC08C 06o 01.79' N 100o 11.52' E 7.4 8.39
7 WC09A 05o 46.88' N 100o 10.57' E 19.0 21.48
8 WC09B 05o 42.43' N 100o 14.98' E 7.1 11.11
9 WC09C 05o 37.96' N 100o 14.11' E 7.8 11.11
10 WC10A 04o 43.54' N 100o 20.98' E 10.4 26.85
11 WC10B 04o 40.52' N 100o 22.78' E 12.9 23.71
12 WC10C 04o 38.01' N 100o 24.96' E 11.8 17.78
13 WC11A 04o 05.85' N 100o 36.12' E 7.6 7.41
14 WC11B 04o 00.25' N 100o 36.99' E 12.6 10.19
15 WC11C 03o 55.57' N 100o 39.03' E 15.2 6.85
16 WC12A 03o 50.78' N 100o 41.49' E 17.3 5.56
17 WC12B 03o 46.98' N 100o 43.94' E 20.2 8.52
18 WC12C 03o 42.38' N 100o 47.59' E 20.8 12.96
19 WC13A 03o 21.30' N 101o 07.22' E 12.0 8.33
20 WC13B 03o 19.43' N 101o 08.97' E 13.2 9.07
21 WC13C 03o 17.13' N 101o 11.10' E 17.3 4.63
22 WC14A 02o 49.16' N 101o 17.22' E 18.0 2.26
23 WC14C 02o 40.806' N 101o 26.867' E 34.2 3.70
24 WC15A 02o 20.46' N 102o 00.54' E 11.9 2.59
25 WC15B 02o 19.56' N 102o 01.98' E 10.4 2.41
26 WC15C 02o 18.10' N 102o 03.00' E 7.9 2.65
27 WC16A 02o 01.06' N 102o 26.00' E 10.2 10.93
28 WC16B 01o 59.89' N 102o 28.07' E 8.6 9.26
29 WC16C 01o 58.54' N 102o 30.54' E 6.4 6.85
30 WC17A 01o 49.18' N 102o 43.91' E 13.0 4.19
31 WC17B 01o 48.84' N 102o 44.85' E 14.6 6.15
32 WC17C 01o 48.00' N 102o 46.0' E 16.0 7.33
33 WC18A 01o 38.25' N 103o 05.30' E 8.0 4.63
34 WC18B 01o 37.27' N 103o 06.90' E 8.9 4.26
35 WC18C 01o 36.41' N 103o 08.40' E 5.2 3.89
Table 2. Comparison of concentrations determined in certified reference material (IAEA-SOIL7) with
certified or recommended values for INAA by comparative method and K0-method.
Element Certified & Measured Value Recovery Measured Value Recovery

Reco. Value mg/kg (ppm) (%) mg/kg (ppm) (%)
mg/kg (ppm) by Comparative by K0-Method
Method
Al 47000 ± 3500 41925 ± 4519 89 50000 ± 1075 106
As 13.4 ± 0.80 13.63 ± 0.56 102 13.84 ± 2.42 103
Ba 159 ± 32.5 - -
Br 7 ± 3.5 - 8.32 ± 0.84 119
Ca 163000 ± 8500 150449 ± 11584 92 205000 ± 17117 126
Ce 61 ± 6.71 77.33 ± 4.83 54.64 ± 3.32 89
Co 8.9 ± 0.89 9.27 ± 0.73 104 -
Cr 60 ± 12.6 69.00 ± 7.33 115 -
Cs 5.4 ± 0.76 - 4.31 ± 0.44 80
Cu 11 ± 1.98 - -
Dy 3.9 ± 1.05 - -
Eu 1.0 ± 0.2 - 1.18 ± 0.02 118
Fe 25700 ± 550 - 27236 ± 1670 106
Hf 5.1 ± 0.36 4.85 ± 0.23 5.94 ± 0.69 116
K 12100 ± 700 11898 ± 199 98 14700 ± 2521 121
La 28 ± 1.12 19.46 ± 3.22 70 28.74 ± 3.68 102
Lu 0.3 ± 0.15 0.27 ± 0.03 90 -
Mg 11300 ± 400 10364 ± 858 92 -
Mn 631 ± 25 605.63 ± 24.35 96 718.50 ± 6.47 114
Na 2400 ± 100 2419 ± 20 101 2770 ± 99.72 115
Nd 30 ± 6 - -
Rb 51 ± 4.59 47.87 ± 4.18 94 -
Sb 1.7 ± 0.2 1.72 ± 0.14 101 2.01 ± 0.30 118
Sc 8.3 ± 1.08 8.47 ± 0.66 102 9.52 ± 0.46 114
Sm 5.1 ± 0.36 5.84 ± 0.51 1145 -
Ta 0.8 ± 0.2 - -
Tb 0.6 ± 0.19 0.45 ± 0.11 75 -
Th 8.2 ± 1.07 8.02 ± 0.53 98 7.55 ± 0.98 92
Ti 3000 ± 550 - - -
U 2.6 ± 0.55 3.15 ± 0.29 121 -
V 66 ± 7.26 64.10 ± 1.85 97 68.62 ± 5.73 104
Yb 2.4 ± 0.36 - - 2.59 ± 0.23 108
Zn 104 ± 6.24 110.08 ± 13.47 106 -
Zr 185 ± 11.1 - - -
Table 3. Comparison of concentrations determined in standard reference material (IAEA-SOIL7) with

certified or recommended values for ICP-MS results
IAEA-SOIL7
Element Cert. or Rec. 95% Con. Average Value Recovery
Value Interval mg/kg (%)
mg/kg (ppm) mg/kg (ppm)
Zn 104 ± 6.24 101-113 144.00 ± 3.84 138
Sr 108 ± 5.4 103-114 83.43 ± 2.00 77
Ti 3000 ± 550 2600-3700 3201 ± 124 106
Ni 26 ± 8 21-37 34.66 ± 1.25 133
Cu 11 ± 1.98 9-13 14.44 ± 0.39 131
As 13.4 ± 0.80 12.5-14.2 14.24 ± 0.27 106
Ag ** ** 0.31 ± 0.01 -
Cd 1.3 ± 0.80 1.1-2.7 0.84 ± 0.01 65
Pb 60 ± 7.8 55-71 68.73 ± 0.63 114
Mo 2.5 ± 2.1 0.9-5.1 1.73 ± 0.03 69
Mn 631 ± 25 604-650 643.60 ± 15.25 102
Ba 159 ± 32.5 131-196 115.23 ± 1.39 72
Table 4. Concentrations of anthropogenic elements (ppm dry-weight) in grab sediment samples from Strait of
Melaka
Location
*Ag (ppm) *Cd (ppm) Cr (ppm) *Cu (ppm) *Mo (ppm) *Pb (ppm) Zn (ppm) *Ni (ppm)
WC07A LOD 0.03 ± 0.01 63.60 ± 17.87 14.90 ± 0.84 1.00 ± 0.03 42.09 ± 1.77 49.50 ± 12.24 31.65 ± 1.11
WC07B LOD 0.08 ± 0.01 69.84 ± 19.96 13.32 ± 0.75 1.50 ± 0.05 39.35 ± 1.78 57.46 ± 12.28 40.10 ± 1.43
WC07C 0.43 ± 0.0 1 0.21 ± 0.0 17 70.61 ± 17 .47 14.65 ± 0 .93 0.90 ± 0.0 3 54.74 ± 2.17 70.68 ± 13.64 39.60 ± 1.38
WC08A 0.21 ± 0.01 0.27 ± 0.01 46.69 ± 5.86 16.60 ± 0.88 1.00 ± 0.03 39.19 ± 1.68 63.32 ± 15.02 28.90 ± 1.09
WC08B 0.40 ± 0.0 1 0.29 ± 0.01 86.49 ± 8.87 18.51 ± 1 .15 1.55 ± 0.0 6 55.85 ± 2.24 72.36 ± 17.70 36.25 ± 1.53
WC08C 1.00 ±0.03 0.27 ± 0.01 65.54 ± 8.71 13.05 ± 0.65 1.00 ± 0.03 51.53 ± 2.35 62.16 ± 14.82 31.05 ± 1.23
WC09A 0.18 ±0.01 0.30 ± 0. 01 65.07 ± 17 .70 16.55 ± 0 .78 1.00 ± 0.03 41. 90 ± 1.92 75.58 ± 9.81 40.50 ± 1.32
WC09B 0.41 ± 0.0 1 0.22 ± 0. 01 61.58 ± 15.67 13.90 ± 0. 71 1.00 ± 0.03 52. 30 ± 2. 16 80.38 ± 12.38 33.00 ± 1.28
WC09C 0.10 ± 0.01 0.28 ± 0.01 47.36 ± 14.99 10.25 ± 0.59 1.15 ± 0.04 47.76 ± 2.32 50.91 ± 11.66 23.05 ± 1.21
WC10A 0.28 ± 0.0 1 0.25 ± 0.01 59.88 ± 5.39 12.21 ± 0 .74 0.95 ± 0.0 3 59.65 ± 2.44 44.75 ±14.17 25.30 ± 1.33
WC10B 0.54 ± 0.01 0.27 ± 0.01 55.87 ± 5.64 12.00 ± 0.86 2.00 ± 0.07 57.73 ± 2.31 60.57 ±14.29 37.00 ± 1.57
WC10C 0.45 ± 0.0 2 0.25 ± 0. 01 57.85 ± 5.44 15.40 ± 0.1.24 1.00 ± 0.0 3 57.04 ± 2. 28 107.80 ± 18.24 36.10 ± 1.36
WC11A 0.30 ± 0.0 1 0.20 ± 0 .01 45.63 ± 4.78 12.90 ± 0.61 0.90 ± 0.0 3 64.50 ± 2. 48 60.42 ± 12.90 34.50 ± 1.45
WC11B 0.44 ± 0.02 0.47 ± 0.01 48.94 ± 5.09 13.88 ± 0.72 1.00 ± 0.03 61.80 ± 2.54 97.47 ± 16.62 34.80 ± 1.44
WC11C 0.33 ± 0.0 1 0.16 ± 0.01 52.96 ± 5.46 15.10 ± 0 .83 1.00 ± 0.0 3 64.77 ± 2.44 92.30 ± 19.37 29.00 ± 1.25
WC12A 0.32 ± 0.01 0.09 ± 0.01 50.07 ± 4.58 15.10 ± 0.87 1.00 ± 0.03 34.85 ± 1.97 79.47 ± 15.52 26.80 ± 1.40
WC12B 0.30 ± 0.0 1 0.10 ± 0. 01 45.32 ± 4.07 13.30 ± 0.74 0.90 ± 0.0 3 34.70 ± 1.81 62.52 ± 13.21 25.90 ± 1.47
WC12C 0.33 ± 0.0 1 0.12 ± 0. 01 45.30 ± 4.62 16.86 ± 1. 20 1.00 ± 0.0 3 52.70 ± 2.09 65.23 ± 14.67 26.40 ± 1.64
WC13A 0.35 ± 0.01 0.10 ± 0.01 57.17 ± 5.76 9.29 ± 0.76 1.30 ± 0.05 34.40 ± 1.80 81.41 ± 12.36 23.10 ± 1.26
WC13B 0.31 ± 0.0 1 0.19 ± 0. 01 65.94 ± 7.08 14.10 ± 0 .84 1.00 ± 0.0 3 42.80 ± 1.79 97.34 ± 14.52 27.00 ± 1.34
WC13C 0.23 ± 0.01 0.06 ± 0.01 31.85 ± 4.33 7.80 ± 0.59 1.00 ± 0.03 37.46 ± 1.77 86.91 ± 12.10 20.30 ± 1.04
WC14A 0.20 ± 0.0 1 0.10 ±0.0 1 77.29 ± 6.75 8.30 ± 0. 56 0.30 ± 0.0 1 32.70 ± 1.35 57.01 ± 7.87 10.00 ± 0.72
WC14C 0.30 ± 0.01 0.09 ± 0.01 29.39 ± 3.76 8.81 ± 0.53 3.00 ± 0.11 34.60 ± 1.37 34.71 ± 7.19 13.80 ± 0.90
WC15A L OD 0.11 ± 0. 01 31.62 ± 4. 07 11.70 ± 0 .71 1.00 ± 0.03 45.46 ± 1. 87 32.04 ± 7.01 23.00 ± 1.21
WC15B 0.50 ± 0.0 1 0.14 ± 0. 01 52.72 ± 5.78 18.70 ± 1 .18 1.00 ± 0.0 3 51.76 ± 1. 91 79.23 ± 11.50 25.30 ± 1.25
WC15C 0.30 ± 0.01 0.20 ± 0.01 35.45 ± 3.91 9.90 ± 0.52 1.10 ± 0.04 34.50 ±1.42 42.43 ± 6.94 18.10 ± 0.95
WC16A 0.39 ± 0.0 1 0.13 ± 0.01 47.76 ± 4.51 12.92 ± 0.86 1.00 ± 0.0 3 40.00 ± 1 .58 74.93 ± 8.85 19.70 ± 0.92
WC16B 0.53 ± 0.02 0.11 ± 0.01 56.11 ± 7.06 21.40 ± 1.39 1.00 ± 0.04 29.60 ± 1.60 89.56 ± 12.81 28.10 ±1.44
WC16C 0.37 ± 0.0 1 LO D 78.64 ± 5. 95 12.10 ± 0.56 1.00 ± 0.0 3 25.64 ± 1.24 119.17 ± 11.23 20.00 ± 0.94
WC17A 0.27 ± 0.01 LOD 57.71 ± 4.58 10.20 ± 0. 67 1.00 ± 0.03 20.60 ± 1.32 97.17 ± 13.17 20.10 ± 0.97
WC17B 0.50 ± 0.02 LOD 57.80 ± 4.45 12.30 ± 0.74 0.90 ± 0.03 24.40 ± 1.11 110.50 ± 16.55 20.50 ± 0.96
WC17C 0.30 ± 0.0 1 LO D 59.26± 5.21 13.67 ± 0. 94 1.00 ± 0.0 3 21.50 ± 1.28 113.54 ± 15.67 21.30 ± 1.11
WC18A 0.25 ± 0.01 0.13 ± 0.01 42.35 ± 4.04 15.36 ± 0.80 0.60 ± 0.02 20.80 ±1.33 62.91 ± 10.38 18.70 ± 1.01
WC18B 0.40 ± 0.0 1 0.02 ± 0. 01 50.00 ± 5.05 12.95 ± 0 .71 1.00 ± 0.0 3 26.49 ± 1. 31 66.75 ± 9.87 24.55 ± 1.05
WC18C 0.31 ± 0.01 LOD 35.48 ± 2.99 9.47 ± 0.59 1.05 ± 0.04 25 .95 ± 1.23 55.67 ± 8.22 24.30 ± 1.42
Mean 0.36 0.17 54.43 13.36 1.09 41.75 72.98 26.79
Max. 1.00 ±0.0 3 0.47 ± 0. 01 86.49 ± 8.87 21.40 ± 1 .39 3.00 ± 0.1 1 64.77 ± 2. 44 119.17 ± 11.23 40.5 ± 1.32
Min. 0.10 ± 0.01 0.02 ± 0.01 29.39 ± 3.76 7.80 ± 0.59 0.30 ± 0.01 20.60 ± 1.32 32.04 ± 7.01 10.00 ± 0.72
*Analysis by ICP-MS; LOD = Lower Than Limit of Detection

Table 4. (cont.) Concentrations of anthropogenic elements (ppm dry-weight) in grab sediment samples
from Strait of Melaka
Location
Sb (ppm) As (ppm) *Sr (ppm) *Ba (ppm) Co (ppm) V (ppm)
WC07A 0.92 ± 0.20 5.61 ± 3.34 61.50 ± 2.61 LOD 5.80 ± 0.85 45.84 ± 7.56
WC07B 1.20 ± 0.21 8.16 ± 5.22 23.10 ± 1.05 LOD 7.04 ± 1.08 52.51 ± 8.35
WC07C 1.42 ± 0.22 11.04 ± 5.79 30.90 ± 1.27 49.00 ± 0.67 7.21 ± 1.12 55.32 ± 9.13
WC08A 1.74 ± 0.16 21.87 ± 3.04 264.30 ± 11.95 33.20 ± 0.30 4.64 ± 0.61 50.82 ± 8.52
WC08B 1.43 ± 0.14 18.06 ± 0.92 26.50 ± 1.21 61.00 ± 0.63 6.80 ± 1.59 72.99 ± 7.93
WC08C 1.56 ± 0.16 14.05 ± 1.15 43.90 ± 2.29 62.00 ± 0.69 5.51 ± 1.49 51.49 ± 6.78
WC09A 1.45 ± 0.15 4.25 ±0.70 378.50 ± 17.26 128.35 ± 0.99 10.16 ±1.50 51.37 ± 7.96
WC09B 1.46 ± 0.19 11.00 ± 5.41 20.80 ± 0.83 41.00 ± 0.31 7.05 ± 1.13 64.57 ± 10.13
WC09C 1.84 ± 0.13 2.72 ± 0.40 344.10 ± 14.48 83.10 ± 0.98 10.01 ± 1.30 50.58 ± 5.97
WC10A 0.77 ± 0.19 10.39 ± 0.89 23.60 ± 1.13 41.50 ± 0.32 7.61 ± 1.36 64.69 ± 9.21
WC10B 0.62 ± 0.19 8.29 ± 0.80 39.40 ± 1.66 64.00 ± 0.69 7.43 ± 1.48 67.71 ± 8.63
WC10C LOD 8.57 ± 0.82 38.90 ± 1.62 62.00 ± 0.65 7.36 ± 1.10 72.98 ± 7.74
WC11A 0.79 ± 0.17 12.29 ± 0.71 79.80 ± 3.63 90.00 ± 0.75 10.53 ± 1.78 53.36 ± 9.23
WC11B 0.63 ± 0.13 7.51 ± 0.62 55.30 ± 2.21 92.00 ± 0.78 7.42 ± 1.08 55.49 ± 9.78
WC11C 0.71 ± 0.22 6.04 ± 0.51 39.90 ± 1.72 51.00 ± 0.39 8.39 ± 1.55 62.81 ±10.13
WC12A 0.54 ± 0.15 5.95 ± 0.49 37.10 ± 1.84 51.00 ± 0.60 10.25 ± 0.89 70.30 ± 9.63
WC12B 0.52 ±0.11 4.14 ± 0.38 51.00 ± 2.13 61.20 ± 0.87 8.06 ± 0.78 58.15 ± 8.73
WC12C 0.41 ± 0.12 5.59 ± 0.49 73.80 ± 3.45 83.00 ± ± 0.75 6.38 ± 0.83 55.15 ± 9.65
WC13A 0.58 ± 0.08 7.44 ± 1.97 17.40 ± 0.91 43.50 ± 0.42 9.55 ± 1.10 97.43 ± 7.68
WC13B 0.68 ±0.07 6.78 ± 2.12 23.70 ±1.09 47.00 ± 0.46 9.46 ± 0.83 79.37 ± 7.96
WC13C 0.55 ± 0.12 4.80 ± 1.37 71.00 ± 3.30 113.00 ± 0.91 8.49 ± 0.72 35.93 ± 6.95
WC14A 0.92 ± 0.10 8.05 ± 0.94 12.40 ±0.74 51.00 ± 0.60 8.39 ± 0.83 39.42 ± 7.98
WC14C 0.47 ± 0.07 4.76 ± 0.77 10.20 ±0.63 33.00 ± 0.27 4.76 ± 0.48 41.36 ± 8.57
WC15A 0.68 ± 0.37 5.98 ± 1.33 104.90 ± 4.58 81.00 ± 1.22 5.65 ± 0.51 54.31 ± 7.96
WC15B 0.71 ± 0.50 8.37 ± 1.80 24.20 ± 1.18 43.00 ± 0.39 9.75 ± 0.82 43.02 ± 7.23
WC15C 0.42 ± 0.21 4.38 ± 1.25 53.00 ± 2.54 89.10 ± 0.88 5.68 ± 0.49 32.58 ± 5.95
WC16A 0.57 ± 0.27 9.25 ± 1.90 23.20 ± 0.96 47.00 ± 0.35 9.47 ± 0.67 93.46 ± 6.37
WC16B 0.99 ± 0.11 8.34 ± 1.08 44.90 ± 2.53 92.90 ± 6.98 12.63 ± 1.11 115.00 ± 5.76
WC16C 1.16 ± 0.14 12.58 ± 3.75 19.70 ± 0.94 47.00 ± 1.15 12.88 ± 0.94 71.43 ± 6.87
WC17A 0.85 ± 0.07 9.46 ± 1.14 16.70 ± 0.94 37.00 ± 0.79 11.69 ± 0.84 83.11 ± 6.31
WC17B 3.99 ± 1.30 10.60 ± 1.17 21.10 ± 10.4 50.30 ± 0.97 11.98 ± 0.99 68.38 ± 7.73
WC17C LOD 9.53 ± 0.83 17.10 ± 0.88 40.00 ± 0.31 12.26 ± 0.89 80.97 ± 6.59
WC18A 1.64 ± 0.49 8.73 ± 0.82 33.30 ± 1.87 62.60 ± 0.48 8.46 ± 0.79 63.28 ± 8.64
WC18B 2.00 ± 1.13 7.60 ± 0.71 30.10 ± 1.42 66.00 ± 0.76 8.68 ± 0.75 65.05 ± 7.89
WC18C 1.06 ± 0.86 10.03 ± 0.78 99.60 ± 4.19 48.70 ± 0.40 9.07 ± 0.82 57.37 ± 8.67
Mean 1.07 8.63 64.43 61.98 8.47 62.22
Max. 3.99 ± 1.30 21.87 ± 3.04 378.50 ± 17.26 128.35 ± 0.99 12.88 ± 0.94 115.00 ± 5.76
Min. 0.41 ± 0.12 2.72 ± 0.40 10.20 ± 0.63 33.00 ± 0.27 4.64 ± 0.61 32.58 ± 5.95

Table 5. Concentrations of non-anthropogenic elements (ppm dry-weight) in grab sediment samples from
Strait of Melaka
Location
Cs (ppm) Fe (%) Hf (ppm) K (%) Mg (%) Mn (ppm) Na (%) Al (%)
WC07A 9.00 ± 1.12 1.84 ± 0.10 6.45 ± 1.12 1.53 ± 0.20 1.49 ± 0.32 253 ± 27.28 2.32 ± 0.24 4.75 ± 0.36
WC07B 11.79 ± 1.25 2.72 ± 0.28 7.79 ± 1.21 1.69 ± 0.11 2.43 ± 0.47 260 ± 27.13 2.61 ± 0.27 7.83 ± 0.53
WC07C 12.25 ± 1.15 2.63 ± 0.22 7.43 ± 0.63 1.60 ± 0.14 2.91 ± 0.41 293 ± 30.38 3.04 ± 0.29 6.80 ± 0.48
WC08A 7.26 ± 0.69 1.88 ± 0.21 5.06 ± 0.49 0.87 ± 0.11 1.66 ± 0.39 148 ± 19.62 1.99 ± 0.20 3.84 ± 0.30
WC08B 14.94 ± 1.96 3.00 ± 0.26 5.21 ± 1.12 1.93 ± 0. 21 2.31 ± 0.45 231 ± 33.20 3.23 ± 0.33 9.39 ± 0.63
WC08C 10.48 ± 1.30 2.39 ± 0.23 9.68 ± 0.69 1.49 ± 0.23 1.69 ± 0.34 188 ± 20.75 2.45 ± 0.23 6.68 ± 0.46
WC09A 7.81 ± 0.69 2.89 ± 0.38 5.26 ±0.57 1.61 ± 0.20 1.64 ± 0.38 1013 ± 99.12 2.12 ± 0.09 7.33 ± 0.56
WC09B 11.88 ± 1.13 3.11 ± 0.28 6.80 ± 0.80 1.59 ± 0.13 1.58 ± 0.33 239 ± 28.63 2.70 ± 0.26 8.29 ± 0.55
WC09C 6.55 ± 0.72 1.89 ± 0.25 9.25 ± 0.97 1.25 ± 0.09 1.31 ± 0.31 233 ± 26.89 2.15 ± 0.21 7.60 ± 0.47
WC10A 11.71 ± 1.50 3.12 ± 0.29 7.67 ± 0.58 1.64 ± 0.12 1.43 ± 0.29 385 ± 39.12 2.73 ± 0.06 8.05 ± 0.55
WC10B 10.82 ± 1.28 2.81 ± 0.29 6.98 ± 0.57 1.57 ± 0.14 1.33 ± 0.29 379 ± 39.58 2.81 ± 0.28 8.85 ± 0.67
WC10C 11.59 ± 1.51 3.09 ± 0.30 6.61 ± 0.66 1.68 ± 0.13 2.08 ± 0.44 420 ± 43.58 3.49 ± 0.34 9.49 ± 0.64
WC11A 9.90 ± 1.29 4.43 ± 0.40 9.14 ± 0.64 1.69 ± 0.15 1.47 ± 0.24 401 ± 38.41 2.13 ± 0.21 8.86 ± 0.57
WC11B 10.43 ± 1.38 2.48 ± 0.25 9.67 ± 0.58 1.66 ± 0.12 2.22 ± 0.73 402 ± 39.03 2.40 ± 0.23 8.14 ± 0.54
WC11C 10.72 ± 1.37 2.68 ± 0.25 5.95 ± 0.57 1.60 ± 0.09 2.31 ± 0.57 555 ± 58.51 3.79 ± 0.09 9.42 ± 0.65
WC12A 11.23 ± 1.39 2.66 ± 0.25 4.56 ± 0.50 1.62 ± 0.15 2.87 ± 0.60 502 ± 61.30 3.12 ± 0.43 9.24 ± 0.71
WC12B 9.49 ± 1.06 2.49 ± 0.84 4.71 ± 0.45 1.57 ± 0.16 2.14 ± 0.45 506 ± 52.72 2.80 ± 0.38 8.42 ± 0.64
WC12C 9.22 ± 1.13 2.20 ± 0.20 4.91 ± 0.36 1.49 ± 0.15 1.08 ± 0.57 409 ± 43.83 2.78 ± 0.35 7.41 ± 0.58
WC13A 12.00 ± 1.12 2.76 ± 0.09 8.47 ± 0.65 1.79 ± 0.11 1.60 ± 0.35 369 ± 40.64 2.47 ± 0.25 11.58 ± 0.84
WC13B 14.15 ± 1.50 3.32 ± 0.11 8.44 ± 0.57 1.59 ± 0.09 1.84 ± 0.50 384 ± 42.81 2.90 ± 0.30 11.79 ± 0.65
WC13C 6.48 ± 0.70 3.65 ± 0.12 14.03 ± 0.77 1.40 ± 0.19 1.47 ± 0.35 302 ± 36.39 1.26 ± 0.18 6.20 ± 0.32
WC14A 7.21 ± 0.86 2.41 ± 0.07 26.91 ± 1.12 0.97 ± 0.35 0.81 ± 0.21 520 ± 49.82 1.01 ± 0.10 4.22 ± 0.32
WC14C 5.34 ± 0.67 1.32 ± 0.04 9.72 ± 0.55 1.11 ± 0.10 0.65 ± 0.28 184 ± 24.42 1.41 ± 0.13 4.03 ± 0.30
WC15A 6.00 ± 0.86 1.74 ± 0.06 8.49 ± 0.48 0.93 ± 0.37 1.44 ± 0.31 310 ± 31.62 1.15 ± 0.12 6.76 ± 0.59
WC15B 10.01 ± 1.27 2.71 ± 0.09 6.37 ± 0.39 1.53 ± 0.12 1.53 ± 0.22 317 ± 35.77 2.37 ± 0.23 6.84 ± 0.43
WC15C 7.41 ± 0.74 1.58 ± 0.05 9.40 ± 0.54 1.24 ± 0.11 1.14 ± 0.30 247 ± 27.48 1.14 ± 0.15 7.15 ± 0.37
WC16A 9.65 ± 0.77 2.44 ± 0.07 6.18 ± 0.33 1.52 ± 0.14 1.92 ± 0.42 297 ± 34.76 1.95 ± 0.23 10.39 ± 0.84
WC16B 11.22 ± 1.32 3.10 ± 0.10 6.08 ± 0.51 1.70 ± 0.21 2.45 ± 0.57 326 ± 37.12 2.32 ± 0.23 12.90 ± 0.93
WC16C 10.83 ± 1.15 3.86 ±0.11 7.42 ± 0.54 1.83 ± 0.19 3.13 ± 0.47 332 ± 38.23 2.49 ± 0.29 8.49 ± 0.59
WC17A 9.31 ± 0.94 3.08 ± 0.11 6.49 ± 0.49 1.30 ± 0.11 2.30 ± 0.39 336 ± 40.90 2.34 ± 0.28 9.69 ± 0.74
WC17B 10.22 ± 1.01 3.42 ± 0.10 6.69 ± 056 0.93 ± 0.08 1.70 ± 0.42 330 ± 39.11 2.24 ±0.26 8.35 ± 0.61
WC17C 10.76± 1.03 3.37 ± 0.11 7.21 ± 0.63 1.85 ± 0.18 2.11 ± 0.63 373 ± 45.97 2.83 ± 0.33 8.90 ± 0.64
WC18A 6.55 ± 0.81 2.68 ± 0.08 7.75 ± 0.46 1.11 ± 0.21 1.56 ± 0.43 272 ± 35.94 1.54 ± 0.18 5.99 ± 0.52
WC18B 7.81 ± 0.93 2.58 ± 0.07 9.34 ± 0.63 1.03 ± 0.22 1.87 ± 0.48 238 ± 31.20 1.90 ± 0.22 7.87 ± 0.63
WC18C 4.94 ±0.56 2.94 ± 0.09 9.93 ± 0.49 0.79 ± 0.23 0.92 ± 0.26 302 ± 34.13 1.03 ± 0.12 6.47 ±0.47
Mean 9.63 2.72 8.06 1.45 1.78 350 2.31 7.94
Max. 14.94 ± 1.96 4.43 ± 4.01 26.91 ± 1.12 1.93 ± 0.21 3.13 ± 0.47 1013 ± 99.12 3.79 ± 0.09 12.9 ± 0.93
Min. 4.94 ±0.56 1.32 ± 0.44 4.56 ± 0.50 0.79 ± 0.23 0.65 ± 0.28 148 ± 19.62 1.01 ± 0.10 3.84 ± 0.30
Table 5. (cont.) Concentrations of non-anthropogenic elements (ppm dry-weight) in grab sediment
samples from Strait of Melaka
Location
Rb (ppm) Ca (%) Br (ppm) Ta (ppm) *Ti (%) Th (ppm) U (ppm)
WC07A 120.79 ± 32.11 1.25 ± 0.23 124.72 ± 2.95 1.23 ± 0.28 0.45 ± 0.01 20.35 ± 2.45 7.30 ± 1.12
WC07B 142.03 ± 38.53 1.18 ± 0.24 215.13 ± 8.70 1.54 ± 0.28 0.32 ± 0.02 22.10 ± 2.87 8.70 ± 1.67
WC07C 115.34 ± 33.65 1.20 ± 0.27 226.83 ± 9.27 1.46 ± 0.30 0.31 ±0.02 21.40 ± 2.76 7.71 ± 1.60
WC08A 73.92 ± 21.09 15.13 ± 0.98 318.99 ± 13.56 1.09 ± 0.48 0.24 ± 0.02 13.46 ± 1.62 14.21 ± 1.87
WC08B 153.17 ± 38.69 0.96 ± 0.27 167.62 ± 5.71 1.93 ± 0.35 0.32 ± 0.02 23.50 ± 2.48 10.03 ± 1.31
WC08C 83.80 ± 21.22 1.54 ± 0.32 188.36 ± 8.76 1.44 ± 0.57 0.28 ± 0.02 18.56 ± 1.72 9.43 ± 0.96
WC09A 109.52 ± 32.54 8.57 ± 0.69 110.85 ± 4.04 1.18 ± 0.51 0.25 ± 0.03 17.22 ± 2.24 17.09 ± 2.19
WC09B 129.05 ± 40.70 1.83 ± 0.26 134.09 ± 4.84 1.85 ± 0.77 0.37 ± 0.02 25.46 ± 2.05 9.86 ± 1.24
WC09C 77.09 ± 27.78 11.16 ± 0.85 82.50 ± 2.00 1.60 ± 0.59 0.25 ± 0.02 17.25 ± 1.78 10.80 ± 1.43
WC10A 104.96 ± 18.59 1.21 ± 0.26 169.49 ± 5.37 1.56 ± 0.39 0.33 ± 0.03 28.56 ± 1.96 4.92 ± 0.93
WC10B 106.45 ± 17.97 1.37 ± 0.26 135.89 ± 4.14 1.72 ± 0.43 0.37 ± 0.03 26.80 ± 1.86 4.19 ± 1.56
WC10C 107.15 ± 18.29 0.87 ± 0.26 178.95 ± 6.06 < 0.90 0.42 ± 0.03 28.04 ± 2.00 6.51 ± 1.54
WC11A 95.85 ± 16.16 2.23 ± 0.38 111.03 ± 6.40 1.58 ± 0.42 0.27 ± 0.02 27.91 ± 1.86 5.51 ± 1.62
WC11B 104.71 ± 15.17 2.33 ± 0.32 128.28 ± 2.98 1.40 ± 0.35 0.30 ± 0.02 24.60 ± 1.84 5.89 ± 1.14
WC11C 97.67 ± 13.33 2.15 ± 0.41 140.96 ± 5.21 LOD 0.31 ± 0.01 26.74 ± 1.77 4.16 ± 1.41
WC12A 88.59 ± 12.66 2.13 ± 0.38 105.08 ± 3.24 LOD 0.31 ± 0.02 25.86 ± 1.83 3.00 ± 0.99
WC12B 92.09 ± 16.24 2.79 ± 0.41 86.50 ± 3.03 1.35 ± 0.35 0.27 ± 0.02 22.76 ± 1.57 3.80 ± 0.89
WC12C 68.77 ± 11.07 LOD 146.80 ± 3.54 LOD 0.31 ± 0.02 19.46 ± 1.34 5.18 ± 0.69
WC13A 134.38 ± 38.21 0.76 ± 0.21 129.22 ± 6.73 1.75 ± 0.27 0.30 ± 0.02 26.65 ± 1.81 4.48 ± 1.10
WC13B 163.71 ± 47.04 0.77 ± 0.25 148.13 ± 7.37 1.78 ± 0.31 0.35 ± 0.03 30.28 ± 2.31 4.10 ± 1.12
WC13C 131.19 ± 32.42 1.91 ± 0.29 54.54 ± 3.30 1.33 ± 0.23 0.23 ± 0.01 23.36 ± 1.65 4.39 ± 1.10
WC14A 71.05 ± 12.41 0.22 ± 0.11 44.62 ± 1.60 1.78 ± 0.32 0.27 ± 0.02 37.38 ± 2.09 9.39 ± 1.69
WC14C 70.96 ± 10.95 0.73 ± 0.24 37.55 ± 1.42 1.10 ± 0.32 1.11 ± 0.07 12.26 ± 0.75 2.73 ± 0.96
WC15A 119.90 ± 31.70 1.93 ± 0.36 45.60 ± 6.40 5.96 ± 0.77 0.68 ± 0.05 18.82 ± 1.14 6.14 ± 1.03
WC15B 177.29 ± 45.10 1.42 ± 0.31 97.10 ± 5.09 1.34 ± 0.19 0.51 ± 0.04 19.26 ± 1.27 3.91 ± 0.93
WC15C 96.19 ± 28.58 1.82 ± 0.29 45.29 ± 2.46 0.95 ± 0.14 0.28 ± 0.02 14.66 ± 1.08 3.35 ± 0.94
WC16A 157.00 ± 41.35 1.31 ± 0.25 90.00 ± 4.03 1.15 ± 0.45 0.33 ± 0.02 18.05 ± 1.14 3.40 ± 0.89
WC16B 156.53 ± 40.61 1.21 ± 0.26 102.57 ± 4.29 1.36 ± 0.45 0.30 ± 0.02 19.80 ± 0.67 4.56 ± 1.12
WC16C 91.99 ± 13.12 1.01 ± 0.25 144.39 ± 8.04 1.54 ± 0.37 0.34 ± 0.03 30.93 ± 1.56 7.01 ± 0.98
WC17A 103.34 ± 18.91 1.34 ± 0.31 94.43 ± 3.51 1.17 ± 0.38 0.29 ± 0.02 21.73 ± 1.27 4.20 ± 1.06
WC17B 147.13 ± 35.04 1.15 ± 0.30 97.53 ± 4.27 1.85 ± 0.92 0.34 ± 0.02 25.06 ± 1.27 6.53 ± 1.07
WC17C 132.14 ± 27.17 0.86 ± 0.27 103.31 ± 5.68 1.75 ± 0.86 0.31 ± 0.02 26.52 ± 1.34 4.59 ± 1.10
WC18A 90.56 ± 24.73 1.59 ± 0.34 82.24 ± 23.74 1.22 ± 0.81 0.27 ± 0.02 18.86 ± 1.27 5.68 ± 0.93
WC18B 100.94 ± 24.93 1.28 ± 0.31 88.91 ± 6.40 1.51 ± 0.75 0.34 ± 0.03 20.12 ± 1.40 6.67 ± 1.21
WC18C 75.25 ± 21.40 3.19 ± 0.47 42.80 ± 13.41 1.30 ± 0.58 0.26 ± 0.02 13.26 ± 0.97 6.23 ± 0.97
Mean 111.07 2.36 120.58 1.61 0.35 22.49 6.45
Max. 177.29 ± 45.10 15.13 ± 0.98 318.99 ± 13.56 5.96 ± 0.77 1.11 ± 0.07 37.38 ± 2.09 17.09 ± 2.19
Min. 68.77 ± 11.07 0.22 ± 0.11 37.55 ± 1.42 0.95 ± 0.14 0.23 ± 0.01 12.26 ± 0.75 2.73 ± 0.96

Table 6. Rare earth elements concentrations (ppm dry-weight) in grab sediment samples from Strait of
Melaka
Sm (ppm) Ce (ppm) Eu (ppm) La (ppm) Lu (ppm) Sc (ppm) Yb (ppm)

WC07A 5.74 ± 0.50 75.92 ± 5.68 0.90 ± 0.55 34.04 ± 4.66 0.88 ± 0.30 7.39 ± 0.50 2.80 ± 0.47
WC07B 6.71 ± 0.60 77.68 ± 4.56 1.05 ±0.11 43.54 ± 4.78 1.29 ± 0.42 10.91 ± 0.82 4.02 ± 0.67
WC07C 6.47 ± 0.57 89.25 ± 6.13 1.07 ± 0.70 43.66 ± 4.87 1.13 ± 0.37 10.87 ± 0.81 4.04 ± 0.85
WC08A 4.40 ± 0.39 51.23 ± 4.40 0.71 ± 0.13 103.45 ± 9.67 0.29 ± 0.08 6.47 ± 0.51 6.15 ± 0.84
WC08B 6.11 ± 0.54 96.91 ± 7.51 1.09 ± 0.49 69.94 ± 5.69 0.08 ± 0.03 11.92 ± 0.96 4.73 ± 0.82
WC08C 6.17 ± 0.55 89.68 ± 7.64 1.07 ± 0.12 70.19 ± 7.38 0.20 ± 0.06 8.90 ± 0.66 5.34 ± 0.87
WC09A 6.32 ± 0.58 65.35 ± 5.30 1.18 ± 0.12 98.66 ± 8.63 LOD 8.48 ±0.72 6.56 ± 0.91
WC09B 6.63 ± 0.59 97.98 ± 8.61 1.28 ± 0.24 68.05 ± 5.97 LOD 11.18 ± 0.84 4.94 ± 0.85
WC09C 4.38 ± 0.39 77.24 ± 6.53 1.00 ± 0.46 64.56 ± 7.39 LOD 6.54 ± 0.52 5.52 ± 0.93
WC10A 8.75 ± 0.78 106.82 ± 6.80 0.99 ± 0.20 48.13 ± 6.97 0.34 ± 0.13 12.71 ± 1.47 6.36 ± 1.36
WC10B 7.75 ± 0.68 90.2 ± 6.54 1.10 ± 0.13 44.58 ± 6.24 0.28 ± 0.11 12.66 ± 1.53 3.33 ± 0.79
WC10C 7.26 ± 0.64 107.58 ± 7.02 1.05 ± 0.14 46.34 ± 6.56 0.43 ± 0.16 11.40 ± 1.33 3.76 ± 1.18
WC11A 7.48 ± 0.66 89.09 ± 6.70 0.98 ± 0.12 42.81 ± 4.10 0.38 ± 0.16 9.19 ± 1.06 3.77 ± 0.92
WC11B 7.04 ± 0.62 105.89 ± 7.86 0.81 ± 0.18 44.18 ± 4.55 0.33 ± 0.11 9.31 ± 0.79 3.75 ± 0.86
WC11C 5.09 ± 0.63 100.25 ± 6.54 0.88 ± 0.18 40.61 ± 3.89 0.52 ± 0.14 9.70 ± 0.80 3.33 ± 0.74
WC12A 4.81 ± 0.43 92.25 ± 7.44 1.07 ± 0.13 37.23 ± 5.26 0.57 ± 0.18 10.70 ± 0.88 3.31 ± 0.80
WC12B 4.46 ± 0.39 90.99 ± 6.17 0.93 ± 0.12 27.93 ± 2.63 LOD 8.89 ± 0.72 3.39 ± 0.74
WC12C 5.74 ± 0.51 68.78 ± 4.90 0.76 ± 0.18 32.25 ± 3.09 0.31 ± 0.11 8.43 ± 0.73 2.64 ± 0.63
WC13A 7.49 ± 1.47 89.24 ± 6.30 1.12 ± 0.11 42.14 ± 4.39 0.57 ± 0.12 11.71 ± 0.89 3.07 ± 0.84
WC13B 6.80 ± 1.43 107.00 ± 8.25 1.30 ± 0.13 40.37 ± 4.42 0.48 ± 0.10 13.23 ± 1.18 3.71 ± 0.72
WC13C 4.72 ± 1.01 52.37 ± 3.61 0.68 ± 0.08 28.63 ± 3.20 0.48 ± 0.08 5.92 ± 0.48 3.91 ± 0.83
WC14A 8.84 ± 0.72 152.60 ± 9.52 0.83 ± 0.07 52.60 ± 5.22 0.90 ± 0.23 8.99 ± 0.72 13.16 ± 2.07
WC14C 2.87 ± 0.26 47.86 ± 4.30 0.61 ± 0.09 18.11 ± 1.86 0.30 ± 0.05 5.38 ± 0.43 2.74 ± 0.53
WC15A 5.77 ± 1.31 69.57 ± 5.38 0.93 ± 0.11 28.49 ± 2.79 1.24 ± 0.24 7.45 ± 0.61 2.85 ± 0.58
WC15B 6.30 ± 1.08 73.83 ± 5.54 1.05 ± 0.11 35.54 ± 3.67 0.40 ± 0.09 11.69 ± 0.99 2.46 ± 0.54
WC15C 4.01 ± 0.96 55.86 ± 4.16 0.92 ± 0.07 25.05 ± 2.65 0.36 ± 0.07 6.89 ± 0.55 1.74 ± 0.37
WC16A 6.32 ± 1.24 65.34 ± 4.34 1.06 ± 0.10 38.41 ± 3.91 0.31 ± 0.07 10.73 ± 0.70 2.50 ± 0.56
WC16B 6.46 ± 0.66 95.64 ± 7.83 1.19 ± 0.11 40.23 ± 4.27 0.48 ± 0.08 12.24 ± 1.06 3.06 ± 0.75
WC16C 8.51 ± 0.65 113.61 ± 6.68 1.45 ± 0.13 48.28 ± 4.04 0.54 ± 0.08 15.53 ± 0.94 3.77 ± 0.62
WC17A 6.21 ± 0.55 88.95 ± 5.92 1.14 ± 0.10 37.63 ± 3.62 0.44 ± 0.07 12.40 ± 0.97 3.95 ± 0.67
WC17B 7.43 ± 0.16 107.95 ± 7.54 1.68 ± 0.17 52.25 ± 8.88 0.43 ± 0.24 14.47 ± 1.11 3.66 ± 0.74
WC17C 7.25 ± 0.64 103.56 ± 6.27 1.52 ± 0.13 49.23 ± 4.69 0.45 ± 0.21 14.29 ±1.23 3.85 ± 0.72
WC18A 6.68 ± 0.15 72.81 ± 5.71 1.01 ± 0.12 30.02 ± 3.79 0.27 ± 0.12 10.34 ± 0.79 3.83 ± 0.78
WC18B 5.85 ± 0.16 83.11 ± 6.13 1.08 ± 0.14 38.69 ± 6.26 0.43 ± 0.17 11.17 ± 0.89 4.24 ± 1.15
WC18C 4.05 ± 0.12 61.21 ± 4.05 0.74 ± 0.09 28.09 ± 2.80 0.26 ± 0.11 7.07 ± 0.57 1.98 ± 0.64
Mean 6.40 86.10 1.04 45.54 0.49 10.15 4.06
Max. 8.84 ± 0.72 152.60 ± 9.52 1.68 ± 0.17 103.45 ± 9.67 1.29 ± 0.42 15.53 ± 0.94 13.16 ± 2.07
Min. 2.87 ± 0.26 47.86 ± 4.30 0.61 ± 0.09 18.11 ± 1.86 0.08 ± 0.03 5.38 ± 0.43 1.74 ± 0.37

Table 7 Comparison of elemental concentration ranges in grab sediments of Strait of Melaka with
those of Strait of Johor and with those in average shale, mean crustal material and guideline of
sediment quality of Canadian and Netherlands[11,12,13,14,15]
Strait of Johor Strait of Melaka Average Shales Mean Crustal Materials Sediment GL
Element Range Average Range Average B, M&M or T&W M&M B Canadian Netherlands
Al 3.03-12.19 8.25 ± 2.49 3.84-12.90 7.94 8.0-8.8 8.13 8.20

