Pakistan Journal of Scientific and Industrial Research Series A: Physical Sciences

EDITORIAL BOARD Dr. Shahzad Alam Chief Editor

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Pakistan Journal of Scientific and Industrial Research ( PJSIR ) was started in 1958 to disseminate research results based on utilization of locally available raw materials leading to production of goods to cater to the national requirements and to promote S&T in the country. Over the past 58 years, the journal conveys high quality original research results in both basic and applied research in Pakistan. A great number of major achievements in Pakistan were first disseminated to the outside world through PJSIR.

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From 54th Volume in 2011, it has been bifurcated into Series A: Physical Sciences & Series B: Biological Sciences. Each series appears three times in a year as follows:

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Due to many global issues, we are encouraging contributions from scientists and researchers from all across the globe with the sole purpose of serving scientific community worldwide on the whole and particularly for our region and third world countries. Pakistan Journal of Scientific and Industrial Research Series A: Physical Sciences Vol. 59, No. 2, May-June, 2016

Contents

(E)-N'-(2,4-dihydroxybenzylidene)nicotinohydrazide and its Metal Complexes: Synthesis, Characterisation and Antitubercular Activity Kehinde Olurotimi Ogunniran, Joseph Adeyemi Adekoya, Cyril Ehi-Eromosele, Olayinka Oyewale Ajani, Akinlolu Kayode and Tadigoppula Narender 63

Quantification of Cr(VI)-Thymoquinone Complex Using Cyclic Voltammetry Farah Kishwar, Khalid Mohammed Khan, Rubina Perween, Anila Anwar and Nasir Akhtar 76

Effect of Processing on Physicochemical Properties and Fatty Acid Composition of Fluted Pumpkin (Telfairia occidentalis) Seed Oil Jacob Olabode Alademeyin and Jacob Olalekan Arawande 83

Liberation Studies of Padhrar Coal by Using Fractionation Method, XRD Analysis and Megascopic and Microscopic Techniques Muhammad Shahzad, Zulfiqar Ali, Yasir Majeed, Zaka Emad, Muhammad Aaqib and Bilal Adeel 90

Modeling the Land Suitability Using GIS and AHP for Cotton Cultivation in Punjab, Pakistan Nabila Naz and Haroon Rasheed 96

Quality Variation Minimizer: A New Approach for Quality Improvement in textile industry Muhammad Amin, Muhammad Amanullah and Atif Akbar 109

Effect of Different Processing Stages on the Crystallinity % and Tensile Strength of 100% Cotton Fabric Zahid Hussain, Muhammad Qamar Tusief , Sharjeel Abid, Muhammad Tauseef Khawer, Nabeel Amin and Mudassar Abbas 114

Short Communication

Biosorption Characteristics of Water Hyacinth (Eichhornia crassipes) in the Removal of Nickel (II) Ion under Isothermal Condition Chidi Obi and Sylvester Eigbiremonlen 118 Pak. j. sci. ind. res. Ser. A: phys. sci. 2016 59(2) 63-75

(E)-N'-(2, 4-dihydroxybenzylidene)nicotinohydrazide and its Metal Complexes: Synthesis, Characterisation and Antitubercular Activity Kehinde Olurotimi Ogunnirana*, Joseph Adeyemi Adekoyaa, Cyril Ehi-Eromoselea, Olayinka Oyewale Ajania, Akinlolu Kayodea and Tadigoppula Narenderb aDepartment of Chemistry, College of Science and Technology, Covenant University, PMB, 1023, Ota, Ogun State, Nigeria bMedicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, Lucknow, India

(received April 15, 2015; revised August 6, 2015; accepted August 7, 2015)

Abstract. Nicotinic acid hydrazide and 2,4-dihydoxylbenzaldehyde were condensed at 20 °C to form an 1 acylhydrazone (H3L ) with ONO coordination pattern. The structure of the acylhydrazone was elucidated by using CHN analyzer, ESI mass spectrometry, IR, 1H NMR, 13C NMR and 2D NMR such as COSY and HSQC. Thereafter, five novel metal complexes [Mn(II), Fe(II), Pt(II) Zn(II) and Pd(II)] of the hydrazone ligand were synthesized and their structural characterization were achieved by several physicochemical methods namely: elemental analysis, electronic spectra, infrared, EPR, molar conductivity and powder X-ray diffraction studies. An octahedral geometry was suggested for both Pd(II) and Zn(II) complexes while both Mn(II) and Fe(II) complexes conformed with tetrahedral pyramidal. However, Pt(II) complex agreed with tetrahedral geometry. In vitro antitubercular activity study of the ligand and the metal complexes were evaluated against Mycobacterium tuberculosis, H37Rv, by using micro-diluted method. The results obtained revealed that (PtL1) (MIC = 0.56 mg/mL), (ZnL1) (MIC = 0.61 mg/mL), (MnL1) (MIC = 0.71 mg/mL) and (FeL1) (MIC = 0.82 mg/mL), exhibited a significant activity when compared with first line drugs such as isoniazid (INH) (MIC = 0.9 mg/mL). H3L1 exhibited lesser antitubercular activity with MIC value of 1.02 mg/mL. However, the metal complexes displayed higher cytotoxicity but were found to be non- significant different (P > 0.05) to isoniazid drug. Keywords: hydrazones, metal complexes, electron spin resonance, thermogravimetry, powder X-ray diffraction, antitubercular agents