Ag 0.10-1.00 0.36 0.07
As 6.40-64.0 19.7 ± 6.90 2.72-21.87 8.63 13 1.8 1.50 - 29
Ba - - 33.0-128 61.98 - 425
Br 37.55-319 120.58 2.5
Ca 0.22-15 2.36 3.63
Cd 0.11-0.36 0.18 ± 0.06 0.02-.47 0.17 0.30 0.20 0.11 0.7 0.8
Ce 34.80-97.20 69.3 ± 17.30 47.86-152 86.10 50-96 60 68
Co 1.80-8.10 5.80 ± 1.50 4.64-12.88 8.47 19 25 20
Cr 21.9-62.8 45.20 ± 11.2 29.39-86.49 54.43 90 100 100 52.3 -
Cs - - 4.94-14.94 9.63 - 3.0 -
Cu 10.8-92.9 30.7 ± 22.5 7.80-21.40 13.36 45 55 50 - 36
Eu 0.49-1.20 0.79 ± 0.20 0.61-1.68 1.04 1.0-1.20 1.2 2.1
Fe 1.53-4.13 3.04 ± 0.67 1.32-4.43 2.72 4.7-4.8 5.0 4.10
Hf - - 4.56-26.91 8.06 3.0
K - - 0.79-1.93 1.45 2.59
La 13.3-65.1 32.1 ± 9.9 18.11-103 45.54 24-92 30 32
Lu 0.25-0.47 0.40 ± 0.07 0.08-1.29 0.49 0.6-0.8 0.5 0.51
Mg - - 0.65-3.13 1.78 2.09
Mn 113-861 265 ±152 148-1013 350 850 950 950
Mo - - 0.30-3.00 1.09 1.50
Na - - 1.01-3.79 2.31 2.83
Ni 21.2-46.8 30.2 ± 6.6 10.0-40.5 26.79 68 75 80
Pb 26.4-69.9 42.3 ± 11.0 20.6-64.77 41.75 20 13 14 30.2 85
Rb 68.8-177 111 90
Sb 0.73-2.73 1.56 ± 0.55 0.41-3.99 1.07 1.5 0.2 0.2
Sc 3.0-13.7 9.9 ± 2.8 5.38-15.53 10.15 13 22 16
Sm 2.1-16.6 7.4 ± 3.4 2.87-8.84 6.40 6.2 6.0 7.9
Sr - - 10.2-378.5 64.43 375
Ta - - 0.95-5.96 1.61 2.0
Th 7.5-64.3 21.2 ± 6.6 12.26-37.38 22.49 12 7.2 12
Ti 0.25-.53 0.40 ± 0.07 0.23-1.11 0.35 0.46 0.44 0.56
U 2.4-7.3 5.2 ± 1.3 2.73-17.09 6.45 3.7 1.8 2.4
V 29.2-118.9 72.5 ± 22.5 32.58-115.0 62.22 130 135 160
Yb 1.3-401 2.7 ± 0.7 1.74-13.16 4.06 2.4 3.4 3.3
Zn 68.5-230.7 132.5 ± 52.6 32.04-119.17 72.98 95 70 75 124 140
B= Bowen (1979); M&M = Mason and Moore (1982); T&W = Turekian and Wedepohl(1961).
References
1. Fukue, M., Nakamura, T., Kato, Y., Yamasaki, S. (1999) “Degree of Pollution for Marine Sediments”
Engineering Geology. 53. 131-137.
2. Fukue, T., Kato, Y., Nakamura, T., Yamasaki, S. (1995) “Heavy Metals Concentration in Bay
Sediments of Japan” STP. 1293. ASTM. 58-73.
3. Morillo, J., Usero, J., Gracia, I. (2004) “Heavy Metal Distribution in Marine sediments fro the
Southwest Coast of Spain” Chemoshere. 55. 431-442.
4. Szefer, P., Szefer, K., Glasby, GP., Pempkowiak, J., Kaliszan, R. (1996) “Heavy Metal Pollution in
Surficial sediments from the southen Baltic Sea off poland” J. Environ Sci Health. A. 31. 2723-2754.
5. Thia-Eng., C., Ross, S., Yu, H. (1997) “Malacca Straits Environmental Profile”. Published by the
GEF/UNDP/IMO Regional programme for Prevention and Management of Marine Pollution in the
East Asian Seas. Quezon City, Philippine.
6. Agusa, T., Kunito, T., Yasunaga, G., Iwata, H., Subramanian, A., Ismail, A., Tanabe, S (2005) “
Concentrations of trace elements in marine fish and its risk assessment in Malaysia” Marine Pollution
Bulletin. Article in Press.
7. Ahmed, H. (1997) “The Straits of Malacca: International Co-Operation in Trade, Funding and
Navigational Safety”, Academe Art & Printing Services Sdn. Bhd.
8. Din, Z. (1992) “Use of Aluminum to normalize heavy-Metal Data from Estuarine and Coastal
Sediments of Straits of Melaka. Marine Pollution Bulletin 24, 484-491.
9. Yuan, C., Shi, J., He, B., Liu, J., Liang, L., and Jiang, G. (2004) “Speciation of Heavy Metals in
Marine Sediments from the East China Sea by ICP-MS with Sequential Extraction” Environmental
International. 30. 769-783.
10. Bokhari,A., Kassim, M. S.(2005) “Operation and Maintenance of 1MW PUSATI TRIGA Reactor”
The 2004 Workshop on the Utilization of Research Reactors. Bangkok, Thailand, 17-21 January 205.
11. Wood, A. K. H., Ahmed, Z., Shazili, N. A. MD., Yaakob, R., and Carpenter, R. (1997) “
Geochemistry of sediments in Johor Strait between Malaysia and Singapore” Continental Shelf
Research. 17. 10. 1207-1228.
12. Bowen, H.J. M. (1979) “Environmental Chemistry of Elements”, London, Academic Press.
13. Mason, B.J., and Moore, C. B. (1982) “Principles of Geochemistry”, 4th ed., New York, J. Wiley and
Sons.
14. Turekian, K. K., and Wedepohl, K. H. (1961) “Distribution of the Elements in Some Units of the
Earth’s Crust”. Bulletin of the Geological Society of America. 72. 175-192.
15. International Atomic Energy Agency (1992) “Sampling and Analytical Methodologies for
Instrumental Neutron Activation Analysis of Airborne Particulate Matter”.IAEA-TCS-4. Vienna.
CHARACTERIZATION OF TWO DIFFERENT LIQUID SCINTILLATION COUNTERS:
EFFECTS ON AGE DETERMINATION IN RADIOCARBON DATING SYSTEM
Noraishah Othman, Kamisah Alias, Nasasni Nasrul, Azwah Jaafar
Tracer Application Group (TAG)

Industrial Technology Division (BTI)
Malaysian Institute For Nuclear Technology Research (MINT)
Bangi, 43000 Kajang, Selangor, Malaysia
Email: noraishah@mint.gov.my
Abstract
Age estimation of the radiocarbon samples like geological, hydrological, environmental and
archaeological samples are conducted by using simple program written in Visual Basic, which we
adopt from UCD Radiocarbon Laboratory, Dublin. Initially, the radioactivity counts were derived
from Liquid Scintillation Analyzer Series TRI-CARB Model 2770TR/SL. The counted activities were
then keyed into the program after calculating three different sections of samples, which are standard
of reference samples, radiocarbon samples and background. The current model available is having
low background, which is a good counter for age estimation. Due to the mechanical failure, we
switched to TRI-CARB Model 2700TR for radioactivity counts which produces higher background as
compared to the previous counter. In this paper the properties of the two different counters were
characterized and the effects to the age calculation were determined. The later counter was
crosschecked and is found capable to produce result as reliable as the former counter after some
modifications are done to the calculated counts.
Keywords: Characterization, Liquid Scintillation Counters, Radiocarbon Dating
Abstrak
Penentuan umur sesuatu sampel dibuat menggunakan teknik tentumur radiokarbon C-14.
Benzin yang diperolehi daripada teknik ini akan dimasukkan ke dalam Pembilang Sintilasi
Cecair untuk menentukan nilai keaktifan yang masih tinggal dalam sampel tersebut. Model
pembilang yang digunakan ialah Liquid Scintillation Analyzer Series TRI-CARB Model
2770TR/SL. Model ini mempunyai bacaan background yang rendah dan amat sesuai
digunakan untuk penentuan umur sesuatu sampel. Kerosakan yang dialami telah mendorong
usaha untuk bertukar kepada Pembilang Sintilasi Cecair TRI-CARB Model 2700TR yang
sedia ada tetapi memberikan bacaan background yang tinggi. Kertas kerja ini menunjukkan
pencirian yang dibuat ke atas kedua-dua pembilang ini dan membuktikan bahawa pembilang
kedua juga mampu bagi tujuan pengiraan yang baik untuk penentuan umur sampel setelah
beberapa rumusan dilakukan.
Katakunci: Pencirian, Pembilang Sintilasi Cecair, Tentumur Radiokarbon.
Introduction
The availability of Liquid Scintillation Counter, LSC, TRI-CARB Model 2770TR/SL in MINT is
used extensively for counting C-14 radioactivity in radiocarbon dating system. The results of sample,
standard and background counts obtained are keyed in simple age calculation program written in
Visual Basic for Percent Modern Carbon (pMC) determination. One major problem occurred when
this LSC broke down; the pMC of the samples cannot be determined and all the subsequent processes
delayed. Thus, the presence of LSC TRI-CARB Model 2700TR is fully utilized for the substitution.
The practice has not been done before, but some characterizations and similarities on these two
counters have been conducted as to predetermine the reliability and possibility of them if only any of
these counters cannot perform well. Moreover, the objective of any counting procedure is to
determine the radioactive content of the sample in question. Whenever a sample is made and counted,
the counter determines the count rate in term of count per minute (cpm) that is a relative value. This is
the number of light flashes, which have been seen by the detector, and must be related to
disintegrations per minute, which is a nuclear decay. Thus, in order to know the absolute value, the
sample’s net cpm value should always be converted to (dpm), which requires the knowledge of the
counting efficiency appropriate for each sample. The relationship between cpm and dpm is given
below:
dpm = cpm / (counting efficiency) (1)
Counting efficiency is obtained from the graph of the tSIE vs efficiency correlation curve. Therefore,
the characterizations have been conducted to the counters in term of disintegrations per minute (dpm),
it’s efficiency, which is the correlation with tSIE values, sensitivity of the instrument in Figure of
Merit (FOM), radioactivity units and also the difference in pMC values.
Experimental
Benzene recovery
The samples obtained are denoted as Arabic numbering from sample one to sample number five. All
the samples are from environmental and experimental samples, that are terrestrial snail shells that
were found from different areas in Malaysia namely, Kedah and calcium carbonate. The samples will
undergo three major stages, namely carbon dioxide recovery, acetylene recovery and benzene
recovery respectively. (Noakes et al. 1965, Tamers et al. 1974, Belluomini et al. 1978, Gupta and
Polach 1985, McCormac et al. 1993, Enerson et al. 1998, Muraki et al. 1998, Pawlyta et al. 1998,
Cook 2002)
heat
Sample Combustion: O2 + C CO2 (2)
HCl
Sample Hydrolysis: Carbonates CO2 (3)
Carbide Hydrolysis: LiC2 + 2H2O C2H2 + 2LiOH (4)
Trimerisation (benzene recovery) 3C2H2 CASA C6H6 (exothermic) (5)
The catalyst to complete the polymerization process is chromium activated silica alumina
catalyst. Then, the benzene yield is counted using the two different counters for
determination of carbon-14 radioactivity.
LSC Counting
In many analytical laboratories, Liquid Scintillation Counter (LSC) has become an indispensable
technique for the analysis of conventional radiocarbon dating samples [K.G Sushi, H. Polach (1985)].
In the liquid scintillation counting technique the scintillation solvent is benzene (C6H6) because of its
excellent light transmission properties and the high chemical conversion yield of sample C to
benzene.
In this experiment, each sample is experiencing 500 minutes counts for 10 cycles to obtain the
stability of cpm counted. In each cassette, the background, standard and samples are placed in a series
for data recording purposes.
Method using an external standard quench is chosen for this research. The purpose is to provide a
means of determining counting efficiency of any homogenous sample, independent of its radioactive
content. This method was devised to overcome problems such as the so-called ‘Wall effect’ present in
some of the external standard method. Recently, the study of the efficiency and background variation
related to liquid scintillation counter measurement sites were extensively discussed and reviewed by
F. Bella et al.
Results and Discussions
Characterizations of LSC TRI-CARB Model 2770TR/SL and LSC TRI-CARB Model 2700TR are
being discussed in a form of table which encompass gross and net cpm, disintegrations per minute
(dpm), efficiency, which is the correlation with tSIE values, sensitivity of the instrument in Figure of
Merit (FOM), radioactivity units and also the difference in pMC values.
Calculation of gross cpm and net cpm (cpmcorrected)
Gross cpm is counted from the average value of the total cpm. In this case, the total cpm is
being divided into 10, the number of cycles per sample run, whereas corrected cpm is the
difference between cpm of samples to cpm of background.
cpm Corrected = cpm samples – cpm background (6)
Type of Gross cpm cpm Corrected

sample
Samples Model Model 2700TR Model Model
2770TR/SL 2770TR/SL 2700TR
Sample 1 Terrestrial 9.4025 23.096 7.88 9.71

snail shell
snail shell
snail shell
Sample 4 Calcium 1.666 21.403 4.98 8.014
carbonate
Sample 5 Oxalic Acid 32.9125 54.742 31.36 41.426
Table 1. Gross cpm and net cpm of every sample for 500 minutes x 10 cycles
Quench Curve
In liquid scintillation counting, the terms quenching and counting efficiency are synonymous.
Quenching refers to any factor, which results in the loss of photon emission from the samples, which
include chemical agents that reduce the transfer efficiency of the beta energy by the solvent and
colored agents, which absorb, within the sample results in a loss of counting efficiency. The level of
quench is determined by tSIE for Model 2770TR/SL and SIS for Model 2700TR. The typical tSIE
quench correction curves are shown in Figure 1.
tSIE vs efficiency
120
100
80
efficiency
60
40
20
0
0 200 400 600 800 1000 1200
tSIE
Figure 1. tSIE curves for carbon –14
Disintegrations per minute, dpm = cpm / (counting efficiency)
Equation (1) is applied to obtain dpm from the value of net cpm for each sample. Interpolation is
conducted to the quench curve for estimation of counting efficiency for each sample.
Counting efficiency Disintegrations per minute (dpm)

Samples Model Model 2700TR Model Model 2700TR
2770TR/SL 2770TR/SL
Sample 1 92.74 95.41 8.50 10.17
Sample 2 92.84 95.38 2.31 4.61
Sample 3 92.22 95.49 5.99 11.30
Sample 4 92.00 96.09 4.58 8.34
Sample 5 92.31 95.04 33.97 43.59
Table 2. Disintegrations per minute (dpm) counted from correlation of cpm and counting efficiency.
Figure of Merit (FOM)
FOM is defined as a measurement of instrument’s sensitivity based on instrument’s counting

efficiency for Carbon-14 and its background.
FOM = (Efficiency)2 (7)

Background
The FOM values recorded for LSC Model 2770TR/SL and Model 2700TR are 7865.35 and 494.20 for
the range of 1 to 156keV respectively. The details are as follow:
Parameters Model 2770TR/SL Model 2700TR
Background 2.19 cpm 12.53cpm

Efficiency 95.93% 95.85%
FOM 494.20 7865.35
Table 3. Figure of Merit values for different counters
Radioactivity Units
The term dpm, is commonly used to quantify the absolute radioactive content of a radiolabel samples.
However, a unit of radioactivity in term of the International System of Unit (SI) is the bacquerel, Bq,
which is equivalent to one disintegration per second.
Radioactivity Units (Bq)

Samples/Standard Model 2770TR/SL Model 2700TR
Sample 1 0.14 0.17

Sample 2 0.04 0.08
Sample 3 0.10 0.19
Sample 4 0.08 0.14
Sample 5 0.57 0.73
Table 4. Radioactivity Units in Bacquerel (Bq)
Percent modern carbon (pMC)
Absolute Percent Modern Carbon Estimated Error

Samples (pMC)
Model 2770TR/SL Model 2700TR
Sample 1 30.58 ± 0.5 29.46 ± 0.25 3.8 %
Sample 2 30.9 ± 1 31.03 ± 0.59 4.0%
Sample 3 32.75 ± 0.63 34.06 ± 0.3 3.8%

Sample 4 26.65 ± 0.34 27.91 ± 0.31 4.5%
Table 5. Absolute percent modern carbon values for different counters
Conclusions
Both counters Model 2770TR/SL and Model 2700TR are reliable to be used in radiocarbon dating
system calculation particularly in determining the radioactivity of carbon-14. They represent likeable
results for pMC and radioactivity values for which the differences are not prominent. Nevertheless,
LSC Model 2770TR/SL is more sensitive due to its low background values. Moreover, the simple
program written in Visual Basic is used to determine the radiocarbon content residue in the samples
by pMC calculation and the radioactivity calculation for radioactivity estimation respectively. It is
also found that the amount of residual C14 activity from the counters will not determine the
radioactivity of the C14 in the sample.
Acknowledgement
The author would like to thank En Ramzah Mohamed from the Instrumentation Group for his
assistance in the counter operations and all the colleagues from Radiocarbon Dating Laboratory
(RDL) in making this paper a worthwhile.
References
1. Liquid Scintillation Analyzer, Reference Manual. Publication No.169-4142. Rev C

2. Liquid Scintillation Analyzer, Manual Reorder No. 1694141. Publication No. 169-4141 Rev.
C
3. Mc Cormac, F.G., 1992, Liquid scintillation counter characterization, optimization and
benzene purity correction, Radiocarbon, 34, 1, 37-45
4. Kojola, H., Polach, et al, 1984, High resolution low-level liquid scintillation beta-
spectrometer, Int. J. Appl. Rad. Isot., 35, 949-952,207
5. Kamisah Hj Alias, Bashillah Baharudin, Juhari Muhd Yusof, Ahmad Raduan Ujang,
Radiocarbon Dating Development and Practices at MINT, Seminar R& D MINT 2002, 25 –
27 June 2002.
6. Radiocarbon Age Calculation Program, www.ucd.ie\c14
7. K.G Sushi, H. Polach., Radiocarbon Dating Practices at ANU, Radiocarbon Laboratory
Research School of Pacific Studies (1985).
8. Noraishah Othman, Kamisah Alias, Norinsan Kamil Othman, Trimerisation of Chromium
Activated Silica Alumina (CASA) in Radiocarbon Dating System, The 17TH Symposium of
Chemical Engineering 2003, 29-30 December 2003.
9. Kamisah Hj Alias, Bashillah Baharudin Juhari, Muhd Yusof, Wan abdul Aziz Wan
Muhammad, MINT Radiocarbon Dating Facilities, Seminar R& D MINNT 2000, 17 – 19
Oktober 2000.
10. Kamisah Hj Alias, Noraishah Othman, Zainudin Othman, M.J.Head, Radiocarbon Dating
Development At MINT, N.Z.
11. Rozanski K, Stichler W. et al, The IAEA Intercomparison Exercise 1990.
CLASSIFICATION OF CHILLI SAUCES : MULTIVARIATE PATTERN
RECOGNITION USING SELECTED GCMS RETENTION TIME PEAKS OF
CHILLI SAUCE SAMPLES
Low Kah Hina, Sharifuddin M. Zaina, Mohd. Radzi Abasa, Mustafa Ali Mohd.b
a
b
Shimadzu-UMMC Centre for Xenobiotics Studies, University of Malaya Medical Centre,
University Malaya, 50603, Kuala Lumpur, Malaysia
Email: smzain@um.edu.my / Tel: 603-79674202 / Fax: 603-79674193
Abstract
As a preliminary work on the possibility of separating classes of chili sauces based on taste or customer
preferences, organic compounds from different kinds of chili sauces of various brands were separated and analyzed by gas
chromatography/mass spectrometry (GC/MS). It was found that these organic compounds do form a basis for separation of
different types of sauces. The similarity and dissimilarity of chromatograms due to the organic composition of the chili
sauces were explored by multivariate pattern recognition techniques based on cluster analysis (CA) and principal component
analysis (PCA). Both CA and PCA results exhibit four linearly separable classes, namely general sauces, hot sauces, sauces
with benzoic acid and sauces with garlic. It was concluded that by using chosen retention peaks in the chromatograms of
various sauce samples as multivariate features, CA and PCA can be successfully used to reveal the natural clusters existing
in chili sauces according to their organic composition.
Keywords: Chemometrics; Chili sauce; Principal component analysis; PCA; Cluster analysis
1. Introduction
Originally, chilli sauces were produced at home. Today, this practice still preservere but most chilli sauces
available nowadays are mass manufactured in factories. Today, much effort is spent to ensure that mass
manufactured sauces resemble as close as possible home made ones. Some brands do this better than others.
The ingredients used in chili sauces are very similar to those used in tomato ketchup. However, the methods
of preparation and the amount of the ingredients used may vary considerably. The primary ingredients in chili
sauce are red tomatoes and chilies. The usual sweetener is sugar (sucrose). The proportions of the various spice
ingredients are not standardized between manufacturers [1].
Humans are very good at perceiving similarities and differences between objects of different shapes. The
goal of pattern recognition in analytical chemistry is finding similarities and differences between chemical
samples based on measurements made on the sample [2]. Therefore, pattern recognition appears to be a useful
tool to authenticate foodstuffs according to their quality/variety/brand, when a set of samples whose
classification is known a priori is available.
This study initially involve data collection via instrumental analysis, basically GC/MS. By using only
chromatographic techniques, we able to separate the chemical compounds in the sauces, which are mainly
capsaicinoid compounds, the pungent principle of capsicum fruits, benzoic acid which is a preservative, and
other compounds due to the ingredients of the sauce.
Capsaicinoid compounds are a group of pungent compounds found mostly in capsicum fruits - the main
components are acid amides of vanillylamine and C9 – C11 branched-chain fatty acids. There are five naturally
occurring capsaicinoids, which have been reported, namely capsaicin, nordihydrocapsaicin, dihydrocapsaicin,
homocapsaicin and homodihydrocapsaicin. Of these, capsaicin and dihydrocapsaicin are the major compounds
of most capsicum species. Therefore these compounds have been considered as major indicators of chili
product quality [3].
Techniques using GC/MS provide accurate and efficient analysis of content and type of capsaicinoids
present in a chili sauce sample. These results are probably the major factors in determining the
qualities/variety/brand of those sauces, and are the most important parameters which are used in this statistical
study.
In this work, different multivariate chemometric methods been employed in the exploration; this primarily
involve the use of cluster analysis (CA) and principal component analysis (PCA). Cluster analysis, which
involves the search for natural grouping among samples is a preliminary way to study data sets and to discover
the structure residing in them. PCA is used to transform the original data matrix (X n × m) into a product of two
matrices, once which contains information about the objects (Scores matrix, S n × m) and the other about the
variables (Loadings matrix, L n × m) [4]. PCA enables one to study data structure in reduced dimensions.
2. Experimental
2.1 Sample preparation

About 1mL of sauce were weighed and extracted with 5mL dichloromethane in a vial for overnight. Then
1mL of sauce extracts was drawn and filtered using a 0.4µm nylon filter membrane with a syringe filter into a
small glass vial. The obtained filtrate was dried under nitrogen gas, and reconstituted with 200µL of chloroform
and was later used for a GC/MS analysis with injection volume of 1µL.
2.2 GC/MS analysis

Samples were assayed using a Gas Chromatograph (Shimadzu GC-17A) with a Mass Spectrometer
(Shimadzu QP-5000). A 30 m × 0.247 mm × 0.25 µm capillary column, with a stationary phase of (5% - phenyl)
– methyl polysiloxane (J & W Scientific, DB-5), was used. For analysis, the initial temperature was 60°C held
for 1 min. The temperature was increased from 60 to 280°C at the rate of 10°C/min and held at 280°C for 22
min. The total run time was 38 min. The injection volume was 1.0µL and the total flow of carrier gas was 39.5
mL/min helium.
3. Data processing
Data preprocessing is a very important part of any chemometric data analysis project. It consist of whatever
mathematical manipulation of data prior to primary analysis. From the obtained GC profiles, 12 marked peaks
were considered as the multivariate variables. They are the compounds eluting at retention times, 7.6 (1), 9.2
(2), 12.2 (3), 17.0 (4), 19.2 (5), 19.5 (6), 21.0 (7), 21.2 (8), 24.4 (9), 25.4 (10), 25.7 (11), and 36.9 (12) minutes.
These peaks were examined for their mass spectra and identification of these peaks were attempted. The peaks’
area per weight of sample were preprocessed, then arranged the in an X-matrix (201 x 12). The values of each
row represent the measurement of different variable for each sample, and the values of each column represent
the measurement of different samples for a particular variable. Each replicate was treated as an individual
sample in the data matrix, which then undergo CA and PCA. All these were done by combination of several
statistical software packages such a Microsoft® Excel, Teach/Me®-Data Analysis, JMP® and JMP IN®.
Figure 1: Typical GC/MS chromatogram of a chili sauce extract.
Retentiom
7.5 9.2 12.2 17.0 19.2 19.5 21.0 21.2 24.4 25.4 25.7 36.9
Time
GCS0036
782155 0 186056 28568683 12228225 0 5353903 9774055 1426550 10714811 6485459 2731411
S01a
GCS0035
1387435 0 597293 23716034 6519874 0 4026651 10841376 1902577 13662227 8002463 4064320
S01b
GCS0035S01c 1613369 0 819216 37251905 11069449 0 6242748 14612206 2076992 19084453 11712994 6083270
GCS0035
801396 0 386139 18332483 4543486 0 923887 8586443 1914108 10339062 6540138 2461134
S02a
GCS0035
968122 0 385629 14260443 3855149 0 1528811 9105060 1396709 11473199 7115043 2353073
S02b
Table 1: Peak areas at particular retention times for a part of sauce samples
In this work, CA was applied to the autoscaled data, the sample were calculated on the basis of Euclidean
distances, while Ward hierarchical agglomerative method was used to establish the clusters. Next, PCA was
performed on the unscaled data, used to provide data structure in a reduced dimension, retaining the maximum
amount of variability present in the data.
4. Results and discussion
4.1 Mass spectra analysis

From the total ion chromatograms of chili sauces extracts, compounds eluting at particular retention times
were examined for their mass spectra. Compounds 1, 3, 6 and 8 were not resolved. The m/z values obtained
from mass spectra of the molecular ions (M+) and fragment ions of selected compounds are shown in Table 2.
Mass/charge, m/z Molecular

Retention time / Compound
Fragment ions Ion, M+
1 7.5 unresolved
2 9.2 Benzoic acid 51 65 77 94 105 122
3 12.2 unresolved
4 17.0 Myristic acid 60 73 85 129 185 228
5 19.2 Palmitic acid 60 73 85 129 213 256
6 19.5 unresolved
7 21.0 Linoleic acid 67 81 95 109 123 280
8 21.2 unresolved
9 24.4 Nordihydrocapsaicin 43 122 137 152 195 293
10 25.4 Capsaicin 43 122 137 152 195 305
11 25.7 Dihydrocapsaicin 43 122 137 152 195 307
12 36.9 Vitamin E 43 57 71 165 205 430
Table 2: m/z values of selected compounds.
Figure 2: The capsaicinoids commonly found in chilies.
As mentioned before, the pungent capsaicinoids have been considered as major indicators of chili product
quality. Typical mass spectra of dihydrocapsaicin is shown in Figure 3. The molecular ions (M+) of the
capsaicinoid compounds were confirmed from their mass spectra. The base peak of each compound was found
to be common at m/z = 137. Other fragment ions had m/z = 43, 122, 152 and 195.
A possible empirical formula for fragment ion m/z = 137 from isotopic abundance calculation is
C8H9O2. An achievable fragment ion is shown below as:-
H3CO H3CO
CH2 CH2
O O
H H
m/z = 137 m/z = 137
The resonance stabilization of this fragment ion suggested its occurrence as the base peak [5].
Figure 3: Mass spectrum of dihydrocapsaicin.
The isotopic abundance calculation of the fragment ion m/z = 195 suggested an empirical formula of
C10H13O3N. A possible fragment ion is shown in the decomposition reaction below [6]:-
R
H CH
O OH
H3CO CH2 H3CO
N C CH2 N C CH2
H H
HO
HO
m/z = 195 +
H2C CHR
Fragment ions with m/z = 153 and 152 were perhaps due to the ion decomposition reaction below:-
H
H C R' HC R'
H3CO H3CO
N C O N C O
H H2
HO HO
H3CO H3CO
NH2 H NH2 + R' CH C O
HO HO
m/z = 152 m/z = 153
4.2 Multivariate correlation analysis
From the correlation matrix for GC peaks, peak no. 1 and no. 3 correlate strongly (r = 0.9861); followed by
inter-correlation between peaks no. 9, nordihydrocapsaicin, no. 10, capsaicin and no. 11, dihydrocapsaicin.
Others correlate moderately or weaker. The strong correlation between nordihydrocapsaicin, capsaicin and
dihydrocapsaicin is expected because these compounds are the major pungent capsaicinoids in capsicum fruits.
In other word, we can easily conclude that capsaicinoids present in chili sauces or natural chili are in a constant
ratio. On the other hand, we suspect that the unknown peaks no.1 and no.3 also show similar characteristics as
the capsaicinoids, but this needs further investigation.
4.3 Cluster analysis

CA is a well known technique of data analysis, commonly applied before other mutivariate procedures
owing to its unsupervised character, that reveals the natural clusters existing in a data set on the basis of the
information provided for the measured variables. The results obtained for the case at hand, using the Ward’s
method and Euclidean distance are presented as a dendrogram in Figure 4. At distance 700, 3 clusters that can
be identified as ‘similar’ were found: from the top, the first cluster is composed of general sauces including a
sub-cluster of hot sauces. The second cluster is a group made up of sauces containing benzoic acid as
preservative. The last cluster is those sauces with the addition of garlic.
General
sauces
Hot sauces
Sauces with
benzoic acid
Sauces with
garlic
Figure 4: Dendrogram of Cluster analysis.
4.4 Principal component analysis

By performing PCA on the dataset, which was initially preprocessed from GC/MS data of the chili sauces
extracts, a series of principal components (PCs) could obtained. Each PC is associated with an eigenvalue. PC1
has the largest eigenvalue and carries the largest variance of the original data, and subsequent PCs carry
variance in a decreasing order.
Loadings graph (Figure 5) is used to determine which variables are important for describing the variation in
the original data set, and there are no variables with loading close to ±1, revealing that no PC is closely aligned
with any peaks. For PC1, peaks no. 1, 2, 3 and 6 do not contribute much to the variance described by PC1. For
PC2, it is the other way round; peaks no. 4, 5, 7, 8, 9, 10, 11 and 12 have near zero loading. In addition, peak no.
2 show high negative loading value, which indicates benzoic acid is a meaningful parameter for PC2.
Figure 5: Loading Plot: PC1 vs. PC2.
The final result of PCA, Figure 6, shows a plane spanned by first two PCs, representing 95.87 of the
variation in the data. There are crowds of points discriminated by PC2 while PC1 primarily determines the
spread of scores. Four major separable classes namely general sauces, hot sauces, sauces with benzoic acid and
sauces with garlic can be observed from the plot.
Sauces Hot Sauce

with sauce s with
benzoic s garlic
acid
General sauces
Figure 6: Score Plot of PC1 vs. PC2.
As had been discussed, peaks no. 1, 2, 3 and 6 are the dominant features in PC2. From scores plot, it is
demonstrated that those sauces with benzoic acid clusters on the negative area of axis PC2 corresponding to
its’ negative loading, while those sauces with garlic are on the positive side. These indicates that the unknown
peaks no. 1 and 3 should correspond to the garlic content of the sauces.
5. Conclusion
In this study, the feasibility of using just GC profiles to distiguish between chili sauces variety was
investigated with multivariate chemometric methods. CA and PCA methods have demonstrated similar results;
that using certain peaks in the GC profiles as multivariate parameters we are able to reveal the natural clusters
existing in the chili sauce samples studied.
Acknowledgements
The authors gratefully acknowledge Shimadzu-UMMC Centre for Xenobiotics Studies Laboratory for providing
the GC/MS instrument and technical support.
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(Capsaicum Species), Ahmad bin Ab. Wahab, MARDI Res. Bull. 12, 3, 1984, 290-297.
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1980.
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Ellis Horwood imprint, England, 2000.
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chromatography, Agnes Kosa, Tibor Cserhati, Esther Forgacs, Helena Morias, Teresa Mota, A. C. Ramos,
Journal of Chromatography A, 916, 2001, 149-154.
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Mohammed Razak Hamdan, Zhari Ismail, and Mohd. Noor Ahmad, Journal of Analytical Chemica Acta,
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and Z. Badriah, MARDI Res. Bull. 13, 2, 1984, 190-193.
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Beijing Tan, James K. Hardy, Ralph E. Snavely, Analytica Chemica Acta, 422, 2000, 37-46.
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Compounds Using the Simplex Method, Rachaneewan Karnka, Mongkon Rayanakorn, Surasak Watanesk
and Yuthsak Vaneesorn, Analytical Sciences, Vol. 18, June 2002, 661-665.
[13] The hottest chilli variety in India, Current Science, Vol. 79, No. 3, 10 August 2000, 287-288
[14] Gas Chromatographic Determination of Capsaicinoids in Green Capsicum Fruits, Anna M. Krajewska and
John J. Powers, J. Assoc. of Anal. Chem., Vol. 70, No.5, 1987, 926-928.
[15] Simultaneous Determination of Capsaicin, Dihydrocapsaicin and Nordihydrocapsaicin in Capsicum Fruits
by Gas Chromatography, Kazuhiko Sagara, Sadao Kakizawa, Kohji Kasuya, Tetsuo Misaki and Hiroshi
Yoshizawa, Chemical and Pharmaceutical Bulletin, 28(9), 1980, 2796-9.
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CORRELATING STUDENTS’ VIEWS ABOUT CHEMISTRY AND THEIR CONCEPTUAL
KNOWLEDGE IN FIRST YEAR CHEMISTRY
Zarila Mohd Shariff & Jaafar Jantan

Applied Science Education Research
Faculty of Applied Sciences,
Universiti Teknologi MARA, 40450 Shah Alam, Selangor
Email: zarila@salam.uitm.edu.my; jjnita@salam.uitm.edu.my
Phone: 03-5544-4606; 03-5544-4593. Fax: 03-5544-4562
Website: http://www.uitm.edu.my/faculties/fsg/drjj1.html
Abstract: Research have shown that learning and academic achievement for students in chemistry has been associated with
not only their perceptions on science but most importantly their existing knowledge, particularly in Chemistry. Samples for
this study are the applied science students doing their degrees and diplomas at UiTM Shah Alam and Arau campuses and the
science students at Matriculation College, Arau Campus. The students’ views about chemistry were determined using the
Views about Science Survey (VASS) in Chemistry (translated into Bahasa Melayu by J. Jantan with permission from the
author) which was developed by Hestenes et. al., at the University of Arizona. The views were categorized into profiles of
either experts (EP), high transitional (HTP), low transitional (LTP) or folk profiles (FP). We found that only 8.5% out of 483
are the EP (matches that of chemistry lecturers), 31.7% the HTP, 41.6% the LTP and 18.2% the folk profile. Conceptual
understanding in basic chemistry were determined using the Chemical Concept Inventory (CCI), a 22-item, multiple choice
question, developed by Mulford (1996). The mean score on the CCI is 30.38% with a standard deviation of 12.7%. This
score is very low considering that CCI probes only the conceptual understanding. The mean CCI score for each of the VASS
profiles are 31.71% for the EP, 33.07% for the HTP, 29.95% for the LTP and 27.94% for the FP. The mean scores between
the HTP and the LTP, and the HTP and the FP are shown to be statistically significant. Thus, we can infer that students,
whose views about chemistry match those of the experts, do have a better understanding about basic chemistry.
Keywords: FCI, Chemical Concept Inventory, VASS, constructivist, chemistry, learning.
Introduction
Many students leave high school chemistry or college chemistry with profound misunderstandings
about the nature of matter, chemical processes, and chemical systems. Due to that there is a large
growing body of research on misconceptions (or “alternate conceptions”) had accumulated. To help
faculty identify the concepts that their students do not understand or misconceptions that they have, a
number of instruments called concept inventories or CI’s had been developed. An oft-cited instrument
is the Force Concept Inventory (Hestenes, Wells, and Swackhamer, 1992 [1]; Hake 1998 [2]) which
was then revised by Hestenes et. al in 1995. The revised version of FCI has 30 qualitative items, with
subscales, dealing only with the Newtonian concept of force and motion. It is extremely effective in
eliciting the “commonsense” notions of students about motion as opposed to the scientific beliefs. It is
an instrument designed to assess students understanding of the most basic concepts in Newtonian
physics. The questions were designed to be meaningful to students without formal training in
mechanics. This FCI looks at six dimensions namely kinematics, Newton's First, Second, and Third
Laws, the superposition principle, and types of forces (such as gravitation, friction). Each question
offers only one correct Newtonian solution, with common-sense distractors (incorrect possible
answers) that are based upon students’ misconceptions about that topic, gained from interviews. The
FCI is now available in nine languages: Chinese, English, Finnish, French, German, Bahasa Melayu,
Spanish, Swedish, and Turkish.
Another widely accepted instrument is the mechanical baseline test (MBT) developed by
Hestenes, D. and Wells, M. [3]. MBT is an advance companion of the FCI. It contains questions that
are designed to probe concepts and principles that cannot be grasped without formal knowledge about
mechanics, and require both qualitative and quantitative approach to answer them, which is more
involved than just plugging in numbers into formulas. The two tests together assess students’
conceptual understanding of basic Newtonian mechanics that are generally covered in an introductory
physics course. The MBT covers concepts in kinematics (linear and curvilinear motion), basic
principles (Newtons' First, Second, and Third Laws, superposition principle, energy conservation,
impulse-momentum, and work) and special forces (gravity and friction). The MBT is available in four
languages: English, German, Spanish, and Bahasa Melayu. These CI have been tested for their
validity and reliability. Prompted by the success of FCI, many CI’s, have been and continues to be
developed especially to evaluate identify students’ initial beliefs and students’ understanding in the
basics of chemistry such as Chemistry Concept Inventory (ChCI) developed by Pavelich, Michael
[4,5], Chemical Concepts Inventory (CCI) developed by Mulford, Douglas R. [6] and California
Chemistry Diagnostic Test (CCDT) developed by Russell, A.A. [7].
Achievement in chemistry classes cannot progress very much to most learners as long as their
alternative conceptions are not confronted and new knowledge is not constructed in due time. In fact,
some of these alternative conceptions were found to be the result of instructions [8]. Much of the
areas of chemistry and the learning difficulties that learners encounter in chemistry were reviewed by
Taber [9]. On the other hand, research into the knowing and learning of chemistry were done by
Halloun et.al., [8] through the use of the Views About Science Survey (VASS), an instrument
developed and validated by the same group.
The Chemistry VASS (Views About Science Survey) is a paper-and pencil instrument
consisting of 30 items and intended to probe the personal beliefs about the nature of science and about
the learning of science. Beliefs about science were probed within three scientific dimensions
pertaining to the structure, methodology and validity of science. Beliefs about the learning of science
were probed within three cognitive dimensions pertaining to learnability, reflective thinking and
personal relevance of science [8].
The aim of this study is to determine the correlation between the students’ views about
chemistry and their conceptual knowledge in first year chemistry among applied science students
pursuing their degrees and diplomas at UiTM Shah Alam and Arau campuses and the science students
at Matriculation College, Arau Campus. We intend to find out if academic background and gender
contribute to students’ beliefs about chemistry and chemical concepts.
Methodology
Perception about chemistry.
Students’ perception about chemistry, were probed by using VASS. VASS profiles provide a
comprehensive index of students’ views about knowing and learning chemistry. VASS contains 30
items, 13 pertaining to the scientific dimensions and 17 to the cognitive dimensions. The items are
formulated in a novel Contrasting Alternatives Design (CAD). Each CAD item requires respondents
to balance primary view against a contrary view on the eight-point scale [7]. This is shown in Figure
1. Contrasting Alternatives Design is used in the construction of VASS in order to overcome major
validity and reliability problems encountered in traditional assessment formats. [8].
Penyelesaian masalah kimia memerlukan saya:
(a) pernah melihat penyelesaian kepada masalah yang sama sebelum ini.
(b) boleh menggunakan teknik umum penyelesaian masalah.
1 2 3 4 5 6 7 8
Samarata Bukan
Kearah “Hanya (a)” Kearah “Hanya (b)”
(a) & (b) (a) atau (b)
Figure 1. A contrasting Alternative Design in one of the VASS items.

In this study the Bahasa Melayu version of VASS was used. VASS was translated by one of
the authors in the year 2001 with the permission from Hestenes et.al. The instrument was piloted by
UiTM chemistry and chemical engineering lecturers to eliminate ambiguities, confusion or
weaknesses in the translation and sentence structure. Following suggestions by the experts, changes
were made to improve the clarity of the questions.
Conceptual Knowledge in Basic Chemistry
Students’ conceptual knowledge in basic chemistry was diagnosed by using Chemical Concepts
Inventory (CCI) developed by Mulford, R. CCI is a multiple choice instrument that contains 22 items.
Questions on CCI are non-mathematical and are based on concepts from several topic areas of first
semester college chemistry covering phase changes; conservation of substance, weight and mass;
chemical symbols, chemical equations and stoichiometry; macroscopic versus microscopic properties;
solutions; and size of atoms. CCI was chosen in this study because the questions in CCI are based on
concepts which form the foundation of several topics of beginning chemistry.
The subjects
In 2004 and early 2005, Chemical Concept Inventory and the Bahasa Melayu Chemistry VASS were
administered to 324 UiTM chemistry students at the Faculty of Applied Sciences from the Shah Alam
campus and from the Arau campus and also to 159 Matriculation College students. The UiTM
subjects are students who are doing their diploma in industrial chemistry and doing degrees in
chemistry and applied chemistry, ranging from Year 1 to Year 3. Our intended population is
chemistry-based students at colleges and universities but for this work, the sample were chosen based
on convenience and the willingness of the lecturers in contact with the students to administer both the
CCI and the Chemistry VASS. The students usually spent about an hour to complete the CCI and
thirty minutes to complete the survey. Those who could not finish on time were allowed to take the
VASS papers home and were requested to send the completed survey to us in due time.
Subjects from the Matriculation College were science students from Arau campus and in their
final semester. One of the authors administered the CCI to the students while Chemistry VASS was
administered by their lecturers.
Subjects’ personal profile and their responses to both CCI and VASS were keyed-in into
spreadsheet MS™ Excel. Statistical analysis was performed using both the SPSS (Statistical Program
for Social Science) version 12 and MS™ Excel for Windows. Some of the responses were coded for
analysis along the scientific and cognitive dimensions.
This section offers a broad characterization of college students’ views about knowing and learning
chemistry and their relation of these views to students’ conceptual knowledge of basic chemistry. The
first part of this section describes the students’ perception of chemistry and how they are classified.
I. Items Response Classification
Students’ views on individual items were classified into three types, namely the expert, mixed and
folk views. An expert answer refers to an answer chosen by teachers or professors in chemistry. Their
answers are polarized toward one end of the scale (scale of 1 through 8). Therefore, on some items,
these experts overwhelmingly chose the extreme option of 1 or 7. On others, a large majority
concentrated on two or all three options 1 through 3, or 5 through 7. A student is classified to be
holding an expert view on a given item if his or her answer falls within the ranges chosen by the
expert. On the contrary, a student is classified as holding a folk view on a given item if he or she
chose a polar opposite to the experts’ options. A student is classified as holding a mixed view on a
given item if he or she shares the middle position with those of the experts who did not express an
expert view. Example of this is shown in figure 2 below.
Item 17 Responses 17. Setelah guru menyelesaikan sesuatu masalah yang

mana penyelesaian saya adalah salah:
30.0
(a) saya buang penyelesaian saya dan
Student Percentage
25.0 saya belajar penyelesaian yang ditunjukkan oleh

20.0 guru
15.0 Series1
(b) saya cuba menentukan perbezaaan penyelesaian
10.0
saya dan penyelesaian guru.
5.0
0.0
1 2 3 4 5 6 7 8
folk mixed expert
Figure 2: Students’ responses on VASS item 17 showing classification of expert, mixed and folk views
Figure 2 shows that 67% of the students responded with the expert view which are options 5 through
7; 17% of the students responded with the mixed view which is option 4 and the 14% responded with
the folk view which is option 1 through 3. For this particular item, a significantly high number of
students share the views of experts but it is not the intention of this paper to comment or analyze
responses to each and every one of the items. Instead, the focus is to classify students’ profile with
respect to VASS encompassing all 30 items.
II. Profile Classification and Description
Further classification was made to the students’ choice of answers to establish the overall industrial
chemistry students profile distribution on the perception of chemistry. Hence, students responses had
been grouped to four types of profiles namely, expert (EP), high transitional (HTP), low transitional
(LTP), and folk (FP). Cutoffs for the profiles are shown in Table 1.
Table 1: General Profile Characteristics.

Profile Code Abbreviation Number of Items out of 30
Expert 1 EP 19 items or more with expert views
High Transitional 2 HTP 15 to 18 expert views
Low Transitional 3 LTP 11 to 14 items with expert views and at most the same number of
items with folk views
Folk 4 FP 10 items or less with expert views
Hence, students scoring more than 19 items corresponding to the expert views, are
categorized as experts and those scoring more than 15 items corresponding to the expert views are
considered as strong potential to become experts while the others are quite far from sharing views of
the experts.
As shown in figure 3, out of 483 matriculation, diploma and degree students, only 8.5%
evinced an Expert Profile (EP), 31.7% in the High Transitional Profile (HTP), 41.6% in the Low
Transitional Profile (LTP) and 18.2% in the Folk Profile (FP). More than 59% of the subjects in our
sample made up the LTP and FP. It is understandable that only a small portion of our sample exhibits
the expert view in knowing and learning of chemistry but the large percentage, 42%, of the sample
belonging to LTP is quite upsetting. Our finding agrees with the study done by Shariff [10] on
Industrial Chemistry students views on the knowing and learning of chemistry.
Students Overall Profile Distribution
50.0
40.0
Percent
30.0
20.0
10.0
0.0
EP HTP LTP FP
VASS Profile
Figure 3: Overall profile distributions of students.

a. Academic Background
In order to better understand the trend, an analysis of the profile according to academic background is
done. Table 2 and figure 4 shows the profile distribution of students by academic background. The
percentage of Diploma students sharing the experts’ views is the highest at 10.5% compared to only
6.3% for the Matriculation students and 8.9% for the Degree students. Even so, a chi-square analysis
revealed that the differences are not significant at the 95% confidence interval.
Table 2: Profile distribution of students based on academic background.

Students Count Students Count Students Count
Matriculation Diploma # % Degree # %
(N=159) # % (N=133) (N=191)
EP 10 6.3 EP 14 10.5 EP 17 8.9
HTP 40 25.2 HTP 45 33.8 HTP 68 35.6
LTP 67 42.1 LTP 53 39.8 LTP 81 42.4
FP 42 26.4 FP 21 15.8 FP 25 13.1
The result shows that more matriculation students belong to the Low Transitional Profile and
the Folk Profile (68.5%) compared to students who are doing their diploma (55.6%) and degree
(55.5%) programs. We believe that students’ knowing and learning of chemistry gets better as they
take more chemistry courses. Since the matriculation students only had a maximum of 4 chemistry
classes as compared to at least 10 courses for the diploma and degree students, hence more of the
matriculation students are in the low transition and folk profiles. Again, a chi-square analysis revealed
that the differences are not significant; hence we infer that whether we are dealing with matriculation,
diploma or degree students, the trend of the profile is equal.
Students Profile Distribution
50.0
40.0
Percentage
30.0 Matriculation
Diploma
20.0
Degree
10.0
0.0
EP HTP LTP FP
VASS Profiles
Figure 4: Profile distribution of students according to academic background.

b. Gender
We also looked at how the male and female differ in ways that they view chemistry. For this purpose
we only looked at the general trend and not analyzing the gender issue for each of the academic
group. Generally, the trend is quite similar for both the gender. Both, have about the same percentage
in the transitional profile (72% - 75%). The difference is the percentage of female (10.5%) sharing the
experts profile is higher than that of the male (4.0%), and the female students also have lower
percentage belonging to the folk profile (17.2%) compared to the male (20.5%). A further analysis
using the chi square method show that the difference is not statistically significant at the 95%
confidence limit. In other words, gender is not a factor in the distribution of the profile.
Table 3: Profile distribution of students according to gender.

Students Count Students Count
Male Female # %
(N=151) # % (N=332)
EP 6 4.0 EP 35 10.5
HTP 50 33.1 HTP 103 31.0
LTP 64 42.4 LTP 137 41.3
FP 31 20.5 FP 57 17.2
Students Profile Distribution
50.0
40.0
Percent
30.0 EP
20.0 HTP
LTP
10.0
FP
0.0
Male Female
VASS Profile
Figure 5: Profile distribution of students according to gender
III. Conceptual Knowledge of Chemistry
The overall mean score of CCI in our study is 30.38% with a standard deviation of 12.7%. The mean
score of chemical concept inventory for various academic backgrounds is shown in Table 4. Our
results show no matter at what level a student is regarding their academic level; their understanding of
fundamental chemical concepts is, on the average, equal. The results show that our science or
chemistry students’ beliefs and reasoning are not scientific in many of the basic chemistry topics. In
other words, these students have not been able to construct and connect the most basic and
fundamental chemical concepts even though they have taken and passed or aced the chemistry courses
they had taken. If we had placed a cutoff score of 60 as the crossover score [1], then it is obvious that
all our subjects failed to qualify as chemical believers. In fact, the mean score is just a bit higher
compared to a score of 25 which a student would obtain by random guessing.
Table 4: Means of CCI by academic background

Academic N Mean (%)
Degree 191 29.8
Diploma 133 30.5
Matriculation 159 30.9
A t-test comparing the mean score for the male students and female students found the
difference to be significantly different at the 99% confidence limit (t = 5.998, p = 0.000) but the score
is still far below the cut-off score. It is as if the students don’t know the answer to each item and are
just randomly guessing or in fact responding as what they believe the answer should be, without
strong conviction and scientific reasoning to justify their answer. Failing to justify their belief indicate
poor knowledge about the topic being assessed.
Table 5: Means of CCI by gender

Gender N Mean (%)
Male 151 35.3
Female 332 28.1
IV. Student Views and Academic Achievement
Educational researchers speculated that students’ views about knowing and learning science affect
their understanding of what they are taught in science courses [9, 10]. In testing this conjecture, we
assessed the relation between VASS profiles and their conceptual knowledge in basic chemistry
CCI Mean (%) versus VASS Profile
34.0
33.0
32.0
31.0
Percent
30.0
29.0
28.0
27.0
26.0
25.0
EP HTP LTP FP
VASS Profile
Figure 6: CCI mean score for various VASS profiles
The mean scores for those belonging to the experts and high transitional profiles are
significantly higher than those whose views about chemistry belong to the low transitional and folk
profile. Since the Chemistry VASS measure along both the scientific and cognitive dimensions we
expect a strong correlation between scores on CCI and the students’ profile on VASS. In fact, the
spearman’s-ρ statistical analysis showed a positive correlation at the 99% level. Upon performing
ANOVA and t-tests for independent samples, we found that the difference in the CCI scores between
the HTP and LTP (t = 2.903, p = 0.004) and the difference between the HTP and FP (t = 2.881, p =
0.004) are statistically significant. The HTP mean score is 33.1%, the mean score for the LTP is
30.0% and the mean score for the FP is 27.9%. It’s quite surprising that the EP group mean score to
be the same as the HTP group. Nonetheless, we could infer that the knowledge and understanding of
the EP and HTP group are at par but are a bit higher than the LTP and FP group. Still, due to the low
scores, the differences we observed do not contribute very much to the advancement of chemical
learning. However, being able to categorize the students into the respective profile will help us
identify those who will have good grasps of chemistry concepts and hence might lead us to predict
those who will excel in chemistry-related courses and career.
Conclusion
We feel that the mean score for CCI is embarrassingly low for students who are taking or have taken
many chemistry or chemistry-related courses. If we can adopt interpretation from the FCI, the
worryingly low score reflects shallow understanding which is then related closely to failure of
instruction. Instructors failed to identify common-sense beliefs that are at odds with the scientific
beliefs that students possess and continue to hold-on to, even after completing few chemistry courses.
It seems that those whose science beliefs are matching or on the verge of matching the experts beliefs,
their CCI score will be much higher, but not high enough to crossover into the chemical beliefs.
Again, instructional strategies play a major role in assuring that students do have the knowledge of
basic concepts in chemistry in order for them to generate new scientific knowledge. More research
and researchers are required to identify, problems in students learning of chemistry and how to best
teach the subject of chemistry. Advancement in the world of chemistry can be achieved in a much
shorter period if all who graduates in chemistry do actually understand the chemical concepts and
processes in chemistry and are able to contribute immediately in the accumulation of new knowledge.
Acknowledgement
This work is supported, in part, by the Institute of Research, Development and Consultancy and by
the Faculty of Applied Sciences, UiTM, Shah Alam.
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10. Shariff, Z. & Jantan, J. (2003). Proceedings Symposium Kimia Analisis Malaysia XVI,
Sarawak, Malaysia.
SYNTHESIS AND FLUORESCENCE CHARACTERISTIC OF 2-SUBSTITUTED
AND 6-SUBSTITUTED PURINES
Zanariah Abdullah, *Maizatul Akmam A. Bakar, Ernie Iryana Awang Din, Nadiah Rifhan
Abd. Rani, Noordini Mohd Salleh, Goh Poh Leong, Liow Pei Ling and Zaharah Aiyub
Chemical Synthesis Group, Department of Chemistry, Faculty of Science, University of Malaya,

50603, Kuala Lumpur
email: zana@um.edu.my
Abstract. 2-Fluoropurine was prepared from 2–aminopurine through a diazotization reaction, followed by
treatment with fluoroboric acid. 2-Aminopurine was obtained from a series of reactions, using 5-nitrouracil as
the starting material. 2-Piperidino and 2-anilinopurines were obtained by treating 2-fluoropurine with piperidine
and aniline respectively. 6-Piperidino and 6-anilinopurines were obtained by treating 6-chloropurine with
piperidine and aniline. Fluorescence studies was carried out in various solvents and maximum fluorescence was
observed in 75% ethanol. 2-Aminopurine showed the highest fluorescence intensity, followed by 2-anilino and
2-piperidinopurines. 6-Substituted purines showed stronger fluorescence intensities compared to 2-substituted
purines.
Abstrak. 2-Fluoropurina disediakan daripada 2-aminopurina melalui tindak balas pendiazoan diikuti dengan
pengolahan dengan asid fluoroborik. 2-Aminopurina diperolehi melalui beberapa peringkat tindak balas bermula
dengan 5-nitrourasil. 2-Piperino dan 2-anilinopurina diperolehi apabila 2-fluoropurina ditindak balas dengan
piperidina dan anilina. 6-Anilino dan 6-piperidinopurina pula diperolehi apabila 6-kloropurina ditindak balaskan
dengan piperidina dan anilina. Kajian pendafluoran dilakukan menggunakan berbagai pelarut dan kadar
pendafluoran maksima diperolehi dalam 75% etanol. 2-Aminopurina menunjukkan kadar pendafluoran yang
tinggi diikuti dengan 2-anilino dan 2-piperidino purina. Purina tertukar ganti di kedudukan 6 menunjukkan
kadar pendafluoran yang lebih tinggi berbanding dengan purina tertukar ganti di kedudukan ke 2 yang setara.
Introduction
The purine system is one of the most important systems present in living systems. The purine system
or rings can be found in many natural products, including nucleotides, co-enzymes and several
compounds which are valuable for treatment of cancer. Even though many of its rings can be found
in the natural occurring products, the parent compound itself cannot be found in nature.
The first pure purine, uric acid, was obtained by the isolation from kidneystones in 1776 [1] but its
structure was confirmed one hundred year later by Medicus [2]. But the chemistry of purines
flourished from 1906 onwards [3-5] until today.
The purine ring can be synthesized using two main synthetic route, either using pyrimidine or
imidazole as the precursor. The most extensively used method is the Traube Sythesis which involved
the use of diaminopyrimidines, whereby the diaminopyrimidines are condensed with a simple
compound to supply a one-carbon fragment to bridge the two pyrimidine amino nitrogen atoms to
form a five-membered ring. In this work, the Traube method was used in the synthesis of 2-
fluoropurine.
The fluorescence characteristic of purines or other heterocycles are not extensively studied,
eventhough a wide variety of heterocyclic compounds are known to be fluorescent [6]. Fluorescence
studies of these compounds are made more difficult because their fluorescence characteristics are
often dependent on the solvents used. The aim of this work is to study the fluorescence charateristic of
selected alkylaminopurines and the effect of solvent on the fluorescence characteristic. In this paper,
only the characteristic of 2- and 6- anilino, 2- and 6-piperidino purines will be discussed.
Experimental
Synthesis of 2-fluoropurine
2,4-Dichloro-5-nitropyrimidine [7]
5-Nitrouracil (26 g), phosphoryl chloride (130 ml), and dimethylaniline (32 ml) were heated with
occasional shaking until the reaction commerced. When this has subsided, the mixture was refluxed
for 1.5 hours, cooled and phosphoryl chloride was evaporated off. The residue was poured onto
crushed ice with vigorous stirring, and extracted with ether (500 ml). The ether extracts were washed
with water and dried over anhydrous sodium sulphate. Removal of ether and vacuum distillation gave
pure product.
89%, IR (cm-1):1670, 1620, 1350, 785; 1H NMR (CDCl3) δ: 9.26, s, 1H, (H6).
2, 4-Diamino-5-nitropyrimidine [8]
2, 4-Dichloro-5-nitropyrimidine (10.3 g) and phenol were heated under reflux while a stream of
ammonia was passed into the mixture for four hours. The phenol was distilled off and the residual
suspension was cooled and filtered. The crystals of 2, 4-diamino-5-nitropyrimidine were washed with
water, followed by ethanol and dried.
90%, IR (cm-1): 3345, 1675, 1630, 1350; 1H NMR (CDCl3) δ: 9.15, s, 1H, (H6), 7.20, b, 2H (NH2 of
C4), 6.50, b, 2H (NH2 on C2).
2, 4, 5-Triaminopyrimidine [8]
Finely powdered 2, 4-diamino-5-nitropyrimidine (8.9 g) was heated to 80 oC with water (15 ml) and
mechanically stirred. Sodium dithionite (3.75 g) was added during 3 - 4 minutes. The resulting
solution was stirred until 60 oC, followed by addition of powdered anhydrous sodium carbonate (5.5
g). The thick suspension was taken to dryness in an open basin on water bath. The solid was machine
ground and extracted for 20 minutes with stirred boiling alcohol (22 ml) which was filtered while hot.
The filtrate was taken to dryness and the product was re-extracted with ethanol (7.5 ml) to remove
sodium carbonate. Evaporation of filtrate gave the pure product.
71%; IR (cm-1): 3363, 1670, 1625; 1H NMR (DMSO-d6) δ: 7.25 s, 1H (H6), 6.00, b, 4H (NH2 of C2
and C4), 5.10, b, 2H (NH2 of C5).
2-Aminopurine [9]
2, 4, 5-Triaminopyrimidine (0.3 g), formyl morpholine (1.2 ml) and formic acid (0.6 ml) was heated
under refluxed for one hour in nitrogen atmosphere. Acetone (2 ml) was added to precipitate 2-
aminopurine which was dissolved in boiling water (3 ml). The solution was passed through a wide
filter, cooled to 50 oC and diluted with 3N nitric acid and refrigerated. The nitrate was filtered off. The
solution was then suspended in boiling water and brought to pH 6.8 with sodium citrate and 6N
sodium hydroxide solution. The solution was boiled with charcoal, giving buff-coloured crystals of 2-
aminopurines. Crystallisation with boiling water gave colourless crystals.
40%, IR (cm-1): 3245, 1670, 1620; 1H NMR (D2O) δ: 8.70, s, 1H (H6), 8.00 s, 1H (H8) 6.25, b, 2H
(NH2), 6.50, b, 1H (NH).
2-Fluoropurine (improved method) [10, 11]

An aqueous solution of sodium nitrite (800 mg in 4 ml of water) was added with stirring to a solution
of 2-aminopurine (200 mg) in 48% fluoroboric acid ( 22 ml) at the rate of 0.1 ml per minute at -11 oC.
After the addition was completed, the mixture was stirred for one hour between
- 10 to 0 oC. The mixture was then neutralized with 50% sodium hydroxide solution. The neutral
slurry was evaporated to dryness. The crude purine was isolated by extraction of dry residue with
ether in a soxhlet extractor. Evaporation of ether gave crude product, which was recrystallised from
water.
35%, IR (cm-1): 3325, 1673, 1623, 1132; 1H NMR (DMSO-d6) δ: 8.70 s, 1H, (H6), 8.00 s, 1H (H8),
6.50, b, 1H (NH).
2-Piperidinopurine
Piperidine (20 mg) in ethanol (4 ml) was added to a solution of 2-fluoropurine (40 mg) in ethanol (7
ml) and the mixture was refluxed for 1 hour at 100 oC. The mixture was cooled and ethanol was
evaporated off. The slurry was extracted twice with ether. The ethereal layer was washed with water
and dried over anhydrous sodium sulphate. Evaporation of ether gave crude product, which was
recystallised from petroleum ether.
50%, decomposed above 215, IR (cm-1): 3115, 1673, 1620; 1H NMR δ: 8.70, s, 1H (H6), 8.00, s, 1H
(H1), 3.35, m, 4H (H2’, H6’), 1.68, m, 6H (H3’, H4’, H5’); M+: 203.1163.
2-Anilinopurine
2-Fluoropurine (80 mg) was added to aniline (3 ml) and warmed at 60 oC for four hours. The mixture
was cooled and refrigerated overnight. 2-Anilinopurine was crystallized out of the reaction mixture.
The crystal was filtered, washed with ice-cold water and dried. Pure product was obtained after
recrystallisation from dichloromethane.
73.5%, decomposed above 200o, IR (cm-1): 3330, 1670, 1630; 1H NMR (DMSO-d6) δ: 8.65, s, 1H
(H6), 8.00, s, 1H (H8), 7.20, m, 3H (H3’, H4’, H5’), 6.90, d, 2H (H2’ and H6’), 5.40, d, 2H, (N-H);
M+:211.0858.
6-Piperidinopurine
Piperidine (0.5 ml) in ethanol (3 ml) was added to 6-chloropurine (0.232 g) [11-12] and the mixture
was refluxed for 2 hours. The mixture was cooled and ethanol was evaporated off. The slurry was
extracted twice with ether; the ethereal layer was washed with water and dried over anhydrous sodium
sulphate. Evaporation of ether gave grey product, which was recystallised from petroleum ether.
30%, decomposed above 270, IR (cm-1): 3115, 1673, 1620; 1H NMR (DMSO-d6) δ: 8.23, s, 1H (H2),
8.15, s, 1H (H8), 2.62, m, 4H (H2’, H6’), 1.66, m, 6H (H3’, H4’, H5’); M+: 203.1163.
6-Anilinopurine
2-Chloropurine (0.161 g) was added to aniline (0.3 ml) in ethanol (3 ml) and refluxed for 3 hours. The
mixture was cooled and the solvent was evaporated. The slurry was extracted with ether, washed with
water and dried over anhydrous sodium sulphate. Evaporation of ether gave light brown solid.
50.3%, decomposed above 280o, IR (cm-1): 3330, 1670, 1630; 1H NMR (DMSO-d6) δ: 9.81, s, 2H (N-
H), 8.43, s, 1H (H2), 8.34, s, 1H (H8), 8.01, d, 2H (H2’ and H6’), 7.38, t, 2H (H3’, H5’), 7.08, t, 1H (H4’);
M+: 211.0858.
Spectroscopic Analysis
All solvents were redistilled before use. Melting points were determined with Electrothermal Melting
Point Appararus and were not corrected. Infrared spectra were recorded using Perkin Elmer 298
Infrared Spectrometer and FTIR Perkin Elmer 1600 Series. 1 H NMR spectra were recorded on Bruker
WP-80 and Bruker AM 250.
Fluorescence studies
2- and 6-Substutitued purines at the same concentration were prepared in various solvents. Quinine
sulphate with the same concentration as the compounds under studied were also prepared and used as
the standard. The fluorescence intensity of quinine sulphate was taken to be 1.00. The fluorescence
measurement was carried out using Fluorescence Spectrometer Model F-2000 Hitachi.
The synthetic route of 2-fluoropurine is as shown below:-
H
N Cl N NH2
N O
POCl3/C6H5NMe2 Phenol/NH3
NH N N
O2N O 2N O 2N
O Cl NH2
1 2 3
sodium dithionite
N NH2
N N
N NaNO2/HBF 4 N HCOOH
N N
F N NH2 N N H2N
H H NH2
6 5
4
Scheme 1
The preparation of 2-fluoropurine was carried out according to the Brown method [4 ], using 5-
nitrouracil (1) as the starting material. Purine ring was obtained through cyclisation reaction of 2, 4, 5-
triaminopyrimidine (4) with formic acid to give 2-aminopurine (5). Compound 5 undergoes
diazotization reaction, followed by treatment with HBF4 to give 2-fluoropurine (6). Low percentage
yield was obtained in the synthesis of 6-chloropurine. Due to low percentage yield obtained, 6-
chloropurine used in this work was obtained commercially.
Treatment of 2-fluoropurine and 6-chloropurine with aniline and piperidine are as shown in Schemes
2 and 3.
N
NH2 N
H
N N N
H
N
N
2-anilinopurine
F N N
H
N
2-fluoropurine N
N
H
N N N
H
2-piperidinopurine
Scheme 2
NH2 H N
Cl N
N
N
N N
N
N H
N
6-anilinopurine
H
6-chloropurine
N
N N
N
H
N N
H
6-piperidinopurine
Scheme 3
The structures of 2- and 6- substituted purines were confirmed by 1H NMR and mass spectra as given
in the experimental section.
The fluorescence studies of 2- and 6-substituted purines were carried out in 75% ethanol. Quinine
sulphate at the same concentration as the compounds studied was used as the standard and its
fluorescence intensity was taken to be 1.00. The fluorescence band of 2- and 6-substituted purines
were given in Table 1.
Purine Solvent Excitation Fluoresence Rel. Fluorescence

wavelength/nm wavelength/nm intensity
2-fluoro 75% EtOH 350 430 0.640
2-amino 75% EtOH 340 380 0.964
2-anilino 75% EtOH 340 473 0.371
2-piperidino 75% EtOH 340 390 0.307
6-chloro 75% EtOH 330 420 0.650
6-anilino 75% EtOH 340 480 0.452
6-piperidino 75% EtOH 340 395 0.351
Table 1: Fluorescence peaks of 2- and 6- substituted purines in 75% ethanol
It can be seen from the table that, the fluorescence intensity of 6-substituted purines is stronger than 2-
substituted derivatives. This is probably due to at the 6- position, the substituent is not sandwich
between the two nitrogen atoms as in the 2nd position. As the result, the electrons can move freely
from the substituent to the rest of the ring.
Anilinopurines showed stronger fluorescence intensities and fluoresced at a higher wavelength

compared to the piperidinopurines. The fluorescence peak observed at a higher wave length is
believed to be due to the increase in the degree of conjugation in the anilinopurines compared to
piperidinopurines. The same phenomena was observed with pyrimidine derivatives studied earlier
[13]. Lower fluorescence intensity was recorded with piperidinopurines in 75% ethanol. This is
probably due to the piperidino ring flipping from one conformation to another, resulted in the loss of
energy in the transition state. As the result, low fluorescence was observed.
N N N
N N N
N N N
N N N N
N N
H H H
Flipping of piperidino ring
2- and 6-Piperidino purines were less rigid compared to 2- and 6- anilino purines. Some energy may
also loss in the transition state, which also resulted in low fluorescence intensity observed.
2-Aminopurines showed the highest fluorescence intensity amongst all the derivatives of purines
studied. This is probably due to the electron donating nature of the amino group which enhances the
mobility of electron in the system. The free mobility of the electron enhances the π to π∗ transitions,
which resulted in high fluorescence intensity.
Conclusion
All the 2- and 6-substituted purines studied are fluorescent compounds. 2-Aminopurine is the most
fluorescent and the fluorescence intensity depending on the type of the substituents. Substituent with
high degree of conjugation, as in the case of anilinopurine showed higher fluorescence intensity and
fluoresced at a higher wavelength. The purine system with unconjugated substituent (in the case of
piperidinopurines) fluoresced at a lower wavelength. Further work on the purine system and other
heterocyclic systems are still under going before any concrete conclusion can be made on solvent-
structure-fluorescence and substituent-fluorescence relationship.
Acknowledgement
Financial support of this work by University of Malaya and Academy of Science is gratefully
acknowledged.
References
1. Scheel K. W., Opuscula, (1776), 2, 73.

2. Medius, V. L. (1875), Annalen, 175, 230.
3. Baxter, R. A. and Spring F. S., (1944), Nature, 154, 462.
4. Brown D. J. and Waring P., (1974), J. Chem. Soc. Perkins Trans. II, 204.
5. Tong Y. C., (1975), J. Heterocyclic Chem., 451.
6. Abdullah Z., PhD. Thesis, (1989), Queen Mary & Westfield College, Uni. of London.
7. Whittaker N., (1951), J. Chem Soc., 1565.
8. Brown D. J., Albert, A. and Cheeseman G. (1951), J. Chem Soc., 474.
9. Albert A. and Brown D. J., (1954), J. Chem Soc., 2060.
10. Montgomery J. A. and Hewson J., (1960), J. Amer. Chem. Soc., 82, 463.
11. Abdullah, Z. and A. Bakar M. A. (2005), sent to Jour. of Organic Chem.
12. Robins R.K., Dille K.J., Willits L.H. and Christensen K., (1975), J.Amer. Chem. Soc., 388.
13. Abdullah, Z., Mohd Tahir N., Abas M. R., Low B. K. and Aiyub Z. (2004), Molecules, 9, 520-
26.
TINDAK BALAS PENGETOKSILAN ASID LAURIK BERASASKAN MINYAK KELAPA
SAWIT
Zarina Edris*, Norhafipah Mohamad, M. Ambar Yarmo
Jabatan Kimia, Pusat pengajian Sains Kimia dan Teknologi Makanan, Fakulti Sains dan Teknologi,
Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia.
e-mail: zarinaedris@yahoo.co.uk
Abstrak. Dalam kajian ini, surfaktan tidak berion berasaskan tindak balas antara asid laurik (C12)
minyak kelapa sawit dengan EO berjaya dilakukan. Sintesis ini menggunakan kaedah pengetoksilan
asid laurik dengan siri mol gas etilena oksida iaitu 3,5 dan 7 untuk mengkaji kesan perbezaan di
antaranya. Tindak balas ini menggunakan 1% KOH sebagai mangkin dan dilakukan pada suhu di
antara 130°C-135°C di dalam reaktor khas. Produk teretoksilat dicirikan menggunakan teknik FTIR
dan hasil menunjukkan kehadiran kumpulen ester melalui regangan C=O pada jarak gelombang 1700
cm-1 dan ikatan O-H pada jarak gelombang 2500 – 3450 cm-1. Analisis menggunakan HPLC pula
menunjukkan peratus penukaran hasil yang tinggi (>90%). Pengaruh bilangan mol gas etilena oksida
terhadap sifat-sifat surfaktan ini dikaji dan didapati bahawa asid lemak teretoksilat (ALT) mempunyai
ciri-ciri surfaktan tidak berion yang boleh digunakan dalam detergen.
Abstract. In these studies, nonionic surfactant based on reaction between lauric acid from palm oil
derivative with ethylene oxide product was produced successfully. Ethoxylation reaction was carried
out between acid lauric with varies moles of ethylene oxide (EO) namely 3, 5 and 7 was investigated
effect of different that. The reaction was performance using 1% KOH as a homogenous catalyst
performs reaction at temperature between 130°C-135°C. The product was characterization using FTIR
technique showed the present of ester group by C=O stretch at wavelength 1700 cm-1 and O-H bond at
2500 – 3450 cm-1. HPLC analysis showed a high percent conversion of product (>90%). The effect of
moles of EO to surfactant properties are investigated and found that this product have a surfactant
properties that can use as a special detergent.
Katakunci: pengetoksilan, asid laurik minyak kelapa sawit, surfaktan tidak berion.
Pengenalan
Kegunaan utama bahan-bahan oleokimia selain daripada sektor makanan adalah menghasilkan
surfaktan. Sifat ampifilik yang terdapat pada asid lemak boleh diubahsuai dengan mengubah
kumpulan karboksil kepada kumpulan hidrofilik lain. Keadaan ini boleh menghasilkan pelbagai jenis
surfaktan sama ada anionik, kationik, amfoterik dan tidak berion [1]. Rantai karbon seperti C12-C14
daripada minyak laurik, C22 daripada minyak ikan dan C16-C18 daripada kebanyakkan sumber lain juga
boleh digunakan dalam menghasilkan surfaktan. Kebanyakkan surfaktan menggunakan asid lemak
dan alkohol lemak yang mempunyai terbitan amida dan amina sebagai bahan pemula [1].
Tindak balas pengetoksilan berasaskan terbitan produk minyak kelapa sawit merupakan
alternatif kepada penghasilan surfaktan yang berasaskan minyak kacang soya dan petroleum seperti
sulfonat dan sulfat [2]. Penghasilan surfaktan ini banyak digunakan dalam sektor tekstil, farmasi,
detergen, penjagaan diri, kosmetik dan lain-lain sebagai agen pelembap, pengemulsi dan egen
pembuih. Dalam industri petroleum pula, surfaktan tidak berion digunakan sebagai pengemulsi bagi
menstabilkan minyak mentah dalam air, penghidrat minyak mentah dan menghalang penyingkiran
paraffin [2].
Penggunaan surfaktan teretoksilat khususnya surfaktan tidak berion telah berkembang sejak
30 tahun yang lalu mewakili lebih 25% daripada hasil pengeluaran surfaktan dunia. Surfaktan jenis ini
tidak menghasilkan ion dalam larutan akues dan bersesuaian dengan jenis agen tensoaktif yang lain
dalam formulasi kompleks. Ianya kurang sensitif terhadap kehadiran elektrolit, terutamanya ikatan
divalen kation dan bahagian kaunter ionnya pula menjadikan surfaktan ini sangat berguna dalam
larutan air bergaram [2]. Sintesis surfaktan tidak berion boleh dilakukan melalui tindak balas di antara
etilena oksida (EO) atau propilena oksida dengan kumpulan organik yang mengandungi hidrogen
aktif atau kumpulan berfungsi lain dalam kehadiran mangkin bes [1,3]. Tindak balas umum ini
ditunjukkan dalam persamaan 1 seperti di bawah [2]
O Mangkin
bes
R-X-H + n H2C CH2 R-X-[CH2-CH2-O]n-H
Persamaan 1
X= -O-, -COO-,-CONH-,-NH.
R= bahagian rantai panjang organik
Surfaktan berasaskan oleokimia seperti minyak kelapa sawit boleh terurai lebih baik daripada
petrokimia dan sumbernya juga boleh diperbaharui. Apa yang paling penting, ianya tidak
menghasilkan bahan cemar seperti NOx, SO2, CO dan hidrokarbon [4]. Dalam kajian ini, proses
pengetoksilan dilakukan ke atas asid laurik daripada sumber minyak kelapa sawit. Asid laurik
merupakan komponen yang paling banyak terdapat dalam isirong buah sawit. Kelebihan Malaysia
sebagai pengeluar utama minyak sawit dunia sehingga tahun 2010 kelak memberikan kemudahan bagi
mendapatkan bahan mentah yang lebih murah untuk menghasilkan surfaktan [8]. Fenomena ini
sekaligus membuka ruang kepada Malaysia bagi menceburi dan menguasai pasaran surfakatan dunia
kelak.
Eksperimen
Bahan kimia
Etilena oksida (EO) (99.8% ketulenan) yang diperolehi dari Fluka. Asid laurik (99% ketulenan)
daripada Cognis, kalium hidroksida (85% ketulenan) daripada Aldrich, metanol (99.99% ketulenan)
daripada BDH dan Asetonitril bergred HPLC (99.99% ketulenan) daripada Ficher. Air ternyahion
(18.2MΩ) digunakan dalam semua penyediaan.
Prosedur Tindak balas
Tindak balas pengetoksilan dilakukan dalam reaktor 1L yang diperbuat daripada besi tahan karat
(daripada Vinci Technology, Perancis). Reaktor ini dilengkapi dengan sistem pengacau, pemanas
reaktor dan pengawal suhu. Sebelum tindak balas dilakukan, 1% KOH dalam metanol disediakan
sebagai larutan mangkin bes. Kemudian mangkin ini dimasukkan ke dalam reaktor dan dipanaskan
pada suhu 95°C dalam keadaan vakum selama 1 jam. 100g asid laurik (Cognis) dicampurkan ke
dalam reaktor dan suhu tindak balas ditingkatkan kepada 135°C. Apabila suhu telah stabil, gas EO
akan disuntik masuk ke dalam sistem reaktor mengikut bilangan mol yang dikehendaki iaitu 3,5 dan 7
mol.
Suhu tindak balas akan meningkat secara mendadak sejurus selepas kemasukkan EO
menunjukkan tindak balas ini adalah jenis eksotermik. Tindak balas pengetoksilan dibiarkan berlaku
selama 2 jam sebelum suhu reaktor diturunkan kepada suhu bilik. Gas nitrogen dilalukan bagi
menyejukkan produk dan menyingkirkan lebihan EO yang mungkin tidak bertindak balas sebelum
produk dikeluarkan.
Pencirian
Produk yang telah dikeluarkan akan dicirikan menggunakan teknik FTIR jenis Perkin Elmer Model
GX dan HPLC jenis Agilent Technologies Siri 1100 dengan pengesan ELSD jenis Altech Model
2000. Bagi mendapatkan spektum FTIR, nombor gelombang yang direkodkan adalah dalam julat
450 cm-1 sehingga 4000 cm-1 dengan resolusi 1 cm-1. Penyediaan untuk analisis FTIR adalah
berdasarkan teknik pepejal KBr dan CH2Cl2.
Kromatogram HPLC diperolehi dengan menggunakan turus jenis Zorbax 300SB-C18 (4.6mm
x 250mm).Pelarut yang digunakan adalah campuran asetonitril dan air pada nisbah 77:23. Amaun
sampel yang disuntik adalah 20 µL dengan kadar alir adalah 1 ml/min.
Keupayaan pembuihan dan kestabilan pembuihan
Analisis penentuan pembuihan telah dilakukan terhadap surfaktan tidak berion. Ia biasanya terbahagi
kepada penentuan keupayaan pembuihan (foaming power) iaitu kemampuan surfaktan menghasilkan
buih dan kestabilan pembuihan (foaming stability) iaitu kemampuan surfaktan mengekalkan tahap
pembuihan dalam masa tertentu. Akan tetapi sesuatu surfaktan yang mempunyai kuasa pembuihan
yang baik tidak semestinya memberikan kestabilan pembuihan yang baik [5].
Kaedah yang digunakan dalam penentuan pembuihan adalah diperolehi daripada AOTD in-
house method. Kaedah ini menggunakan silinder 500ml yang diisi dengan larutan surfaktan (0.2g
dalam 200ml air ternyahion). Larutan surfaktan tersebut akan dikocak dengan menggunakan rod
selama 30 kali pada kadar yang tetap. Selepas pembentukan buih, bacaan akan diambil daripada takat
atas dan takat bawah silinder. Bagi keupayaan pembuihan, ia diukur selepas 30 kali kocakan
manakala kestabilan pembuihan diukur selepas 5 minit kocakan. Kedua-dua parameter yang
digunakan adalah seperti:
Keupayaan pembuihan = [(Takat atas – Takat bawah)/Jumlah ketinggian] Persamaan 2
(selepas 5 minit),
Kestabilan pembuihan = [(Takat atas – Takat bawah)/Jumlah ketinggian] Persamaan 3
Keupayaan Detergen
Keupayaan detergen surfaktan yang dihasilkan dilakukan dengan menggunakan fabrik cotton AS9
pigment/oil. Kaedah pencucian ini menggunakan 0.1g surfaktan tidak berion. Kemudian surfaktan
tersebut akan dicairkan dengan 100 ml air suling dan dicampur dengan 50 ppm air liat (hardness
water) selama 3 minit. Selepas proses pencampuran, fabrik cotton AS9 pigment/oil dimasukan ke
dalam bekas untuk pencucian selama 10 minit. Akhir sekali, fabrik cotton AS9 pigment/oil dibilas
sebanyak dua kali dengan 1000 ml dan 500 ml air liat (hardness water).
Spektrofotometer CM36000 telah digunakan untuk menentukan peratus penyingkiran kotoran

melalui persamaan di bawah:
% penyingkiran kotoran = [(AW-BW)/ (BS-BW)] × 100 Persamaan 4
dengan
AW = selepas cucian terhadap cotton AS9 pigment/oil
BW = sebelum cucian terhadap cotton AS9 pigment/oil
BS = sebelum cucian dalam standard clothes
Keseimbangan hidrofilik-lipofilik
Keseimbangan hidrofilik-lipofilik menunjukkan hubungan antara hujung hidrofilik dan hujung

lipofilik bagi suatu surfaktan. Biasanya, surfaktan yang larut minyak mempunyai nilai keseimbangan
hidrofilik-lipofilik yang tinggi. Nilai tersebut juga berkaitan dengan peratus kandungan etilena oksida
dalam surfaktan. Jadual di bawah menunjukkan nilai HLB, kandungan etilena oksida dan
penggunaannya.
Jadual 1: Nilai HLB , kandungan etilena oksida dan penggunaannya
Nilai HLB Kandungan Etilena Oksida Penggunaan

(% berat)
4–6 20 – 30 Pengemulsi air dalam minyak
7 – 15 35 – 75 Agen pembasahan
8 –18 40 – 90 Pengemulsi minyak dalam air
10 – 15 50 – 75 Bahan pencuci
10 – 18 50 – 90 Solubilizer
Nilai keseimbangan hidrofilik-lipofilik bagi molekul surfaktan ditunjukkan dan dikira secara teori
melalui persamaan:
Asid laurik teretoksilat:

HLB = 20 x Berat molekul bahagian hidrofilik / Berat molekul bagi molekul Persamaan 5
Rajah 1: Profil suhu bagi tindak balas pengetoksilan asid laurik (ALT-3EO).
Sintesis surfaktan tidak berion jenis asid laurik teretoksilat (ALT) telah berjaya dihasilkan
berdasarkan Persamaan 1. Bahan pemula yang digunakan adalah asid laurik daripada minyak kelapa
sawit dengan menggunakan 1% KOH sebagai mangkin. Tindak balas dilakukan dalam reaktor besi
tahan karat pada suhu 135°C. EO disuntik ke dalam reaktor sehingga nisbah molar EO terhadap asid
laurik tercapai iaitu 3,5 dan 7 mol.Rajah 1, menunjukkan profil suhu bagi tindak balas eksotermik
ALT-3EO. (a) menunjukkan penurunan suhu akibat kemasukkan asid laurik ke dalam reaktor.
Kemudian suhu reaktor dinaikkan kepada 130°C -135°C.
Setelah mencapai kestabilan suhu, EO disuntik masuk. (b) menunjukkan sedikit penurunan
suhu akibat kemasukkan EO kemudian terus mendadak naik. Pada keadaan ini, tindak balas
eksotermik telah bermula kerana berlaku peningkatan suhu melebihi suhu yang ditetapkan (130°C -
135°C). Masa tindak balas dibiarkan selama 2 jam (c) dan didapati terdapat penurunan dan
peningkatan suhu pada tindak balas tersebut. Menurut kajian yang pernah dilaporkan, proses
pengetoksilan asid lemak berlaku dalam dua peringkat. Peringkat pertama berlaku pembentukan
produk sampingan. Kemudian secara perlahan-lahan tindak balas di antara asid dan etilena oksida
akan bermula selepas masa induksi untuk memberikan lebih banyak etilena glikol monoester. Pada
peringkat kedua, tindak balas akan meningkat iaitu selepas kemasukkan lebih kurang 1 mol EO [6].
Rajah 2 : Spektrum FTIR bagi ALT-3EO, ALT-5EO dan ALT-7EO.
Selepas tindak balas, pepejal asid laurik yang asalnya berwarna putih telah bertukar kepada
cecair yang berwarna kuning keemasan. Hasil analisis daripada FTIR menunjukkan kehadiran
kumpulan ester melalui regangan C=O pada jarak gelombang 1700 cm-1 dan ikatan O-H pada jarak
gelombang 2500 – 3450 cm-1. Analisis ini telah mengesahkan persamaan tindak balas pengetoksilan
asid laurik di sebelah
O Mangkin O
Bes ||
C11H23COOH-H + n H2C CH2 C11H23 -CO-[CH2-CH2-O]n-H
Asid laurik EO Asid laurik teretoksilat

Persamaan 6
Peratus penukaran produk telah ditentukan dengan menggunakan teknik HPLC. Daripada
analisis kromatogram, peratus penukaran produk bagi asid laurik teretoksilat hampir mencapai 100 %
seperti yang tercatat dalam Jadual 1 di bawah dan pada Rajah 3 menunjukkan kromatogram HPLC
bagi asid laurik teretoksilat. Puncak pada masa penahanan 6.174, 6.222 dan 6.237 minit adalah
komponen monoester yang merupakan komponen utama dalam ALT manakala puncak pada masa
2.777, 2.779 dan 2.780 minit adalah komponen diester.
Jadual 2 : Peratus penukaran produk melalui analisis HPLC.

Produk Bilangan mol EO Masa penahanan Peratus penukaran
(min) produk
Asid laurik - 5.644 -
teretoksilat 3EO 6.237 99.32 %
(ALT) 5EO 6.222 99.24 %
7EO 6.174 81.49 %
(a) (b)
(c) (d)
Rajah 3 Kromatogram HPLC bagi (a) asid laurik, (b) ALT-3EO (c) ALT-5EO dan (d) ALT-7EO
Keupayaan pembuihan dan kestabilan pembuihan
Buih dikenali sebagai penyerakan yang dihasilkan daripada gas buih dalam sesuatu cecair. Dalam
detergen, jumlah buih yang terbentuk biasanya dianggap sebagai penentuan terhadap keberkesanan
detergen tersebut [7]. Buih yang stabil terbentuk apabila surfaktan terjerap ke dalam permukaan
udara/air dengan molekul tersusun membentuk struktur lamella. Interaksi antara kumpulan hidrofilik
surfaktan dalam susunan dual lapisan (bilayer arrangement) juga memberikan kesan terhadap
kestabilan pembuihan selain interaksi terhadap permukaan surfaktan [8]. Secara umum, surfaktan
tidak berion memberikan sifat pembuihan yang rendah. Ia juga dapat memberikan tahap pembuihan
yang berbeza dengan ciri yang berbeza. Sebagai contoh, terdapat surfaktan yang memberikan
kestabilan buih yang baik dan ada juga surfaktan yang mempunyai sifat buih yang mudah runtuh
dengan cepat.
0 minit 5 minit
180
160
140
K etinggian buih (m l)
120
100
80
60
40
20
0
3 EO 5 EO 7 EO
Laurik asid teretoksilat
Rajah 4 : Keupayaan pembuihan dan kestabilan pembuihan bagi asid laurik teretoksilat
Hasil ujian ke atas ALT-3EO, ALT-5EO dan ALT-7EO menunjukkan bahawa keupayaan pembuihan
meningkat mengikut peningkatan bilangan mol EO. Dengan menggunakan Persamaan 2 dan 3,
kestabilan buih bagi setiap surfaktan didapati berbeza. ALT-5EO adalah lebih stabil berbanding ALT-
3EO dan ALT-7EO. Keadaan ini berbeza seperti apa yang pernah dilaporkan sebelum ini bahawa
kelikatan cecair yang tinggi akan memberikan kestabilan buih yang tinggi . Ini kerana kelikatan cecair
yang tinggi akan mengurangkan pembentukan buih-buih besar yang mudah pecah melalui
penggabungan buih-buih kecil [9]. Namun begitu ujian ini agak sukar dijelaskan kerana kestabilan
buih turut dipengaruhi oleh banyak faktor seperti ketegangan permukaan, taburan saiz gelembung
udara, kemasukkan udara dalam gelembung, kelikatan permukaan, elastik Gibbs, pengaliran cecair
dari lemella, reologi penjerapan lapisan, tekanan luar dan suhu [10].
Keupayaan Detergesi ALT
29
% Penyingkiran kotoran
28
27
26
25
24
23
22
0
3 EO 5 EO 7 EO
Surfaktan tidak berion
Rajah 5 : Peratus keupayaan penyingkiran kotoran oleh ALT
Keupayaan detergen adalah merujuk kepada kemampuan sesuatu surfaktan dalam membantu proses
penyingkiran kotoran daripada fabrik. Ia melibatkan pengemulsian, pemelarutan dan daya penolakan
antara fabrik dan partikel kotoran [11].
Keupayaan detergen boleh dibahagikan kepada pembasahan ke atas fabrik yang dibasuh
dengan menggunakan larutan detergen atau sabun, penyingkiran kotoran daripada permukaan fabrik
dan mengawal kotoran dalam larutan stabil. Dalam air pembasuhan, sabun atau detergen mampu
meningkatkan keupayaan pembasahan dan juga dengan mudah menembusi fabrik untuk mencapai
kotoran. Oleh yang demikian, penyingkiran kotoran telah bermula [12].
Ujian ke atas sampel ALT-3EO, ALT-5EO dan ALT-7EO,menunjukkan bahawa ALT-5EO
mempunyai keupayaan detergen yang lebih baik. Keadaan ini dipengaruhi oleh panjang rantai EO dan
struktur molekul surfaktan. Bagi ALT-3EO dan ALT-5EO, struktur molekul surfaktan adalah
berbentuk monomer dan molekul ini akan menjerap ke permukaan kotoran dengan bahagian hidrofilik
mengarah ke permukaan air. Oleh kerana bilangan rantai EO lebih tinggi bagi ALT-5EO, kotoran
mudah ditanggalkan kerana keterlarutannya yang lebih berbanding ALT-3EO.
Walaupun ALT-7EO mempunyai bilangan mol EO yang lebih tinggi, namun kehadirannya
dalam 100ml air menjadikan ia agak tepu dan menyebabkan pembentukan banyak misel. Keadaan ini
menyebabkan hanya sedikit sahaja molekul surfaktan yang terjerap ke permukaan kotoran untuk
ditanggalkan. Oleh itu pencucian yang lebih baik adalah yang menggunakan kepekatan yang sedikit
[13]. Dalam keadaan ini, ALT-5EO memberikan pencucian yang lebih baik.
Keseimbangan hidrofilik-lipofilik (Hydrophilic-Lipophilic Balance, HLB)
Residu yang tertinggal semasa proses pencucian boleh memberikan kesan terhadap kebolehan sesuatu
bahan pencuci. Dalam sistem akueus, nilai HLB yang tinggi akan memberikan kehadiran residu yang
sedikit selepas melalui proses pembilasan. Tambahan lagi, pembilasan yang baik juga boleh
diperolehi daripada surfaktan yang mempunyai bilangan EO yang tinggi. Dalam ujian ini, nilai HLB
dikira berdasarkan Persamaan 5 dan didapati nilainya meningkat berdasarkan peningkatan bilangan
mol EO daripada 3-7.
Jadual 3 menunjukkan asid laurik teretoksilat dengan nilai EO 3, 5 dan 7 mempunyai nilai
HLB masing-masing 10.66, 12.62 dan 13.90. Nilai HLB ini menunjukkan bahawa surfaktan ini
cenderung untuk bertindak sebagai agen pembasahan, pengemulsi minyak dalam air dan bahan
cuci/detergen.
Jadual 3 : Asid laurik teretoksilat dan nilai HLB

Berat molekul Berat molekul bagi HLB
Surfaktan tidak berion bahagian hidrofilik molekul
Laurik asid teretoksilat 177 332 10.66
3 EO
5 EO
7 EO
Kesimpulan
Kajian ini telah berjaya menghasilkan surfaktan tidak berion berasaskan tindak balas pengetoksilan
terhadap asid laurik daripada minyak kelapa sawit. Hasil pencirian menggunakan FTIR dan HPLC
menunjukkan wujudnya komponen surfaktan yang dikehendaki dan juga nilai penukaran produk yang
tinggi. Ujian-ujian surfaktan seperti keupayaan pembuihan, keupayaan detergen dan HLB melaporkan
bahawa surfaktan ini mempunyai kuasa pembuihan yang baik dan stabil, kuasa pencucian yang
sederhana dan sesuai digunakan sebagai agen pembasahan, pengemulsi minyak dalam air dan bahan
cuci/detergen.
Penghargaan
MOSTE ‘Ministry of Science Technology and Innovation of Malaysia’ geran penyelidikkan IRPA 09-
02-02-0033 dan Advanced Oleochemical Technology Division AOTD.
Rujukan
1. Ovalles, C., Bolivar, R., Cotte, E., Aular, W., Carrasquel, J., Lujano, E. (2000) “Novel
Ethoxylated Surfactans from Low-value Refinery Feedstockc” Fuel. 80. 575-582.
2. Shahidi, F. (2005) “Bailey’s Industrial Oil and Fat Products”. 6th Ed, John Wiley and Sons,
Inc.New York. Ms 27-29.
3. O’Lenick, A.J., Jr., dan Parkinson, J.K. (1996) “A Comparison of The Ethoxylation of A
Fatty Alcohol, Fatty Acid and Dimethiconol” J. Am Oil Chem Soc. 73. 63-66.
4. Liana, L., (2003 Okt 7-8) 3rd “Global Oils and Fats Business Forum 2003 : Green Palmbased
Baby Products” California.
5. Wilson. A.J. (1989) Foams: Physics, Chemistry and Structure. Springer-Verlag. Great
Britain.
6. http://www.zenitech.com/documents/castor_oil.pdf capaian tahun 2005
7. Davidsohn. A & Milwidsky. B.M. 1978. Synthetic Detergents. 6th edition. John Wiley &
Sons Inc, New York.
8. Lim W.H. and Salmiah A. (2001) Surface Tension, Foaming and Detergency Properties of Mixed
Alpha-sulphonated Methyl Esters Derived From Palm Stearin with Some Commercial Sufactants.
Journal of oil palm research. Vol. 13 No. 1, p 75-83.
9. http://www.tramfloc.com/antifoam-select.doc capaian tahun 2005
10. Karsa D.R. (1988) Industrial Applications of Surfactant. The Royal Society of Chemistry,
London. Ms 148.
11. Piispanen. P. S. (2002) Synthesis and Characterization of Surfactants Based on Natural
Product. Vetenskep Och Konst. Kungl Tekniska Hogskolan (KTH).
12. Austin. G. T. (1984) Shreve’s Chemical Process Industries. 5th Ed. Mc Graw-Hill, Inc.
13. http://www.chemistryquestion.com capaian tahun 2005
14. Holmberg, K. (2000) “Natural Surfactants” Curr Opin Colloid Interface Sci. 6. 148-159.
15. Behler, A. (2000 Jun 20-21) “Final Conference CTVO-net : New Oleochemical Surfactants”.
Bonn. Germany.
16. McMurray, J., (2000) “Organic Chemistry” 5th Ed, USA, Brooks/Cole. Ms 441-458.
NITRATE AND PHOSPHATE CONTENTS AND QUALITY OF WELL WATER IN
NORTH-EASTERN DISTRICTS OF KELANTAN
Noor Wahida Mahasim1, Ahmad Saat2*, Zaini Hamzah1, Rita Rohaizah Sohari2,
Khaniza Hasliza Abdil Khali2
1
Faculty of Applied Sciences
Universiti Teknologi MARA, 40450 Shah Alam
2
International Education Centre (INTEC)
Universiti Teknologi MARA, Kampus Seksyen 17
40200 Shah Alam
*
Author for correspondence
Fax: 03-55227065, e-mail: ahmad183@salam.uitm.edu.my
Abstract. In the north-eastern district of state of Kelantan almost 40% of the populations are still relying on well water for
their domestic uses and consumption. Generally, the wells are located within the vicinity of agricultural activities, such as
tobacco, paddy, rubber plantation and orchards. There is possibility that the activities, especially related to application of
chemical fertilizers, may pollute the well either through runoff or underground transportation. A study was carried out on
fifteen wells in the district of Bachok, Pasir Puteh and Machang to determine the well water quality as well as the nitrate and
phosphate contents of the water samples. The sampling was done twice, one in early October 2004 (representing dry season
sampling), and the end of January 2005 (representing rainy season sampling). Data for turbidity, temperature, DO, and pH
were determined in-situ, while the nitrate and phosphate contents, BOD and COD were determined in laboratory. Water
Quality Index (WQI) of the samples was determined based on the method suggested by the Interim Water Quality Standard
(INWQS) for Malaysia. The study found that nitrate contents ranged between 0.01 to 2.42 mg/L in rainy season and between
non detectable to 0.3 mg/L in dry season. The corresponding phosphate contents for rainy and dry seasons are ranging
between 0 – 2.2 mg/L and 0.5 – 1.5 mg/L respectively. In general, WQI of all the wells studied fall into class III or IV.
However, WQI values of all the wells improved (increased) in wet season as compared to dry season. Based on the INWQS,
these classes of water required extensive treatment before being regarded as suitable for public water supply.
Key words: Well-water, WQI, phosphate, nitrate.
Abstrak. Di timur laut negeri Kelantan Darulnaim, hampir 40% penduduk masih bergantung kepada air telaga untuk
kegunaan domestik dan minuman. Amnya, kebanyakkan telaga terletak berdekatan dengan kawasan aktiviti pertanian seperti
penanaman tembakau, padi, getah dan dusun. Dengan in terdapat kemungkinan aktiviti pertanian tersebut akan mencemar air
telaga, terutamanya berkaitan penggunaan baja kimia, yang akan masuk ke telaga melalui aliran air atau perpindahan bawah
tanah. Satu kajian telah dijalankan ke atas lima belas buah telaga di daerah Bachok, Pasir Puteh dan Machang, untuk
menentukan kanduangan nitrat dan fosfat dan kualiti air telaga-telaga berkenaan. Pensampelan dijalankan dua kali, iaitu pada
awal Oktober 2004 ( mewakili musim kering) dan akhir Januari 2005 (mewakili musim hujan). Data untuk kekeruhan, suhu,
DO dan pH ditentukan in-situ, manakala kandungan nitrat, fosfat, BOD dan COD ditentukan di dalam makmal. Indeks
Kualiti Air (WQI) bagi sample-sampel yang dikaji ditentukan berdasarkan Interim Water Quality Standard (INWQS) for
Malaysia. Kajian ini mendapati kandungan nitrat berjulat dari 0.01 ke 2.42 mg/L semasa musim hujan, dan antara tidak
dapat dikesan dan 0.3 mg/L di musim kering. Kandungan fosfat di musim hujan dan musim kering pula masing-masing
berjulat 0 – 2.2 mg/L dan 0.5 – 1.5 mg/L. Pada amnya nilai WQI bagi semua telaga dikaji jatuh dalam Kelas III dan IV.
Walau bagaimanapun, nilai-nilai WQI bertambah baik (meningkat) di musim hujan di banding musim kering. Berdasarkan
saranan INQWS air dalam kelas-kelas ini memerlukan rawatan yang menyeluruh sebelum boleh dianggap sesuai untuk
kegunaan bekalan air awam.
Kata kunci: Air telaga, WQI, fosfat, nitrat.
Introduction
In line with the increasing awareness on the importance of good health among Malaysian, drinking
water quality is becoming an increasing debating subject and important issue in Malaysia [1]. Demand
for good quality water is ever increasing. Moreover the global demand in water consumption has
doubled since 1940, and expected to be doubled again within twenty years [2]. With this kind of
demand and the inability of the relevant authorities to supply processed cleaned water, majority of
people in less developed areas resort to consuming untreated raw well water.
The majority of people living in these less developed areas involved in agriculture and farming
activities. Each year they cover their field in chemicals in order to obtain higher yields and faster
growing crops. They spray their fields with increasing amount of fertilizers – containing phosphates
and nitrates. The excess fertilizers eventually will find their way into pools and ponds and wells. [3,
4]. Besides fertilizers, phosphate salts used in detergents may also contribute to phosphate
accumulation in well water. Animal wastes also contribute to phosphate and nitrate in runoff water
[5].
Phosphate and nitrate are nutrient for plant growth. However they can also be the primary cause of
lake (well) enrichment leading to the growth of algae and weeds. This process is known as
eutrophication. The presence of algae and weeds will affect the water quality index. In fact the
presence of 10 ppm nitrate-nitrogen in drinking water can cause methemoglobinemia (inability to use
oxygen) in infants [6] and other health related risks may happen at higher concentration [7]. Thus, this
may me the reason to concern about the concentration of phosphate and nitrate in water.
Although the presence of nitrate and phosphate in groundwater in agricultural areas has become the
main concern in various countries [5, 8] not many study has been carried out in Malaysia. Since many
households in agricultural areas in Malaysia still utilized untreated well water for their drinking and
other domestic purposes, it is interesting to study the concentrations of phosphate and nitrate in well
waters and their relation to the water quality index.
Methodology
Sampling Area
The study covered three districts, Bachok, Pasir Puteh and Machang, in Kelantan. From each district,
five wells were selected. The depth of the wells ranged between 30 to 40 feet. However, the water
volume in each well depends on various factors, such as season, location from river, height (elevation)
from sea level. In general, water volume increases during rainy season. Table 1 summarizes the well
location, usage and type of plantation around the wells. Majority of the wells were used as drinking
water as well as other domestic usage such as washing, bath and cooking.
Sampling
Sampling was done twice, one in early October 2004 (representing dry season), and the other at the
end of January 2005 (representing wet season). Water samples were collected using ‘water theft’
coupled to a 1-liter plastic sample container, at a point about 15 cm from surface. The plastic
container was first rinsed with the respective well water before being used in the collection
procedures. After in-situ analysis, the water sample in the container is kept in closed cooler box at ice
temperature, before being transported to chemical analysis laboratory. While in the laboratory the
samples were kept at 4 oC until further analyses.
Analysis
In-situ measurements were carried out on the water samples parameters of pH, temperature, dissolve
oxygen (DO) and turbidity. Other parameter measurements were carried out in laboratory. The Water
Quality Index (WQI) was calculated using the method used by Mohd Talib Hj Latiff et al., [9], based
on the formula suggested by the Department of Environment, Malaysia [10]. Table 2 summarizes the
instruments and methods used in determining the magnitudes of the respective parameter.
Table 1. Sampling points.