Introduction tubercle bacillus after initial infection. As a consequence, Human tuberculosis (TB) has re-emerged with devastating the present level of tuberculosis comprises both indivi- consequences on global public health and it is currently duals with “new” exogenous infections and those with one of the most widespread infectious diseases. In a reactivation of “old” endogenous disease (De Backer addition, it is the leading cause of death due to a single et al., 2006). In terms of absolute number of TB cases, infectious agent among human adults in the world (Jenkins 22 countries of the world have the highest TB burden et al., 2011). Mycobacterium tuberculosis is one of the with at least 270 cases per 100,000 populations. Among most harmful pathogens of mankind, infecting one- the top five ranking countries are India, China, Indonesia, third of the global population and claiming two million South Africa and Nigeria (Harper, 2007; Laughon, lives every year (Stewart et al., 2003). Tuberculosis 2007). The situation has become more deplorable than spreads by aerosols from patients with pulmonary disease it appeared as 0.5 million new cases due to multidrug- (Phillip and Graham, 2004). Mycobacterial infection resistant (MDR) TB were recorded in 2010 (WHO, has increased in number worldwide due to a global 2013). The alarming estimates exposes that 0.22 billion increase in the number of patients with HIV infection people may acquire TB and 79 million could die due and AIDS disease, increase in number of elderly patients to TB by the year 2030. and the emergence of resistant tuberculosis. Tuberculosis Effective TB treatment is difficult, due to the unusual arises in two different ways: either from a recent infection structure and chemical composition of the Mycobacterium with M. tuberculosis or from the reactivation of dormant cell wall, which makes many antibiotics ineffective and *Author for correspondence; hinders the entry of drugs (Jia et al., 2005). TB disease E-mail: [email protected] can be treated by taking several drugs for 6 to 9 months