District Code Location Elevation Usage Plantation
(m)
B1 N06o 07.314’ 8 Drinking, Domestic, Tobacco
E102o 21.245’ Watering
B2 N06o 08.165’ 5 Drinking, Domestic, Tobacco
Bachok B3 N06o 08.118’ 5 Drinking, Domestic, Tobacco
B4 N06o 06.634’ 6 Drinking, Domestic, Paddy
B5 N06o 06.488’ 5 Abandon Paddy
E102o 19.412’
P1 N05o 55.961’ 16 Drinking, Domestic Paddy
E102o 18.587’
E102o 19.009’
Pasir Puteh P3 N05o 55.513’ 25 Watering Orchard
E102o 19.134’
E102o 19.134’
P5 N05o 55.500’ 23 Abandon Paddy
E102o 19.053’
M1 N05o 51.951’ 32 Drinking, Domestic Paddy
E102o 12.781’
M2 N05o 51.932’ 45 Watering Rubber
E102o 12.453’
Machang M3 N05o 51.700’ 36 Drinking, Domestic Paddy
E102o 13.804’
E102o 13.987’
E102o 13.519’
Table 2. Methods used to determine nitrate and phosphate, and the various parameters in the study.
Parameter/Ions Instruments and Method

Nitrate HACH quick programme 355, DR 2000 Spectrometer
Phosphate HANNA Phosphate High Range ISM, HR HI 93717
pH Portable pH meter, model WP-81, TPS
Temperature, oC Portable Temperature meter, model WP-81, TPS
Dissolved Oxygen (DO), mg/L HANNA portable DO probe
Turbidity, NTU HANNA portable turbidity meter
Total Suspended Solid (TSS), mg/L Total non-filterable residue, dried at 103 – 105 oC, using
Whitmann GF/C filter
Chemical Oxygen Demand (COD), Digestion. COD Reactor MERCK TR-420. Measured using
mg/L HACH DR 2010 Spectrometer
Biochemical Oxygen Demand HACH BOD track sample and HACH incubator
(BOD), mg/L
N-NH3, mg/L HACH quick programme 380, coupled to HACH DR 2000
Spectrometer
Results of the study for the fifteen wells are shown in Table 3 and Table 4 for wet and dry season
respectively. Although, there was temporal variation between wet season and dry season sampling,
the average temperature of the samples between the two sampling periods are identical, 27.8 oC.
However, there is observable variation of temperature between samples ranging between 25.5 oC to
30.7 oC during wet season and between 26.9 oC and 28.8 oC in dry season. This may be attributed to
the location of the wells, either under shaded area or in the open areas, as well as the time of
sampling, morning or afternoon.
Nitrate
Nitrate contents ranged between 0.01 to 2.42 mg/L in wet season. Generally well water samples in
Bachok district contained higher nitrate than samples from any other districts. The locations of the
wells studied in Bachok district are within the vicinity of the tobacco plantations and paddy fields.
Being very soluble in water, nitrate from chemical fertilizers used in the plantation and fields can be
transported into the wells either vertically or horizontally with the rainwater during rainy season.
These concentration, however are lower than the limit of 10 mg/L suggested by the Interim Water
Quality Standard of Malaysia (INWQS). During dry season, the concentrations of nitrate are relatively
lower in the well water studied. In some samples nitrate are even non-detectable.
Table 3. Wet Season (Sample collected at the end of January 2005)
B1 B2 B3 B4 B5 P1 P2 P3 P4 P5 M1 M2 M3 M4 M5
Nitrate, 0.23 1.28 0.22 0.32 2.42 0.07 0.03 0.01 0.76 0.01 0.03 0.10 0.01 0.35 0.21
mg/L
Phosphate, nd 0.9 0.4 0.9 2.2 0.4 nd nd 1.4 nd nd 0.9 1.0 1.5 1.1
mg/L
pH 6.9 7.8 7.0 8.1 6.3 6.7 6.6 5.9 6.1 6.8 6.5 5.9 5.2 5.8 5.5
Class I I I I IIA I I III IIA I I III III III III
DO, mg/L 1.41 1.37 1.25 0.65 1.03 3.14 1.83 1.31 1.84 1.02 1.11 2.24 1.26 1.52 1.91
Class IV IV IV V IV III IV IV IV IV IV IV IV IV IV
COD, mg/L 40.9 35.1 30.5 27.2 37.1 41.9 31.1 25.5 29.4 33.6 32.2 33.0 36.5 39.2 33.1
Class III III III III III III III III III III III III III III III
BOD, mg/L 0.6 5.0 4.0 6.2 13.0 1.4 1.7 1.8 5.8 2.4 5.8 3.1 3.8 0.8 3.6
Class I III III IV V IIA IIA IIA III IIA III IIB IIB I IIB
N-NH3, 0.19 0.32 0.45 0.79 0.30 0.33 0.67 0.40 0.16 0.44 0.29 0.41 0.02 0.22 0.25
mg/L
Class IIA IIB III III IIA IIB III IIB IIA IIB IIA IIB I IIA IIA
TSS, mg/L 15.5 13.4 6.5 6.32 9.21 3.7 5.9 13.5 5.3 14.6 3.4 4.6 2.8 2.4 3.1
Class I I I I I I I I I I I I I I I
TDS, mg/L 76.2 78.0 53.0 100.7 56.9 82.5 38.7 33.3 26.2 62.3 16.1 27.6 20.4 35.7 14.5
Turbidity, 0.51 0.55 0.60 0.63 0.54 0.61 1.11 0.93 0.63 0.74 0.53 0.92 0.61 0.50 0.53
NTU
Temp. (oC) 28.9 30.7 28.7 28.8 28.3 25.5 27.7 27.2 27.1 27.0 26.5 27.9 27.8 27.3 27.5
WQI Value 65.5 63.0 49.6 49.2 66.9 57.1 47.7 48.4 48.6 58.8 57.1 49.3 49.6 49.9 47.5
OVERALL III III IV IV III III IV IV IV III III IV IV IV IV

CLASS
Table 4. Dry season (Sample collected at the end of October 2004)
B1 B2 B3 B4 B5 P1 P2 P3 P4 P5 M1 M2 M3 M4 M5
Nitrate, nd 0.1 nd nd 1.0 nd nd nd nd 2.0 0.1 0.1 0.3 0.2 0.2
mg/L
Phosphate, 1.0 1.0 0.5 1.1 1.2 0.9 1.5 1.1 1.5 0.6 0.6 1.4 1.4 1.7 1.3
mg/L
pH 5.5 6.8 5.5 6.9 5.5 6.4 5.9 5.4 5.1 6.4 4.7 6.8 4.3 5.1 5.0
Class III I III I III IIA III III III IIA V I V III IV
DO, mg/L 3.45 1.93 1.03 4.61 1.86 5.64 2.92 1.50 2.25 1.70 0.99 3.55 3.18 2.06 1.70
Class III IV IV III IV IIB IV IV IV IV V III III IV IV
COD, mg/l 26.4 20.1 17.9 15.7 21.7 9.1 6.8 5.9 7.9 7.1 7.3 5.4 13.6 11.6 8.4
Class IIB IIA IIA IIA IIA I I I I I I I IIA IIA I
BOD, 6.8 11.8 8.7 10.3 9.2 27.4 17.3 9.5 12.7 19.4 11.3 12.8 11.5 13.7 13.0
mg/L
Class III III III III III V V III IV V IV IV IV V V
N-NH3 0.42 0.58 0.06 1.23 0.48 0.11 0.46 0.51 0.72 0.85 0.51 0.59 0.35 0.47 0.16
(mg/L)
Class IIB IIB I IV IIB IIA IIB IIB IIB IIB IIB IIB IIB IIB IIA
TSS, mg/L 13.6 12.9 4.2 5.3 12.2 4.3 7.8 13.8 7.9 13.2 3.6 2.1 1.6 3.7 2.4
TDS, 113.5 121.0 42.6 98.3 90.1 86.0 65.4 24.2 23.0 53.1 20.3 67.1 16.8 29.6 14.7
mg/L
Turbidity 0.51 0.58 0.50 0.71 0.61 0.57 1.56 1.76 0.83 0.93 0.64 1.23 0.76 0.61 0.59
(NTU)
Temp. (oC) 28.1 28.3 28.3 28.0 27.2 28.3 28.4 27.8 27.3 27.4 27.3 28.8 26.9 27.6 27.1
WQI 56.2 54.1 37.4 44.0 50.9 53.0 41.2 36.6 36.5 48.7 28.5 31.3 34.5 36.6 30.6
Value
OVERALL III III IV IV IV III IV IV IV IV V IV IV IV V

CLASS
Phosphate
The maximum phosphate concentration in the samples studied is 2.2 mg/L during wet season and 1.7
mg/L during dry season. In some samples phosphate are non-detectable. There is no correlation trend
between concentration during dry and wet season. However, generally samples from Machang district
contained higher concentration of phosphate. Phosphate is not soluble in water, but when applied to
soil it will get binds to the soil particles, thus it will not be transported easily through leaching or
washing together with water in the soil profiles. Therefore, although phosphate may originated from
chemical fertilizers applied to the plantation soil, the concentration trends shown by nitrate is not
observable in the case of phosphate.
PH Value
In wet season pH of the well water studied ranged between 5.2 to 8.1, with an average of 6.5. While
during dry season the well water samples become more acidic, with pH ranging between 4.7 and 6.9,
with an average of 5.7. Based on the INWQS the wet season average falls into Class I, while the dry
season falls into Class III category. This observation might be attributed to the great reduction in
water volume in the wells during dry season.
Dissolved Oxygen, DO
Dissolved oxygen the water samples studied ranged between 1.02 mg/L to 3.14 mg/L and averaged to
1.53 mg/L during wet season. The average increased to 2.56 mg/L in dry season, ranging between
0.99 to 5.64 mg/L. Generally this falls into Class IV category. High DO for both seasons was for
sample P1, with 3.14 mg/L and 5.64 mg/L in dry and wet season respectively.
Chemical Oxygen Demand, COD

Generally the COD measurements were higher in all the wells during rainy season than during dry
season. The average COD for the fifteen wells during rainy season was 33.8 mg/L, and improved to
12.3 mg/L in dry season. Although the maximum value of 40.9 mg/L was recorded for water sample
from well B1 during rainy season, this value is still below the 50 mg/L limit of Class III standardized
in INWQS.
Biochemical Oxygen Demand, BOD
In wet season, average BOD of the fifteen wells is 3.9 mg/L, with a maximum value of 13.0 mg/L at
well B5. Well B5 is within the vicinity of a paddy field, and is abandoned. The average BOD
increases in dry season to 13.0 mg/L. Results show that the BOD of all the well water samples in dry
season exceeded the standard value of 6 mg/L (Class III) of INQWS.
Nitrogen-Ammonia, N-NH3
On average during wet season the nitrogen-ammonia concentration ranged between 0.02 – 0.67 mg/L.
These concentrations are below the INQWS limit (for Class III) of 0.9 mg/L. Except for well B4,
identical trend is also observed during dry season. However, for B4 the concentration of 1.23 mg/L is
beyond the INQWS limit. Generally, the nitrogen-ammonia concentration of the wells shows
reduction in value during wet season.
Total Suspended Solids, TSS

All of the well water studied either during wet or dry season fall into TSS Class I, with wet and dry
season average of 8.4 mg/L and 7.2 mg/L respectively. This shows that the well water studied posed
no problems in terms of TSS.
Turbidity
As turbidity is closely correlated to TSS, the study found that all the well water (either during wet or
dry season) fall into Turbidity Class I. Physical observation also showed that all the water samples
were very transparent in nature. In wet season the turbidity range between 0.50 – 1.11 NTU, while in
dry season the range is 0.50 – 1.76 NTU.
Water Quality Index, WQI

Based on the WQI values the study found that the majority of the well water samples fall into Class
IV or V. The general trend showed that the WQI values decreased during dry season. Only wells B1
and B2 maintained the Class III category during wet as well as dry season. According to the INWQS
classification only Class III and below are suitable as water supply, and that needs extensive
treatment. From Table 3 and Table 4, it can be seen that the main contributing and affecting
parameters on the WQI values are the DO, COD and BOD. The pH values contribute a significant
role during dry season in influencing the WQI values.
A comparative analysis for wet and dry season parameters was also carried out. Generally, it was
found that the quality of well water studied improved during wet season. This can obviously be seen
in Figure 1 where the averages of the respective parameters studied were compared for wet and dry
season.
100.0
48.1 57.7 53.9 41.3
33.8
27.8 27.8
12.3 13.0 14.7
10.0 7.2
6.5 5.7
3.9
2.56
Units
1.53
0.83 1.07 1.12
1.0 0.66
0.50 0.50
0.35 0.40
Wet
Dry 0.1
i ty
3
O
e
pH
ue
e
S
H
at
O
at
m
D
TD
TS
id
al
-N
ph
C
itr
Te
rb
Iv
N
os
Tu
Q
Ph
W
Figure 1. Comparison on the average of the parameters studied during wet and dry season.
Conclusion
The nitrate and phosphate contents of all the samples studied are still below the suggested limit,
indicating the effect of fertilizers on the wall water is minimal. In general, the study has shown that
only six out of the fifteen wells studied are suitable as water supply based on the INWQS index and
classification, and that need further extensive treatment. The low DO level and high COD and BOD
are the main contributing factors attributed to the classification of the well water samples to between
Class III and Class V. In dry season the water become more acidic, and thus play an important role as
well in determining the class. The water quality improved during wet season.
Acknowledgement: The authors wish to thanks IRDC of UiTM for providing the research grant for the study (Grant No.:
600-IRDC/ST 5/3/938).
References
1. Pillay, M.S., Talha, M.Z., 2003; Drinking Water Quality Issues; Water and Drainage Conference, K.L.
2. Karr, J., Mannusen, J., McKnight, D., Naiman, R., Stanford, J., 1995; Freshwater Ecosystems and Their
Management: A National Initiative; Science, Vol. 270, 27.
3. Barber, L., Beneke, A., Breedlove, B., 2002; Nitrate and Phosphates Levels in Pfeffer Park Stream; Natural
System I, Fall 2002, Miami University.
4. Hooda, P.S., Edwards, A.C., Anderson, H.A., Miller, A., 2000; A Review of Water Quality Concerns in Livestock
Farming Areas; The Science of the Total Environment, Vol. 250, Issues 1 – 3, 143 – 167.
5. Gymer, R.G., 1977; Chemistry in the Natural World; D.C. Heat & Company, USA; 571p.
6. Rosen, C.J., White, D.B., 1999; Preventing Pollution Problems from Lawn and Garden Fertilizers; Note FO-2923-
GO, College of Agricultural, Food, and Environmental Sciences, University of Minnesota, USA.
7. NECi, 2000; Nitrate: Health Risks to Consumers; The Nitrate Elimination Co., Inc, USA, 3p.
8. Thornburn, P.J., Biggs, J.S., Weier, K.L., Keating, B.A., 2003; Nitrate in Ground waters of intensively agricultural
areas in coastal Northeastern Australia; Agriculture, Ecosystem & Rnvironment, Vol.94, Issue 1, 49 –58.
9. Mohd. Talib Hj Latiff, Zulfahmi Ali Rahman, Jasni Yaakob, Ruzaidah Yusof, 2004; Cirian Fiziko-kimia air perigi
di kawasan Selandar, Melaka; MJAS, Vol. 8, No. 1; 87 – 92.
10. jabatan Alam Sekitar (JAS), 1991; Environment quality report, 1990; Kementerian Sains, Teknologi dan Alam
Sekitar, Kuala Lumpur.
AKTIVITI 226Ra DAN 228Ra PADA PERMUKAAN SEDIMEN BAGAN LALANG,
SELANGOR.
NIOO SIEW YEW, ZAHARUDDIN AHMAD2 dan CHE ABD. RAHIM MOHAMED1
Pusat Pengajian Sains Sekitaran Dan Sumber Alam

Fakulti Sains Dan Teknologi
43600 Bangi, Selangor, Malaysia
corresponding author: carmohd@pkrisc.cc.ukm.my1
Malaysian Institute for Nuclear Technology2
Key words: 226Ra, 228Ra, sediment
ABSTRAK
Sebanyak enam stesen sampel sedimen telah dipilih di Sungai Sepang Kecil, Selangor pada bulan Mac,
September dan Oktober 2004. Hasil kajian mendapati bahawa terdapat perbezaan yang bererti antara masa
persampelan bagi aktiviti 226Ra dan 228Ra (P < 0.01). Purata aktiviti pada bulan Mac mencatatkan nilai tertinggi,
iaitu masing-masing dengan aktiviti 161.30 Bq kg-1 dan 466.88 Bq kg-1 bagi 226Ra dan 228Ra. Manakala aktiviti
isotop radium pada bulan September dan Oktober pula masing-masing mencatatkan nilai purata 24.77 Bq kg-1
dan 69.22 Bq kg-1 bagi 226Ra serta 73.13 Bq kg-1 dan 1284.58 Bq kg-1 bagi 228Ra. Kesan Monsun Timur Laut
mungkin merupakan faktor yang mendorong aktiviti isotop radium yang lebih tinggi ketika persampelan bulan
Mac. Hasil kajian memperolehi bahawa sedimen di Bagan Lalang adalah jenis berpasir dan mempunyai variasi
saiz yang terhad. Di samping itu, didapati bahawa isotop radium adalah lebih terjerap dengan sedimen yang
bersaiz antara 63 – 125 µm yang biasanya mengandungi kandungan feldspar alkali yang tinggi.
ABSTRACT
Six stations had selected from Sungai Sepang Kechil, Selangor during Mac, September and October 2004 for
sediment analysis. The results showed a significant difference among the 226Ra and 228Ra activities (P < 0.01)
with sampling period. The highest activities of 226Ra and 228Ra have obtained during the sampling on Mac,
which show the average activities 161.30 Bq kg-1 and 466.88 Bq kg-1, respectively. Meanwhile, 226Ra activities
obtained on September and October are in the average of 24.77 Bq kg-1 and 69.22 Bq kg-1, respectively.
Average activities of 228Ra are 73.13 Bq kg-1 and 1284.58 Bq kg-1, respectively for September and October.
Northeast Monsoon effect might cause the higher activities of radium isotopes during sampling on Mac.
Sediments from Bagan Lalang are sandy type with a limited size variation. Furthermore, radium isotopes are
more adsorbed on the particles sediments between size 63 – 125 µm, which usually rich in alkaline feldspar.
PENGENALAN
Sifat radiologi isotop radium yang membahayakan kesihatan awam merupakan salah satu
faktor ia sering kali dikaji oleh para saintis. Di samping itu, setengah hayat isotop radium
semulajadi (223Ra, 224Ra, 226Ra dan 228Ra) yang berbeza dari beberapa hari hingga 1600 tahun
mencetuskan ia sesuai digunakan sebagai penyurih semulajadi dalam pelbagai bidang seperti
proses percampuran dalam sistem akues serta kajian kronologi dalam lautan dan tasik (1).
Selain itu, hubungan antara isotop-isotop radium juga sesuai untuk mengkaji fluks dan
kadar percampuran air daripada daratan dengan air laut dan estuari serta menyiasat
pertukaran antara air bawah tanah dengan air permukaan (2, 3). Manakala perubahan pekali
jerapan (adsorption coefficient) di antara radium dengan air tawar dan air masin telah
mengakibatkan radium terikat kuat dengan butiran sedimen dalam air tawar, malah cenderung
terlarut dalam air masin. Keadaan ini disebabkan oleh radium lebih cenderung terjerap dalam
bahan terampai dengan kekuatan ionik menigkatkan dan kepekatan partikel menurun. Oleh
yang demikian, keterlarutan radium adalah lebih tinggi di kawasan estuari dan persisir pantai
(4). Tujuan kajian ini dilakukan adalah untuk melihat taburan aktiviti isotop 226Ra dan 228Ra
di permukaan sedimen kawasan Bagan Lalang, Selangor serta mengkaji kesan saiz partikel
terhadap isotop radium.
BAHAN DAN KAEDAH

Lokasi kajian dan persampelan
Pantai Bagan Lalang terletak berdekatan dengan kawasan Lembah Kelang dan diperkenalkan
oleh Kerajaan Selangor pada tahun 1990-an (5). Projek “Sepang Gold Coast” dan “Sepang
Water City” yang diperkenalkan oleh Permodalan Negeri Selangor Berhad (PNSB) dengan
CNI dijangka akan menggunakan tanah seluas 4621 ekar, iaitu pinggir Sungai Sepang Kecil
dan Sungai Sepang Besar serta pinggir pantai Bagan Lalang hingga ke Tanjung Sepat (6).
Oleh yang demikian, salah satu sebab kajian ini dilakukan di Sungai Sepang Kecil adalah
untuk menghasilkan satu data pangkalan sebelum ia diperbangunkan.
Sebanyak tiga kali persampelan (Jadual 1) dijalankan di enam stesen yang ditentukan
seperti yang ditunjukkan dalam Rajah 1 dan Jadual 1. Sungai Sepang Kecil dipilih sebagai
kawasan kajian kerana ia dapat memaparkan sistem aliran yang membezakan anntara air
payau dengan air laut.
JADUAL 1 Stesen-stesen persampelan sedimen di sekitar Sungai Sepang Kecil, Bagan Lalang.
Stesen Kod Latitud (U) Longitud (T)

o
1 BL1 02 36’55’’ 101o41’07’’
2 BL2 02o36’35’’ 101o40’20’’
3 BL3 02o36’30’’ 101o39’19’’
4 BL4 02o36’05’’ 101o38’05’’
5 BL5 02o36’11’’ 101o36’56’’
6 BL6 02o36’06’’ 101o35’38’’
Sampel sedimen permukaan dasar laut diambil pada setiap stesen kajian dengan
Ponar Grab. pH setiap sampel diukur dengan pH meter (model 210Aplus, Thermo Orion) di
lapangan. Seterusnya, sampel sedimen disimpan dalam beg plastik dan dibawa ke makmal
untuk analisis lanjutan.
Analisis kajian
Ketika di makmal, sampel sedimen dikeringkan dengan ketuhar pada suhu 60oC. Porositi (Ф)
sampel sedimen dicatatkan. Setreusnya ketumpatan sedimen dikira dengan menggunakan
persamaan 1(7).
101o34’T 101o35’T 101o36’T 101o37’T 101o38’T 101o39’T 101o40’T 101o41’T
U 2o38’U
➲
0 1
KM
2o37’U
①
②
③
⑥ ⑤ 2o36’U
④
2o35’U
RAJAH 1 Stesen-stesen kajian di Bagan Lalang, Selangor.

248
Ketumpatan sedimen (ρs) = 2.6__ (1)

(1-Ф)
Seterusnya, sampel sedimen kering ditumbuk dengan menggunakan mortar dan

diayak dengan pengayak elektronik bagi menentukan partikel saiz sedimen. Diameter (d)
sampel sedimen yang bersaiz: 125 <d<250 µm, 63<d<125 µm dan d < 63 µm disimpan untuk
analisis yang seterusnya. Pengelasan partikel sedimen dilaklukan menggunakan skala
Wentworth. Skala ini digunakan secara meluas dan partikel saiz sedimen ditukar kepada unit
phi (Ø) (8).
Ø = - log2 mm (2)
Pengukuran statistik yang lazim digunakan dalam skala Wentworth adalah:
Purata ( X_ ) = (Ø 16 + Ø 50 + Ø 84)
3 (3)
Ralat ( σ ) = Ø 84 –Ø 16 + Ø 95 –Ø 5
4 6.6 (4)
Skewness (sk) = Ø 16 + Ø 84 - 2 Ø 50 + Ø 5 + Ø 95 - 2 Ø 50
2(Ø 84 –Ø 16) 2(Ø 95 –Ø 5) (5)
Sampel sedimen kering dicerna dengan asid hidroklorik. Lebih kurang 1 g sampel
sedimen dimasukkan ke dalam bikar kaca bersama dengan 1 ml pembawa Ba2+ dan 20 ml 8
M asid hidroklorik ditambahkan. Campuran ini ditutup dengan penutup kaca dan dipanaskan
atas piring pemanas selama 2 – 3 jam. Seterusnya, hasil hadaman dibiar sejuk dan dituras
dengan kertas turas yang berdiameter 47 mm dengan saiz liang 0.45 µm (G/FC filter,
Whatman). Hasil turasan dikeringkan atas piring panas. Bahan yang tertinggal dalam bikar
dilarutkan dengan lebih kurang 50 ml asid perklorik 1% untuk proses penulenan radium.
Radium ditulenkan dengan menggunakan turus pemisah kation AG 50W-X4 Resin (200-
400 mesh, Bio-rad). Di mana sampel (≈ 50 ml) dialir dalam turus pemisah kation untuk
penulenan. Kemudiannya, 200 ml asid hidroklorik 1.5 M dialirkan melalui turus. Efluen
disimpan dalam bikar dan dikeringkan atas plat pemanas. Hasil pengeringan dilarutkan
dengan asid nitrik 0.5 M (≈ 100 ml). Seterusnya, 2 ml asid sulfurik pekat ditambahkan.
Mendakan barium sulfat (BaSO4) yang terbentuk dituraskan dengan mengguna kertas turas
berdiameter 25 mm (GF/C, Whatman). Berat bersih BaSO4 dicatatkan untuk mendapatkan
249
perolehan semula kimia sampel. Seterusnya, BaSO4 diletakkan atas disk stainless steel dan
dilabut dengan kertas plastik. Disk yang disiap balut dibilang dengan Spektrometer Gross
Alfa/Beta (model LB5100-W, Tennelec) setelah keseimbangan sekular radium dengan
anak-anaknya dapat dicapai (dibiarkan selama tiga bulan).
HASIL DAN PERBINCANGAN
Taburan isotop radium pada permukaan sedimen
Rajah 2 menunjukkan taburan isotop radium pada permukaan sedimen di Bagan

Lalang. Stesen 4 terletak pada mulut muara sungai mencatatkan aktiviti 226Ra dan 228Ra
tertinggi pada bulan September dan Oktober. Hal sedemikian mungkin disebabkan oleh
pengumpulan partikel sedimen yang dibawa dari sungai (semasa air surut) serta laut (semasa
air pasang). Partikel sedimen yang terkumpul mungkin mengalami penjanaan semula dan
mengakibatkan aktiviti radium pada permukaan sedimen yang tinggi di Stesen 4. Oleh sebab
228
Ra mempunyai setengah hayat yang pendek (5.75 tahun), jadi penjanaan semula bagi
aktiviti 228Ra adalah lebih cepat (≅ 280 kali) berbanding 226Ra. Maka, nisbah aktiviti
228
Ra/226Ra di Stesen 4 juga tinggi, iaitu lebih kurang lima.
Manakala taburan aktiviti isotop radium pada permukaan sedimen pada bulan Mac
2004 adalah hampir sekata, kecuali aktiviti 228Ra yang abnormal (870 Bq kg-1) di Stesen 6.
Faktor yang menyumbangkan aktiviti 228Ra yang tinggi di Stesen 6 adalah tidak jelas.
Aktiviti isotop 226Ra dan 228Ra (P < 0.01) pada permukaan sedimen pula
menunjukkan perbezaan yang sangat bererti dengan masa persampelan. Rajah 3 memaparkan
taburan aktiviti isotop radium pada permukaan sedimen mengikut masa peersampelan. Hasil
kajian mendapati bahawa purata aktiviti 226Ra dan 228Ra adalah maksimum pada
persampelan bulan Mac, iaitu masing-masing dengan purata aktiviti 161.30 Bq kg-1 dan
466.88 Bq kg-1. Manakala aktiviti isotop radium pada bulan September dan Oktober
adalah lebih rendah, iaitu dengan masing -
250
1000
(a)
800 Ra-226
Ra (Bq kg-1)
600 Ra-228
400
200
0
1 2 3 4 5 6
Stesen
300
(b)
250
Ra (Bq kg-1)
200
150
100
50
0
1 2 3 4 5 6
Stesen
500
(c)
400
Ra (Bq kg-1)
300
200
100
0
1 2 4 5 6
Stesen
RAJAH 2 Taburan 226Ra dan 228Ra pada permukaan sedimen ketika persampelan bulan (a) Mac, (b) September,
dan (c) Oktober.
masing mencatatkan nilai purata 24.77 Bq kg-1 dan 69.22 Bq kg-1 bagi 226Ra serta 73.17 Bq
kg-1 dan 184.58 Bq kg-1 bagi 228Ra.
251
500
Ra-226
400
Ra-228
Aktiviti (Bq kg )
-1
300
200
100
0
Mac Sept Okt
Persampelan
RAJAH 3 Purata aktiviti 226Ra dan 228Ra pada permukaan sedimen mengikut masa persampelan.
Pantai barat Semenanjung Malaysia mengalami musim hujan ketika Monsun Timur
Laut (Mac). Partikel sedimen dari daratan dan sungai turut dibawa oleh air hujan ke estuari
dan lautan. Justeru itu, terdapat input tambahan bagi aktiviti isotop radium pada permukaan
sedimen. Jadi, aktiviti isotop radium pada bulan Mac adalah lebih tinggi berbanding dengan
persampelanpada bulan September dan Oktober.
Kesan partikel saiz sedimen terhadap isotop radium
Jadual 2 menunjukkan partikel saiz dan jenis sedimen bagi ketiga-tiga persampelan. Secara
keseluruhannya, permukaan sedimen Bagan Lalang dikelaskan dalam kumpulan pasir. Purata
saiz sedimen ( X_ ) adalah daripada saiz -0.83Ø (pasir sangat kasar) hingga 3.12 Ø (pasir sangat
halus). Ralat partikel sedimen (σ) yang besar merujuk kepada pengelasan sedimen Bagan
Lalang adalah kurang baik, iaitu variasi partikel saiz sedimen yang diangkut atau termendak
adalah kurang. Nilai skewness (sk) pula digunakan sebagai penunjuk bagi sejarah taburan
partikel saiz sedimen. Sampel yang mempunyai nilai sk yang positif menunjukkan bahawa
stesen tersebut mengandungi lebihan partikel yang halus. Sumber kemasukan partikel halus
ini mungkin disebabkan oleh peningkatannya partikel halus yang termendak ataupun partikel
yang kasar telah terpindah daripada stesen tersebut. Manakala bagi sampel yang mempunyai
nilai sk yang negatif merujuk kepada pemendakan partikel kasar telah berlaku di kawasan
252
tersebut. Faktor yang mendorong kepada perubahan nilai sk adalah angin dan kesan ombak
(8).
JADUAL 2 Pengelasan sedimen di Bagan Lalang.
Purata saiz Skew Porositi

Stesen _ Ralat (σ) pH Kelas sedimen
( X), Ø (sk) (Φ)
Mac
1 1.95 0.84 0.18 0.41 9.51 Pasir sederhana
2 2.18 1.44 -0.16 0.52 8.65 Pasir halus
3 2.13 0.64 0.07 0.74 8.76 Pasir halus
4 -0.60 1.23 -0.39 0.83 8.36 Pasir sangat kasar
5 0.26 0.63 -0.32 0.81 5.58 Pasir kasar
September
1 1.91 1.98 -0.23 0.54 7.06 Pasir sederhana
2 1.67 0.54 -0.17 0.76 7.18 Pasir sederhana
3 0.32 0.61 0.19 0.81 7.58 Pasir kasar
4 -0.83 0.90 0.21 0.77 7.54 Pasir sangat kasar
Oktober
1 1.169 1.85 0.39 0.29 6.72 Pasir sederhana
2 1.09 2.20 0.05 0.39 7.22 Pasir sederhana
6 0.01 0.75 -0.18 0.81 7.83 Pasir kasar
253
16
(a)
Ra (x10-2 Bq g-1)
12 R2 = 0.124
P > 0.05
8
4
226
0
0 20 40 60 80 100
% Partikel Sedimen
16
(b)
Ra (X 10 Bq g )
-1
12
-2
8
2
R = 0.691
4
P < 0.001
226
0
0 5 10 15 20 25 30 35
% Partikel sedimen
30
(c)
Ra (X 10-2 Bq g-1)
25
2
20 R = 0.160
15 P > 0.05
10
5
226
0
0 10 20 30 40 50 60
% Partikel sedimen
RAJAH 4 Kolerasi aktiviti 226Ra dalam sedimen Bagan Lalang dengan peratusan partikel sedimen dengan saiz
partikel (a) +2<Ø<+3, (b) +3<Ø<+ 4, dan (c) Ø>+4.
254
200
(a)
Ra (X 10 Bq g )
-1
150
-2
2
100 R = 0.017
P > 0.05
50
228
0
0 20 40 60 80 100
% Partikel sedimen
70
(b)
Ra (X 10 Bq g )
60
-1
2
50 R = 0.388
P < 0.05
40
-2
30
20
10
228
0
0 5 10 15 20 25 30 35
% Partikel sedimen
100
(c)
Ra (X 10 Bq g )
-1
80
60 2
R = 0.019
-2
40 P > 0.05
20
228
0
0 10 20 30 40 50 60
% Partikel sedimen
RAJAH 5 Kolerasi aktiviti 228Ra dalam sedimen Bagan Lalang dengan peratusan partikel sedimen dengan saiz
partikel (a) +2<Ø<+3, (b) +3<Ø<+ 4, dan (c) Ø >+4.
Rajah 4 dan 5 memaparkan kesan saiz partikel terhadap isotop radium. Hasil kajian
mendapati bahawa aktiviti 226Ra (P < 0.01) dan 228Ra (P < 0.05) lebih terjerap dalam partikel
sedimen bersaiz +3<Ø<+ 4 (63-125µm). Menurut Zhang et al (9), logam alkali bumi, iaitu
barium dan strontium (mempunyai sifat yang menghampiri dengan radium) juga
255
menunjukkan kolerasi possitif degan partikel saiz yang lebih kasar. Keadaan ini mungkin
disebabkan oleh feldspar alkali yang biasanya wujud dalam partikel yang lebih kasar
mengandungi unsur barium, strontium, rubidium dan cesium yang tinggi (10).
KESIMPULAN
Secara kesimpulannya, aktiviti isotop radium pada permukaan sedimen adalah lebih tinggi
ketika musim hujan. Air hujan merupakan agen pembawa partikel sedimen yang
mengandungi isotop radium dari daratan dan sungai ke estuari dan lautan. Sampel sedimen di
Bagan Lalang dikelaskan sebagai jenis berpasir dengan mengandungi variasi saiz yang
kurang. Di samping itu, aktiviti isotop radium juga boleh disimpulkan bahawa ia lebih
terjerap dalam partikel yang bersaiz 63 – 125 µm yang biasanya mengandungi kandungan
feldspar alkali yang tinggi.
PNEGHARGAAN
Terima kasih kepada MINT dan ahli makmal yang memberi bantuan dalam kajian ini.
RUJUKAN
1. Kim, Y.J., Kim, C.K., Kim, C.S., Yun, J.Y., Rho, B.H. 1999. Determination of 226Ra
in environmental samples using high-resolution inductively coupled plasma mass
spectrometry. J. Radioanal. Nucl. Chem. 240: 613-618.
2. Nour, S., El-Sharkawy, A., Burnett, W.C., Horwitz, E.P. 2004. Radium-228
determination of natural waters via concentration on manganese dioxide and
separation using Diphonix ion exchange resin. Appl. Radiat. Isot. 61: 1173-1178.
3. Eikenberg, J., Tricca, A., Vezzu, G., Stille, P., Bajo, S., Ruethi, M. 2001.
228
Ra/226Ra/224Ra and 87Sr/86Sr isotope relationships for determining interactions
between ground and river water in the upper Rhine Valley. J. Environ. Radioact.
54: 133-162.
4. Rama, Moore, W.S. 1996. Using the radium quartet to estimate water exchange and
ground water input in salt marshes. Geochim. Cosmochim. Act. 60: 4645-4652.
5. Dharmender, S. 2003. Rare sights at Bagan Lalang. (atas talian)
http://www.allmalaysia.info/news/story. (17 Januari 2005).
6. Sepang Gold Coast dan Sepang Water City. (atas talian)
http://www.pnsb.com.my/n_jan03.htm. (17 Januari 2005).
256
7. DiToro, D.M. 1999. Sediment Flux Modelling. New York: John Wiley & Sons.
8. Pethick, J. 1984. An introduction to coastal geomorphology. London: Edward Arnold.
9. Zhang, C., Wang, L., Li, G., Dong, S., Yang, J., Wang, X. 2002. Grain size effect on
multi-element concentrations in sediments from the intertidal flats of Bohai Bay,
China. Appl. Geochem. 17: 59-68.
10. Gotze, J. 1998. Geochemistry and provenance of the Atlendrof feldspathic sandstone
in the Middle Bunter of the Thuringian basin (Germany). Chemical Geol. 150:
43-61.
257
KERADIOAKTIFAN TABII DAN KANDUNGAN LOGAM BERAT DALAM IKAN DARI

SUNGAI ATOK DAN SUNGAI TAHAN ,TAMAN NEGARA PAHANG.
Muhamad bin Othman, Redzuwan bin Yahaya & Hiew Ching Ching
Program Sains Nuklear,Pusat Pengajian Fizik Gunaan,Fakulti Sains dan Teknlogi

Universiti Kebangsaan Malaysia, 43600 BANGI, Selangor Darul Ehsan.
nina@pkrisc.cc.ukm.my
Abstrak. Dalam kajian ini keradioaktifan tabii dan kandungan logam berat dalam sampel ikan species
Puntius gonionotus, Leptobarbus hoevenii, Mytus nemurus Puntius schwanwnfeldii dan Cyprinus
carpio dari Sungai Atok dan Sungai Tahan, Taman Negara, Pahang telah ditentukan. Keradioaktifan
U-238 dan Th-232 telah ditentukan menggunakan Spektrometri Sinar Gama manakala kandungan
logam berat telah dianalisis menggunakan Spektrometri Emissi Optik Plasma Gandingan Teraruh
(ICP-OES). Sampel ikan telah dipisahkan kepada bahagian tulang dan otot, dikering dan
dihomogenkan. Sampel seterusnya dihadamkan menggunakan asid nitrik sehingga jernih. Hasil
analisis mendapati hanya U-238 sahaja yang ditemukan didalam ikan. Kepekatan U-238 adalah
dalam julat 1.47 ± 0.006 – 6.17 ± 0.007 Bq/kg. Bagi logam berat pula 13 unsur telah dapat dikesan,
unsur-unsur tersebut adalah aluminium, argentum, arsenik, ferum, magnesium, mangan, kobalt,
kromium, kuprum, plumbum, selenium, strontium dan zink. Perbandingan dengan Piawaian
Antarabangsa (FAO) menunjukkan kandungan unsur-unsur tersebut adalah pada paras yang selamat
untuk dimakan.
Abstract. This study was conducted to determine the natural radioactivity and the concentration of
heavy metal in fish of species Puntius gonionotus, Leptobarbus hoevenii, Mytus nemurus Puntius
schwanenfeldii dan Cyprinus carpio from Sungai Atok and Sungai Tahan of National Park, Pahang.
The radioactivities of U-238 and Th-232 were analysed using gamma spectrometry whereas the
concentrations of heavy metals were determined by Inductively Coupled Plasma Optical Emission
Spectrometry (ICP-OES). Fish samples were separated into muscle and bone, dried and homogenized.
Samples were then digested using nitric acid until a clear solution was obtained. Result showed that
only U-238 was found in all fish species. The activity of U-238 was in the range of 1.47 ± 0.006 –
6.17 ± 0.007 Bq/kg. Thirteen heavy metals were detected, namely aluminium, silver, arsenic, iron,
magnesium, manganese, cobalt, chromium, copper, lead, selenium, strontium and zinc. A comparison
with the International Standard (Food and Agriculture Organization) showed that the level of heavy
metal in fish from both rivers was safe for human consumption.
Key words: fish, muscle, bone, U-238, River Atok, River Tahan , heavy metals.
Pendahuluan
Pencemaran air kini semakin meluas akibat daripada proses pembangunan, perindustrian dan kegiatan
pertanian yang kian meningkat (Kulikova et. al., 1985). Pembuangan sisa-sisa daripada kilang terus ke
dalam sungai menyebabkan air sungai mengandungi bahan kimia beracun (Hargrave, 1991).
Kemasukkan bahan kimia ke dalam air sungai akan menyebabkan perubahan kimia air dan
mengganggu sistem akuatik. Hidupan akuatik yang tidak dapat menyesuaikan diri dengan perubahan
ini akan mati dan pupus sementara bagi hidupan yang dapat menyesuaikan diri kandungan bahan
kimia beracun dalam organisme akan meningkat.
Bahan kimia beracun ini terdiri dari bahan organik dan tak organik termasuk logam berat dan bahan
radionuklid. Logam berat merupakan pencemar tak organik yang berbahaya dan dapat dikumpulkan
oleh jasad akuatik melalui rantai makanan. Kajian terhadap sungai yang tercemar menunjukkan
kandungan logam berat tertentu meningkat didalam plankton, moluska dan algae. Kajian pada ikan di
dalam sungai yang telah lama tercemar dengan logam berat pula menunjukkan peningkatan
258
kandungan zink, kuprum, plumbum, raksa dan kadmium dalam tisu otot. Kepekatan kadmium dan
zink yang tinggi ditemukan pada ginjal sementara hati menunjukkan kepekatan kuprum yang tinggi
(Wachs, 1985). Oleh kerana ikan merupakan sumber protin utama bagi manusia, peningkatan logam
berat dalam ikan akan menjejaskan kesihatan manusia.
Kajian terhadap kandungan logam berat didalam ikan di kawasan yang masih bersih menjadi penting
bagi dijadikan piawaian untuk menentukan aras pencemaran. Taman Negara merupakan kawasan
perlindungan bagi tumbuhan dan haiwan didalam persekitaran semulajadi. Di dalam kawasan ini
segala bentuk aktiviti pembangunan tidak dibenarkan. Oleh yang demikian kajian kepekatan logam
serta keradioaktifan tabii dikawasan ini boleh memberikan ukuran latar atau rujukan bagi kawasan
yang belum tercemar. Selain itu nilai yang diperolehi dapat dibandingkan dengan kawasan lain
samada yang belum tercemar ataupun yang telah tercemar. Tambahan pula kajian seumpama ini
belum pernah dijalankan di kawasan tersebut (komunikasi peribadi). Selain daripada itu beberapa
tempat dikawasan Taman Negara telah dibenarkan untuk dilawati oleh orang ramai. Kawasan seperti
Sungai Tahan dibenarkan untuk dikunjungi sehingga ke kawasan Lata Berkoh. Di kawasan ini
pengunjung dibenarkan untuk mandi manda dan memancing. Di kawasan Sungai Keniam pula pada
musim-musim tertentu orang ramai dibenarkan untuk memancing. Pengangkutan untuk sampai kedua-
dua kawasan tersebut kebanyakkannya menggunakan bot-bot yang menggunakan minyak petrol. Oleh
itu kajian ini jika dilakukan secara berkala selain daripada menyediakan bacaan latar juga dapat
digunakan untuk menilai kesan kemasukkan orang ramai terhadap kandungan unsur-unsur surihan di
dalam kawasan tersebut.
Bahan dan Kaedah
Kawasanan kajian
Kajian telah dilakukan dikawasan Sungai Atok dan Sungai Tahan, Taman Negara, Pahang (lihat rajah
1). Di Sungai Atok pensampelan telah di buat di dua lokasi, lokasi pertama (SA 1) terletak 2 km dari
muara sungai sementara lokasi ke 2 (SA 2) berada kira-kira 10 km dari muara sungai. Di Sungai
Tahan, pensampelan juga dilakukan pada dua lokasi. Lokasi pertama (ST 1) terletak 2 km dari muara
sungai sementara lokasi ke 2 (ST 2) terletak 8 km dari muara sungai. Kedua- dua buah sungai
bermuara ke dalam Sungai Tembeling yang merupakan sempadan Taman Negara di sebelah Timur
Laut.
Kaedah pensampelan
Sampel ikan telah ditangkap dengan menggunakan jaring nilon. Jaring direntangkan mengikut
kelebaran sungai dan dibiarkan selama satu malam (11jam). Ikan yang terlekat pada jaring diasingkan
mengikut spesies dan dimasukkan ke dalam beg plastik. Setiap spesies diambil sebanyak tiga ekor.
Ikan yang diambil dipilih berdasarkan beratnya (900 – 1000 gm). Sampel yang telah dibungkus
kemudian dimasukkan kedalam bekas penyejuk (Coleman) berisi ais (0° C).
259
Rajah 1: Lokasi pengambilan sampel di Sungai Atok dan Sungai Tahan Taman Negara, Pahang.
260
Rawatan sampel
Apabila sampai di makmal ikan-ikan dicuci bersih dengan menggunakan air suling. Ikan kemudian
disiang dan dibilas dengan air suling. Sampel tersebut diwapkan selama satu jam pada suhu 100°C
untuk memisahkan bahagian otot dan bahagian tulang. Setelah dipisahkan, kedua-dua bahagian
dikeringkan sehingga berat menjadi tetap. Sampel otot dan tulang kemudian dihaluskan dengan
pengisar dan di ayak untuk mendapatkan sampel yang homogen. Sampel yang telah dihomogen ini
siap untuk digunakan.
Penentuan radioaktiviti dalam sampel
Botol polietilina kosong ditimbang dan dicatatkan beratnya. Sampel otot dan tulang seperti persediaan
diatas dimasukkan ke dalam botol tersebut. Sampel dimasukkan ke dalam botol mengikut banyaknya
bahan piawai yang digunakan (bahan piawai terdapat dalam botol polietilina dengan isipadu tertentu).
Botol dimaterikan dan ditimbang semula untuk mendapatkan berat sampel. Botol yang telah
dimaterikan dibiarkan selama satu bulan untuk mencapai kesaimbangan sekular. Piawai yang
digunakan untuk kajian ini adalah IAEA Standard – 134 cockle flesh.
Aktiviti sampel dianalisis dengan menggunakan sistem pembilang spektrometri sinar gama.
Keradioaktifan uranium-238 ditentukan menerusi puncak bismuth-214 sementara kehadiran torium-
232 dikenalpasti melalui talium-208. Keradioaktifan uranium-238 dan torium-232 ditentukan dengan
menggunakan formula berikut:
WS = AS x MP x WP / AP x MS
Dimana WS = keradioaktifan unsur dalam sampel (Bq/kg)

WP = keradioaktifan unsur dalam piawai (Bq/kg)
AS = aktiviti unsur dalam sampel (b/s)
AP = aktiviti unsur dalam piawai (b/s)
MS = jisim sampel (g)
MP = jisim piawai (g)
Penentuan logam berat dalam sampel
Ambil 200 mg sampel dan ditambah dengan 3 ml asid nitrik (HNO3) dan 3 ml hidrogen peroksida
(H2O2). Sampel dipanaskan di dalam ketuhar mikro gelombang sehingga larutan menjadi jernih.
Setelah disejukkan sampel dicairkan dengan air suling sehingga larutan menjadi 15 ml. Larutan
tersebut kemudian dituraskan. Hasil turasan dimasukkan ke dalam alat Spektrometri Emissi Optik
Plasma Gandingan Teraruh (Inductively Coupled Plasma-Optical Emission Spectrometry ICP-OES)
untuk mendapatkan bacaan kandungan logam berat Larutan piawai yang digunakan adalah ICP Multi
Element Standard Solution (IV) (Merck)
Keradioaktifan uranium-238 dan torium-232 mengikut lokasi
Dalam kajian ini lima spesies ikan telah dapat ditangkap daripada kedua-dua buah sungai tersebut.
Ikan-ikan tersebut adalah terdiri daripada species Puntius gonionotu (Lampan Jawa), Leptobarbus
hoevenii (Ikan Jelawat), Mytus nemurus(Ikan Baung) Puntius schwanwnfeldii (Lampan Sungai) dan
Cyprinus carpio (Ikan Lee Koh). Secara relatif didapati kandungan uranium-238 dalam ikan di
Sungai Atok adalah lebih tinggi jika dibandingkan dengan Sungai Tahan. Perbezaan keradioaktifan
diantara kedua buah sungai tersebut adalah 1.68 Bg/kg (lihat Jadual 1) Perbezaan ini mungkin
disebabkan oleh uranium-238 yang hadir dalam bahan organik tidak banyak termendap dalam Sungai
260
261
Tahan kerana pengaliran air di sungai ini sangat deras dan menghalang pemendakan bahan organik
yang banyak di dasarnya. Sementara di Sungai atok pengalirannya yang perlahan memberi peluang
untuk berlakunya pemendakan bahan organik didasarnya dan sekaligus meningkatkan kandungan
uranium-238. Selain daripada itu terdapat perbezaan pH diantara kedua-dua buah sungai yang dikaji.
Air Sungai Atok dikawasan kajian berada pada pH antara 7. 2 – 7.4 sementara air di Sungai Tahan
pada kawasan kajian berada dalam julat 6.5 – 6.8. Menurut Drummond et. al. (1974) pengumpulan
metal mekuri dalam insang dan sel darah pada ikan brook trout adalah lebih tinggi pada pH 6 jika
dibandingkan pada pH 9. Apakah perbezaan pH yang kecil dalam kajian ini juga turut menyumbang
dalam penyerapan uranium-238 tidak dapat dipastikan.
Unsur torium-232 pula tidak dapat dikesan di dalam ikan di kedua-kedua sungai tersebut.Ini mungkin
di sebabkan torium-232 sukar diserap oleh ikan jika dibanding dengan uranium-238. Kajian terhadap
kandungan torium-232 didalam tanah dikawasan tersebut juga mendapati kepekatan torium-232
adalah jauh lebih rendah daripada uranium-238 (Norizan, 2002). Ini menyebabkan kandungan torium-
232 dalam air mahupun bahan organik jauh lebih rendah daripada uranium-238 dan sukar untuk
diambil oleh ikan. Torium juga bukanlah merupakan unsur yang diperlukan.
Jadual 1: Kandungan U-238 dan Th-232 didalam sampel ikan mengikut lokasi
Keradioaktifan (Bq/kg)
Lokasi
Uranium-238 Torium-232
Sungai Atok 4.42 ± 0.012 TH
Sungai Tahan 2.74 ± 0.008 TH

TH – unsur tidak dapat dikesan
Kandungan uranium-238 dan torium-232 mengikut spesies ikan
Kajian mengikut spesies ikan pula mendapati Puntius schwanwnfeldii mempunyai kepekatan
uranium-238 yang tertinggi, iaitu sebanyak 6.17 ± 0.007 Bq/kg manakala Puntius gonionotus
menunjukkan kepekatan uranium-238 yang terendah iaitu 1.47 ± 0.006 Bq/kg (Jadual 2). Kandungan
uranium-238 dalam Leptobarbus hoevenii dan Mytus nemurus adalah hampir sama iaitu masing-
masing 4.13 ± 0.006 Bq/kg dan 4.74 ± 0.006 Bq/kg. Sementara dalam Cyprinus carpio kepekatan
uranium-238 adalah 3.32 ± 0.008 dan nilai ini merupakan nilai kedua terendah.
Logam berat dan bahan radionuklid dalam air dapat masuk kedalam tubuh ikan melalui penjerapan
substrat ke atas permukaan badan, penyerapan melalui insang dan dinding usus atau kedua-duanya
sekali (Luoma, 1983) Perbezaan kepekatan uranium-238 dalam kelima-lima spesies mungkin
disebabkan oleh perbezaan penjerapan logam melalui permukaan, badan atau perbezaan dalam
penyerapan insang dan dinding usus. Selain daripada itu faktor yang menyebabkan perbezaan dalam
kandungan uranium-238 adalah cara pemakanan dan jenis makanan yang dimakan. Ikan-ikan yang
mencari makan didasar sungai berkemungkinan mempunyai kepekatan uranium-238 yang lebih tinggi
disebabkan terdapat pemendakan bahan organik yang tinggi di dasar sungai. Ikan-ikan yang memakan
moluska juga akan tinggi kandungan logam beratnya kerana moluska mempunyai kebolehan yang
tinggi untuk menumpuk logam berat di dalam tubuhnya jika dibanding dengan organisme akuatik
yang lain (Kulikova et. al., 1985).
Dalam kajian ini tiada torium-232 ditemukan dalam semua spesies ikan.Hasil kajian ini agak berbeza
dengan kajian yang telah dijalankan oleh Norsaliza (2004). Kajian beliau di Sungai Keniam dan
261
262
Sungai Terengan, Taman Negara, Pahang mendapati hanya torium -232 ditemukan di dalam Puntius
schwanenfeldii dan Cyclocheilichthys apogon (Temperas) dengan kepekatan masing-masing 0.57 ±
0.081 Bq/kg dan 0.67 ± 0.094 Bq/kg. Sementara di dalam Osteochilus spilurus (Ikan Rom) dan
Labiobarbus festivus (Ikan Kawan) didapati kepekatan uranium-238 adalah dalam julat 2.63 ± 0.84
Bq/kg hingga 10.24 ± 2.84 Bq/kg dan kepekatan torium-238 adalah dalam julat 4.01 ± 0.94 Bq/kg
hingga 20.01 ± 2.84 Bg/kg. Perbezaan ini mungkin disebabkan ikan-ikan yang dikaji adalah berbeza
kecuali Puntius schwanenfeldii.
Jadual 2: Kandungan uranium-238 dan torium-232 mengikut spesies ikan
Keradioaktifan (Bq/kg)
Spesies
Uranium-238 Torium-232
Puntius gonionotus (Lampan Jawa) 1.47 ± 0.006 TH
Leptobarbus hoevenii (Ikan Jelawat) 4.13 ± 0.006 TH
Mytus nemurus(Ikan Baung) 4.74 ± 0.007 TH
Puntius schwanenfeldii (Lampan Sungai) 6.17 ± 0.007 TH
Cyprinus carpio (Ikan Lee Koh). 3.32 ± 0.008 TH

TH tidak dapat dikesan
Kandungan logam berat mengikut lokasi
Hasil kajian mengikut lokasi menunjukkan 12 unsur ditemukan dalam ikan di Sungai Atok manakala
13 unsur pula ditemukan di Sungai Tahan(Jadual 3). Logam aluminium hanya dijumpai dalam ikan di
Sungai Tahan. Secara keseluruhan unsur yang terdapat di dalam ikan di kedua buah sungai
menunjukkan nilai yang hampir serupa kecuali unsur kuprum yang didapati lebih tinggi di Sungai
Atok dan unsur ferum didapati lebih tinggi di Sungai Tahan. Secara keseluruhan kepekatan argentum
dalam ikan dari kedua-dua sungai adalah palingi tinggi dan diikuti oleh magnesium dan kobalt
menunjukkan kepekatan paling rendah.
Menurut Post (1983) kepekatan maksimum aluminium pada pH 7 adalah 3.75 mg/kg manakala pada
pH 9 kepekatan aluminium adalah sekurang-kurangnya 375 mg/kg. Tidak diketahui kenapa nilai
aluminium di Sungai Tahan agak tinggi sedangkan pH dikawasan tersebut adalah diantara 6.5- 6.8.
Secara relatif ikan-ikan dari Sungai Atok menunjukkan kepekatan argentum, zink, kuprum,kobalt
kromium, plumbum dan strontium yang lebih tinggi. Sementara kepekatan arsenik, selenium, ferum,
mangan dan magnesium adalah lebih tinggi dalam sampel ikan dari Sungai Tahan. Tidak dapat
dipastikan apakah perbezaan ini disebabkan oleh perbezaan pH diantara kedua-dua buah sungai.
Jadual 3.; Kandungan logam berat dalam ikan mengikut lokasi
Kepekatan (mg/kg berat kering)

Jenis logam
Sungai Atok Sungai Tahan
Aluminium BHP 47.84 ± 1.84
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263
Argentum 3761.21 ± 585.32 3750.69 ± 582.29
Arsenik 1.69 ± 0.60 1.71 ± 0.62
Selenium 1.52 ± 0.53 1.77 ± 0.44
Zink 49.95 ± 1.61 46.35 ± 1.52
Kuperam 55.03 ± 0.50 46.77 ± 0.65
Kobalt 0.25 ± 0.03 0.21 ± 0.02
Ferum 4.07 ± 0.13 9.44 ± 0.63
Mangan 26.83 ± 0.33 30.39 ± 0.33
Kromium 1.23 ± 0.02 0.91 ± 0.02
Plumbum 0.39 ± 0.11 0.26 ± 0.21
Magnesium 751.45 ± 3.59 766.78 ± 8.06
Strontium 5.88 ± 0.06 5.82 ± 0.07
Kandungan logam berat dalam otot dan tulang ikan
Jadual 4 menunjukkan kandungan logam berat didalam otot dan tulang ikan. Dari jadual tersebut
dapat dilihat aluminium mempunyai kepekatan yang sangat tinggi di dalam otot/daging ikan iaitu
sebanyak 163. 51 ± 3.28 mg/kg berat kering. Sementara zink, kromium dan strontium dalam tulang
ikan adalah lebih kurang dua kali ganda kepekatannya jika dibandingkan dengan kepekatan dalam
otot.
Kepekatan unsur selenium dan plumbum dalam tulang adalah separuh daripada yang terdapat dalam
daging ikan. Manakala kepekatan argentum, arsenik dan kobalt adalah hampir sama baik di dalam
daging mahupun di dalam tulang. Kuprum, ferum mangan dan magnesium mempunyai kepekatan
yang lebih tinggi di dalam sampel tulang.
Kandungan logam berat dalam daging dan tulang ikan berasal daripada pengambilannya melalui
makanan semasa proses pencernaan dan proses asimilasi makanan tercemar. Logam berat diserap oleh
dinding usus dan dibahagikan ke tisu lain. Pembahagian ini dipengaruhi oleh spesies dan keadaan
kimia logam, kualiti makanan dan tempoh masa selepas pencernaan.
Jadual 4: Kandungan logam berat didalam otot dan tulang ikan

Jenis logam
Otot Tulang
Aluminium 163.51 ± 3.28 27.82 ± 4.06
Argentum 3752.77 ± 4.72 3759.11 ± 694.63
Arsenik 1.95 ± 0.52 1.45 ± 0.69
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264
Selenium 2.04 ± 0.45 1.24 ± 0.52
Zink 61.18 ± 2.12 46.35 ± 1.52
Kuperam 56.71 ± 0.61 46.77 ± 0.65
Kobalt 0.23 ± 0.03 0.21 ± 0.02
Ferum 32.86 ± 1.37 9.44 ± 0.63
Mangan 30.50 ± 0.39 30.39 ± 0.33
Kromium 1.42 ± 0.02 0.91 ± 0.02
Plumbum 0.15 ± 0.05 0.26 ± 0.21
Magnesium 878.32 ± 7.97 766.78 ± 8.06
Strontium 7.94 ± 0.08 5.82 ± 0.07
Kandungan logam berat dalam pelbagai spesies ikan
Kandungan logam berat dalam pelbagai spesies ikan tercantum pada jadual 5. Secara
keseluruhan didapati argentums menunjukkan kepekatan yang sangat tinggi di dalam semua
spesies. Julat kepekatan logam tersebut adalah 3703.01±707.91mg/kg berat kering hingga
3859.44±589.29 mg/kg berat kering. Kepekatan tertinggi terdapat pada Puntius gonionotus
sementara kepekatan terendah adalah pada Leptobarbus hoevenii. Unsur kedua tertinggi adalah
magnesium dengan julat 644.14 ± 10.79mg/kg berat kering iaitu terdapat pada Leptobarbus hoevenii
sehingga 861.99 ± 7.21 mg/kg berat kering yang terdapat pada Puntius schwanenfeldii.Logam
aluminium pula hanya ditemukan pada Mystus nemurus. Kepekatan yang agak tinggi ini mungkin
disebabkan sifat permukaan badan ikan tersebut yang tidak bersisik dan memudahkan unsur
aluminium untuk dijerap.
Bagi unsur zink dan kuprum kepekatan terendah ditemukan pada Cyprinus carpio masing-
masing adalah 39.87 ± 1.03 mg/kg berat kering dan 27.54 mg/kg berat kering. Kepekatan
tertinggi bagi zink adalah pada Puntius gonionotus dan bagi kuprum pula adalah pada
Leptobarbus hoevenii. Spesies Mystus nemurus pula menunjukkan kandungan mangan dan
ferum terendah iaitu masing-masing 26.43 ± 0.34 mg/kg berat kering dan 1.83 ± 0.19 mg/kg
berat kering. Kandungan ferum dan mangan tertinggi pula dijumpai pada Puntius gonionotus,
kepekatannya adalah masing 32.49 ± 0.21 mg/kg berat kering. dan 29.06 ± 0.32 mg/kg berat
kering. Bagi unsur kobalt, plumbum dan arsenik didapati kepekatannya hampir sama dalam
semua spesies. Unsur kromium dan strontium menunjukkan kepekatan terendah dalam
Cyprinus carpio iaitu masing 0.42 ± 0.02 mg/kg berat kering dan 3.80 ± 0.04 mg/kg berat
kering. Nilai tertinggi bagi kedua-dua unsur tersebut masing-masing ditemukan pada Puntius
gonionotus dan Puntius schwanenfeldii.
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265
Jadual 5: Kandungan logam berat dalam ikan mengikut spesies

Unsur
Puntius Leptobarbus Cyprinus Mystus Puntius
gonionotus hoevenii carpio nemurus schwanenfeldii
Al TH TH TH 47.84±1.84 TH
Ag 3859.44±589.29 3703.01±707.91 3743.72±401.01 3784.16±749.15 3711.80±425.00
As 1.53±0.43 1.27±0.40 1.95±0.76 1.81±0.52 1.59±0.74
Se 1.23±0.71 1.08±0.37 2.04±0.34 1.57±0.53 2.06±0.47
Zn 62.31±1.68 48.26±2.17 39.87±1.03 43.35±1.86 48.33±1.24
Cu 61.60±0.63 84.58±0.83 27.54±0.43 66.28±0.89 31.35±0.30
Co 0.19±0.03 0.11±0.03 0.22±0.02 0.22±0.03 0.23±0.03
Fe 32.49±0.21 TH 12.51±0.24 1.83±0.19 12.66±0.29
Mn 29.06±0.32 28.94±0.56 32.13±0.26 26.43±0.34 28.40±0.30
Cr 2.94±0.04 0.82±0.02 0.42±0.02 0.62±0.01 1.30±0.02
Pb 0.15±0.07 0.19±0.04 0.15±0.11 0.19±0.10 0.19±0.07
Mg 748.35±5.42 644.14±10.79 734.46±7.53 696.91±7.88 861.99±7.21
Sr 6.24±0.05 10.15±0.14 3.80±0.04 5.02±0.06 6.62±0.07
TH dibawah had pengesanan
Hasil kajian ini jika dibandingkan dengan kajian yang telah dijalankan di Vietnam dan dengan
piawaian antarabangsa mendapati semua logam dalam ikan di kedua buah sungai berada pada paras
yang selamat atau lebih rendah daripada paras selamat kecuali mangan dan strontium. Kedua-dua
logam ini didapati dalam semua spesies adalah lebih tinggi daripada keputusan yang ditemukan di
Vietnam (Wagner & Boman, 2003). Selain itu dalam kajian ini kandungan unsur dinyatakan
berdasarkan berat kering sampel, bagaimanapun kajian lain yang dijalankan menyatakan
hasil berasaskan berat basah. Kepekatan logam dalam tisu ikan yang dinyatakan dalam berat
kering boleh ditukar kepada berat basah dengan membahagikan nilai tersebut dengan faktor 4
hingga 6 (Khan et. al., 1987; Parsons, 1999; Romeo et. al., 1999; Lewis et. al., 2002).
Kesimpulan
Keradioaktifan uranium-238 dalam ikan di kedua buah sungai berada pada paras yang normal
sementara unsur torium-232 tidak ditemukan di dalam kesemua spesies ikan yang dikaji.
Bagi kandungan logam berat pula semua spesies menunjukkan paras yang selamat atau lebih
rendah daripada paras yang selamat kecuali unsur mangan dan unsur strontium. Kedua-dua
unsur tersebut mungkan tersedia lebih tinggi secara alamiah menyebabkab ia ditumpuk lebih
tinggi didalam ikan. Kehadiran unsur yang rendah didalam ikan menunjukkan kawasan
sungai tersebut masih merupakan kawasan yang bersih
Rujukan
Drummond, R. A., Olsen, G. F. & Batterman, A. R. (1974). Cold response and uptake of mercury by
brook trout, Salvelinus fontinalis, exposed to mercuric compounds at different hydrogen-ion
concentration. Trans. Am Fish. Soc. 103: 244-256.
265
266
Hargrave, B. T. (1991). Impacts of man’s activities on aquatic systems. DlmBarnes, R. S. K.

& Mann, K. H. (pnyt.). Fundementals of aquatic Ecology. U.S: Blackwell Science.
Khan, A. H., Ali, M., Biaswas, S. K. & Hadi, D. A. (1987). Trace element in marine fish from the Bay
of Bengal. Sci. Total Environ. 65: 121–130.
Kulikova, I., Seisuma, Z., & Legsdina, M. (1985). Heavy metals in marine organisms.Dlm Salanki,
J.(pnyt.). Heavy Metals in Water Organisms. Akademiai Kiado, Budapest, 141-151.
Lewis, M. A., Scott, G. I. Bearden, D. W., Quarles, R. L. Moore, J., Strozier, E. D., Sivertsen, S.
K., Dias, A. R. & Sanders, M. (2002). Fish tissue quality in near-costal areas of the Gulf of
Mexico receiving point source discharges. Sci. Total Environ. 84: 249-262.
Louma, S. N. (1983). Bioavailability of trace metalsto aquatic organisms – a review. Sci. Total
Environ. 38: 1-22
Norizan Abd. Majid .(2002). Keradioaktifan tabii dan kandungan logam berat dalam tanah dan
sedimen di kawasan Sungai Atok dan Sungai Tahan, Taman Negara, Pahang. Tesis Sm. Sn.
(Kep.). UKM. Bangi, 32-34.
Norsaliza Zakaria (2004). Radioaktiviti dan kandungan logam berat dalam ikan di Sungai Keniam
dan Sungai Terengan, Taman Negara, Pahang. Tesis Sm. Sn. (Kep.), UKM, Bangi, 21–23.
Parson, E. C. M. (1999). Trace element concentration in whole fish from North Lantau waters, Hong
Kong. ICES J. Mar. Sci. 56: 791–794.
Post, G. W. (1983). Textbook of Fish Health. T. F. H. Publication Inc. Ltd., Hong Kong.
Romeo, M., Siau, Y. Sidoumou, Z. & Gnassia-Barelli, M. (1999). Heavy metal distribution in
different fish species from the Mauritania coast. Sci. Total Environ. 232: 169–175.
Wachs, B. (1985). Bioindicators for the heavy metal load of river ecosystem. Dlm. Salanki, J. (pnyt.).
Heavy Metals in Water Organisms. Akademiai Kiado, Budapest,179–180.
.Wagner, A. & Boman, J. (2003). Biomonitoring of trace element in muscle and liver tissues of
freshwater fish. Spectrochemica Acta Part B: Atomic Spectroscopy. 58(12): 2215–2226.
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TINDAK BALAS PENGKOMPLEKSAN ASID-3-(3-BENZOILTIOUREDO) PROPIONIK

DENGAN BIS[ASETATOTRIFENILFOSFINARGENTUM(I)] DAN KOBALT(II)KLORIDA
NURZIANA NGAH, BOHARI M. YAMIN & MOHAMMAD BIN KASSIM

Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains & Teknologi
Universiti Kebangsaan Malaysia, Bangi
ABSTRAK
Tindak balas sebatian kompleks bis[asetatotrifenilfosfinargentum(I)] dengan asid-3-(3-
benzoiltiuredo)propionik dalam pelarut kloroform telah menghasilkan kompleks [(PPh3)2Ag(µ-
C11H12O3N2S)2Ag(PPh3)2]. Dalam tindak balas ini, kumpulan asetat telah mengalami tindak balas
gantian oleh sebatian asid-3-(3-benzoiltiouredo)propionik. Kajian kristalografi sinar-X mendapati
produk ini mempunyai sistem triklinik dengan kumpulan ruang Pī, a = 12.691(3) Ǻ, b = 13.080(3) Ǻ,
c = 14.267(3) Ǻ, α = 94.888(5) o , β = 93.818(6) o dan γ = 97.254(5) o. Dua molekul asid-3(3-
benzoiltiouredo)propionik berkoordinat kepada kedua-dua atom argentum melalui atom sulfur
kumpulan tiono dan oksigen kumpulan karboksilik membentuk kompleks dimerik. Tindak balas asid-
3-(3-benzoiltiouredo) propionik dengan kobalt(II) klorida dalam pelarut etanol telah menghasilkan
kompleks [Co(C11H11N2O3S)2(C2H5OH)2]. Kajian mendapati atom kobalt berkoordinat kepada 2
molekul asid-3-(benzoiltiouredo)propionik melalui atom S dan 2 molekul etanol melalui atom O
dalam sekitaran 4 koordinatan. Kumpulan karboksilik telah mengalami nyahproton menjadikan
sebatian tiourea sebagai ligan negatif ekacas. Kajian kristalografi sinar-x mendapati hablur ini
mempunyai sistem triklinik dengan kumpulan ruang Pī, a = 8.779(2) Ǻ, b = 9.094(2) Ǻ, c = 9.118(2)
Ǻ, α = 78.592(4) o , β = 72.443(4) o dan γ = 75.895(4) o.
PENGENALAN
Sebatian asid-3-(3-benzoiltiouredo)propionik (Rajah 1) merupakan sebatian benzoiltiourea terbitan

asid amino β-alanin. Sebatian ini menarik kerana terdapat beberapa tapak untuk berlakunya tindak
balas pengkompleksan dengan logam, iaitu kumpulan tiono (C=S), karbonil (C=O) dan karboksilik
(COOH). Kertas ini membincangkan tindak balas pengkompleksan sebatian asid-3-(3-
benzoiltiouredo)propionik dengan kompleks bis[asetatotrifenilfosfinargentum(I)] dan
kobalt(II)klorida.
S
1 2
1
C NH
8
9 2 6
O H2 C NH C 3
7
C11 CH2 O 4
10
5
HO
Rajah 1: Struktur asid-3-(3-benzoiltiouredo)propionik
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268
EKSPERIMEN
Asid-3-(3-benzoiltiouredo)propionik disediakan mengikut kaedah yang dilaporkan oleh Yusof &

Yamin [1]. Asid-3-(3-benzoiltiouredo)propionik dan bis[asetatotrifenilfosfinargentum(I)] dipanaskan
dalam pelarut kloroform dan dibiarkan memekat pada suhu bilik. Sedikit aseton dan etanol ditambah
untuk mencairkan larutan. Larutan ini dituras dan dibiarkan menghablur pada suhu bilik. Dalam
tindak balas dengan kobalt(II)klorida pula, larutan akues CoCl2 ditambah setitis demi setitis ke dalam
larutan asid 3-(3-benzoiltiouredo)propionik yang telah dicampurkan sedikit larutan KOH. Mendak
merah jambu yang terbentuk dituras dan dikeringkan.
PERBINCANGAN
Tindak balas asid-3-(3-benzoiltiouredo)propionik dengan sebatian

bis[asetatotrifenilfosfinargentum(I)], filtrat jernih terhasil didapati memberikan hablur bersinar tanpa
warna setelah dibiarkan selama ½ jam pada suhu bilik. Mikroanalisis unsur CHNS mendapati hablur
yang terbentuk mempunyai unsur karbon, hidrogen, nitrogen dan sulfur. Analisis mikrounsur
menunjukkan peratus unsur yang hampir sama, kecuali peratus unsur karbon yang agak berbeza
(Jadual 1) berdasarkan formula [(PPh3)2Ag(µ-C11H12O3N2S)2Ag(PPh3)2]. Spektrum inframerah hablur
menunjukkan regangan bagi υ(C=O), υ(C-O), υ(C=S) dan υ(C-N) berbanding ligan bebas asid-3-(3-
benzoiltiouredo)propionik (Jadual 2). Analisis spektrum 1H RMN menunjukkan perubahan yang agak
ketara bagi δ(N1) dan δ(N2), manakala spektrum 13C RMN pula menunjukkan perbezaan yang besar
bagi δ(C8) dan δ(C11) berbanding ligan bebas (Jadual 3). Dalam tindak balas dengan
kobalt(II)klorida, penambahan larutan aques kobalt(II)klorida kepada asid-3-(3-
benzoiltiuredo)propionik yang telah dicampur dengan larutan KOH telah memberikan mendakan
merah jambu secara perlahan-lahan. Analisis mikrounsur CHNS mendakan merah jambu
menunjukkan persetujuan peratus karbon, hidrogen, nitrogen dan sulfur dengan jangkaan (Jadual 1).
Analisis spektrum inframerah menunjukkan perubahan nombor gelombang bagi υ(C=S) berbanding
ligan bebas (Jadual 2) menunjukkan kemungkinan koordinatan berlaku pada atom sulfur. Kajian
RMN terhadap sebatian ini tidak menunjukkan sebarang isyarat. Oleh yang demikian, ia bersifat
paramagnet.
JADUAL 1: Takat lebur dan analisis mikrounsur sebatian

Takat Peratus unsur
Sebatian
lebur(oC) C H N S
[(PPh3)2Ag(µ- 56.36 4.65 2.86 2.90
180.7-181.5
C11H12O3N2S)2Ag(PPh3)2] (63.87) (4.79) (3.17) (3.62)
48.98 5.66 7.62 8.78
[Co(C11H11N2O3S)2(C2H5OH)2] 174.5-176.3
(49.19) (5.90) (8.19) (9.38)
*(nilai jangkaan)
JADUAL 2: Mod getaran spektrum inframerah sebatian

Sebatian [(PPh3)2Ag
[Co(C11H11N2O3S)2
C11H12O3N2S1 (µ-C11H12O3N2S)2
(C2H5OH)2]
Frekuensi, cm-1 Ag(PPh3)2]
υ(C=S) 660.0 612.4 619.0
Υ(C=O) 1671.6 1677.7
1397.6
υ(C-O) 1029.4 1094.2
υ(C-N) 1227.4 1272.2 1240.1
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JADUAL 3: Anjakan kimia 1H dan 13C RMN bagi [(PPh3)2Ag(µ-C11H12O3N2S)2Ag(PPh3)2]

Sebatian
[(PPh3)2Ag(µ-C11H12O3N2S)2
C11H12O3N2S1
Ag(PPh3)2]
Anjakan kimia, ppm
δ(N1) 11.33 7.74
δ(N2) 11.05 7.46
δ(C1-C6) 128.5-133.08 129.45-135.96
δ(C7) 168.19 167.52
δ(C8) 180.35 207.22
δ(C9) 40.69 42.81
δ(C10) 32.39 34.63
δ(C11) 173.10 179.45
Kajian kristalografi sinar-X
Kajian kristalografi sinar-X sebatian [(PPh3)2Ag(µ-C11H12O3N2S)2Ag(PPh3)2] mendapati ia

mempunyai sistem triklinik dengan kumpulan ruang Pī, a = 12.691(3) Ǻ, b = 13.080(3) Ǻ, c =
14.267(3) Ǻ, α = 94.888(5) o , β = 93.818(6) o dan γ = 97.254(5) o. Kedua-dua atom argentum
terkoordinat kepada 2 atom fosforus kumpulan trifenilfosfin, oksigen dan sulfur sebatian asid-3-(3-
benzoiltiouredo)propionik, menjadikan geometri di sekitar atom argentum tetrahedron terherot dengan
sudut di antara 85.10(14) dan 116.67(6)o. Rajah 1 menunjukkan struktur molekul dengan skema
penomboran.
RAJAH 2 Struktur [(PPh3)2Ag(µ-C11H12O3N2S)2Ag(PPh3)2] dengan skema penomboran

yang digunakan
Kajian kristalografi sinar-X mendaapti hablur kompleks [Co(C11H11N2O3S)2(C2H5OH)2] bersistem

triklinik dengan kumpulan ruang Pī, a = 8.779(2) Ǻ, b = 9.094(2) Ǻ, c = 9.118(2) Ǻ, α = 78.592(4) o ,
β = 72.443(4)o dan γ = 75.895(4)o ]. Molekul ini bersifat sentrosimetri berpusat pada atom kobalt, di
mana atom kobalt berkoordinat kepada 2 molekul C11H12O3N2S1 melalui atom S1 dan 2 molekul
C2H5OH melalui atom O4 (Rajah 2). Geometri atom kobalt adalah segi empat planar dengan sudut
S1-Co1-S1A dan O4-Co1-O4A sebanyak 180.0(1)o. Dalam penyelesaian struktur ini, ketaktertiban
atom C12 belum dapat diselesaikan sehingga kini.
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RAJAH 3 Struktur [Co(C11H11N2O3S)2(C2H5OH)2] dengan skema penomboran yang

digunakan
KESIMPULAN
Kompleks [(PPh3)2Ag(µ-C11H12O3N2S)2Ag(PPh3)2] terhasil dalam tindak

bis[asetatotrifenilfosfinargentum(I)] dengan asid-3-(3-benzoiltiouredo)propionik. Dalam tindak balas
ini, kumpulan asetat sebatian bis[asetatotrifenilfosfinargentum(I)] telah diganti oleh sebatian asid-3-
(3-benzoiltiouredo)propionik membentuk kompleks dimerik. Pengkompleksan pada kedua-dua atom
argentum melibatkan atom oksigen dan sulfur satu ligan secara jejambat. Tindak balas dengan
CoCl2.6H2O pula, sebatian asid-3-(benzoiltiouredo)propionik telah mengalami deprotonasi pada
kumpulan karboksilik. Pengkompleksan 2 molekul anion tersebut dan 2 molekul etanol dengan 1
molekul ion Co2+ ini menjadikan keseluruhan kompleks [Co(C11H11N2O3S)2(C2H5OH)2] ini bercas
neutral.
PENGHARGAAN
Ucapan terima kasih kepada Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains
Teknologi UKM di atas kemudahan penyelidikkan yang disediakan. Jutaan terima kasih kepada
Kementerian Sains, Teknologi & Inovasi (MOSTI) kerana bantuan kewangan yang dihulurkan
melalui Skim Biasiswa National Science Fellowship (NSF).
RUJUKAN
[1]Yusof, M. S. M. & Yamin, B. M. 2003. 3-(3-benzoylthiouredo)propionic acid. Acta Cryst. 2003.

o828-
829
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SYNTHESIS, STRUCTURAL CHARACTERIZATION AND BIOLOGICAL ACTIVITY OF

ORGANOTIN(IV) DERIVATIVES WITH CARBOHYDRAZIDE -
BIS(SALICYLALDEHYDE) (H4CBS): X-RAY CRYSTAL STRUCTURE OF [Me2Sn(H2CBS)]
M. A. Affan1*, Y. Z. Liew 1, Fasihuddin B. Ahmad1, Bohari M. Yamin 2 and Mustaffa B. Shamsuddin

3
1
Resource Chemistry, Faculty of Resource Science and Technology
Universiti Malaysia Sarawak, 94300 Kota Samarahan,
Sarawak, Malaysia
2
School of Chemical Sciences and Food Technology
Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia
3
Universiti Teknologi Malaysia, 81310 UTM Skudai
Johor Darul Takzim, Malaysia
Abstract
Five new organotin(IV) complexes of carbohydrazide-bis(salicylaldehyde) [H4CBS, (1)] ligand with

RnSnCl4-n (n = 1, 2) have been synthesized in the presence of base and refluxing in 1:2:1 molar ratio
(metal:base:ligand). All organotin(IV) complexes (2-6) have been characterized by different physico-
chemical techniques like molar conductivity measurements, elemental analysis, UV-visible, IR and 1H
NMR spectral studies. All organotin(IV) complexes (2-6) are non electrolytic in nature. Among them,
dimethyltin(IV) complex (2) is also characterized by X-ray crystallography diffraction analyses. In the
solid state, the carbohydrazone ligand (1) exist as the keto tautomer but in solution in the presence of
base and organotin(IV) chloride(s), it converted to the enol tautomer and coordinated to the tin(IV)
atom in its deprotonated enolate form. X-ray crystallographic analysis shows that the dimethyltin(IV)
complex [Me2Sn(H2CBS)] (2), is five-coordinate and has a distorted trigonal-bipyramidal geometry
with the ligand coordinated to the tin(IV) as a tridentate dinegative fashion through its phenolic-O,
enolic-O and imine-N atoms. The general bond length order is oxo < phenolato < enolato. The Sn-O
(enolato) bond is longer than Sn-O (phenolato) bond by ~0.08 Å and is identical with Sn-O
(carboxylate) bond. The crystal structure of [Me2Sn(H2CBS)] (2) is monoclinic with space group
P2(1)/c with a = 13.163(3) Å, b = 11.143(2) Å, c = 12.203(2) Å, α = 90°, β = 106.063(3)°, γ = 90°, V
= 1750.4(6), Z = 4 and Dcalc = 1.689 Mg/m3. The IR and 1H NMR data are consistent with all the
organotin(IV) derivatives having similar geometry. The biological activities of the ligand [H4CBS,
(1)] and its organotin(IV) complexes have been studied by screening the compounds against several
bacteria. The complexes showed certain degree of biological activities.
Keywords: Carbohydrazone ligand; Organotin(IV) compounds; Crystal structure;

Biological activities.
Corresponding author: Tel.: +6082-679235; fax: +6082-672275

E-mail address: maaffan@frst.unimas.my
Introduction
Hydrazides and hydrazones have interesting ligational properties due to the presence of several
potential coordination sites. The chemistry of carbohydrazide compounds has been studied by Swamy
and Siddalingaiah [1]. Variety of metal complexes of symmetrical dihydrazones derived from
thiocarbohydrazides have been synthesized and their stereochemistry have been reported in the
literatures [2-3] and potentially useful biological activities [3]. However, the coordination chemistry
of the corresponding carbohydrazide derivatives is less explored [4]. Complexes of the
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carbohydrazide with non-transition metal ions such as organotin(IV) have not received as much
attention. In view of the importance of tin compounds in medicinal chemistry and biotechnology [5]
and as part of our on going work on tin-hydrazones [6-7], we report here the synthesis and
characterization of the new carbohydrazide-bis(salicylaldehyde) ligand (Scheme 1) and of its
organotin(IV) complexes (Scheme 2) and together with the X-ray crystal structure of dimethyltin(IV)
complexes [Me2Sn(H2CBS)] (2). The antimicrobial properties of these compounds are evaluated.
O
H
H C H
+
2 O C
N N
H2N NH2 HO
Carbohydrazide Salicylaldehyde
Abs. ethanol
Keto form
+ 2H2O
Enol form
Scheme 1
Experimental
Synthesis
Chemicals were obtained from Fluka and Aldrich and were used without further purification. Solvents
were analytical grade and purified by standard methods [8]. The C, H, N elemental analyses were
performed on a Carlo Erba EA 1108 model analyser. Infrared spectra were recorded as KBr disc using
Shimadzu 8201 PC Fourier-Transform Spectrometer. 1H NMR spectra were recorded in DMSO-d6
solution on a Bruker 300 FT-NMR spectrophotometer. Electronic spectra were recorded on a
Shimadzu 2401 PC UV-Visible spectrophotometer.
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Carbohydrazide-bis(salicylaldehyde) ligand (H4CBS) [C15H14N4O3] (1)
A mixture of carbohydrazide (0.005 mole, 0.450 g) and salicylaldehyde (0.0010 mole, 1.221 g) in
absolute ethanol (30 mL) were heated under reflux for 3-4 hours. The reaction mixture was allowed to
cool down to room temperature for half an hour. Then the white precipitate was filtered off and
washed several times using absolute ethanol. The crystalline white solid obtained was purified by
recrystallization from hot absolute ethanol and dried in vacuo over P2O5 overnight. Yield = 70.05%.
M. p. = 178-180 °C. Found: C, 59.20; H, 4.59; N, 18.40%. Calc. for C15H14N4O3: C, 59.21; H, 4.61;
N, 18.42%. λmax (nm) (DMF): 262, 292, 328. IR (νmaxcm-1 (KBr): 1681(-C=O), 1616 (C=N)+(C=C),
1272 (C–O, phenolic), 946 (N–N). 1H NMR (300 MHz, DMSO-d6): δ 10.86 (br, OH), δ 8.44 (s,
N=CH), δ 7.69 (br, CONH), δ 6.62–7.27 (m, aromatic-H).
Me2Sn(H2CBS) [Me2Sn(C15H12N4O3)] (2)
Carbohydrazide-bis(salicylaldehyde) (0.002 mole, 0.596g) was dissolved in hot absolute methanol (20
mL) under nitrogen atmosphere with potassium hydroxide (0.0042 mole, 0.236g) previously dissolved
in methanol (15 mL). The colour of the solution changed from off-white to yellow. The resulting
mixture was refluxed for one hour and a solution of Me2SnCl2 (0.002 mole, 0.439g) in methanol (10
mL) was added dropwise to the potassium salt of ligand solution, the color of the solution became
darker. The resulting solution was refluxed for four hours and allowed to cool. The precipitated
potassium chloride (KCl) was removed by filtration and the filtrate was evaporated to obtain the
yellow solid. The yellow micro-crystals were filtered off and washed with hexane and dried in vacuo
over P2O5 overnight. Single crystals suitable for X-ray diffraction studies were obtained by slow
evaporation of chloroform-hexane solution (1:1). Yield = 69.51%. M. p. = 218-220 °C. Found: C,
45.91; H, 4.03; N, 12.56%. Calc. for C15H12N4O3Me2Sn: C, 45.88; H, 4.05; N, 12.59%. λmax (nm)
(DMF): 266, 343, 398. IR (νmaxcm-1 (KBr): 1604 (C=N)+(C=C), 1318 (C–O, phenolic), 960 (N–N),
608 (Sn–C), 566 (Sn–O), 470 (Sn–N). 1H NMR (300 MHz, DMSO-d6): δ 11.41 (br, OH), δ 10.89 (s,
NH) δ 8.14 & 8.43 (s, N=CH)), δ 6.63–7.93 (m, aromatic-H). δ 0.68 (s, Sn-CH3).
Bu2Sn(H2CBS) [Bu2Sn(C15H12N4O3)] (3)
Complex (3) was synthesized in a similar way as in (2), using dibutyltin(IV) dichloride (0.002 mole,
0.688 g), instead of dimethyltin(IV) dichloride. Yield = 68.51%. M. p. = 225-227 °C. Found: C,
52.00; H, 6.03; N, 10.55%. Calc. for C15H12N4O3Bu2Sn: C, 52.02; H, 6.02; N, 10.55%. λmax (nm)
(DMF): 267, 340, 396. IR (νmaxcm-1 (KBr): 1604 (C=N)+(C=C), 1323 (C–O, phenolic), 952 (N–N),
590 (Sn–C), 560 (Sn–O), 440 (Sn–N). 1H NMR (300 MHz, DMSO-d6): δ 11.45 (br, OH), δ 11.05 (s,
NH) δ 8.15 & 8.41 (s, N=CH)), δ 6.65–7.89 (m, aromatic-H), δ 0.79 (t, 6H, (-CH3) of butyltin), δ 1.18
(m, 4H, (-CH2) of butyltin), δ 1.26-1.45 (m, 4H, (-CH2) of butyltin) δ 1.55-1.64 (m, 4H, -CH2-Sn).
Ph2Sn(H2CBS) [Ph2Sn(C15H12N4O3)] (4)
Complex (4) was prepared similarly to complex (2), using diphenyltin(IV) dichloride (0.002 mole,
0.688 g) instead of dimethyltin(IV) dichloride. Yield = 64.33%. M. p. = 208-210 °C. Found: C, 56.65;
H, 3.88; N, 9.87%. Calc. for C15H12N4O3Ph2Sn: C, 56.69; H, 3.87; N, 9.85%. λmax (nm) (DMF): 266,
342, 398. IR (νmaxcm-1 (KBr): 1600 (C=N)+(C=C), 1325 (C–O, phenolic), 960 (N–N), 600 (Sn–C),
520 (Sn–O), 460 (Sn–N). 1H NMR (300 MHz, DMSO-d6): δ 11.48 (br, OH), δ 11.15 (s, NH) δ 8.17 &
8.38 (s, N=CH)), δ 6.70–7.62 (m, aromatic and Sn-C6H5 protons).
MeSnCl(H2CBS) [MeSnCl(C15H12N4O3)] (5)
The procedure described above for (2) was followed for the preparation of (5), with methyltin(IV)
trichloride being used instead of dimethyltin(IV) dichloride and the refluxing time for this preparation
was 2-3 hours. Yield = 65.46%. M. p. = 205-206 °C. Found: C, 41.19; H, 3.25; N, 12.05 %. Calc. for
C15H12N4O3MeSnCl: C, 41.23; H, 3.22; N, 12.02%. λmax (nm) (DMF): 267, 341, 398. IR (νmaxcm-1
(KBr): 1610 (C=N)+(C=C), 1340 (C–O, phenolic), 952 (N–N), 590 (Sn–C), 560 (Sn–O), 472 (Sn–N).
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1
H NMR (300 MHz, DMSO-d6): δ 11.42 (br, OH), δ 10.96 (s, NH) δ 8.14 & 8.44 (s, N=CH)), δ 6.64–
7.90 (m, aromatic-H). δ 1.14 (s, Sn-CH3).
PhSnCl(H2CBS) [PhSnCl(C15H12N4O3)] (6)
The procedure described above for (5) was followed for the preparation of (6), with phenyltin(IV)
trichloride being used instead of methyltin(IV) trichloride. Yield = 64.23%. M. p. = 213-215 °C.
Found: C, 49.10; H, 3.63; N, 10.22 %. Calc. for C15H12N4O3PhSnCl: C, 49.15; H, 3.66; N,
10.25%.λmax (nm) (DMF): 268, 340, 399. IR (νmaxcm-1 (KBr): 1608 (C=N)+(C=C), 1330 (C–O,
phenolic), 956 (N–N), 615 (Sn–C), 520 (Sn–O), 451 (Sn–N). 1H NMR (300 MHz, DMSO-d6): δ 11.44
(br, OH), δ 11.10 (s, NH) δ 8.18 & 8.39 (s, N=CH)), δ 6.72–7.62 (m, aromatic and Sn-C6H5 protons).
X-ray Crystallography
Yellow single-crystal of (2) (size 0.34 × 0.32 × 0.15 mm) was grown from chloroform-hexane mixture
at the room temperature. The measurements were performed at 273 (2) K on Siemen SMART CCD
diffraction using graphite-monochromated Mo-Kα radiation (λ = 0.71073 Å). Orientation matrix and
unit cell parameters were obtained from the setting angles of 25-centered reflection. The crystals are
monoclinic, space group P2(1)/c with a = 13.163(3), b = 11.143(2), c = 12.203(2) Å, α = 90°, β =
102.064(3)°, γ = 90°, V = 1750.4(6) A3, Z = 4, Dcalc = 1.689 Mg/m3, µ = 1.484 mm-1. The diffraction
intensities were collected by ω scans (2.4 to 27.0°). A total of 9750/3798 reflections were collected (-
16< =h< =16, -14< =k< =14, -15< =l< =11). The structure was solved using direct methods and
refined using the full-matrix least-square method on F obs2 using the SHELXTL [9] software
package. All non-H atoms were anisotropically refined. The hydrogen atoms were located in a
difference Fourier map and then were fixed geometrically and treated as riding atom on the parent C
atoms, with C-H distances = 0.97 Å.
Antimicrobial Activity Test

The antimicrobial activities of the ligand and complexes were assayed by plate diffusion method [10]
against Gram-positive bacteria, Gram-negative bacteria and fungi. The Gram-positive bacteria
consisting of Staphylococcus aureus and Bacillus subtilis and the Gram-negative bacteria, which
consist of Escherichia coli. The ligand and complexes were dissolved in DMSO and concentration of
3 mg/mL and 5 mg/mL were tested in triplicate plates. Chloramphenicol at 3 mg/mL was employed as
control.
Results and discussion
The carbohydrazone ligand [H4CBS, (1)] was synthesized by the condensation reaction of
carbohydrazide with salicylaldehyde in 1: 2 ratio in absolute ethanol (Scheme 1). Five new
organotin(IV) complexes of carbohydrazide-bis(salicylaldehyde) ligand (1) have been synthesized by
direct reaction of the appropriate organotin(IV) halide(s) with the ligand in the presence of a base
(Scheme 2). All the cases, a base is added to the reaction mixture in order to force the deprotonation
of the ligand. The low molar conductance values (10-40 ohm-1cm2 mol-1) (see Table 1) indicating non-
electrolytic nature for all the organotin(IV) complexes [11].
RnSnCl4-n + Ligand [H4CBS, (1)] + 2KOH
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H
H N H
N C
C N N C
O
abs. MeOH
OH Sn
O
R
R
R = Me, Bu and Ph
OR
H
H N C N H
C N N C
O
OH Sn
O
R
Cl
R = Me and Ph
Scheme 2
Table 1. Molar conductance values for organotin(IV) complexes of ligand (1)
Compound Molar conductance, Λm (ohm-1 cm 2 mol -1)