63 64 Kehinde Olurotimi Ogunniran et al. which includes the first 2 months of isoniazid, rifampicin, in the range of 200-700 nm. The molar conductivities pyrazinamide, and ethambutol in the intensive phase of the metal complexes at ambient temperature in DMF and after that period isoniazid and rifampicin in the solution (10-3 M) were measured using systronics-304 continuous phase (Jindani et al., 2004). Most of the drugs conductivity meter at Chemistry Department, Covenant in the current tuberculosis regime result from the research University, Canaan Land, Sango-Ota, Ogun State, Nigeria. performed over 50 years ago (Sacchettini et al., 2008). The magnetic measurements of paramagnetic metal With the global emergence of multidrug-resistant complexes in powder form were measured at room tuberculosis (MDR-TB) and extensively drug-resistant temperature by using vibrating susceptibity magnometer tuberculosis (XDR-TB) there is an urgent need to develop (PAR 155) with magnetic field of -10 to +10 kOe at new anti-mycobacterial agents. Instrumentation Center, Indian Institute of Technology, Roorkee, India. TGA/DTA thermograph of the metal In the search for new compounds, isoniazid derivatives complexes were obtained by heating the complexes at have been found to possess potential tuberculostatic the rate of 10 °C/min under inert atmosphere by using activities (Aboul-Fadl et al., 2011). Hydrazones are thermogravimetric analyzer TGA Q500 V20.8 Build important compounds for drug design, as possible 34 model at Indian Institute of Science and Technology, ligands for metal complexes, organocatalysis and also Hyderabad, India. The EPR spectra of the metal for the syntheses of heterocyclic compounds. These complexes at 77 K were recorded on a Varian E-112 compounds have interesting biological properties, spectrometer using TCNE as the standard, with 100 such as anti-inflammatory, analgesic, anticonvulsant, KHz modulation frequency, modulation amplitude 2 G antituberculous, antitumor, anti-HIV and antimicrobial and 9.1 GHz microwave frequency at sophisticated activity (Nataliya et al., 2010). In the present work, an analytical instruments facility, Indian Institute of acylhydrazone obtained in the reaction of nicotinic Technology, Bombay, India. Powder X-ray diffraction hydrazide with 2,4-dihydroxybenzaldehyde and its data for one of the metal complexes were collected by Mn(II), Fe(II), Pt(II) Zn(II) and Pd(II) complexes were using Powder X-ray diffractometer at Instrumentation characterised and tested for their antimycobacterial center, Indian Institute of Technology, Roorkee, India. activity. The presence of the hydrazine pharmacophore Powder diffraction data was recorded on a Bruker AXS in these compounds is expected to contribute to high D8 Advance diffractometer operating in the q:q mode, antimycobacterial activity. equipped with a secondary beam graphite monochromator, a Na(Tl)I scintillation counter, and pulse-height amplifier Materials and Methods discrimination. CuKa radiation (l=1.5418 Å) was used. Measurements. All the chemicals and solvents used The X-ray generator and diffractometer settings were were reagent grade and used without further treatment 40 kV, 40 mA, DS 0.5°, AS 0.5°, and RS 0.1 mm. unless otherwise noted. Nicotinic acid hydrazide and Experimental conditions were step scan mode, with 2,4-dihydroxylbenzaldehyde were purchased from 5 < q <105°, Dq = 0.02°, and t = 30 s/step. Silicon NBS Sigma-Aldrich. ESI-MS spectrum of the ligand was 640b was used as an external standard. obtained using Agilent 6520 Q-TOF mass spectrometer. Synthesis of (E)-N¢-(2,4-dihydroxybenzylidene) The % of carbon, nitrogen and hydrogen in the synthesized nicotinohydrazide. The synthetic methods previously hydrazones and metal complexes were determined by described by Cui et al. (2012) were modified and adopted. using Vario EL CHNS analyzer. 1H, 13C and 2D NMR The nicotinic acid hydrazide (10 mmol, 1.37 g) was (COSY, HSQC and DEPT 135) spectra of the hydrazone were recorded by using Bruker AMX 300 FT-NMR dissolved in 20 mL of absolute ethanol by heating gently spectrometer with DMSO-d6 at sophisticated analytical on water bath. The solution obtained was mixed with instruments facility, Central Drug Research Institute, ethanolic solution of 2,4-dihydroxylbenzaldehyde (10 Lucknow, India. The infrared spectroscopy of the hydrazone mmol ) in a round bottom flask. The mixture was stirred and the metal complexes were recorded on Perkin-Elmer at 20 °C for 6 h after which it was allowed to stand at RX-1 Fourier Transform Infrared Spectrometer using ambient temperature for 24 h. The precipitate formed KBr pellets in the range of 4000-400 cm-1. The electronic was filtered and washed several times with ethanol. data of the hydrazone and the metal complexes were The precipitate was recrystallized in mixture of methanol obtained in methanol/DMSO by using Perkin Elmer and chloroform (1:1). It was filtered off, washed with Spectro UV-visible Double Beam UVD spectrometer ether and dried in vacuum. The purity of the hydrazone Antitubercular Activity of Acylhydrazone 65 was confirmed by single spot displayed by TLC using bottom flask, after which 10 mL of distilled water was methanol: chloroform (2:8) mixture. added. The mixture was stirred at 60 °C for 30 min before adding ethanolic solution of H L1 (10 mmol, Yield 1.82 g (70.8%); light pink solid; mp: 273-275 °C; 3 2.57 g) in 10 mL of absolute ethanol. The mixture was R = 0.83 (CHCl /CH OH, 4:1, at RT.). 1H-NMR f 3 3 then refluxed for 3 h. The green precipitate formed was (DMSO-d6) d: 12.16 (s, 1H, NH), 11.36 (s,1H, Ar- 1 1 filtered after cooling the solution in ice blocks, washed OH(2)), 10.06 (s, H, Ar-OH(3)), 9.10 (s, H, H(4)), 8.80 1 1 with 30 mL of cold ethanol and dried over P4O10 in (d, J= 3.84 Hz, H, H(5)), 8.55 (s, H, H-CN), 8.30 ( dt, 1 vacuo. J = 7.9 Hz, H, H(7)), 7.60 (dd, J1 = 4.80 Hz, J2 = 7.9 1 J 1 1 Hz, H, H(8)), 7.39 (d, = 8.43 Hz, H, H(9)) 6.42 (dd, [Zn(H3L )(CH3COO)2H2O] (4). 10 mmol (1.83 g) of J J 1 J 1 1 = 2.1 Hz, 2 = 8.43 Hz. H, H(10)) 6.36 (d, = 2. Hz, Zn(CH3COO)2 was dissolved in 20 mL of mixture of 1 13 H, H(11)) ppm. C-NMR (DMSO-d6) d: 161.3 (C1), absolute ethanol and distilled water (1:1). The solution 161.2 (C2), 159.8 (CO), 159.3 (C4), 149.9 (C5), 148.8 obtained was added gradually to ethanolic solution of 1 (NCH), 135.6 (C7), 131.6 (C8), 129.1 (C9), 123.9 H3L (10 mmol in 20 mL of ethanol) in a round bottom (C10), 110.7 (C11), 108.1 (C12), 102.9 (C13) ppm. IR flask after which two drops of TEA were added. The (KBr) cm-1: 3432 (ArOH), 3158 (NH), 1639 (C=O), solution was refluxed at 80 °C for 4 h. The yellow preci- 1508 (C=N), 1466 (N-N), 1353 (C-O), 1165(C-N). MS pitate formed was allowed to cool to ambient temperature (ESI+): in m/z: 258.0 [M + H]+. Anal. calcd. for after which it was filtered, washed with 30 mL of ethanol