[Me2Sn(H2CBS)] (2) 10
[Bu2Sn(H2CBS)] (3) 23
[Ph2Sn(H2CBS)] (4) 29
[MeSnCl(H2CBS)] (5) 40
[PhSnCl(H2CBS)] (6) 35
Electronic absoption spectra
The UV-Vis electronic spectra of ligand (1) and its organotin(IV) complexes (2-6) were measured at
room temperature in DMF (10-4 M) solutions over 200 – 800 nm range are given in Table 2.
Carbohydrazone ligand (1) exhibited three main bands at 262, 292 and 328 nm. The first and second
bands were attributed to benzene π – π* and imino (C=N) π – π* transition, which were not affected by
the chelation. The third band is assigned to n – π* transition. The appearance of two new bands at 266-
343 nm and at 396-399 nm regions showed the metal-ligand coordination in all the complexes (2-6).
The high wavelength bands in the region 396-399 nm can be assigned due to charge transfer transition
involving the tin atom [12].
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Table 2. The λmax (nm) peaks of ligand (1) and its organotin(IV) complexes (2-6).
Compounds λmax (nm)

(1) H4CBS 328, 292, 262
(2) [Me2Sn(H2CBS)] 398, 343, 266
(3) [Bu2Sn(H2CBS)] 398, 342, 266
(4) [Ph2Sn(H2CBS)] 396, 340, 267
(5) [MeSnCl(H2CBS)] 398, 341, 267
(6) [PhSnCl(H2CBS)] 399, 340, 268
Infrared spectra
The IR data for the ligand (1) and its all organotin(IV) complexes (2-6) are described in the synthesis
section. The IR spectra of the complexes (2-6), the disappearance of the ν(C=O) band indicating
enolization of the amide group into C-OH enolic form through the amide-imidol tautomerism process
and also indicating deprotonation of the functional group during coordination to tin atom [13]. The
ν(C=N) band in the complexes (2-6) appeared as a strong band in the region 1600–1610 cm-1, is
considerably shifted towards lower frequencies with respect to that of the free ligand (1620 cm-1),
confirming the coordination of the azomethine nitrogen to organotin(IV) moiety [13]. This is also
apparent from the ν(Sn–N) band at 440–472 cm-1 in the IR spectra of all the complexes (2-6). The
hydrazinic stretching ν(N–N) band observed at 950 cm-1 for the ligand (1) is shifted in the higher
region at 952–962 cm-1 in the complexes (2-6) further supporting that azomethine nitrogen is
coordinated to Sn(IV) ion [12]. The high intensity band observed at 1272 cm-1 in the ligand (1)
attributed due to phenolic ν(C–O), appears as a medium band at 1319-1340 cm-1 in the IR spectra of
all the complexes (2-6). These observations favour the formation of Sn–O bond via deprotonation.
The medium and weak bands observed at 590-651, 520–560 and 440–472 cm-1 in the IR spectra of all
the complexes (2-6) are attributable to the ν(Sn–C), ν(Sn–O) and ν(Sn–N) vibration bands
respectively, indicating coordination of the free ligand to the central Sn(IV) atom via deprotonated
enolic oxygen and azomethine nitrogen in the complexes (2-6).
1
H NMR spectra
The 1H NMR data for the ligand (1) and its all complexes (2-6) are presented in the experimental
section. The 1H NMR spectrum of ligand (1) is characterized by four signals at 10.86, 8.44, 7.69 and
6.62–7.27 ppm, which are assigned to the protons associated with –OH, –N=CH, –CONH and
aromatic ring protons, respectively. In 1H NMR spectra of organotin(IV) complexes (2-6), azomethine
N=CH signal is splited into two signals at 8.14-8.17 and 8.38-8.43 ppm. This indicates that only one
of the azomethine nitrogen in ligand (1) could be bonded to the Sn(IV) ion in the complexes (2-6),
another one HC=N group could be free in the complexes (2-6). The disappearance of the signal due to
–CONH proton in the complexes (2-6) indicated the enolization of the form of the ligand (1) in the
complexes (2-6) [7]. The new signal at 10.89-11.15 ppm in the complexes (2-6) is due to the free NH
group of ligand (1) which is not involved in the coordination to tin(IV) in the complexes (2-6). A
signal appeared at 11.41-11.48 ppm, which is indicated that phenolic proton present in the complexes
(2-6). The sharp signal attributed to methyl group attached to tin atom is appeared as a singlet at 0.68
ppm in the dimethyltin(IV) complex (2). Another sharp resonance signal attributed to methyl group
attached to tin(IV) atom is appeared as a singlet at 1.14 ppm in methyltin(IV) complex (5). In the
complex (3), a multiplet in the region 0.79-1.64 ppm is assigned to the butyl group attached to the
tin(IV) atom Complexes (4 and 6) showed a multiplet in the region 6.70-7.62 ppm, which may be
assigned to aromatic ring protons and Sn-Ph protons, respectively. The signals could not properly
assigned due to overlap of corresponding signals of Ph-Sn and aromatic ring protons.
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Crystal structure of [Me2Sn(H2CBS)] (2)
The molecular structure along with atom numbering scheme for [Me2Sn(H2CBS)] (2) is shown in
Figure 1. The crystal data and structure refinement for compound [Me2Sn(H2CBS)] (2) are
summarized in Table 3. The selected bond distances and angles for the molecules [Me2Sn(H2CBS)]
(2) are given in Table 4. The X-ray structural investigations of [Me2Sn(H2CBS)] (2) confirm that the
carbohydrazone ligand, [H4CBS, (1)] behaves as a tridentate dibasic coordinating agent via
azomethine nitrogen, one enolic oxygen and one phenolic oxygen atoms and elimination of two
potassium chloride. The two methyl groups and azomethine nitrogen atom of ligand take the
equatorial positions, while enolic and phenolic oxygen atoms are in the axial position [O(1)–Sn(1)–
O(2) 153.12(8)º]. The ligand forms six- and five-membered chelate rings. The ligand is not
completely planar in [Me2Sn(H2CBS)] (2) and a distorted trigonal–bipyramidal geometry around the
tin centre is found. The angles subtended at tin(IV) by two oxygen atoms are compressed to
153.12(8)º [O(1)-Sn(1)-O(2)]. The bite angles O(1)–Sn(1)–N(1) [83.58(8)º] and O(2)-Sn(1)-N(1)
[72.61(7)º] are also compressed from 90º. These distortions arises from the rigidity of chelate rings,
compounded by the large tin(IV) covalent radius. These bit angles are comparable to those reported
for other diorganotin(IV) complexes containing both five- and six–membered chelate rings with
oxygen and nitrogen donor atoms [14].
Table 3. Crystal data and the structure refinement for the complex [Me2Sn(H2CBS)] (2).
Formula C17H18N4O3Sn
Formula weight 445.04
Crystal system Monoclinic
Space group P2(1)/c
Z 4
a (Å) 13.163(3)
b (Å) 11.143(2)
c (Å) 12.203(2)
α (°) 90
β (°) 102.064(3)
γ (°) 90
V (Å3) 1750.4(6)
Dcalc (mg m-3) 1.689
Absorption coefficient (mm-1) 1.484
Temperature (K) 273(2)
Wavelength (Å) 0.71073
Final R indices [I>2sigma(I)] R1 = 0.0275, wR2 = 0.0643
R indices (all data) R1 = 0.0327, wR2 = 0.0670
Goodness-of-fit on F^2 1.095
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278
Figure 1. Molecular structure of [Me2Sn(H2CBS)] (2).

Thermal ellipsoids at the 50% level.
Table 4. Selected bond lengths (Å) and angles (°) of dimethyltin(IV) complex [Me2Sn(H2CBS)] (2).
Bond lengths
Sn(1)-O(1) 2.077(2) Sn(1)-N(1) 2.176(2)
Sn(1)-O(2) 2.164(19) Sn(1)-C(16) 2.096(3)
Sn(1)-C(17) 2.111(3) O(1)-C(1) 1.324(3)
O(2)-C(8) 1.271(3) N(1)-C(7) 1.293(3)
N(1)-N(2) 1.389(3) N(2)-C(8) 1.321(3)
N(3)-N(4) 1.364(3) N(3)-C(8) 1.366(3)
C(6)-C(7) 1.441(4) N(4)-C(9) 1.278(3)
C(10)-C(9) 1.444(4)
Bond angles
O(1)-Sn(1)-C(16) 94.19(10) O(1)-Sn(1)-C(17) 101.90(11)
C(16)-Sn(1)-C(17) 127.26(13) O(1)-Sn(1)-O(2) 153.12(8)
C(16)-Sn(1)-O(2) 90.10(10) C(17)-Sn(1)-O(2) 96.43(11)
O(1)-Sn(1)-N(1) 83.58(8) C(16)-Sn(1)-N(1) 126.21(10)
C(17)-Sn(1)-N(1) 105.46(11) O(2)-Sn(1)-N(1) 72.61(7)
C(1)-O(1)-Sn(1) 128.80(17) C(8)-O(2)-Sn(1) 113.57(16)
C(7)-N(1)-N(2) 116.2(3) C(9)-N(4)-N(3) 117.7(2)
C(7)-N(1)-Sn(1) 126.23(17) N(2)-N(1)-Sn(1) 117.20(15)
O(1)-C(1)-C(6) 122.3(2) C(1)-C(6)-C(7) 124.3(2)
N(1)-C(7)-C(6) 126.3(2) O(2)-C(8)-N(2) 126.8(2)
The asymmetry of ligand [H4CBS, (1)] is strongly reflected in the Sn-O distances. In the
complex [Me2Sn(H2CBS)] (2), the Sn(1)-O(1) bond, 2.077(2) Å, is shorter than the Sn(1)-O(2) bond,
2.164(19) Å. This is due to O(2) being a carbonyl and O(1) being bound to a benzene ring. It is worth
noting that in complex [Me2Sn(H2CBS)] (2) the Sn-N bond length is a little longer (2.176 (2) Å) than
those reported in analogous diorganotin(IV) complex (2.146(6) Å) [15], but shorter than that of
[Me2Sn(2-OC6H4CH=NC6H4COO)] (2.230(5) Å) [16] and is considerably less than the sum of the
van der Waals radii of tin and nitrogen atom, 3.75 Å [17]. Due to the involvement of N(1) atom in tin
binding, the bond length of N(1)-C(7) is significantly increased to 1.293(3) Å as compared with the
imine function N(4)-C(9) (1.278(3) Å) which is having a double bond character. The Sn-C (methyl)
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279
bond lengths [2.096(3) Å and 2.111(3) Å] compare well with the reported values for diorganotin(IV)
complexes derived from ONO donor tridentate Schiff bases [18].
Antimicrobial activity of the ligand (1) and its organotin(IV) complexes (2-4)
Table 5 below shows the antibacterial activity of the ligand (1) and organotin(IV) complexes (2-4)
studied.
Table 5. Antibacterial activity of carbohydrazide-bis(salicylaldehyde) ligand (1) and its organotin(IV)

complexes (2-4)
Diameter of inhibition zone (mm)
Bacteria H4CBS (1) [Me2Sn(H2CBS)] (2) [Bu2Sn(H2CBS)] (3) [Ph2Sn(H2CBS)] (4)

E. coli 10 15 20 16
B. subtilis 12 17 22 19
Stap. aureus 14 20 25 23
The preliminary screening showed that the ligand is less active compared to the organotin(IV)
complexes (2-4), which indicates metallation increases activity. Other biological studies on these
compounds are still under investigations.
Conclusions
The synthesis and physical properties of a new series of organotin(IV) compounds with
carbohydrazone ligand (1) have been described. The ligand behaved as a tridentate dinegative fashion
towards to tin(IV). The complexes (2-6) are monometallic. The coordination around the tin(IV) ion is
established by means of single crystal X-ray diffraction analysis on [Me2Sn(H2CBS)] (2).
Acknowledgement
The authors are very grateful to Universiti Malaysia Sarawak (UNIMAS), for the financial support
(Grant # -01(101)/453/2004(190). The author also would like to thank the School of Chemical
Sciences and Food Technology, Universiti Kebangsaan Malaysia (UKM), for the CHN analyses and
also X-ray single crystal determination. We would also like to thank the Ibnu Sina Institute, UTM, for
the help in obtaining the 1H NMR spectra.
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[5] Tergioglu, N., Gurso, N. (2003) “Synthesis and anticancer evaluation of some new hydrazone
derivatives of 2,6-dimethylimidazo[2,1-b][1,3,4]thiadiazole-5-carbohydrazide” Euro. J. Med.
Chem. 38. 781-786.
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[6] Affan, M. A., Fasihuddin,B. A., Ramli, B. H. Mustaffa, B. S., Bohari, M. Y. (2003) “In:
Analytical Chemistry” eds. B. A. Zaini, B. A. Fasihuddin, B. J. Ismail, S. Lau, and M.
Murtedza, Published by UNIMAS & ANALIS, Sarawak, Malaysia. 214-221.
[7] Affan, M. A. (1999) “Novel complexes of molybdenum carbonyls, organotin(IV) and
lanthanide(III) ions with hydrazones and azines” PhD Thesis, Universiti Teknologi Malaysia.
[8] Armarego, W. L. F., Perrin, D. D. (1998) “Purification of Laboratory Chemicals” 4th edition,
Butterworth Heinemann, Oxford, USA.
[9] Sheldrick, G. M. (1997) “SHELXTL V5.1. Software Reference Manual. Bruker AXS” Inc.,
Madison, WI, USA.
[10] Tarafder, M. T. H., Jin, K. T., Krouse, K. A., Ali, A. M., Yamin, B. M., fun, H. K. (2002)
“Synthesis and Characterization of Cu(II), Ni(II) and Zn(II) metal complexes of bidentate NS
isomer Schiff bases derived from S-methyldithiocarbazate (SMDTC): bioactivity of the
bidentate NS isomeric Schiff bases, some of their Cu(II), Ni(II) Zn(II) complexes and the X-
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complex” polyhedron 21. 2547-2554.
[11] Geary, W. G. (1971) “The use of conductivity measurement in organic solvent for the
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[12] Khalil, T. E., Labib, L., Iskandar, M. F. (1994) “Organotin(IV) complex with tridentate
ligand” Polyhedron 13. 2569-2578.
[13] Casas, J. S., Sanchez, A., Sordo, J., Lopez, V. A., Castellano, E. E. (1994) “Diorganotin(IV)
derivatives of salicylaldehyde thiosemicarbazone” Inorg. Chim. Acta 216. 169-172.
[14] Khoo, L. E., Xu, Y., Goh, N. K., Chia, L. S. and Koh, L. L. (1997) “Molecular adducts of
diorganotin dichloride with N- (2-oxidoarylideneaminoacidato) diorganotin(IV) complexes.
Crystal structure of [Ph2Sn(2-OC10H6Ch=NCH2COO)]SnPh2Cl2” Polyhedron 16. 573-576.
[15] Yin, H. D., Wang, Q. B. and Xue, S. C. (2004) “Synthesis and structural characterization of
diorganotin(IV) esters of salicylidene-amino acids” J. Organomet. Chem. 689. 2480-2485.
[16] Dey, D. K., Saha, M. K., Gielen, M., Kemmer, M., Biesemans, M., Willem, R., Gramlich, V.
and Mitra, S. (1999) “Synthesis, spectroscopy and structure of [N-(2-carboxyphenyl)
salicylideneiminato]dimethyltin(IV)” J. Organomet. Chem. 590. 88-92.
[17] Ma. C., Jiang, Q. and Zhang, R. (2004) “Synthesis, characterization and crystal structures of
new organotin complexes with 2-mercapto-6-nitrobenzothiazole” Polyhedron 23. 779-786.
[18] Goh, N. K. and Khoo, L. E. (1993) “Synthesis of structural characterization of dimeric
diorganotin(IV) N-2-hydroxynaphthalidene-β-amino acid complexes: x-ray structure of
[Me2Sn(OC10H6CH=NCH2CH2COO)]2” Polyhedron 12(8). 925-930.
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Bi-Fe DOPED VANADIUM PHOSPHATE (VPO) CATALYSTS FOR THE SELECTIVE

OXIDATION OF n-BUTANE TO MALEIC ANHYDRIDE
Y.H. Taufiq-Yap*, Chee Keong Goh, Irmawati Ramli and Mohd Zobir Hussein
Department of Chemistry, Universiti Putra Malaysia,

43400 UPM Serdang, Selangor D.E., Malaysia.
Email: yap@fsas.upm.edu.my
Abstract. In this study, the Bi-Fe doped catalysts were found decreased the crystallite structure and surface area of the
catalysts. Furthermore, the Bi-Fe doped catalysts promoted the formation of αII -VOPO4 phase. The Bi-Fe doped catalysts
were also found increased the amount of oxygen removed at higher temperature in H2-TPR analysis. So, the Bi-Fe doped
catalysts significantly decreased the n-butane conversion. However, the Bi-Fe doped catalysts increased the selectivity of
maleic anhydride due to an increase of V5+ phase and surface acidity of the catalysts.
Abstrak. Dalam kajian ini, mangkin dop Bi-Fe didapati mengurangkan struktur kristal dan luas permukaan mangkin.
Tambahan pula, mangkin dop Bi-Fe menggalakkan penghasilan fasa αII -VOPO4. Mangkin dop Bi-Fe juga didapati
menambahkan amaun pengeluaran oksigen pada suhu yang lebih tinggi. Maka, mangkin dop Bi-Fe telah mengurangkan
penukaran n-butana. Walaubagaimanapun, mangkin dop Bi-Fe telah meningkatkan selektiviti maleik anhydrida disebabkan
oleh penambahan fasa V5+ dan keasidan di permukaan mangkin.
Keywords: vanadium phosphate, Bi-Fe, butane oxidation, maleic anhydride
Introduction
Vanadium phosphate catalysts are well known for the oxidation of n-butane to maleic anhydride
(MA) [1]. (VO)2P2O7 is claimed to be the active phase of the catalyst which obtained by topotactic
transformation of vanadyl hemihydrate precursor, VOHPO4·½H2O [2, 3]. The activation of the
vanadyl hemihydrate precursor, under the n-butane/air stream, proceeded in two parallel routes, either
a reoxidation to VOPO4 phases or direct dehydration to (VO)2P2O7. This is followed by a reduction of
the residual VOPO4 phases to (VO)2P2O7 [4]. The beneficial of catalysts performances were
associated with a (VO)2P2O7 structure in which phosphorus atoms were associated with V4+ in a
crystalline matrix and with V4+ in a disordered matrix together with V5+, implying the importance of
V5+/V4+ dimeric species in the (VO)2P2O7 structure [2, 5-7]. In order that the V5+/V4+ balance is
importance in controlling the catalytic performance of n-butane oxidation to MA, thus the addition of
small amount Bi-Fe as dopants can influence this balance.
Experimental
For the preparation of doped precursors, the VOPO4·2H2O (4.0 g) and the desire quantity of
Fe(NO3)3·9H2O (from Hamburg Chemical) (Fe/V = 1, 2 and 3%) as well as bismuth nitrate,
Bi(NO3)3·5H2O from Sigma (Bi/V = 1, 2 and 3%) were suspended by rapid stirring into isobutanol (80
cm3 from BDH). The mixtures were refluxed for 21 h with continuous stirring. Then, the blue solids
were recovered by filtration, washed and dried in an oven at 383 K for 16 h. The undoped precursor
was prepared in the same method abovementioned, but without adding Bi and Fe. The resulting
undoped and doped precursors above were then undergone calcination in a flow of n-butane/air
mixture (0.75 % n-butane in air) for 75 h at 673 K to generate the active catalysts. The prepared
catalysts were characterised by using a Shimadzu Diffractometer model XRD-6000 for X-ray
diffraction (XRD) analyses, a ThermoFinnigan Sorptomatic 1990 for B.E.T. surface area
measurements, and a ThermoFinnigan TPDRO 1100 for temperature-programmed reduction in
hydrogen (H2-TPR) and temperature-programmed desorption of ammonia (NH3-TPD). The oxidation
of n-butane was carried out in a microreactor with a standard mass of catalyst (250 mg) with online
gas chromatography, as shown in Figure 1.
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Figure 1. Schematic diagram of the plug flow reactor.
The introduction of Bi-Fe into VPD system had reduced the surface area from 16.1 m2 g-1 (undoped)
to 13.7 m2 g-1 for VPDBi1Fe1 and 12.0 m2 g-1 for VPDBi1Fe2. However, the VPDBi2Fe1 has slightly
higher surface area of 17.6 m2 g-1. Chemical analysis using ICP showed that the addition of Bi-Fe into
VPD system increased the phosphorus contents of the catalysts with P/V atomic ratio of 1.00
(undoped) to 1.15. Chemical analysis also confirmed the presence of Bi and Fe with Bi/V and Fe/V
atomic ratios which is similar to the calculated value, suggesting that the homogeneous existence of
Bi-Fe promoters into the VPD system. Bi-Fe was found to increase the average oxidation number
from 4.21 to 4.56 due to an increment of V5+ oxidation state from 21 to ~55%. The XRD patterns of
all undoped and Bi-Fe doped catalysts (Figure 2) show only the characteristic reflection of vanadyl
pyrophosphate, (VO)2P2O7. However, the addition of Bi-Fe into VPD lattice promoted the formation
of αII-VOPO4 phase (2θ = 29.2˚), which is identified according to [8]. Besides, the Bi-Fe doped
catalysts reduced the crystallite structure of (VO)2P2O7. The H2-TPR experiments were used to
investigate effect of Bi-Fe on the reducibility of the catalysts, as shown in Figure 3. The undoped
catalyst gave a characteristic of two reduction peaks at the region from 400-1100 K. These peaks
occurred at 779 and 990 K. The first peak associated to the reduction of V5+ phase whereas the latter
peak is assigned to the removal of lattice oxygen from V4+ phase. The amount of oxygen removed
from both peaks is 3.85 x 1020 and 1.20 x 1021 atom g-1, respectively. The ratio of V5+/V4+ is about
0.32. However, the Bi-Fe doped catalysts show a different patterns with the first reduction peak
appeared as major peak whereas the size of second reduction peak decreased remarkably. The first
peak shifted to a higher temperature whereas the latter peak exhibited at lower temperature compared
to the undoped material. This observation indicated that the amount of oxygen related to V5+
significantly increased to 1.49 x 1021, 1.45 x 1021 and 1.24 x 1021 atom g-1 for VPDBi1Fe1,
VPDBi1Fe2 and VPDBi2Fe1, respectively. The amount for oxygen removed for the second reduction
peak, which assigned to the reduction of V4+ reduced to 4.59 x 1020, 3.95 x 1020 and 5.57 x 1020 atom
g-1 for the three doped catalysts. The V5+/V4+ ratio for Bi-Fe doped VPO catalysts increased to 3.25,
3.67 and 2.23 for VPDBi1Fe1, VPDBi1Fe2 and VPDBi2Fe1, respectively. A suitable V5+/V4+ ratio
around 0.25 is suggested by [9] for the best catalyst activity. The high amount of V5+ in the Bi-Fe
doped catalysts significantly reduced the n-butane conversion and induced the MA selectivity. The
addition of Bi-Fe into VPD matrix also doubled the amount acid site from 7.77 x 1020 atom g-1
(undoped) to 1.68 x 1021 atom g-1. Figure 4 shows the catalytic performance of undoped and Bi-Fe
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doped catalysts at reaction temperature of 673 K and gas hourly space velocity (GHSV) of 2400 h-1.
The n-butane conversion for Bi-Fe doped catalysts shown to be reduced to 32-46% compared to 83%
for the undoped material. The deleterious of the catalytic performance of Bi-Fe doped catalysts is due
to poor crystallite of (VO)2P2O7, lower surface area and large excess amount of V5+ phase. However,
the Bi-Fe doped catalysts increased selectivity to MA as compared to undoped catalyst because of
existence of αII-VOPO4 phase and higher amount acid sites.
2500
∇ (VO)2P2O7 ♦ αII−VOPO4
∇ ∇
∇ VPDBi2Fe1
♦
∇ ∇ ∇
2000
VPDBi1Fe2
1500
Intensity / a.u.
VPDBi1Fe1
1000
VPD
500
0
10 20 30 40 50 60
2θ / degree
Figure 2. XRD patterns of undoped VPD Figure 3. H2-TPR spectra of undoped VPD
and Bi-Fe doped catalysts. and Bi-Fe doped catalysts.
Butane conversion MA selectivity
90
80
70
60
Percentage (%)
50
40
30
20
10
0
VPD VPDBi1Fe1 VPDBi1Fe2 VPDBi2Fe1
Sample
Figure 4. Catalysts performance of vanadium phosphate (VPO) for the oxidation of n-butane.
Conclusions
Doping with Bi-Fe decreased the crystallite structure of (VO)2P2O7 phase as well as the surface area
of the catalysts. Furthermore, the Bi-Fe doped catalysts significantly increased the ratio of V5+/V4+,
which is beyond to the optimal ratio. Hence, the n-butane conversion for Bi-Fe doped catalysts shown
to be significantly reduced. However, the formation of αII-VOPO4 and higher amount of acid sites led
to an increase of MA selectivity for Bi-Fe doped catalysts.
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Acknowledgement
Financial assistance from Malaysian Ministry of Science, Technology and Innovation is gratefully
acknowledged.
References
1. Centi, G. (1993) "Vanadyl pyrophosphate- a critical overview" Catal. Today 16. 5-26.
2. Centi, G., Trifiro, F., Ebner, J. R. and Franchetti, V. M. (1988) "Mechanistic aspects of maleic anhydride synthesis
from C4 hydrocarbons over phosphorus vanadium oxide" Chem. Rev. 88. 55-80.
3. Hodnett, B. K. (1985) "Vanadium-phosphorus oxide catalysts for the selective oxidation of C4 hydrocarbons to
maleic anhydride" Catal. Rev. -Sci. Eng. 27. 373-424.
4. Abon, M., Bere, K., Tuel, A. and Delichere, P. (1995) "Evolution of a VPO Catalyst in n-Butane Oxidation
Reaction during the Activation Time" J. Catal. 156. 28-36.
5. Shen, S., Zhou, J., Zhang, F., Zhou, L. and Li, R. (2002) "Effect of Ce-Fe oxides additives on performance of VPO
catalyst for n-butane oxidation to maleic anhydride in the absence of gas-phase oxygen" Catalysis Today 74. 37-
43.
6. Granados, M. L., Conesa, J. C. and Fernandez-Garcia, M. (1993) "Physicochemical Study of Structural Disorder in
Vanadyl Pyrophosphate" J. Catal. 141. 671-687.
7. Sananes, M. T., Tuel, A., Hutchings, G. J. and Volta, J. C. (1997) "The V4+/V5+ balance as a criterion of selection
of vanadium phosphorus oxide catalysts for n-butane oxidation to maleic anhydride: A proposal to explain the role
of Co and Fe dopants" J. Catal. 166. 388-392.
8. Abdelouahab, F. B., Olier, R., Guilhaume, N., Lefebvre, F. and Volta, J. C. (1992) "A study by in situ Laser
Raman Spectroscopy of VPO catalysts for n-butane oxidation to maleic anhydride. 1. Preparation and
characterization of pure reference phases" J. Catal. 134. 151-167.
9. Aït-Lachgar, K., Abon, M. and Volta, J. C. (1997) "Selective oxidation of n-butane to maleic anhydride on vanadyl
pyrophosphate" J. Catal. 171. 383-390.
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POLYCYLIC AROMATIC HYDROCARBONS IN URBAN SOILS OF KEMAMAN,

TERENGGANU
Norhayati Mohd Tahir*1, Lee Boon Jeen1, Hasra Masrifah Abd. Rahim1, Marinah Ariffin1, Suhaimi
Suratman1 and Mhd Radzi Abas2
1
Department of Chemical Sciences, Faculty of Science and Technology
Kolej Universiti Sains dan Teknologi Malaysia (KUSTEM), Mengabang Telipot,
21030 Kuala Terengganu, Terengganu
2
Department of Chemistryt, Faculty of Science, Universiti Malaya (UM), 50603 Kuala Lumpur
e-mail: hayati@kustem.edu.my
Abstract. A study has been carried out to determine the concentration and distribution of polycyclic aromatic
hydrocarbons (PAHs) in urban soils of Kemaman, Terengganu. Surface soil samples (< 500 µm) were
ultrasonicated using dichloromethane as solvent and the extracts fractionated on silica-alumina column.
Detection and quantification of 16 priority PAHs compounds were carried out using GC-FID. With the
exception of two stations, results generally indicated that the sum of 16 priority PAHs concentration (total
PAHs) in soils ranged from to 6.3 to 176 µg/kg (dry weight); the two stations which exhibited significantly
higher levels of total PAHs was at the main road junction located at the heart of the commercial centre of the
town (535 µg/kg) and at an industrial estate, adjacent to a sawmill (547 µg/kg). Statistical analysis suggests that
there is a significant difference in total PAHs concentration (p<0.05) with sampling sites. Most common PAHs
compound observed in almost all the soil samples was BgP indicating the importance of vehicular emission as a
source of PAHs in these soils. In addition, contribution of biomass burning to the presence of PAHs in these
soils was also observed as indicated by a positive correlation between Benzo[a]pyrene with total PAHs.
Abstrak. Satu kajian bagi menentukan kepekatan dan agihan hidrokarbon polisiklik aromatic (PAH) dalam
tanah di Bandar Kemanan, Terengganu telah dijalankan. Sampel tanah permukaan (<500µm) diekstrak
menggunakan kaedah pengekstrakan ultrasonik dengan DCM sebagai pelarut dan hasil ekstrak dipisahkan
menggunakan turus silika-alumina. Pengenalpastian dan pengkuantitian 16 sebatian PAH keutamaan dilakukan
menggunakan GC-FID. Keputusan secara amnya menunjukkan bahawa jumlah PAH dalam tanah adalah dalam
lingkungan 6.3µg/kg ke 176µg/kg (berat kering) kecuali pada dua stesen dimana kepekatan adalah jauh lebih
tinggi; satu stesen terletak di simpang jalan utama di kawasan pusat Bandar (535µg/kg) dan satu lagi terletak di
kawasan estet perindustrian, berhampiran kilang kayu (547µg/kg). Analisis statistik menunjukkkan bahawa
terdapat perbezaan ketara dalam kepekatan total PAH diantara stesen persampelan. Hampir semua sampel tanah
mengandungi BgP dimana kehadirannya sering dikaitkan dengan sumber kenderaan bermotor manakala
sumbangan daripada sumber pembakaran biojisim terhadap kandungan PAH dalam tanah juga dikenalpasti
berdasarkan korelasi positif antara BaP dan total PAH.
Keywords: Polycyclic aromatic hydrocarbons, urban soils, ultrasonic agitation method, vehicular emissions, Kemaman
Introduction
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the global environment and soil remains
as one of the most important sinks for these compounds. Due to their mutagenic and carcinogenic
potentials as well as their persistence, the presence of PAHs in the environment is of great global
concern. PAHs are formed from the incomplete combustion of fossil and biomass fuels. Main sources
of these compounds in soil include wildfires and biomass burning for agricultural land clearing,
municipal incinerators, vehicular emission, residential heating and through the atmospheric deposition
as a result of long-range atmospheric transport [1-6]. In this context, knowledge on the PAHs level
and its dispersion on regional and national scale are essential. At present in Malaysia, even though
studies to determine the concentration, composition and possible sources of PAHs in soils are being
reported in the literature [7,8], however, this type of studies is still limited, particularly in the east
coast states of Peninsular Malaysia. In view of this, a study has been initiated to address this gap in
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knowledge. This paper presents the results of a study to determine the concentrations, composition
and possible sources of PAHs in soils of Chukai, the capital of the district of Kemaman, Terengganu.
Experimental
Study area and sampling

Chukai with a population of 45,873 density, lies on the southeast end of the Terengganu state (Figure
1.1a) and is sandwiched between the booming oil town of Kerteh, Terengganu and the fast growing
industrial area of Gebeng, Pahang. It acts as the center for business activities and public agencies for
the fastest growing district in the state of Terengganu, largely due to the booming petroleum and
related industries in Paka-Kerteh belt. Owing to its strategic location, Chukai also acts as a residential
town to those who work in Paka-Kerteh belt as well as Gebeng and Kuantan, the capital of Pahang.
Soil sampling was carried out around the urban area of Chukai (Table 1 and Figure 1.1b). These sites
were chosen based on traffic density, residential and industrial activities and were generally
diffuse source oriented. Where possible soil samples were collected ca. one metre from the
roadside. Twenty sampling sites were ascertained and classified to three zones. Zone A was located in
the centre part of this town, where most of the economic activities were found with major road
network, Zone B emphasized on residential areas whereas Zone C is located in the industrial zone of
the town. Sampling involved the collection of 20 surfacial soil samples (0-10cm) using metal spades,
wrapped in pre-cleaned aluminum foil and transported to the laboratory in an icebox to minimize
sample degradation. Once in the laboratory, soil samples were homogenized and sieved through a 500
µm sieve.
Figure 1.1a: Chukai Town Map Figure 1.1b : Sampling site map
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Table 1 : Longitude and latitude of the sampling site

Zone Site No Longitude Latitude Sampling Site
1 E 103°25’29.5’’ N 04º15’07.4’’ Jln Bakau Tinggi
2 E 103º25’29.5’’ N 04º14’50.8’’ Sek. Men. Sultan Ismail Kemaman
3 E 103º25’19.0’’ N 04º14’21.4’’ Jln Penghiburan
4 E 103º25’02.5’’ N 04º14’21.4’’ Center Road Junction of Jln Da Omar
9 E 103º25’17.3’’ N 04º13’56.4’’ Jln Da Omar
10 E 103º25’11.2’’ N 04º13’27.0’’ Jln Kubang Lurus
Zone A 15 E 103º25’25.8’’ N 04º13’39.1’’ Bukit Jakar
16 E 103º25’43.5’’ N 04º13’49.6’’ Jln Jakar
17 E 103º25’10.1’’ N 04º14’09.1’’ Bus station
18 E 103º25’38.8’’ N 04º13’54.2’’ Jln Sulaimani
19 E 103º25’33.5’’ N 04º13’48.7’’ Jln Abd. Rahman
20 E 103º25’25.8’’ N 04º13’39.1’’ Jln Mak Donyang
5 E 103°24’31.7’’ N 04º14’44.8’’ Kg Gong Limau

6 E 103°25’02.5’’ N 04º14’10.2’’ Jln Air Putih
Zone B 7 E 103°23’24.4’’ N 04º14’16.0’’ Kompleks Quarters Pend. Kemaman
8 E 103°23’06.0’’ N 04º13’31.7’’ Kg Mentok
11 E 103°24’49.6’’ N 04º13’32.6’’ Jln Pengkalan Lama
12 E 103°25’20.8’’ N 04º13’10.8’’ Jln Jakar (Fire Station)

Zone C 13 E 103°25’31.3’’ N 04º12’59.6’’ Jakar II Industrial Estate (electronic)
14 E 103°25’48.2’’ N 04º13’22.6’’ Jakar I Industrial Estate (sawmills)
Analytical procedure
PAHs were extracted from soils (< 500 µm fraction) with dichloromethane (DCM) as solvent using
ultrasonic method. The extracts were then fractionated on partially deactivated (5%) silica-alumina
columns. PAHs compounds were eluted using a combination of 20ml of 10% DCM in hexane
followed by 20ml of 50% DCM in hexane [9].
Identification and quantification of the 16 priority PAHs compounds were carried out using gas
chromatography fitted with flame ionization detector (GC-FID) based on the retention times
compared to that of external PAHs standards. These compounds were as follows: naphthalene (NAP),
acenaphtylene (ACY), acenaphtene (ACE), fluorene (FLU), phenanthrene (PHEN), anthracene
(ANT), fluoranthene (FTH), pyrene (PYR), benzo(a)anthracene (BaA), chrysene (CHR),
benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF), benzo(a)pyrene (BaP),
dibenz(a,h)anthracene (DA), benzo(g,h,i)perylene (BgP) and indeno(1,2,3,cd)pyrene (IP). Sums of
these 16 compounds were collectively known as total identified PAHs (TIP).
The GC-FID operating conditions were as follows: fused silica column (30m x 0.25 mm i.d; 0.25µm
filmed thickness); injection temperature was set at 290°C using a splitless mode; column temperature
was programmed in the following manner: hold at 50°C for 1 min, first temperature ramp of 50 -
140°C at 5°C min-1 followed by the second temperature ramp of 140 - 290°C at 3°C min-1 and then
maintained at 290°C for 13 min resulting in a total run time of 82 mins; helium was used as the carrier
gas with a flow rate at 1.2ml. min-1; detector temperature was set at 300°C.
Total organic carbon (TOC) in soils was analysed using Walkley and Black’s rapid titration method
[10].
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Soil Organic Carbon (SOC)

Numerous studies have shown that PAHs are strongly retained by the soil matrix [11,12] The
partitioning concept of soil sorption of organic contaminants implies that the sorption of hydrophobic
organic molecules is determined by the organic carbon content of the substrate [13-14]. The organic
matter content is considered to be a very important variable related to PAHs pollution of soils [15]. In
this study, the soils exhibited rather low organic carbon content with values ranging from 0.09% to
0.53% with a mean of 0.27% (Table 2 and Fig. 2). A regression analysis (Fig. 3) showed a negative
and weak relationship between the concentrations of the TIP and the amounts of soil organic carbon
(SOC, %). Because of the very low SOC in soils, it is probable that this parameter does not play an
important role in influencing the concentration of TIP in these soils.
0.6
0.5
0.4
%
0.3
0.2
0.1
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Stations
Figure 2: Distribution of SOC in soil samples
Correlation between TIP and SOC%

600
500
y = -473.25x + 255.11
400
TIP(ug/kg)
R2 = 0.1734
300
200
100
0
0 0.1 0.2 0.3 0.4 0.5 0.6
SOC%
Figure 3: Correlation between TIP and SOC

Total identified Polycyclic Aromatic Hydrocarbon ( TIP)
The distribution of TIP obtained in this study is shown in Figure 4. With the exception of two stations
(Stations 4 and 14) which exhibited significantly higher TIP than the other remaining stations, the range
of TIP values found in this study was between 6.27 µg/kg to 176.3 µg/kg (Table 2); the mean
concentration of TIP values obtained for the 20 sites was 129.7 µg/kg whilst the median was 101.4 µg/kg.
Statistical analysis showed significant differences (p<0.05) of TIP values between the stations monitored.
These values are generally in great excess of the reported natural concentration of PAHs in soil (1-10
µg/kg) [16] but within similar range to those reported in soils of Kuala Lumpur [7, 17] and Kuala
Terengganu [8]. Comparison between the three sampling zones showed that TIP values in Zone A ranged
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from 6.27µg/kg to 534.6µg/kg with mean value of 127.2 µg/kg and a median of 106.1µg/kg, Zone B
ranged 43.66µg/kg to 176.3µg/kg with mean value of 82.08µg/kg and a median of 44.62µg/kg and Zone
C ranging from 71.13µg/kg to 547.3µg/kg with mean value of 219.0 µg/kg and a median of 71.13µg/kg.
The stations which recorded exceptionally high TIP values were located in Zone A and Zone C,
respectively and it is conceded that their values does exhibit an influence on the mean values calculated
for the respective zone, particularly in the case of Zone C where there were only three stations monitored;
Station 4 was located at the busiest road junction in Chukai (Zone A) whereas Station 14 is located in the
vicinity of a sawmill in Zone C. In addition, as expected, it was also observed that most of the sampling
sites located by major roadside generally exhibited relatively higher TIP values (e.g. Stations 3, 4, 8, 9,
10) than sites located in the residential areas (Stations 5, 7, 11, 16).
600
500
concentration (ug/kg)
400
300
200
100
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Stations
Figure 4: Distribution of TIP in soil samples
The distribution of the 16 PAHs compounds monitored at each sampling site were found to differ with
stations, with station 4 showing the presence of all compounds except the very low molecular weight
PAHs (with two benzene rings), viz. NAP, ACY and ACE. In fact, NAP was absent from all stations
whilst ACY and ACE were found in only few stations at rather low concentrations (< 3 µg/kg). This
observation is not surprising as these compounds are generally more volatile compared to the other higher
ring number PAHs [18, 19]. Almost all stations exhibited the presence of BgP, the heavy molecular
weight PAHs commonly associated with vehicular emissions resulting from internal engine combustion
of gasoline [20]; BgP concentration found in this study ranged from 22.08µg/kg to 126.44µg/kg with
station 4 exhibiting highest BgP concentration relative to other stations. Other high molecular weight
PAHs that were present in almost all stations monitored includes DA and IP. BaP, a high molecular
weight PAHs generally taken as a signature of an incomplete burning of biomass or organic matter was
also present but only at selected stations with station 14 exhibiting extremely high concentration of this
compound (423.5 µg/kg) accounting > 70% of the PAHs recorded for this site. These observations are
consistent with the fact that Station 4 is located at the busiest road junction in Chukai whilst Station 14 is
located in the vicinity of a sawmill with obvious signs of wood burning activities.
Table 2 : TIP and selected parameters

Site No TOC % TIP (µg/g)
1 0.42 19.53
2 0.43 19.32
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290
3 0.47 109.2
4 0.12 534.6
5 0.23 44.47
6 0.39 176.3
7 0.41 43.66
8 0.53 101.4
9 0.21 170.4
10 0.18 146.8
11 0.24 44.62
12 0.16 71.13
13 0.19 38.66
14 0.11 547.3
15 0.32 6.27
16 0.24 49.92
17 0.19 130.1
18 0.23 157.1
19 0.09 80.48
20 0.14 102.9
Phenanthrene to antracene ratio (PHEN/ANT) has often been used to investigate possible sources of
PAHs compounds in the environment where a low ratio (PHEN/ANT < 10) is generally considered as
indicative of a predominance of pyrolytic sources (i.e. combustion sources) over petrogenic sources
(i.e. oil spill) [21-24]. In this study, the PHEN/ANT ratios in soil samples of Chukai ranged from 1.2
to 7.5 suggesting that the PAHs compounds found in these soils were derived from pyrolytic sources.
Differentiating between the two major pyrolytic source, viz. internal engine or biomass combustion
require the use of other molecular markers; as indicated above, presence of BaP in the environment is
generally indicative of incomplete combustion sources, in particular combustion of organic whilst the
association of BgP with vehicular emission has long been established [20]. A linear correlation
analysis between TIP and BaP and between TIP and BgP for the 20 stations monitored gave an r-
value of 0.68 and 0.58, respectively (Figs.5 and 6) which suggest that both sources contribute to the
presence of PAHs in these soils. Station 14 is interesting in that BaP contributes over 70% of the TIP
observed at the station, thus to eliminate possible bias, a second linear correlation analysis was
calculated between TIP and BaP and between TIP and BgP by excluding this station giving an r value
of 0.44 and 0.86. Similarly, since Station 4 is suspected to have a high contribution from vehicular
emission, another correlation analysis was carried out by removing the contributions of the two
extreme stations; the r value obtained was 0.41 and 0.63 for correlation between TIP and BaP and
between TIP and BgP, respectively. The results of the latter two correlation analyses clearly show a
stronger correlation exist between BgP and TIP suggesting that vehicular emissions is the more
dominant contributor to the PAHs in the soils around Chukai. The contribution from the vehicular
emission is expected as samplings were conducted in an urban area with a number of stations located
close to major trunk road. The presence of a relatively weaker but positive correlation between BaP
and TIP suggests that biomass burning also contributes to the presence of PAHs in these soils. Apart
from Station 14, it is most likely that these PAHs were contributed from activities of open burning of
rubbish and garden refuse by local residents [17] which are still prevalent in the east coast states
particularly in Terengganu and Kelantan.
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291
Correlation between TIP and BaP

600
500
y = 5.3299x + 88.71
400
TIP (ug/kg)
R2 = 0.1906
300
200
100
0
0 10 20 30 40 50
Bap (ug/kg)
Figure 5: Correlation between TIP and BaP (20 stations)
Correlation betw een TIP and BgP

600
500
y = 2.7365x + 58.938
400
R2 = 0.3322
TIP (ug/kg)
300
200
100
0
0 40 80 120 160
BgP (ug/kg)
Figure 6: Correlation between TIP and BgP (20 stations)
Conclusion
Total concentration of PAHs found in urban soils of Chukai, Kemaman ranged from 6.27 µg/kg to
547.3 µg/kg with two stations exhibiting exceptionally high TIP values. Almost all stations showed the
presence of BgP, a signature PAHs compounds known to be associated with vehicular emissions resulting
from gasoline combustion in engines. A strong correlation between BgP and TIP and low PHEN/ANT
ratio (<10) provide further evidence for the importance of vehicular emission sources of PAHs in these
soils. In addition, contribution of biomass burning to the presence of PAHs in these soils, through
open burning of rubbish and garden refuse by local residents, was also observed as indicated by a
positive albeit a weaker correlation between BaP with TIP relative to the correlation between BgP and
TIP.
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292
Acknowledgement
Financial supports from KUSTEM through a short term grant vote no. 54085 is kindly
acknowledged.
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ASSESSMENT OF NATURAL RADIOACTIVITY IN WATER AND SEDIMENT FROM

AMANG (TIN TAILING) PROCESSING PONDS
Mohsen Nasirian, Ismail Bahari, Pauzi Abdullah
Faculty of science and technology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selengor,
Malaysia
Abstract:
Gamma spectroscopy was performed to determine the concentrations of uranium-238 and thorium-232
concentrations in the environment as a consequence of amang processing. In this study 33 water
samples and 26 sediment samples were collected from 7 amang processing areas. The concentrations
of uranium-238 and thorium-232 were determined by direct counting using a hyper pure germanium
(HPGe) detector inter phased with a multi channel analyzer (MCA) . Results showed that the
maximum mass and activity concentrations of uranium in water samples were 6.64 ppm and 78.53 Bq/l
respectively, while in sediment samples were 69.75 mg/kg and 860.57 Bq/kg respectively. The
maximum mass and activity concentrations of thorium in water samples were 1.71 ppm and 6.90 Bq/l,
while in sediment samples were 157.73 mg/kg and 637.61 Bq/kg respectively. Concentrations of
uranium-238 and thorium-232 in sediment samples were higher than concentrations of uranium-238
and thorium-232 in water samples, and this may be attributed to insolubility of these radionuclides in
water. The concentrations of both radionuclides were higher in sediments collected from ponds
involved in the close water recycle system compared to those ponds involved in the open water system.
Results also showed that the concentrations of these radionuclides were higher than background
indicating that amang processing activity has enhanced the natural radionuclides contents in water and
sediment.
Keywords: Amang processing, natural radionuclides, uranium-238, Thorium-232, gamma

spectrometry
Introduction:
Tin mining has been a major activity of Malaysia since 1848. Up till 1980, Malaysia contributed 30.7
% of the world’s produce of tin. However her contribution to the world’s tin dropped sharply since
1983. By 1996 Malaysia’s contributed only 3.9 % and by then there were only 63 mines in operation
(Malaysian Department of Mines, 1997). With the drop in tin production and the cost of world’s tin,
attention shifted toward processing amang (a tin by product) for valuable minerals [1]. Amang is a
local (Malaysian) slang word used by the tin mining community to describe tin tailing consisting of a
mixture of tin ore, sand and minerals initially discarded by tin miners [5, 16]. Amang or by- product
of tin minerals reprocessing, has been found to contain valuable minerals such as ilmenite, zircon,
monazite, xenotime, columbite and struvirite that has high demand in production industry [2]. Studies
done by the Atomic Energy Licensing Board have shown that the uranium and thorium concentrations
vary in monazite, xenotime and ilmenite respectively [3]. Valuable minerals such as monazite
([Ce,La,Y,Th]PO4 ) are radioactive because they contain naturally occurring thorium. Zircon becomes
radioactive when cations, such as Zr+4, are replaced with uranium or thorium [2, 14]. Other minerals
may be contaminated with minerals that are radioactive. Amang consists of natural occurring
radioactive materials (NORM) such as 238U and 232Th that are technologically enhanced natural
occurring radioactive materials (TENORM) during the mining and amang processing activities.
Amang also consists of heavy metals. The mining of tin has been blamed for upsetting the ecosystem.
Beside the obvious scaring of large and beautiful landscape and turning it into barren lands, tin mining
together with amang processing have also been blamed for changing concentration distribution of
elements in the ecosystem, namely the distribution of heavy metals as well as NORM in soil and
water [15].
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In amang processing separation and concentration of valuable minerals are based on three physical
properties, i.e. different specific gravities, magnetic and electrostatic properties. In this process large
volume of water is used in wet gravity separation process and has become a potential source of
environmental pollution depending how the water is managed. The water may be released directly
into the environment (open water management system) or recycle (close water management system).
Such activities have been associated with giving rise to radiological environmental problems [16].
The risk of such problem is high due to the fact that legally, amang plants in Malaysia are categorized
as small amang factory and is exempted from licensing by the Atomic Energy Licensing Board
(small amang factory) Order 1994 [4].
In Malaysia, there are 113,700 hectares (281,000 acres) of former mining land and 14.4 percent of
it was in the form of water pond, used extensively for aqua culture. About four percent has been
turned into food production areas, when tin mining collapsed in the 1980s [6].
Using energy dispersive X-ray fluorescence (EDXRF) A.F.Oluwole [10] measured the
concentrations of radionuclides and toxic heavy metals in the soil around a lead / tin smelter and also
air particulate and mining wastes collected from some tin mines and a tin mill. The concentrations of
thorium and uranium reported ranged between 0.01 - 2.94 % and 0.002 - 0.11% in the tailing and
between 2.25 - 9.09% and 0.25 - 5.65% in the monazites respectively. Studies by Hu [12] and
Kandaiya have also shown the presence of naturally occurring radionuclides in the valuable minerals
of amang. Ismail B. [15, 16] reported that amang processing reduces the pH of water and
radionuclides contaminates the water and consequently decrease quality of water.
Materials and Methods

Sampling location:
Seven different amang processing plants employing three kinds of water management systems (i.e.
open water system, close water natural and close water man made systems) were chosen for this
study. Thirty three water samples and 26 sediment samples were taken from seven amang plants. All
water and sediment samples were taken from Selangor and Perak State in Malaysia. Hand made water
sampler was used for taking water sample from surface level (top), mid and bottom levels of the lakes
and ponds. If the depth of pond was less than three meters, only one water sample from top was taken,
if the depth was more than three meter and less than four meter, two water samples from top and
bottom were taken. If the depth of the lake was more than four meters, three water samples (top,
middle and bottom levels) were taken. Water samples were collected and stored in extra clean
polyethylene bottle. Water samples collected were labeled as SXLy , where S indicates sampling
station and L indicates depth at which the water samples were collected. X represents station number
from 1-18, and y represents depth of the water samples from 1-3. For example water sample S1L1
means station number 1 and top level of water.
Twenty six sediment samples were collected from two different amang processing plants
employing close water natural system. Sediment samples were collected in special PVC container.
Sediment sampler model Ejkelkamp with PVC transparent tubes (60,100,150 cm length and 63 mm
diameter) was used for taking the sediment samples. Sediment samples collected were labeled as SXLy
. Where S indicates sampling station and L indicates depth at which the sediment samples were
collected.
Treatment of samples:
The determination of uranium-238 and thorium-232 concentrations in water samples were based on
2000 ml of water samples collected and subsequently evaporated to 200 ml and stored, capped and
sealed in Merinelli containers. In sediment samples, large stones and other objects were removed, then
were dried in oven at 105ºC for 24 hours to constant mass, then sieved through mesh 500 µm. All
sediment samples were weighted and sealed in Merinelli containers. All water and sediment samples
were kept for at least four weeks before counting in order to allow the in-growth of uranium and
thorium decay products and achievement of secular equilibrium for 238U and 232Th with their
respective progenies.
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Gamma spectroscopy:
A stand-alone high-resolution gamma spectrometric system was used for the measurement of the
energy spectrum of the emitted gamma rays in the energy range between 50 kev and 3000 kev [12].
The gamma spectroscopy system consists of the high purity germanium (HPGe) detector from Oxford
Company with an efficiency of 15%. Detector model number is CNVDS30 with crystal characteristics
of diameter 45.3 MM, length 47.3 mm, active volume 75 and germanium dead layer thickness 0.3
microns and detector to window distance less than or equal to 5 mm. Also end cap outside diameter is
76-mm aluminum 1 mm thick. The spectra were fed through the Amplifier Canbera model 2020 to the
multi channel analyzer with two analog to digital converters and the memory containing 8192
channels. The multi channel analyzer was directly connected to a personal computer where the spectra
were processed and stored. In this system bias supply is from Ortec Company. The detector is
mounted on a cryostat which is dipped in to a 30 liters dewar filed liquid nitrogen. The detector is
surrounded by a cylindrical shield consisting of lead with thickness of 5 cm, which provides an
efficient suppression of background gamma radiation present at laboratory site. Also Soil-IAEA-375
was used as standard reference for sediment samples and uranium and thorium mix stock standard
solutions were used as standard reference for water samples.
Analysis and instrumentation:
Gamma spectroscopy was used to determine the concentrations of uranium-238 and thorium-232 in
water and sediment samples. Water sample was put into the shielded HPGe detector and the activity
concentration present was counted for 86400 seconds (24 hours), while sediment samples were
counted for 43200 seconds (12 hours). Prior to the sample measurement, the environmental gamma
background at the laboratory site was determined using a blank Merinelli under identical
measurement conditions. The laboratory background reading was averaged from four readings taken.
Based on the measured gamma ray photo peaks, emitted by specific radio nuclides in the thorium-
232 and uranium-238 decay series, their radiological concentrations in samples collected can be
determined. Calculations relied on establishment of secular equilibrium in the samples, due to the
much smaller lifetime of daughter radionuclides in the decay series of thorium-232 and uranium-238.
More specifically, the thorium-232 concentration was determined from the concentrations of Tl -208
in the samples, and the concentration of U-238 was determined from concentrations of the Bi-214
decay products.
Energy 1120.3 kev belonging to radionuclide Bi-214 was used for measuring mass concentration
and activity of uranium-238 in water samples. Energy 2614.4 kev belonging to radionuclide Tl-208
was used for measuring concentration and activity thorium-232 in water samples. Energy 609.3 kev
belong to radionuclide Bi-214 was used for measuring mass concentration and activity of uranium-
238 in sediment samples. Energy 2614.4 kev belonging to radionuclide Tl-208 was used for
measuring mass concentration and activity concentrations of Th-232 in sediment samples.
The mass and activity concentrations of radionuclides were obtained using related formula [3].
Results and discussion:
Before detail discussion is made in this study, the overall finding of this study is prepared first.
Figures 1 and 2 show the mass and activity concentrations of uranium-238 and thorium-232 in water
samples. S1- S16 are water sampling stations in different amang ponds, S17 and S18 are water
sampling stations along a river (S17 being upstream and S18 down stream). L1, L2 and L3 are
different depth where water samples were taken. L1 being near the surface and L3 being near the
bottom. Figures 3 and 4 show the mass and activity concentrations of uranium and thorium
respectively in sediment samples in amang plant number 1. Figures 5 and 6 show the mass and
activity concentrations of uranium and thorium in sediment samples in amang plant number 2. Amang
plants number 1 and 2 represent two different amang plants. S1-S4 are sediment sampling stations
around the ponds, and L1-L4 are the sediment layer (L1 means top sediment layer and L4 means
bottom sediment layer).
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Figure 1 and 2 show the mass and activity concentrations of uranium-238 and thorium-232 in
water samples respectively. Maximum mass concentration of uranium in water samples was 6.64 ppm
and maximum activity concentration was 78.53 Bq/l belonging to sample taken at station 8 (ie.
S8L1). Maximum mass and activity concentrations of thorium-232 in water samples were 1.71 ppm
and 6.90 Bq/l respectively. These readings were recorded in station 15 (ie. S15-L1).
Figures 3 and 4 show the mean mass and activity concentrations of uranium-238 and thorium-232
in sediment samples in amang plant number 1 respectively. Maximum mass concentration of
uranium-238 in sediment samples was 69.75 mg/kg and maximum activity concentration was 860.57
Bq/kg . These readings were recorded at station 3 (ie. S3L1). Maximum mass concentration of
thorium-232 in sediment samples in amang plant 1 was 157.73 mg/kg and maximum activity
concentration was 637.61 Bq/kg, recorded at station 2 (S2L1).
Figures 5 and 6 show the mass and activity concentrations of uranium-238 and thorium-232 in
sediment samples sampled at amang plant number 2. Maximum mass concentration of uranium-238
in sediment samples was 27.59 mg/kg and maximum activity concentration was 340.40 Bq/kg, these
values were observed in station 1 (S1L1). Also maximum of mean mass concentration of thorium in
sediment samples in amang plant 2 was 150.8 mg/kg and maximum activity concentration was 609.60
Bq/kg. These readings were recorded at station 1 (S1L1).
Table 1 shows the summary and statistical calculations of data collected from all water and
sediment samples. Results in table 1 showed that the mean concentration of uranium-238 in water
samples was 4.34 ± 1.58 ppm and with a range 0.12 - 6.64 ppm. Also results showed that the mean
concentration of thorium-232 in water samples was 0.37 ± 0.37 ppm and the range was between 0.01
– 1.71 ppm. Mean maximum concentrations of uranium-238 and also thorium-232 were observed in
stations 8 (S8-L1) and station 15 (S15-L1) respectively. Both stations were close to water discharge
point of the plant. Both stations involve amang plant using the close water management system
Results from table 1 showed that the mean concentration of uranium-238 in the sediment samples
in amang plant 1 was 18.00 ± 17.55 mg/kg and the range was between 6.82 - 69.75 mg/kg. The mean
concentration of thorium-232 in sediment samples taken from amang plant 1 was 62.05 ± 39.34
mg/kg and the range was between 26.00 – 157.73 mg/kg. Maximum mean concentrations of uranium-
238 and also thorium-232 were observed in stations 3 (S3-L1) and 2 (S2-L1) respectively. These two
stations were close to water discharge point, and where the water management in these amang plants
is close water system type.
Table 1 also shows the statistical calculations of uranium-238 and thorium-232 in the sediment
samples taken from amang plant 2. The mean concentration of uranium in sediment samples in amang
plant 2 was 9.62 ± 6.47 mg/kg and was in the range 4.96-27.95 mg/kg. The mean concentration of
thorium-232 in sediment samples in amang plant 2 was 40.49 ± 39.41 mg/kg and was in the range
11.92 - 150.80 mg/kg. Maximum mean concentrations of uranium-238 and also thorium-232 were
observed in station 1 (S1-L1), i.e. discharge point in this amang plant, where the management system
in this amang plant is close water system type.
Table 2 shows statistical calculations and activity concentrations of uranium-238 and thorium-232
in water collected from seven amang plants highlighting the difference water samples collected at
point of discharge and those collected elsewhere. This table shows their mean ± standard error mean,
median, range and standard deviation. Based on these results the highest uranium-238 and thorium-
232 in all amang plants were recorded near or at the point of water discharge (except amang plant 5).
The median concentrations of uranium-238 in discharge points were 56.53, 64.93, 71.55, 20.40,
64.00, 54.61and 7.62 Bq/l respectively and for thorium-232 were 1.19, 1.39, 2.55, 1.33, 1.46, 6.9 and
0.12 Bq/l respectively. It should be mentioned that near station S3 in amang plant no 5 there were
several mounds and valuable minerals next to the point rainfall could have washed down these
minerals and carry them into the pond.
296
Activity concentrations (Bq/l) Mass concentrations (ppm)
0
1
2
3
4
5
6
7
0
10
20
30
40
50
60
70
80
90
S 1-L1
S 1-L1
S 1-L2
S 1-L2
S 2-L1
S 2-L1
S 2-L2
S 2-L2
S 3-L1
S 3-L1
S 3-L2 S 3-L2
S 4-L1 S 4-L1
S 4 –L2 S 4 –L2
S 4-L3 S 4-L3
S 5-L1 S 5-L1
S 5-L 2 S 5-L 2
S 5-L3 S 5-L3
S 6-L1 S 6-L1
S 7-L1 S 7-L1
S 7-L 2 S 7-L 2
S 8-L1
Uranium-238
S 8-L1
Uranium-238
297
S 8-L 2 S 8-L 2
Stations
Stations
S 9-L1 S 9-L1
S 10-L1 S 10-L1
S 11-L1 S 11-L1
Thorium-232
Thorium-232
S 12-L1 S 12-L1
S 12-L 2 S 12-L 2
S 12-L 3 S 12-L 3
S 13-L1 S 13-L1
S 13-L 2 S 13-L 2
S 13-L 3 S 13-L 3
S 14-L1 S 14-L1
S 14-L 2 S 14-L 2
S 14-L 3 S 14-L 3
Figure 1. Mass concentrations of U-238 and Th-232 in water samples
Figure 2. Activity concentrations of U-238 and Th-232 in water samples

S 15-L1 S 15-L1
S 16-L1 S 16-L1
S 17-L1 S 17-L1
S 18-L1 S 18-L1
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298
Uranium-238 Thorium-232
180
160
140
Mass concentrations (mg/kg)
120
100
80
60
40
20
0
S 1-L1 S 1-L2 S 1-L3 S 1-L4 S 2-L1 S 2-L2 S 2-L3 S 3 –L1 S 3-L2 S 3-L3 S 3-L4 S 4-L1 S 4-L2 S 4-L3 S 4-L4
Stations
Figure 3. Mass concentrations of U-238 and Th-232 in sediment sample (amang plant 1)
1000
900
800
Activity concentration (Bq/kg)
700
600
500
400
300
200
100
0
S 1-L1 S 1-L2 S 1-L3 S 1-L4 S 2-L1 S 2-L2 S 2-L3 S 3 –L1 S 3-L2 S 3-L3 S 3-L4 S 4-L1 S 4-L2 S 4-L3 S 4-L4
Stations
Figure 4. Activity concentrations of U-238 and Th-232 in sediment samples (amang plant 1)
298
299
160
140
120
Mass concentrations (mg/kg)
100
80
60
40
20
0
S 1-L1 S 1-L2 S 1-L3 S2 -L1 S 2-L2 S 2-L3 S 2-L4 S 3–L1 S 3-L2 S 3-L3 S 3-L4
Stations
Figure 5. Mass concentrations of U-238 and Th-232 in sediment samples (amang plant 2)
700
600
Activity concentration (Bq/kg)
500
400
300
200
100
0
S 1-L1 S 1-L2 S 1-L3 S2 -L1 S 2-L2 S 2-L3 S 2-L4 S 3–L1 S 3-L2 S 3-L3 S 3-L4
Stations
Figure 6. Activity concentrations of U-238 and Th-232 in sediment samples (amang plant 2)
299
300
Table 1. Statistical calculations of water and sediment samples
Sample Lower Upper

Std error Minimum Media Maximum
Sample size Mean
of mean
95% 95%
( 50 percentile)
Conf limit conf limit
U in Water 33 4.34 0.275 3.785 4.909 0.12 4.750 6.64

ppm ppm ppm ppm
sample
Th in water 33 0.37 0.064 0.231 0.501 0.01 0.331 1.71

ppm ppm ppm ppm
sample
U in 15 18.00 4.533 8.285 27.730 6.82 8.84 69.75

mg/kg mg/kg mg/kg mg/kg
Sediment
sample
(amang plant 1)
Th in 15 62.05 10.159 40.261 83.844 26.00 46.80 157.73

sediment
sample
(amang plant 1)
U in 11 9.62 1.952 5.275 13.975 4.96 7.13 27.59

sediment
Sample
(amang plant 2)
Th in 11 40.49 11.882 12.024 66.970 11.920 22.57 150.80

sediment
sample
(amang plant 2)
300
301
Table 2. Activity concentrations of 238U and 232Th in water collected from 7 amang plant.
Uranium-238
Thorium-232
Mean ± Media Range St. Mean ± Media Range St.
Amang Plants/Sample
sem n(Bq/ (Bq/L) dev. sem n (Bq/L) dev.
(Bq/L) L) (Bq/L) (Bq/L)
Plant 1
S1-Point of 56.53±9.645 56.53 46.88- 13.6 1.19±0.27 1.19 0.92-1.46 0.38
discharge 66.17 4
S2 46.74±4.68 46.74 42.06- 6.62 0.37±0.15 0.37 0.35-0.38 0.21
0 51.42
Plant 2
S1-Point of 64.93±1.18 64.93 63.75- 1.66 1.39±0.09 1.39 1.3-1.47 0.12

discharge 66.10
S2 47.64±2.05 48.11 43.88- 3.54 0.532±0.6 0.51 0.43-0.63 0.10
50.92
S3 70.28±4.67 73.21 61.14- 8.09 0.16±0.04 0.12 0.12-0.24 0.07
76.48
Plant 3 (pond 1)
S1-Point of 71.55 71.55 71.55 0.00 2.55 2.55 2.55 0.00

discharge
S2 60.44±3.33 60.44 57.11- 4.71 1.89±0.28 1.89 1.61-2.16 0.39
63.77
S3 71.17±7.36 71.17 63.82- 10.4 0.8±0.39 0.80 0.41-1.19 0.55
78.53 0
Plant 3 (pond-2) 37.75 37.75 37.75 0.00 1.65 1.65 1.65 0.00
301
302
Plant 4
S1-Point of 20.40 20.40 20.40 0.00 1.33 1.33 1.33 0.00

discharge
S2 1952 19.52 19.52 0.00 1.27 1.27 1.27 0.00
Plant 5
S1-Point of 57.03±7.75 64.00 41.56- 13.4 1.42±0.33 1.46 0.82-1.97 0.58

discharge 65.40 2
S2 60.866±1.5 61.31 58.04- 2.63 1.50±0.10 1.47 1.34-1.70 0.18
2 63.25
S3 66.87±5.19 5.19 56.53- 9.0 2.42±0.37 2.77 1.69-2.81 0.64
72.90
Plant 6
S1-Point of 54.61 54.61 54.61 0.00 6.9 6.9 6.9 0.00

discharge
S2 34.05 34.05 34.05 0.00 6.07 6.07 6.07 0.00
Plant 7
S1-Down stream 7.62± 7.62 7.62 0.00 0.12 0.12 0.12 0.00
S2-Up stream 1.48± 1.48 1.48 0.00 0.03 0.03 0.03 0.00
S1-S3 are sampling station
Sem: Standard error mean
St.D: Standard deviation
302
The higher uranium-238 and thorium-232 concentrations in water samples collected at down
stream related to upstream, suggested that amang processing enhances their concentrations. In the
case of plants employing close water system, such enhancement is expected with every recycling
process.
The enhancement of NORM in water may also be attributed to the acidity of the recycling water.
Such acidity is caused by the acidic nature of amang [16]. Acid conditions caused the radionuclides to
dissolved in water.
Another finding from this study is that, the mean mass and activity concentrations of thorium-232
in all sediment samples (amang plants no 1 and 2) were higher than the mass and activity
concentrations of uranium-238 in sediment samples (table 1). However this was the opposite in water
samples.
Results from this also showed that average concentrations of uranium-238 and thorium-232 in both
water and sediment samples were higher than those measured from areas that were not involved in
amang processing or tin mining activities. According to R.M.R. Almedia [6], natural uranium-238
concentration in ground water range from 0.1 to 10 ppb, in this study the mean maximum
concentration in water sample reported was 6.64 ppm (in S8-L1), or 6600 times more than the
maximum concentration of uranium-238 in natural ground water. Natural uranium is the only
radioactive substance for which chemical toxicity is the limiting factor in risk assessment the
maximum contaminant level for uranium is 20 µg/l [6]. As mentioned mean concentration of
uranium-238 in amang water samples was 4.34 ppm, it means the average concentration of uranium in
amang water samples was around 220 times more than maximum contamination level of uranium. A.
Martin Sanchez [18] reported low concentration of uranium series in water samples in Extramadura
(Spain), ranging from 0.024 to 2.69 ppb and most of them were below 1.0 ppb. Likewise the uranium
concentration of Slovenian spas area ranged from 0.2 to 2.7 ppb [Kobal, 9]. According to Boyle
(1982) the mass concentration of thorium in natural water is around 0.005 – 0.5 ppb. In this study the
concentration of thorium-232 in water samples ranged from 0.03 – 1.7 ppm. The maximum mean
concentration of thorium-232 in water taken at station S15-L1 was 3400 times higher than those
reported by Boyle in ground water. I. G. E. Ibeanu [8], showed that the measured concentration levels
of uranium and thorium in tin tailing samples and the measured dose rates in Nigeria were found to
be elevated with values up to approximately 100 times above background levels of control soils.
Higher concentrations of uranium-238 and thorium-232 in sediment relative to water observed in
this study supported other earlier reports [15, 16]. Higher concentrations of both radionuclides in
sediment is attributed to the insolubility of minerals bearing radionuclide in this water, such minerals
include monazite, zircon and ilmenite.
Ismail et al. [15, 16] and Redzuwan et al. [2] carried out similar studies in Perak and Selangor in
Malaysia respectively. Ismail et al reported uranium-238 and torium-232 mass concentrations ranging
from 6.93- 11.45 mg/kg and 27.72-120.88 mg/kg respectively. Redzuwan reported activity
concentrations of uranium-238 and thorium-232 ranging from 6.27-435.95 Bq/kg and 12.90-301.59
Bq/kg respectively. Our finding were in correlations with those of Ismail et al. and Redzuwan et al. in
both the mass and activity concentration of both radionuclides and their differences between uranium-
238 and thorium-232.
Conclusion:
Gamma ray spectrometry definitely appeared to be a useful and sensitive method for obtaining actual
information on radionuclides in the environments. A total of 33 water samples and 26 sediment
samples taken from amang processing plants/ river and ponds where analyzed for uranium-238 and
thorium-232 concentrations. Results further confirm other earlier limited studies that amang
processing enhances NORM into TENORM. Concentrations of uranium-232 were higher in water
than thorium-232.However it was the opposite in sediment. Overall uranium-238 and thorium-232
concentrations were higher in sediment than water indicating the insolubility of these NORM in water
and suggesting that they remained in mineral form in the sediment.
303
Acknowledgment:
The authors would like to acknowledge the Atomic Energy licensing Board of Malaysia (AELB) and
also the Universiti Kebangsaan Malaysia for funding this work through research grant -D-036-2002.
References:
1. Ismail, B.Y., Othman, M. Soong, H. F., (2000) “Effect of tin dredging on the environmental concentrations
of arsenic, chromium and radium-226 in soil and water”, J. Sains nuklear Malaysia, Vol 18, No1.
2. Redzuwan Yahaya, Ismail Bahari , Amran Ab. Majid, Muhamad Samudi Yasir, Lin Cheng lee, (2002) “The
impact of amang processing activity on the water quality and sediment of open water system”, 15th Analytical
chemistry symposium, Penang, Malaysia.
3. AELB. (1991) “Radiological hazards assessment at mineral processing plants in Malaysia “ Atomic energy
Licensing Board of Malaysia, LEM/LST/16/pind. 1.
4. M.J. azlina, B.Ismail, M. Samudi Yasir, Syed Hakimi Sakuma, M.K. Khairuddin, (2003) ”Radiological
impact assessment of radioactive minerals of amang and ilmenite on future land use using RESRAD computer
code, “ J. of Applied radiation and isotopes, 58, 413-419 .
5. M.J.Azlina , B.Ismail , M. Samudi Yasir , Taiman , K. (2001) ”Work activity , radiation dosimeters and
external dose measurement in amang processing plant, j. sains nuclear Malaysia , vol.19, no 1&2 ,31-39 .
6. R. M.R. Alemedia, D.C. Lauria, A. C. Ferreira, O. Sracek, (2004) “ Ground water radon, radium and uranium
concentrations in Regiao dos Lagos, Rio de Janerio State, Brazil”, J. of environmental radioactivity, 73, p. 323-
334.
7. J. Al-Jundi, E.Werner, P. Rot, V. Hollriegl, I. Wendler, P.Schramel, (2004) ”Thorium and uranium in human
urine” , J. of environmental radioactivity, 71, p. 61-70.
8. I. G. E. Ibeanu, (2002) “Tin mining and processing in Nigeria: Cause for concern” , J. of environmental
radioactivity, Vol, 64, Issue 1, P.59-66.
9. Mantazul I, Chowdhury, M. N. Alam, S.K.S.hazari, (1999) ”Distribution of radio nuclides in the river
sediments and coastal soils of Chttagong , Bangladesh and evaluation of the radiation hazard”, J. Applied
radiation and isotopes, 51, P. 747-755.
10. Mohd Tadza Abdul Rahim, Shamsulbahrin Ludin , Mohd Yusof Harun , Amran Kamarudin , Abdul hamid
Latip , Mohd Azwar Hashim , (21-22 June,1994) ”Radiological assessment at mineral processing plants in
Malaysia “ , Radiological hazards in the tin mining and heavy mineral processing , seminar, Ipoh, Malaysia.
11. W.W.S.Yim, (Aug 1976) ”Heavy metals accumulation in estuarine sediments in a historical mining of
Cornwall “ , Marine pollution Bulletin , Vol 7, issue 8, p. 147-150 .
12. Michalis Tzortzis, Haralabos Tsertos, Stelious Christofides, George Christodoulides, (2003) ”Gamma
radiation measurement and dose rates in commercially – used natural tiling rocks (granites)”, J. of
environmental radioactivity, 70, p. 223-235.
13. Hu, S.J., Kandaiya, (1985) “ Radium 226 and Th 232 concentration “, J. Health physics, 49, p. 1003-1007 .
14. Ismail B., Mokhtar M.B. , tan B.H., (1999), “ Impact of amang processing on the water quality of an
immediate water body : A case of a recycling water system “, sci. int . ( lahore) 11(1), p 1-4. 19
15. Ismail Bahari, redzuwan Yahaya , Muhamad Samudi Yasir , Amran Ab. Majid, Lin cheng Lee, (2003) “
The impact of open water management system in amang processing on the water quality and 238U and 232Th
activity concentration in sediment and water “ , J. of Biological science, 3(11), p. 1063-1069.
16. B.Ismail , M.S. Yasir, Y. Redzuwan , A.M.Amran, (2003) ”Radiological environmental risk associated with
different water management system in amang processing in Malaysia“ , Pakistan j. of biological science, 6 (17),
September, p. 1544-1547 .
17. D.Malczewski, L. Teper, J.Dorda, (2004) “Assessment of natural and anthropogenic radioactivity levels in
rocks and soils in the environs of Swieradow Zdroj in Sudetes , Poland , by in situ gamma – ray spectrometry”,
J. of environmental radioactivity, 73, P. 233-245.
18. A. Martin Sanchez, F. Vera Tome R. M. Orantos Quintana, V. Gomes Escobar, M. Jurado Vargas, (1995)
“Gamma and alpha spectrometry for natural radioactive nuclides in the Spa waters of Extramadua-Spain” J. of
environmental radioactivity, Vol 28, No. 2, p. 209-220.
304
COATING THICKNESS MEASUREMENT FOR GOLD BY USING EDXRF
Meor Yusoff M.S., Masliana Muslimin and Fadullah Jili Forsani*
Materials Technology Group,Industrial Technology Division,

Malaysian Institute for Nuclear Technology Research,
Bangi, 43000 Kajang, Selangor,
*Department of Chemistry,School of Science and Technology,
UiTM, Shah Alam
e-mail: meor@mint.gov.my
Abstract
The paper relates a study on the development of an analysis procedure for measuring the gold coating
thickness using EDXRF technique. Gold coating thickness was measured by relating the counts under
the Au Lα peak its thickness value. In order to get a reasonably accurate result, a calibration graph
was plotted using five gold-coated reference standards of different thicknesses. The calibration graph
shows a straight line for thin coating measurement until 0.9µm. Beyond this the relationship was not
linear and this may be resulted from the self-absorption effect. Quantitative analysis was also
performed on two different samples of gold-coated jewelry and a phone connector. Result from the
phone connector analysis seems to agree with the manufacturer’s gold coating value. From the
analysis of gold-coated jewelry it had been able to differentiate the two articles as gold wash and gold
electroplated.
Abstrak
Kertas kerja ini menerangkan kajian ke atas pembangunan prosedur analisis pengukuran ketebalan
penyalutan emas dengan menggunakan teknik EDXRF. Ketebalan penyalutan emas dapat diukur
dengan menghubungkan bacaan keamatan di bawah puncak Lα bagi Au. Bagi mendapatkan
keputusan yang tepat, graf kalibrasi diplotkan mengguna 5 jenis emas salutan rujukan piawai dari
pelbagai ketebalan. Graf kalibrasi menunjukkan garis lurus bagi pengukuran penyalutan nipis iaitu
sehingga 0.9 µm ketebalan. Melebihi nilai ini, perhubungan adalah menjadi tidak linear dan ini
mungkin disebabkan oleh kesan penyerapan tersendiri. Analisis kuantitatif juga dilakukan ke atas 2
sampel berlainan iaitu barang kemas bersalut emas dan penyambung telefon. Keputusan menunjukkan
analisis bagi nilai penyalutan emas penyambung telefon memberikan nilai yang sama dengan nilai
yang diperolehi daripada pihak pengeluar. Bagi analisis barang kemas pula didapati ianya dapat
dibezakan kepada 2 jenis salutan iaitu gold wash dan gold electroplated.
Keywords: EDXRF, gold, coating thickness, electroplated, gold wash
305
Introduction
Coatings represent a variety of metallic mixtures and compound typically used to strengthen certain
features of a product to make it better fit its purpose. Gold is one of the common metal used as coating
[1]. It is applied in many usages especially as imitation jewelry and its related articles. In electronic,
gold coating is largely used in connectors and contacts due to its high electrical conductivity,
excellent corrosion and wear resistant. Electrical connectors are used in a wide range of equipment
such as computers, phones and cars. Measuring the coating thickness of a coated product is an
important quality control procedure. Manufacturers need an excellent quality control of coatings
during their entire processes in order to avoid production loss, waste of coating material and achieve
fastest possible control over their process. Car manufacturers for example need to quickly analyze
different coating weights in order to keep quality at highest standards.
Gold-coated articles can be classified as gold filled, rolled gold plate, gold electroplate and
gold wash [2]. Gold filled is produced when a layer of at least 10-karat gold is permanently bonded by
heat and pressure to one of more surfaces of a supporting metal. These coatings are used as watch
casings, cigarette casings, high fashion jewelry, and numerous other decorative items. Gold-coated
articles can also be produced by mechanically bonding on to a base metal. This technique is called
rolled gold plate. Another technique that is widely used in the industry is by electrolysis. Here, a gold
electroplate is perform on the base metal and among the advantages of this technique is it able to
provide good durability and can be used on complex designs. Gold electroplate is also said to be less
expensive than that of gold-filled products. Finally if the electroplated layer is very thin and less than
0.175 µm, the product is called gold wash.
Energy dispersive x-ray fluorescence (EDXRF) is a popular tool in the determination of

coating thickness and this had been used widely in many industries [3,4,5]. The analysis of metal
coating by using x-ray is an ideal application because x-rays achieve higher penetration depths than
comparable techniques. In this aspect, EDXRF have several advantages compared to the other
analytical techniques. EDXRF systems are used for the precise non-destructive measurement of
coating thickness as well as for alloy coatings and in the incoming material inspection. Besides that
EDXRF also proves to be a relatively fast technique.
306
Experimental
EDXRF analysis was carried out using the Baird Ex-310 instrument at MINT. Gold coating standards
produced by Helmut-Fischer GMBH, Germany with thicknesses of 0.46, 0.90, 2.65 and 5.90 µm were
used for constructing the thickness calibration curve. The standards were analyzed by placing each of
these standards in a sample cup that is enclosed with an x-ray penetrative prolene plastic film. Each
standard was analyzed repeatedly for five times to ensure its accuracy. The Au thickness calibration
curve was constructed by measuring the intensity of Au under Lα peak with its respective thickness.
Similar experimental procedure was then applied during the measurement of gold-coated necklaces
and electrical connector. The Au coating thickness of these samples were then determined by using a
regression quantitative procedure.
In an EDXRF analysis, the transition of a higher energy level electron to its lower level will produce
characteristics x-ray that will be channeled to the detector [6]. The presence of a multi-channel
analyzer card in the computer enables the identification of the elements present through their energy
spectrum. Quantitative coating thickness measurement can be normally performed by two ways [1].
The first is called excitation and is the most common for single layer determination. When a sample is
exposed to an x-ray beam, the x-rays in the incident beam will interact with the coating material and
cause it to emit its own characteristic x-rays. In the absorption method, the interest is on the
characteristic x-rays produced by the substrate material. The x-rays emitted by the substrate are
absorbed as they passed through the coating material and the amount of absorption is dependent on
the type of coating and its thickness.
A typical EDXRF spectrum of a standard Au coating sample is as that shown in Figure 1. As

there was considerably high signal counts registered by using this x-ray energy, a Mo filter was placed
in front of the detector to reduce any error resulted from the dead time effect [7]. The EDXRF
spectrum shows characteristics energy peaks belonging to the L lines attributed by Au. The spectrum
also shows the presence of Fe substrate by the two smaller peaks (FeKα and FeKβ) located at left of
Au peaks.
307
Figure 1. EDXRF spectrum of Au coating standard
The energy spectral intensity data for standards and samples were processed by least-square
regression analysis with statistical evaluation carried out after repeating each analysis 5 times. In this
study the intensity of Au Lα was then correlated with the certified thickness of the different gold
standards using the excitation method (Figure 2).
Figure 2. Calibration graph for Au thickness coating
The excitation curve for very thin coating (<0.90 µm Au) is linear, that is, there is an
approximate straight-line relationship between x-ray intensity and thickness with a correlation value,
308
R2 of 0.998 (Figure 3). Beyond this an exponential relationship is establish for the intensity and
coating thickness. A quantitative analysis regression equation as that shown below was then applied
to measure the coating thickness [8];
Ci = Ao + AiIi (1)
where;
Ii is the radiation intensity of gold (kilocounts per second)
Ai is the slope
Ao is the intercept
Ci is the coating thickness of gold
Gold coating thickness measurement on three different samples was then performed using the
above-developed quantitative procedure. These samples include 2 gold-coated necklaces that are
purchased from a night market and a direct selling company respectively. The other sample is a phone
connector produced from a factory in Sg. Petani, Kedah. The analytical procedure was done similar to
that performed on the standards with each sample analysed repeatedly for five times. Table 1 shows
the results obtained from these three samples.
Table 1. Gold coating thickness analysis of different samples
Gold coating thickness (µm)

Analysis no. Gold necklace 1 Gold necklace 2 Phone connector
(night market) (direct selling)
1 0.17136 3.3402 1.1876
2 0.17022 3.2153 1.3459
3 0.17135 3.2610 1.3911
4 0.16909 3.2384 1.2780
5 0.17135 3.4458 1.3911
Mean 0.1707 ± 0.0010 3.3001 ± 0.0940 1.3187 ± 0.0867
309
The analysis for thin layer of gold coating is very good by the EDXRF technique. This can be
demonstrated by the highly consistent results and very small standard deviation obtained for gold
necklace 1 sample. The low price gold necklace 1 sample purchase from the night market has a
coating thickness of 0.1707 ± 0.0010 and this thickness is within the linear region of the calibration
graph (Figure 2). From the classification of gold-coated articles, this sample can be classified as a
gold wash as the coating thickness is less than 0.175 µm [2]. A gold coating thickness of 3.3001 ±
0.0940 µm was obtained in the analysis of another jewelry sample (gold necklace 2) purchased from a
direct selling company. The result shows that at this thickness the standard deviation value is bigger
and this relates to the bigger differences in the five analysis runs as compared to the previous sample.
A major factor that may attribute for this is the thicker gold coating of this sample. This results to the
readings to fall at the exponential relationship between intensity and coating thickness as shown in
Figure 2.
Gold necklace 2 can also be classified as a gold electroplated article as the coating thickness
is way above the 0.175 µm limit for gold wash. And an interesting point to note about this jewelry
analysis result is that it can be co-related with the cost and durability of the sample. Gold necklace 2 is
much more expensive than that of gold necklace 1 and the former was purchased from a direct selling
company. It had been pointed in an earlier article that there is a relationship between coating thickness
and the time estimated for its durability. For jewelry articles that has a thickness of 2.5 – 8.0 µm, it is
estimated that time for its durability is 3 – 7 years while those having 0.5 – 2.5 µm has a lower
durability time of 1 – 3 years [2]. From the physical inspection of these two necklaces it show that the
colour for gold necklace 1 has deteriorated even though both are purchase at the same time.
Another sample that was analysed for its gold coating thickness is the phone connector. The
phone connector gives a mean result of 1.3187 ± 0.0867 µm (or 49.7150 µinches) that is almost
similar to the manufacturer’s value of 50 µinches [9]. The result also shows the developed procedure
is highly accurate to measure gold coating thickness and is comparable with similar analysis
performed by the manufacturer.
Conclusion
EDXRF proves to be a very valuable tool for the measurement of gold coating thickness. Besides
being non-destructive and fast, the analysis done on different samples show that the quantitative
technique developed is also highly accurate and comparable with similar analysis.
310
Acknowledgement
The authors wish to extend their gratitude to all parties that had supported the project in particular to
the MTEC staff and manager, BTI director and MINT management.
References
1. Veeco Instruments. (1996) User Manual XRF-400 And XRF-500 Series XRF Coating
Thickness Measurement System, New York.
2. An Overview Of Gold Filled Processes And Their Legal Classifications:
www.artisanplating.com
3. Nensel B. (2004) “Accuracy Of The Analysis Of Gold Alloy With EDXRF”, Proceedings
Denver X-rays Conference 2004.
4. Havrilla G.J., Miller T. and Doering E. (2003) “Characterization Of A Metal Alloy And Thin
Film Coating Using Elemental Imaging µXRF”, Proceedings Denver X-ray Conference 2003.
5. Kloos D. (2000) “EDXRF Analysis Of Gold Carat Alloys Using Proportional Counter Based
µXRF”, Proceedings 24th International Prcecious Metals conference, USA.
6. Bertin E.P. (1975) “Principles And Practices Of X-Ray Spectrometric Analysis”, 2nd Ed.,
Plenum Press, New York.
7. Yokhin B. and Tisdale R.C. (1993) “High-Sensitivity Energy-Dispersive XRF Technology
Part 1: Overview Of XRF Technique” Am. Lab., 36G.
8. Baird. (1985) “Operation manual of EDXRF spectrometer”, Baird Corporation, Boston.
9. Tang Chee Guan, Huan Hsin Co. (M) Sdn Bhd. (2001) Private Communication,.
311
APPLICATION OF SOLID-PHASE MICROEXTRACTION FOR THE DETERMINATION
OF PESTICIDES IN VEGETABLE SAMPLES BY GAS CHROMATOGRAPHY WITH AN
ELECTRON CAPTURE DETECTOR
Chai Mee Kin1, Tan Guan Huat2 and Asha Kumari3

1
Dept. of Science and Mathematics, College of Engineering,
Universiti Tenaga Nasional, Km 7, Jalan Kajang-Puchong,
43009 Kajang, Selangor.
Mkchai@uniten.edu.my Fax:03-89263506
2,3
Dept. of Chemistry, Faculty of Science,
Universiti Malaya, Lembah Pantai,
50603 Kuala Lumpur.
ABSTRACT. A solid-phase microextraction (SPME) method has been developed for the determination of 9 pesticides in 2
vegetables - cucumber and tomato - samples, based on direct immersion mode and subsequent desorption into the injection
port of a gas chromatograph with an electron capture detector (GC-ECD). The main factors affecting the SPME process such
as extraction time and temperature, desorption time and temperature, the effect of salt addition and fiber depth into the liner
were studied and optimized. The analytical procedure proposed consisted of a 30-minute ultrasonic extraction of the target
compounds from 1.0 g vegetable samples with 5 mL of distilled water. Then, the samples were filtered and topped up with
distilled water to 10 mL. The analytes in this aqueous extract were extracted for 15 minutes with a 100 µm thickness
polydimethylsiloxane SPME fiber. Relative standard deviations for triplicate analyses of samples were less than 10%. The
recoveries of the pesticides studied in cucumber and tomato ranged from 52% to 82% and the RSD were below 10%.
Therefore, the proposed method is applicable in the analysis of pesticides in vegetable matrices. SPME has been shown to be
a simple extraction technique, which has a number of advantages such as solvent free extraction, simplicity and
compatibility with the chromatographic analytical system.
Keywords: Solid-Phase Microextraction, GC-ECD, pesticides, vegetables
INTRODUCTION
One of the major fields in analytical chemistry is the development of faster and easier methodologies
for characterization and quantification of trace compounds in complex matrices. A special attention is
given to substances that can compromise food safety, such as pesticide.
Pesticides are widely used for agricultural and non-agricultural purpose throughout the world.
Although various methods, using highly efficient instruments such as gas chromatography (GC), high
performance liquid chromatography (HPLC), Liquid chromatography (LC) and their combination
with mass spectrometry (MS), have been developed for pesticide analysis, most analytical instruments
cannot handle the sample matrices directly. In general, the analytical method involves processes such
as sampling, sample preparation, separation, detection and data analysis and more than 80% of the
analysis time is spent of sampling and sample preparation steps that include homogenization of
samples, extraction of the analytes with an organic solvent, and clean up of the final organic extract.
Therefore, it is not an exaggeration to say that the choice of an appropriate sample preparation method
greatly influences the reliable and accurate analysis of food.
In contrast to conventional techniques, Solid Phase MicroExtraction (SPME) is a solvent-free

extraction that minimizes sample preparation allowing the extraction and concentration steps to be
focused into a single step. This technique, whose initial concepts were developed by Pawliszyn and
co-workers in 1990 [1], is based on absorption of analytes onto a polymeric-coated fused-silica fiber,
312
usually housed in a modified syringe. The total analytes retained in the fibers are thermally desorbed
in the injector port and deposited at the head of the GC column. Due to its advantages over classic
extraction methods, SPME has received increasing attention since its commercial introduction in the
nearly 1990s [2]. SPME has been applied to the determination of several organic compounds
especially in gas and liquid samples, but also in a few solid samples, in combination with both GC
and HPLC determination. Two modes of application of SPME have been extensively reported [1]:
Direct Immersion (DI-SPME) and Headspace (HS-SPME) extraction. In DI-SPME, the fiber is
directly immersed in the liquid sample or in the sample suspension and the analytes are transported
from the sample matrix to the fiber coating. In headspace extraction mode, the analytes are extracted
in a three-phase system: sample (liquid or solid), headspace, and fiber coating.
SPME has been successfully applied to the determination of pesticide residue analysis in water, soil,
food and biological samples as reported in recent reviews published by Beltran et al. [3] and Kataoka
et al. [1]. Water samples are by far the most widely analyzed by this technique [4-6].
The number of applications of SPME to complex matrices such as biological fluids is still limited; the
headspace mode is the most attractive approach in this field [7-12]. Analysis of other samples such as
soil [13-16] or food commodities [1, 17-18] is generally based on a solvent extraction of the analytes
before application of SPME.
The low number of references about pesticide determination in food samples by SPME derives from
the complexity of these matrices, which makes an extraction of the sample prior to determination by
direct immersion SPME necessary in most of cases. This problem can be overcome if headspace
SPME is applied, as described in several papers dealing with pesticide residue determination in fruits
[19-21] or in a large number of papers related to determination of volatile compounds in food
commodities [22-25].
Determination of non-volatile pesticides has received increasing attention in the recent years in order
to solve some of the problems related with the application of DI-SPME in complex matrices. Several
papers deal with direct immersion of the SPME fiber into a slurry of fruit with water [19,26].
Complex matrix problems can be solved by prior extraction of pesticide and the subsequent
application of DI-SPME over the separated aqueous extract. Once the aqueous extract is obtained, the
presence of interfering substances can reduce the efficiency of SPME. This problem can be overcome
by simply diluting the extract in order to simplify the matrix complexity [27-28]. Still another
problem, closely related with pesticide residue determination in fruits by SPME, is the difficulty of
quantification; in most cases it is necessary to use calibration curves prepared using blank matrix,
standard addition calibration, and internal standards [20].
The aim of this work is to investigate the feasibility of developing a single-step clean up enrichment
procedure for pesticides extracted from vegetables based on SPME prior to gas chromatography with
electron capture detection (GC-ECD). Nine pesticides: Carbaryl, Diazinon, Chlorothalonil, Malathion,
Chlorpyrifos, Quinalphos, Profenofos, Alpha-Endosulfan, Beta-Endosulfan were selected as the
model compounds because residues of these compounds are very often detected in vegetable samples.
Table 1 showed the properties of nine selected pesticides. In this study, SPME-GC-ECD conditions
have been optimized for the target compounds. The developed procedure was then successfully
applied to the analysis of vegetable samples such as cucumber and tomato.
313
Table 1: Name, Molecular Formula, Molecular Weight, Chemical Class of the selected
Pesticides.
Name Molecular Formula Molecular Weight Chemical Class

Carbaryl C12H11NO2 201.22 Carbamate
Diazinon C12H21N2O3PS 304.35 OP
Chlorothalonil C8Cl4N2 265.92 OC
Malathion C10H19O6PS2 330.36 OP
Chlorpyrifos C9H11Cl3NO3PS 350.62 OP
Quinalphos C12H15O3N2PS 298.18 OP
Profenofos C11H15BrClO3PS 373.60 OP
α-Endosulfan C9H6Cl6O3S 406.96 OC
β-Endosulfan C9H6Cl6O3S 406.96 OC
EXPERIMENTAL
Chemicals and Reagents
All solvents used were HPLC grade. Methanol was purchased from Fischer Scientific, Loughborough,
U.K. Deionized water and methanol were filtered through a 0.45 µm membrane filter purchased from
Millipore. The use of high purity reagents and solvents help to minimize interference problems,
pesticide standards (carbaryl, diazinon, chlorothalonil, malathion, chlorpyrifos, quinalphos,
profenofos, α-endosulfan, β-endosulfan) were > 95% pure and purchased from AccuStandard Inc.
New Haven CT, USA. Stock solutions of each pesticide at different concentration level, 50-2000
mg/kg were prepared in methanol and stored at 4 oC. Preparation of different concentration level of
stock solution is due to their sensitivity to the ECD detector. Working standard solutions of pesticides
mixture were prepared by volume dilution in distilled water. In order to avoid the influence on the
results from the possible degradation of pesticides, the working solution was freshly prepared
everyday. 1-chloro-4-fluorobenzene (2mg/kg) was used as internal standard to compensate for sample
and was added to the vial prior to GC analysis.
Gas Chromatography – Electron Capture Detector (GC-ECD)
A Shimadzu GC 17A version 2.21 gas chromatograph with an electron capture detector ECD was
used. A SGE BPX5, 30m x 0.32 mm id capillary column with a 0.25 µm film was used in
combination with the following oven temperature program: initial temperature 120 oC, then 7 oC/min
ramp to final temperature at 250 oC, held for 4.5 min. The total run time was 23.07 min. The injector
temperature was at 240 oC and the detector temperature was at 300 oC. Nitrogen gas (99.999%) was
used as the carrier gas with a gas flow at 24.4 cm/sec linear velocity and the pressure at 94 kPa and
the split mode ratio of 1:36.
SPME Procedure
The SPME fiber holder for manual extraction and the fibers of polydimethylsiloxane (PDMS, 100 µm
film thickness) were purchased from Supelco (Bellefonte, PA, USA). SPME fibers were conditioned
by heating at 250 oC for 0.5 hour in the gas chromatography (GC) injection port according to the
manufacturer recommendations in order to remove contaminants and to stabilize the polymeric phase.
Preliminary experiments were carried out to optimize the main parameters affecting the SPME of the
pesticides investigated from aqueous solution (i.e. extraction time and temperature, desorption time
and temperature, the effect of salt addition, stirring speed of the solution and fiber depth into the
314
liner). In these studies, distilled water samples spiked with the appropriate amount of the standard
solution was used.
After optimization, a typical experiment consisted of the direct immersion of the conditioned fiber
into the spiked water sample, 10mL in a 15 ml clear glass vial and capped with a PTFE-faced silicone
septum (Supelco). The SPME holder needle was inserted though the septum and the fiber was directly
immersed in the sample solution for 15 minutes under magnetic stirring at room temperature (25 oC)
in order to improve mass transfer from the aqueous sample into the fiber coating. After extraction, the
fiber was withdrawn into the holder needle, removed from the vial and immediately introduced into
the GC injector port for 7 min at 240 oC for thermal desorption in a split mode injector.
Calibration curves were constructed by SPME of the target compounds from aqueous samples spiked
at 7 concentration levels and the constant volume of internal standard under above experimental
condition. Three extractions were made for each concentration level of mixture solution. The
calibration graph was plotted by the ratio of the peak area of the analyte against the peak area of the
internal standard from the spiked samples versus the concentration of the analyte. These calibration
lines were used for quantification in subsequent experiments.
Vegetable Samples
In order to evaluate the pesticide recoveries, 2 types of vegetables, cucumber and tomato were
obtained from pesticide free farms under study. 1.0 g of vegetable samples was finely chopped and
placed in a 15ml clear glass vial and capped with a PTFE-faced silicone septum. 5 mL of distilled
water was added and spiked with three concentration levels of stock solution. The mixture was shaken
for 30 minutes in an ultrasonic bath. Then, the samples were filtered, added with internal standard and
topped up with distilled water to 10 mL. Pesticides were then extracted by direct dipping of the
PDMS fiber in the solution. Recoveries of pesticides were determined by comparison of the ratio of
the peak area of the analyte against the peak area of the internal standard from the spiked samples
with that of the standard calibration solutions.
RESULTS AND DISCUSSION
Method Optimization
Preliminary experiments were performed by direct injection of pesticides for GC-ECD conditions
optimization; the temperature program developed was capable of a good separation of the investigated
analytes. In order to developed the SPME described method for pesticides selected extraction in
vegetables, several parameters such as extraction time and temperature, desorption time and
temperature, the effect of salt addition, stirring speed of the solution and fiber depth into the liner
were studied.
On the basis of the results previously published for the target compounds, a 100 µm PDMS fiber was
chosen as this material has been reported to have satisfactory extraction efficiency for a variety of
compounds, including pesticides selected in this study. The 100 µm PDMS fiber (a non-polar phase)
is recommended in the literature because it is a rugged liquid coating able to withstand high injector
temperature up to 300 oC. Fibers coated with thicker films required a longer time to achieve extraction
equilibrium, but might provide higher sensitivity due to the greater mass of the analytes that can be
extracted.
Effects of Extraction Temperature and Time
In order to study the effect of temperature on the extraction process, vials were immersed in a water
bath heated by the magnetic stirring unit. A thermometer was used to monitor the water temperature.
The temperature effect was evaluated by varying the temperature from 25 to 70 oC. An increase in
315
extraction temperature causes an increase in the extraction rate and a simultaneous decrease in the
distribution constant between the analytes and the fiber [27]. The analysis of the 9 pesticide
compounds was performed at room temperature for the subsequent experiments.
The sorption time profile for the selected fiber was obtained by plotting the detector response (peak
area) versus the extraction time for each pesticide in order to obtain the partition equilibrium curve
(figure 1). Blank aqueous samples (10mL) spiked at 0.1 mL standard solution were analyzed at
experimental conditions described in the SPME procedure. Sorption time profiles indicated that a
sampling time higher than 30 minutes is necessary to reach the equilibrium. According to the
literature [1,12,29-31], the sorption time can be shortened by working in non-equilibrium condition
because the amount of analyte adsorbed from the sample onto the fiber is proportional to the initial
concentration in the sample matrix, if the agitation and the sampling time are held constants amongst
samples. Thus, considering a compromise between the extraction time and the chromatographic
analysis time, an extraction time of 15 minutes was selected for further experiments. This time still
allowed a good, reproducible extraction response for all pesticides while minimizing analysis time.
45
Carbaryl
40
Peak Area (x 100000)
Diazinon
35
Chlorothalonil
30
Malathion
25
Chlorpyrifos
20
Quinalphos
15
Alpha-Endo
10
Profenofos
5
Beta-Endo
0
5 8 10 12 15 20 25 30
Extraction Time (min)
Figure 1: Peak area versus extraction time for 9 investigated pesticide
Effects of Desorption Temperature and Time
The temperature of GC injector and desorption time were tested in order to guarantee the complete
desorption of pesticides to avoid carryover. For the PDMS fiber, temperatures ranging between 200
and 280 oC were tested (selected according to the recommended temperature range indicated by the
manufacturer). High desorption temperature can enhance the process but they can also degrade
analytes. Desorption at 200 and 230 oC was not capable of desorbing completely the analytes; they
were completely removed from the coating at 240 – 280 oC and not much significant differences were
observed within this range of temperature. Hence a temperature of 240 oC was selected since high
temperatures can shorten the coating lifetime and can result in the bleeding of the polymer, causing
problems in the separation and quantification [29].
316
12
Carbaryl
10
Peak Area (x 100000)
Diazinon
8 Chlorothalonil
Malathion
6
Chlorpyrifos
4 Quinalphos
Alpha-Endo
2
Profenofos
0 Beta-Endo
0 2 4 6 8 10 12
Desorption Time (min)
Figure 2: Peak area versus desorption time for 9 investigated pesticide.
Desorption profiles of the pesticides were obtained by plotting the detection response versus different
desorption times, 1 – 10 minutes. Desorption profiles showed that a 6 minute-period was sufficient to
desorb pesticides in the GC injector port (Figure 2); therefore a 7 minute-period was chosen to
guarantee a reproducible desorption.
Effects of Salt Addition
The addition of salts into the samples can modify the extraction efficiency, because the partition
coefficients are partially determined by matrix-analyte-fiber interactions [29]. Pesticides that are more
soluble in water have a lower affinity for the fiber coating. The amount of these analytes extracted by
the fiber can be increased if the solubility of the analytes in water is decreased by adding sodium
chloride to alter the ionic strength [27]. The effect of increasing the ionic strength of the sample was
determined with samples containing no salt, 5, 10, 15, 20 and 25% (w/v) of sodium chloride. Result
showed that the amount of compounds extracted decreased when the salt concentration increased. The
best results were obtained when no salt was added; this could possibly be due to the formation of a
thin layer of salt around the fiber, which decreases the extraction efficiency [29].
Effects of Stirring Speed
The use of a magnetic stirrer allows the control of the stirring speed as well as the mode of stirring,
and hence a cyclic change in stirring direction. The results showed the responses increased if the
stirring speed is increased which agrees with the fact that SPME is a technique based on equilibrium
and that good diffusion through the phases is essential to reach equilibrium faster. Although the
equilibrium time progressively decreases with increasing agitation rate, faster agitation tends to be
uncontrollable and the rotational speed might cause a change in the equilibrium time and poor
measurement precision. A constant gentle stirring speed was selected in this study to increase the rate
of extraction.
Effects of Fiber Depth into the Liner
The effect of the fiber depth into the liner was also checked, and the results showed that peak areas
increased when the depth of the fiber into the injector glass-liner was higher, which is closer to the
column entrance and the center of the hot injector zone.
317
Method performance
After optimization of all the variables considered, the recommended procedure was established as
follows: extraction of 10 mL of water sample containing no salt under magnetic stirring for 15 min at
room temperature using a PDMS, 100 µm fiber coating and subsequent desorption at 240 oC over 7
min. The optimum procedure developed was applied to the extraction of nine pesticides in spiked
water samples. With the selected conditions for the SPME procedure, quality parameters of the
SPME-GC-ECD method such as linearity, limits of detection and quantitation, and recovery were
calculated.
The linearity of the method was tested using a series of aqueous solution (distilled water) in the
difference concentration range (7 levels, three replicates for each level). After plotting the ratio of
analyte peak area relative to that of the peak area of internal standard versus the analyte concentration
to generate the calibration curves, a statistical regression model was applied to obtain the
corresponding values for slope and intercept for each compound. The SPME procedure showed a
linear behavior in the ranges tested with r2 values > 0.9900. Linear ranges and determination
coefficients (r2) obtained for each pesticide are given in Table 2. The loss of linearity observed at
higher concentrations can be justified due to overloading of the SPME fiber capacity.
The detection limit (LOD) was calculated by comparing the signal-to-noise ratio (S/N) of the lowest
detectable concentration to a S/N=3. A S/N of 10 was applied for the calculation of the quantification
limit (LOQ). The results obtained are shown in Table 2.
Table 2: Coefficients (r2), linear range, limits of detection (LOD) and limits of quantification(LOQ) of
the investigated pesticides using the optimized SPME extraction method.
Name R2 Linear Range (mg/kg) LOD (mg/kg) LOQ (mg/kg)

Carbaryl 0.9976 0.2 – 200 0.01 0.05
Diazinon 0.9968 0.05 – 50 0.005 0.02
Chlorothalonil 0.9988 0.02 – 20 0.001 0.005
Malathion 0.9965 0.05 – 50 0.005 0.02
Chlorpyrifos 0.9986 0.005 – 5 0.0005 0.001
Quinalphos 0.9985 0.05 – 50 0.005 0.02
Profenofos 0.9941 0.01 – 10 0.001 0.005
α-Endosulfan 0.9952 0.005 – 5 0.0005 0.001
β-Endosulfan 0.9972 0.005 – 5 0.0005 0.001
Vegetable Samples
The developed method has been applied to the vegetable samples, cucumber and tomato, treated as
described in the Experimental section. From Figure 3, it is clear to show that all the target analytes
were detectable in the sample and appeared completely separated from interfering peaks. The
extraction efficiencies were calculated by comparing the chromatogram (Figures 3) obtained from the
extracts of the spiked samples by SPME with those obtained by direct GC injection of non-extracted
(Figure 4). It is shown that SPME is effective in the extraction of all the pesticides investigated
without absorbing any other unwanted compounds from the samples.
318
Intensity
Internal standard : 5.477 min (2 mg/kg) 19.698 22.231

400000 Carbaryl : 10.136 min (40 mg/kg)
Diazinon : 13.794 min (10 mg/kg)
20.026
Chlorothalonil : 14.835 min (4 mg/kg)
300000 Malathion : 16.667 min (10 mg/kg) 16.667
Chlorpyrifos : 16.888 min (1 mg/kg)
Quinalphos : 18.543 min (10 mg/kg)
Alpha Endosulfan: 19.698 min (1 mg/kg) 18.543
200000 Profenofos : 20.026 min (2 mg/kg) 13.794
Beta Endosulfan : 22.231 min (1 mg/kg) 16.888
14.835
100000
5.477 15.415
10.136
6.100
0 10 15 20
5 min
Figure 3 : Chromatogram on recovery of spiked cucumber and extracted by SPME.
Recovery tests were performed in order to study the accuracy. These tests were based on the addition
of known amounts of standard solution of pesticides to the vegetable samples. The study was carried
out in triplicate at three concentration levels. The peak areas obtained when these samples were
analyzed by the same procedure were compared with the standard calibration curves. Mean recoveries
and RSD obtained in the analysis of fortified cucumber and tomato samples are listed in Table 3. For
fortified cucumber samples, the recoveries were between 53 – 75 % and for the tomato, the recoveries
were between 53 – 82 % with is quite similar to that of the fortified cucumber. The precision
determined using the same conditions was good, with the vast majority yielding relative standard
deviations (RSDs) below 10 %.
Intensity
6.105 19.690
30000
5.484
16.884
20000
16.669 22.226
11.386 20.021
10000
18.551
0
5 10 15 20 min
Figure 4: Chromatogram on recovery of spiked cucumber and direct injection
Table 3: Average recoveries and relative standard deviations (RSDs) from three representative
commodities fortified vegetable samples using the optimized SPME extraction method.
319
Name Cucumber (n=3) Tomato (n=3)
Recovery (%) RSD (%) Recovery (%) RSD (%)
Carbaryl 74.10 8.22 82.17 1.26
Diazinon 53.96 1.43 54.13 1.57
Chlorothalonil 58.26 1.27 56.44 1.00
Malathion 75.03 2.11 71.79 0.47
Chlorpyrifos 55.36 8.93 56.90 9.30
Quinalphos 54.76 2.53 52.57 0.99
Profenofos 56.47 3.94 58.41 5.99
α-Endosulfan 60.09 6.05 53.08 3.83
β-Endosulfan 65.27 4.39 58.40 6.92
CONCLUSION
A fast, simple, solvent free screening method based on a 30 min ultrasonic extraction of a 1.0 g
vegetable samples with distilled water and subsequent SPME of the analytes from the aqueous
solution and detected by gas chromatography with an electron capture detector has been developed for
the determination of nine pesticides. The main experimental parameters affecting the SPME step were
optimized. This method offers very low detection limits for all nine pesticides. The recoveries of the
pesticides studied in cucumber and tomato ranged from 53% to 82% and the RSD were below 10%.
Therefore, the proposed method is applicable in the analysis of pesticides in vegetable matrices.
SPME has been shown to be a simple extraction technique, which has a number of advantages such as
solvent free extraction, simplicity and compatibility with the chromatographic analytical system.
ACKNOWLEDGEMENT
The authors acknowledge the financial support provided by the Universiti Tenaga Nasional and
Ministry of Education to pursue the research work. The authors would like to thank Universiti of
Malaya for providing the opportunity and facilities to undertake the research.
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322
APPLICATION OF SOLID PHASE MICROEXTRACTION (SPME) IN PROFILING
HYDROCARBONS IN OIL SPILL CASES
Zuraidah Abdullah Munir*, Nor’ashikin Saim, Nurul Huda Mamat Ghani
Department of Chemistry, Faculty of Applied Sciences, Universiti Teknologi MARA

40450 UiTM Shah Alam, Selangor, Malaysia.
e-mail: zurai394@salam.uitm.edu.my
Abstract. In environmental forensic, it is extremely important to have a fast and reliable method in identifying sources of
spilled oil and petroleum products. In this study, solid phase microextraction (SPME) method coupled to gas
chromatography-mass spectrometry was developed for the analysis of hydrocarbons in diesel and petroleum contaminated
soil samples. Optimization of SPME parameters such as extraction time, extraction temperature and desorption time, was
performed using 100-µm polydimethylsiloxane (PDMS) fiber. These parameters were studied at three levels by means of a
central composite experimental design and the optimum experimental conditions were determined using response surface
method. The developed SPME method was applied to determine the profiles of hydrocarbons in several oil contaminated
soil sample. The SPME method was also used to study the effects of weathering on the profiles of hydrocarbons in unleaded
gasoline, diesel and kerosene contaminated soil samples. After several days, significant losses of the lighter hydrocarbons
were observed compared to the heavier ones. From these data, SPME method can be used to differentiate possible candidate
sources in oil spill cases.
Keywords: Profile of hydrocarbon, solid phase microextraction (SPME), oil spill
Introduction
Gasoline, diesel and kerosene are all created from crude oil by a variety of refining and distillation
processes. Each product is produced by the combination of multiple individual hydrocarbon
compounds all of which have slightly different vaporization and boiling temperatures. Gasoline is the
combination of many lower boiling range compounds while the middle boiling range compounds are
used in differing proportions to create kerosene and diesel. The profile of hydrocarbons in oil may
hence be used to characterize the oil. This enables the identification of the candidate source of oil spill
cases. In forensic chemistry, ability to identify the sources of an oil spill is very important and
hydrocarbon fingerprinting method is now realized as one of the fastest and reliable method for
identifying the origin of oil spill cases.
In this study, a solid phase microextraction (SPME) method coupled to gas chromatography with
mass spectrometry detector (GC-MSD) was developed for analyzing the profile of several types of
oils. The simplicity of operation, sensitivity, selectivity, portability, and the solvent-free nature of the
SPME method makes it a powerful tool for sample introduction method for gas chromatographic
analyses of organic chemicals [1]. It is based on the enrichment of analytes on a polymer or
adsorbent-coated fused-silica fiber either directly to the sample or its headspace. The extraction
efficiency of SPME technique is dependent on several experimental parameters such as the extraction
time, extraction temperature and desorption time [2,3]. These operating parameters were optimized
using an experimental design approach that consisted of three stages; identifying the factors which
may affect the result of an experiment, designing the experiment so that the effects of uncontrolled
factors are minimized, and using statistical analysis to separate and evaluate the effects of the various
factors involved. The optimized SPME method was used to analyze the effect of weathering on three
types of oils: unleaded gasoline, diesel and kerosene.
323
EXPERIMENTAL
Preparation of spiked sample
Three types of oil (unleaded gasoline, diesel and kerosene) were used in this study. About 1 L of each
type of oil was poured into three separate plots of soil. After 2 weeks, the contaminated soil for each
plot was mixed thoroughly, sieved and stored in an amber bottle at -4 ˚C until analysed.
For the weathering study, three plots of soil measuring 2’ x 1.5’ each were chosen for three types of
oil (unleaded gasoline, diesel and kerosene). Each plot was divided into 20 small sections. About 1 L
of oil sample was poured into each dedicated plot. For each analysis, 100 g of two small sections of
soil were thoroughly mixed and a 5 g sample was placed in a headspace vial (5 mL volume) and
capped for SPME analysis. Soil was analysed 1 day, 2 days, 1 week, 2 weeks and 1 month after oil
spillage.
Solid phase microextraction (SPME)
A 100-µm polydimethylsiloxane (PDMS) fiber (Supelco, Bellafonte, Pennsylvania, USA) was

conditioned in a hot GC injection port at 250 °C for 30-60 min prior to sample extraction. In the
optimization study, the SPME needle was inserted through the septum of the vial and the fiber was
released and exposed to the headspace of the sample at a specified temperature (maintained using a
water bath) for a specified extraction time. The fiber was then withdrawn, SPME needle was removed
from the headspace and immediately injected into the gas chromatography (GC) with a desorption
time of 35 sec. For the weathering study, the sample was extracted at 90 °C for 45 min. The
compounds were then transferred into the GC with desorption time of 35 sec.
Experimental design
Statistical software package Design-Expert 6.0.6, an expert system for the design and analysis of
experiments was purchased from Stat-Ease Inc., Minneapolis. Preliminary work on the optimization
of SPME for the extraction of hydrocarbons in soil using three experimental variables (extraction
temperature, extraction time and desorption time) showed that the desorption time was not significant
within the range 20 to 50 seconds and from the response surface analysis, optimum extraction was
achieved at desorption time of 35 sec. Optimization using central composite design was then focused
on two experimental variables, namely extraction temperature and extraction time. The response
variables selected were the GC area count for several common compounds of unleaded gasoline,
diesel and kerosene. The response variables selected were the gas chromatograph area count for
tridecane, hexadecane, and octadecane. The design matrix of the central composite design is shown in
Table 1. The order of these experiments was randomized to remove any systematic error.
GC-MS analysis
GC-MS were performed on Agilent Technologies 6890 Network GC System with Agilent
Technologies 5973 inert Mass Selective Detector. The flow rates of gases were set to manufacturer’s
specifications. The column used was a HP-5MS fused silica capillary column, 30.0 m x 250 µm I.D
and 0.25 µm capillary thickness. Injections were made in the splitless mode. The temperature
programmed was set at an initial 60 °C for 2 min, followed by an increase by 10 °C min-1 to 200 °C
and held for 15 min. Both the injector temperature and the detector temperature are set at 250 °C.
324
Table 1: Central composite design of the optimization experiment
Run Order Extraction Time (min) Extraction Temperature (ºC)

1 50.0 60.0
2 80.0 60.0
3 50.0 90.0
4 80.0 90.0
5 65.0 75.0
6 65.0 75.0
7 43.8 75.0
8 86.2 75.0
9 65.0 53.8
10 65.0 96.2
11 65.0 46.0
12 65.0 40.0
Optimization of SPME conditions
The results from the central composite design were fitted to a quadratic model. Analysis of variance
(ANOVA) was performed on the design in order to determine which variables (A:extraction time and
B:extraction temperature), if any, had a significant effect on the recovery of each compound. From
the statistical analysis of the experimental design, it was found that extraction temperature was the
important factor influencing the amount of hexadecane and octadecane extracted from soil. The
influence of temperature on the extracted amount of hexadecane and octadecane was further studied
using spiked sample with SPME extraction time of 45 min and desorption time of 35 sec. It was
found that in both cases, the amount extracted increases as the extraction temperature increases, but at
temperature more than 90 °C the amount of hexadecane and octadecane extracted decreases. Based on
these analyses, the optimum operating conditions for SPME were: extraction time 45 min, extraction
temperature 90 °C and desorption time 35 sec.
Profile of hydrocarbons in spiked sample
The optimized SPME method was applied in the extraction of unleaded gasoline, diesel and kerosene
from spiked soil samples.
Gasoline is a complex mixture of hydrocarbons compounds predominantly in the range of C3 – C12. It

is the light distillate product of petroleum containing more lower molecular weight hydrocarbons but
higher fraction of both light hydrocarbons and aromatics. It contains about 41% alkanes with
undecane, dodecane, tridecane and tetradecane as major compounds. In the profile of hydrocarbons in
gasoline [Figure 1(a)], the hydrocarbons were eluted in the range between 5.50 min to 12.00 min.
Kerosene is a light end middle distillate of petroleum. It is composed of hydrocarbons mostly in the
range of C9 – C16. It contains 50.5% aliphatic hydrocarbons and 30.9% naphthenes, the rest being
aromatics hydrocarbons. Major compounds in kerosene are dodecane, tridecane, tetradecane and
pentadecane. The hydrocarbons of kerosene were eluted in the range of 7.00 min to 13.50 min [Figure
1(b)].
325
Diesel is a higher boiling point fraction composed of essentially C10 to C25 aliphatic hydrocarbons. It
has a wide range of polyaromatic hydrocarbons such as naphthalenes and phenanthrenes. Diesel
contains 55% paraffins, 24% aromatics, 12% naphthenes, and 5% olefins. It is similar in chemical
composition to kerosene with the exception of additives. As shown in Figure 1, the hydrocarbon
profile of diesel [Figure 1(b)] is quite similar to that of kerosene [Figure 1(c)], with additional
compounds eluted up to 18.00 min.
5 6
3 7
2
4
Unleaded gasoline Diesel
4 3 8
9
2
1
5
(a) (b)
4
3
Figure 1. GC-MS total ion chromatogram of
(a) unleaded gasoline (b) diesel and (c) kerosene
recovered from spiked soil sample.
Identification of compounds:
Kerosene
1: undecane 2: dodecane 3: tridecane
4: tetradecane 5: pentadecane 6: hexadecane
7: heptadecane 8: octadecane 9: nonadecane
6
1
7
(c)
Effects of weathering on oil hydrocarbon fingerprinting
When oil or petroleum products are accidentally released to the environment, they are immediately
subjected to a wide variety of weathering process that can affect their chemical properties [4]. In the
short term after a spill (hours to days), evaporation is the single most important and dominant
weathering process, in particular for the light petroleum products. The lower boiling points
components tend to volatilize more rapidly than the components of higher boiling points. The
hydrocarbon profile of the spiked soil was obtained using SPME method using the optimized
conditions. Figure 2 shows the profile of unleaded gasoline in soil after differing levels of
weathering. It was found that after 1 week, the more volatile compounds, dodecane and tetradecane
326
were reduced by 46% and 45% respectively while naphthalene 1, 6, 7-trimethyl was reduced by less
than 1%. After 2 weeks, dodecane and tetradecane were reduced by 98% while naphthalene 1, 6, 7-
trimethyl was reduced by 96% and after one month the more volatile compounds (dodecane and
tetradecane) were undetected while the amount of naphthalene
1, 6, 7-trimethyl was further reduced by 97%.
(a)
(b)
(c)
Figure 2. Hydrocarbon fingerprint of unleaded gasoline in soil sampled (a) 2 days, (b) 1 week and (c) 2 weeks
after spillage.
Figure 3 shows the profile of diesel in soil after differing levels of weathering. These chromatograms
clearly showed a decreased in peak intensity due to volatilization. It was found that after 1 week, the
most volatile compound (dodecane) was evaporated by 80% while tetradecane, pentadecane,
hexadecane and heptadecane were reduced by 69%, 34%, 26% and 21% respectively. However, after
one month the more volatile compounds (dodecane, tetradecane and pentadecane) had disappeared
while the amount of hexadecane and heptadecane had reduced to 98%. The least volatile component
in diesel (octadecane) was reduced by 89%.
327
(a)
(b)
(c)
Figure 3. Hydrocarbon fingerprint of diesel in soil sampled (a) 2 days, (b) 1 week and (c) 2 weeks after spillage.
Figure 4 shows the profile of kerosene in soil after differing levels of weathering. After 2 weeks,
most of the compounds in kerosene had almost disappeared. Tetradecane and pentadecane had
disappeared by 96% while heptadecane had 86% evaporated. After 1 month these compounds were
almost insignificant in the soil sample analysed.
CONCLUSIONS
The use of SPME-GC-MSD provides a reliable method for hydrocarbon fingerprinting of oil from
soil. SPME technique, which enables the simultaneous extraction and pre-concentration steps, has
been the method of choice for the analysis of these compounds in soil because it is fast, solvent-less
extraction method, inexpensive and can handle the matrix sample directly. Therefore, it reduces
analysis time (allowing processing a higher number of samples) and avoids loss of analytes. Study of
weathered oils of spiked soil samples using SPME was able to show the change in the profiles of their
chemical properties.
328
(a)
(b)
(c)
Figure 4. Hydrocarbon fingerprint of kerosene in soil sampled (a) 2 days, (b) 1 week and (c) 2 weeks after spillage.
ACKNOWLEDGEMENT
The authors would like to acknowledge financial support from IRDC, Universiti Teknologi MARA
for funding this project (IRDC Project number: 600-IRDC/ST 5/3/789).
References
1. Hook, G.L., Kimm, G.L., Hall, T., and Smith, P.A. (2002) “Solid-phase microextraction (SPME) for rapid field
sampling and analysis by gas chromatography-mass spectrometry (GC-MS). Trends in analytical chemistry, 21,8, 534-
543.
2. Pawliszyn, J. (1997) Solid Phase Microextraction, Theory and Practice. New York, Wiley-VCH Inc., 97.
3. Supelco. (2001) “Control Your “T”s for Better Quantitation with Solid Phase Microextraction (SPME)”. The
Reporter, vol 19.7
4. Wang, Z., Fingas, M.F. (2003) “ Development of oil hydrocarbon fingerprinting and identification techniques”,
Marine Pollution Bulletin. 47. 423-452.
329
SQUARE WAVE CATHODIC STRIPPING VOLTAMMETRIC TECHNIQUE FOR
DETERMINATION OF AFLATOXIN B1 IN GROUND NUT SAMPLE
Mohamad Hadzri Yaacob2, Abdull Rahim Hj. Mohd. Yusoff2 and Rahmalan Ahamad2
1
School of Health Sciences, USM, 16150 Kubang Krian, Kelantan, Malaysia
2
Chemistry Dept., Faculty of Sciences, UTM, 81310 Skudai, Johor, Malaysia
Abstract An electroanalytical method has been developed for the detection and determination of the
2,3,6a,9a-tetrahydro-4-methoxycyclo penta[c] furo[3’,2’:4,5] furo [2,3-h][l] benzopyran-1,11-dione
(aflatoxin B1, AFB1) by a square wave cathodic stripping voltammetric (SWSV) technique on a
hanging mercury drop electrode (HMDE) in aqueous solution with Britton-Robinson Buffer (BRB) at
pH 9.0 as the supporting electrolyte. Effect of instrumental parameters such as accumulation potential
(Eacc), accumulation time (tacc), scan rate (ν), square wave frequency, step potential and pulse
amplitude were examined. The best condition were found to be Eacc of -0.8 V, tacc of 100 s, ν of 3750
mV/s, frequency of 125 Hz, voltage step of 30 mV and pulse amplitude of 50 mV. Calibration curve
is linear in the range of 0.01 to 0.15 µM with a detection limit of 0.125 x 10-8 M. Relative standard
deviation for a replicate measurements of AFB1 (n = 5) with a concentration of 0.01 µM was 0.83%
with a peak potential of -1.30 V (against Ag/AgCl). The recovery values obtained in spiked ground
nut elute sample were 94.00 +/- 0.67 % for 3.0 ppb, 91.22 +/- 1.56 % for 9 ppb and 92.56 +/- 2.00 %
for 15.0 ppb of AFB1. The method was applied for determination of the AFB1 in ground nut samples
after extraction and clean-up steps. The results were compared with that obtained by high
performance liquid chromatography (HPLC) technique.
Abstrak Satu kaedah elektroanalisis telah dibangunkan untuk mengesan dan menentukan 2,3,6a,9a-
tetrahydro-4-methoxycyclo penta[c] furo[3’,2’:4,5] furo [2,3-h][l] benzopyran-1,11-dione (aflatoxin
B1, AFB1) menggunakan teknik voltammetri perlucutan katodik denyut pembeza di atas elektrod
titisan raksa tergantung (HMDE) di dalam larutan akuas dengan larutan penimbal Britton-Robinson
(BRB) pada pH 9.0 bertindak sebagai larutan penyokong. Kesan parameter peralatan seperti
keupayaan pengumpulan (Eacc), masa pengumpulan (tacc), kadar imbasan (ν), frekuensi gelombang
bersegi, kenaikan keupayaan dan amplitud denyut telah dikaji. Keadaan terbaik yang diperolehi
adalah Eacc; -0.8 V, tacc; 100 s, ν; 3750 mV/s, frekuensi; 125 Hz, kenaikan keupayaan; 30 mV dan
amplitud denyut; 50 mV. Keluk kalibrasi adalah linear pada julat di antara 0.01 ke 0.15 µM dengan
had pengesan pada 0.125 x 10-8 M. Sisihan piawai relatif untuk 5 kali pengukuran AFB1 dengan
kepekatan 0.01 µM ialah 0.83 %. Nilai perolehan semula di dalam larutan elusi sampel kacang yang
disuntik dengan 3.0 ppb, 9.0 ppb dan 15.0 ppb AFB1 adalah 94.00 +/- 0.67 %, 91.22 +/- 1.56 % dan
92.56 +/- 2.00 % masing-masingnya. Kaedah ini telah digunakan untuk menentukan kandungan AFB1
di dalam sampel kacang tanah selepas proses pengekstraksian dan pembersihan dijalankan. Keputusan
yang diperolehi telah dibanding dengan keputusan dari kaedah kromatografi cecair berprestasi tinggi.
Keywords: square wave stripping voltammetry, HMDE, aflatoxin B1, ground nut
Introduction
Aflatoxins (AF), the mycotoxin produced mainly by Aspergillus flavus and parasiticus and
display strong carcinogenicity [1]. They are dangerous food and contaminants and represent a
worldwide threat to public health. AFB1, B2, G1 and G2 and their metabolites M1 and M2 are the
most common, and of these, AFB1 and AFG1 are observed most frequently in foodstuffs [2]. Of
these, researches have shown AFB1 (Fig 1) exhibits most toxic [3] with the order of toxicity is AFB1
2
Corresponding address:
Chemistry Dept, Faculty of Science, UTM,
81310 Skudai, Johor, Malaysia
tel: 07-553 4492
330
> AFG1 > AFB2 > AFG2; indicating that the terminal furan moiety of AFB1 is a critical point for
determining the degree of biological activity of this group of mycotoxins [4]. Many countries
including Malaysia have stringent regulatory demands on the level of aflatoxins permitted in imported
and traded commodities.
O
O
O O
Figure 1 Chemical structure of AFB1
One of the foodstuffs which is most occurrence of AFB1 is ground nut. In Malaysia, the
AFB1 level in peanut is regulated with maximum level that cannot be greater than 15.0 ppb [5].
Several analytical techniques for quantitative determination of the AFB1 in ground nut have been
proposed such as thin layer chromatograhpy [6], high performance liquid chromatography [7,8,9] and
an enzyme-linked immunosorbent assay, ELISA [10], All these methods, however, require specialist
equipment operated by skilled personal, expensive instruments and high maintenance cost [11].
Due to all these reasons, a voltammetric technique which is fast, accurate and require low cost
equipment [12,13] is proposed. A square wave cathodic stripping voltammetry (SWCV) which is
presented in this paper is one of the voltammetric technique that in particularly, has a few advantages
compared to other voltammetric technique such as high speed, increased analytical sensitivity and
relative insensitivity to the presence of dissolved oxygen [14]. Previous experiment using cyclic
voltammetric technique showed that AFB1 reduced at mercury electrode and the reaction is totally
irreversible [15]. This work aimed to study and develop a SWSV method for determination of AFB1
at trace levels and to determine this aflatoxin in ground nut samples. No such report has been
published regarding this experiment until now.
Experiment
Apparatus
Square-wave voltammograms were obtained with Metrohm 693 VA Processor coupled with a
Metrohm 694 VA stand. Three electrode system was used, consisting of a hanging mercury drop
electrode (HMDE) as the working electrode, Ag/AgCl/3 M KCl as the reference electrode and a
platinum wire as the auxiliary electrode. A 20 ml capacity measuring cell was used for placing
supporting electrolyte and sample analytes. All measurements were carried out at room temperature.
All pH measurements were made with Cyberscan pH meter, calibrated with standard buffers at room
temperature.
331
Reagents
AFB1 standard (1 mg per bottle) was purchased from Sigma Co. and was used without further
purification. Stock solution (10 ppm or 3.21 x 10-5 M) in benzene:acetonitrile (98:2) was prepared
and stored in the dark at 14 0C. The diluted solution were prepared daily by using certain volume of
stock solution, degassed by nitrogen until dryness and redissolved in Britton-Robinson buffer (BRB)
solution at pH 9.0. Britton-Robinson buffer (BRB) solution was prepared from a stock solution 0.04
M phosphoric (Merck), boric (Merck) and acetic (Merck) acids; and by adding 1.0 M sodium
hydroxide (Merck) up to pH of 9.0. All solutions were prepared in double distilled dionised water (~
18M Ω cm). All chemicals were of analytical grade reagents.
Procedure
For voltammetric experiments, 10 ml of BRB solution with pH 9.0 was placed in a

voltammetric cell, through which a nitrogen stream was passed for 600 s before recording the
voltammogram. The selected Eacc = - 800 mV was applied during the tacc = 100 s while the solution
was kept under stirring. After the accumulation time had elapsed, stirring was stopped and the
selected accumulation potential was kept on mercury drop for a rest time (tr = 10 s), after which a
potential scan was performed between -1.00 V as initial potential (Ei) and finished at -1.400 V as the
inal potential (Ef ) by SWSV technique.
Procedure for the determination in ground nut samples
AFB1 was extracted according to the standard procedure developed by Chemistry

Department, Penang Branch, Ministry Of Science, Technology and Innovation, Malaysia [16]. 1ml
of the final solution from extraction and clean-up steps in chloroform was pipette into amber bottle,
degassed with nitrogen and redissolved in 1ml of BRB solution. 200 ul of this solution was spiked
into 10 ml supporting electrolyte in volumetric cell. After that, the general procedure was applied and
voltammogram of sample was recorded. This experiment was followed by a standard addition of 10
ppb of AFB1 standard addition and voltammogram of sample with AFB1 standard was recorded.
Result and discussion
Using previous differential pulse stripping voltammetry (DPCSV) optimum parameters [17], SWSV
was run to determine 0.1 µM AFB1 in BRB pH 9.0. It gave a single reduction signal with peak height
(Ip) of 250 nA at peak potential (Ep) of -1.26 V (against Ag/AgCl). The SWSV voltammograms shows
improved Ip compared to that obtained by DPCSV where the Ip was increased almost 4 times as
shown in Figure 2.
The effect of the pH of the BRB on the stripping was studied in a pH range of 6 – 13 (Figure
3). The Ip increased slowly with increasing pH up to 8.0 followed with sharply increased for pH 8.0
to 9.0, then decreased in pH 10.0 and continuously decreased in a range of 10 – 13. Thus pH 9.0 was
chosen for the analysis. This result is in aggrement with that found by Smyth et al. (1979) when they
performed polarographic study of AFB1 [18].
Figure 4 shows a dependence of the Ep on pH. Shifting of the Ep towards the negative
direction at higher pH implies that the reduction process takes up hydrogen ions [19]. A double bond
in aromatic ring conjugates with ketone group, in general, undergoes a reduction at mercury electrode.
The suggested mechanism of this reaction in BRB pH 9.0 is illustrated in Figure 5 as reported by
Smyth et al. (1979) [20].
332
Figure 2 Voltammograms of 0.1 uM AFB1 obtained by (a) DPCSV and (b) SWSV techniques.
Experimental condition; for DPCSV: Ei = -1.0 V, Ef = -1.4 V, Eacc = -0.6 V, tacc = 80 s, υ = 50 mV/s
and pulse amplitude = 80 mV and for SWSV: Ei = -1.0 V, Ef = -1.4 V, Eacc = -0.6 V, tacc = 80 s,
frequency = 50 Hz, voltage step = 0.02, amplitude = 50 mV and υ = 1000 mV/s.
300
250
200
I p (nA)
150
100
50
0
5 6 7 8 9 10 11 12 13 14
pH of BRB
Figure 3 Influence of pH of BRB on the Ip of 0.10 uM AFB1 using SWSV technique. The SWSV
instrumental parameters are the same as in Figure 2.0.
1.34
1.32
1.3
1.28
Ep (-V)
1.26
1.24
1.22
1.2
1.18
5 6 7 8 9 10 11
pH
Figure 4 Relationship between Ep of AFB1 with pH of BRB
333
O O
O O
O
O
2e-
2 2H + 2
CH3
CH3
2 H 2O
D im e r + 2 OH-
Figure 5 Mechanism for reduction of AFB1 at mercury electrode in BRB pH 9.0
Optimisation of condition for the stripping analysis
The voltammetric determination of analytes at trace levels normally involves very small
current response. For that reason it is important to optimise all those parameters which may have an
influence on the measured current. The effect of Ei , Eacc , tacc, frequency, voltage step and amplitude
were studied. SWSV technique was used with stirring. For this study, 0.1 uM of AFB1 was spiked
into supporting electrolyte.
A study of the influence of Ei showing a peak height (Ip) was obtained for Ei = -1.0 V (Figure
6). The Ip was slowly decreased for Ei more negative than -1.0 V. This value was chosen for
subsequent studies further optimisation step. Influence of Eacc to Ip of AFB1 was investigated where
the Eacc was varied between 0 to -1.4 V. The maximum value of Ip obtained at -0.8 V (366 nA) as
shown in Figure 7. This value was selected for subsequent experiments.
The dependence of Ip on tacc was studied where tacc was varied from 0 to 160 s. The result is
shown in Figure 8 which reveals that there is linearly up to 100s according to equation y = 3.8557x +
21.383 (n=6) with R2 = 0.9936, then it increased rather slowly leveling off at about 140 s. At 160 s, Ip
decreased which may be due to the electrode saturation [21]. Thus, 100 s was appeared to be an
optimum tacc for the pre-concentration prior to stripping.
300
250
200
Ip (nA)
150
100
50
0
0.2 0.4 0.6 0.8 1 1.2 1.4
Ei (-V)
Figure 6 Effect of Ei on the Ip of 0.1 uM AFB1 in BRB pH 9.0
334
400
350
300
250
Ip (nA)
200
150
100
50
0
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6
Eacc (-V)
Figure 7 Effect of Eacc on the Ip of 0.1 uM AFB1 in BRB pH 9.0
450
400
350
300
Ip (nA)
250
200
150
100
50
0
0 40 80 120 160 200
T acc (s)
Figure 8 Effect of tacc on the Ip of 0.1 uM AFB1 in BRB pH 9.0
For other instrumental condition such as square wave frequency, step potential and pulse
amplitude were examined, varying one of them and maintaining constant others. The variable ranges
were: 25 to 125 Hz for the frequency, 0.01 to 0.04 for the voltage step and 25 – 100 mV for pulse
amplitude. Generally, the Ip increase by increasing all of these instrumental parameters [21]. At
higher potential values the peak width increase; at higher frequency values the current background
increases. Finally, the condition selected were: frequency = 125 Hz, voltage step = 0.03 V and pulse
amplitude = 50 mV (Figures 9 to 11). Under optimised parameters, Ip of AFB1 was 956 nA which is
16 times higher compared to that obtained by DPCSV. Figure 12 shows voltammograms of 0.1 uM
AFB1 obtained by SWSV and DPCSV techniques.
335
1000
800
Ip (nA) 600
400
200
0
0 25 50 75 100 125 150
Fre que ncy (Hz)
Figure 9 Dependence of the Ip of AFB1 on SWSV frequency
1200
1000
800
Ip (nA)
600
400
200
0
0 0.01 0.02 0.03 0.04 0.05
Voltage step (V)
Figure 10 Dependence of the Ip of AFB1 on SWSV voltage step
1200
1000
800
Ip (nA)
600
400
200
0
0 0.025 0.05 0.075 0.1 0.125
Amplitude (V)
Figure 11 Dependence of the Ip of AFB1 on SWSV pulse amplitude
336
Figure 12 Voltammograms of AFB1 obtained by (a) DPCSV and (b) SWSV techniques in BRB pH
9.0.
Voltammetric determination of AFB1 and analytical characteristics of the method
Using the selected conditions already mentioned, a study was made of the relationship
between Ip and concentration. There is a linear relationship in the concentration range 0.01 to 0.15
uM as shown in Figure 13. Limit of detection (LOD) was 0.389 ppb (0.125 x 10-8 M) which was
determined by standard addition of low concentration of AFB1 until obtaining the sample response
that is significantly difference from blank response [22]. Relative standard deviation (RSD) of the
analytical signals at several measurement (n=5) of 0.10 uM AFB1 was 0.83%.
1400
1200
1000
Ip(nA)
800
600
y = 62.373x + 246.15
400
R2 = 0.9956
200
0
0 5 10 15 20
[AFB1] / 10-8 M
Figure 13 Calibration plot of AFB1 in BRB pH 9.0 obtained by SWSV technique.

Determination of AFB1 in ground nut samples
The proposed method was applied to the analysis of the AFB1 in ground nut samples.
Recovery studies were performed by spiked with difference concentration levels of AFB1 standard
into eluate of ground nut sample. In this case the concentrations used were 3.0 ppb (0.963 x 10-8 M),
9.0 ppb (2.889 x 10-8 M) and 15.0 ppb (4.815 x 10-8 M). The results of these studies are shown in
Table 1. For analysis of AFB1 in ground nut samples, the standard addition method was used in order
to eliminate the matrix effects. Figure 14 show voltammograms of real sample together with spiked
AFB1 standard. Table 2 listed the content of AFB1 in 6 samples obtained by proposed technique
compared with that obtained by HPLC. The results show that there are no significant different of
AFB1 content obtained by both techniques.
337
Table 1 Percent recovery of AFB1 spiked in real samples (n=3)
Amount added (ppb) Amount found (ppb) Recovery (%) n=3
3.00 2.82 +/- 0.02 94.00 +/- 0.67
9.00 8.21 +/- 0.14 91.22 +/- 1.56
15.00 13.88 +/- 0.30 92.53 +/- 2.00
Figure 14 SWS voltammograms of real sample (b) and spiked AFB1 (c) obtained in BRB pH 9.0 as
the supporting electrolyte (a)
Table 2 AFB1 content in ground nut samples by proposed technique compared with those obtained
by HPLC.
No of sample AFB1 content in real sample
By SWSV By HPLC
1 ND ND
2 ND ND
3 ND ND
4 9.21 8.25
5 13.92 14.34
6 36.00 36.00
338
Conclusion
A SWSV technique was successfully developed for the determination of AFB1 in ground nut
as an alternative method for determination of AFB1 which is sensitive, accurate and fast technique.
The results are not significant different with that obtained by accepted technique used for routine
analysis of AFB1.
Acknowledgement
One of the author would like to thank University Science Malaysia for approving his study
leave and financial support to carry out his further study at Chemistry Dept., UTM. We also indebt to
UTM for short term grant (Vot No 75152/2004) and to Chemistry Department, Penang Branch,
Ministry of Science, Technology and Innovation for their help in analysis of aflatoxin in ground nut
samples using HPLC technique.
References
1. Akiyama, H. Goda, Y. Tanaka, T and Toyoda, M. (2001). “Determination of aflatoxinsB1,

B2, G1 and G2 in spices using a multifunctional column clean-up” J. Of Chromatography A,
932. 153-157.
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Lisan - Prosiding Simposium Kimia Analisis Malaysia Ke-18, 2005

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SKAM 18

SIMPOSIUM KIMIA ANALISIS MALAYSIA

Sains Analisis Cemerlang Negara Terbilang

SESI KERTAS KERJA LISAN

1B Kimia Tak Organik dan Pemangkinan I

1B-2 CHARACTERIZATION OF NOVEL ORGANOTIN(IV) COMPLEXES WITH

SESI KERTAS KERJA LISAN

1D-2 TOXICITY AND ANTITERMITE ACTIVITIES OF THE ESSENTIAL OILS

SESI KERTAS KERJA LISAN

2C-6 ADSORPTION OF CHROMATE ANION FROM AQUEOUS SOLUTION BY

SESI KERTAS KERJA LISAN

3A Alam Sekitar III

SESI KERTAS KERJA LISAN

3D Kimia Tak Organik/Pemangkinan II

SESI KERTAS KERJA LISAN

SESI KERTAS KERJA LISAN

SESI KERTAS KERJA LISAN

5C Tak Org/Pemangkinan III

5C-5 Bi-Fe PROMOTED VANADIUM PHOSPHATE (VPO) CATALYST FOR

SESI KERTAS KERJA LISAN

ANALISIS UNSUR DAN KERADIOAKTIFAN DALAM SAMPEL SEDIMEN TASIK CHINI,

Keywords: Sedimen, radionuklid tabii, logam berat.

Laut Kenawar Pulau Besar

Laut Batu Busuk

Laut Serodong Laut Jemberau

PETA TASIK CHINI

RAJAH 1 : Peta lokasi persampelan sediment Tasik Chini

Penyediaan dan Rawatan Sampel.

Penyediaan Sampel untuk Teknik Pendarfluor Sinar-X

Penyediaan Sampel untuk Teknik Analisis Keradioaktifan.

Sebanyak 15 sampel yang diketahui berat masing-masing dimasukkan kedalam botol

Hasil Dan Perbincangan

Keputusan Analisis Unsur

Analisis keradioaktifan tabii

Selain kandungan unsur, analisis keradioaktifan juga telah ditentukan menggunakan

Jadual 1 : Keputusan analisis unsur bagi setiap stesen persampelan

Stesen Kepekatan (ppm)

Jadual 2: Kepekatan radionuklid tabii sampelsediemn Tasik Chini

PYROLYSIS AND LIQUEFACTION OF ACETONE AND MIXED

Fuel Combustion Research Laboratory

Keywords: Swelling, liquefaction, pyrolysis, coal conversion, oil yield.

Materials And Methods

Solvent Swelling Procedure

Coal Liquefaction Experiments

Table 1: Characteristics of MB coal sample. Table 2: Results of swelling ratio (Q),

activation energy (Ei, kJ/mol) and

Sulphur 0.5 Ace-sw 1.3 230.5 40.4

Calorific value = 24.60 MJ/kg Tet-sw = Tetralin-swelled;

Results And Discussion

Effect of Swelling On Coal by Pure and Mixed Solvents During Pyrolysis

Figure 1: The DTG curves of untreated, acetone-swelled and mixed

Effect of Swelling on Coal Conversion and Oil Yield

Table 3: Liquefaction results of untreated, acetone- and mixed acetone/tetralin-swelled coal.

Temp. (°C) % Coal conversion % Oil+gas % Asphaltene %

Unt. = Untreated; Ace. = Acetone-swelled; Mix. = Mixed acetone/tetralin-swelled.

KANDUNGAN LOGAM BERAT DAN RADIONUKLID TABII DALAM IKAN, AIR,

Keywords: Heavy metal, natural radiobuclides, ex-mining lake, fish, sedimen

Bahan dan kaedah.

c. Perawatan dan analisis sampel

L1, L2 dan L3 Tasik bekas lombong.

RAJAH 3.1 Lokasi tasik kajian

Hasil dan perbincangan

Jadual 1. Pelbagai parameter air dan sedimen tasik