C14H12N3O3 (256.26): C, 65.62; H, 4.68; N, 16.39. and then with 10 mL of ether. The precipitate was then Found: C, 65.89; H, 4.53; N, 16.16 (Scheme 1). dried by using vacuum rotary evaporator. 1 [Pd(H3L )Cl] (5). 10 mmol (1.77 g) of PdCl2 was 1 HN2 HO OH HN O dissolved in 10 mL of DMF while 10 mmol of H L HO 3 EtOH, 20 °C N N was dissolved in 10 mL of absolute ethanol. The solutions O + O N 6 h H H were mixed together in a round bottom flask and refluxed OH H N at 80 °C for 6 h after which the solution was left at room temperature for 48 h. The dark brown precipitate Scheme 1. Synthetic pathway for H L1. 3 formed was filtered and washed with cold ethanol and

then dried over P4O10 in vacuo. Preparation of Mn(II), Fe(II), Pt(II), Zn(II) and Antimycobacterial activity study. MIC determination. Pd(II) complexes of (E)-N¢-(2,4-dihydroxybenzyli- All the compounds were screened for their in vitro dene) nicotinohydrazide (H L1). [Mn(H L1) ].H O 3 3 2 2 antimycobacterial activities against isoniazid (ATCC (1). To an ethanolic solution of H L1 (10 mmole, 2.57 g), 3 35822) resistant strains of M. tuberculosis, using the 10 mmole equivalent of [Mn(CH COO) .4H O] 3 2 2 micro plate Alamar Blue assay (MABA) (Sivakumar dissolved in absolute ethanol were added slowly after and Rajasekaran, 2013). A serial dilution of the com- which 2 drops of TEA was added. The mixture was pounds was made directly on the plate. The final drug refluxed at 80 °C for 4 h. The brown product obtained concentrations tested were 0.01- 20.0 µg/mL. The plates was allowed to stand at ambient temperature for 24 h were covered and sealed with parafilm and incubated after which it was filtered, washed with absolute ethanol at 37 °C for 5 days. After this time, 25 µL of a freshly followed by ether and dried over P O in vacuo. 4 10 prepared 1:1 mixture of Alamar Blue reagent and 10% 1 [Fe(H3L )2Cl2] (2). 10 mmol (2.57 g) of ethanolic tween 80 was added to the plate and incubated for 24 h. 1 solution of H3L was dissolved in absolute ethanol and A blue colour in the well indicated no bacterial growth, mixed directly ethanolic solution of anhydrous FeCl2 and a pink colour indicated growth. The minimal (10 mmol, 1.27 g). The mixture was refluxed at 80 °C inhibition concentration (MIC) was defined as the lowest for 5 h. The solution obtained was allowed to stand at drug concentration, which prevented a colour change ambient temperature for 48 h. The black precipitate from blue to pink. formed was filtered and washed thrice with 30 mL of 1 Cytotoxicity study. Cytotoxicity of H3L and some of cold ethanol, followed by ether and dried in vacuo. its synthesized metal complexes were determined with 1 [Pt(H3L )Cl]Cl (3). 10 mmole (2.66 g) of PtCl2 was the Vero cell line ATCC CCL-81 using an MTS assay heated to dryness in 1 mL of conc. HCl in a round (Protopopova et al., 2005). 66 Kehinde Olurotimi Ogunniran et al.

Results and Discussion Other aromatic protons resonated within the region 8.80-6.36 ppm. The doublet of doublet peak which Mass spectrum of H L1. The molecular mass of the 3 integrates as one proton at 7.60 ppm was assigned to hydrazone was ascertained with the use of mass spectrum proton H(8) which is at the meta position to nitrogen obtained by the use of Agilent 6520 Q-TOF mass atom in the pyridine moiety. The splitting observed is spectrophotometer (ESI). The spectrum (Fig. 1) showed as a result of the coupling effect from adjacent protons. the molecular ion peak at m/z 258.0 which is in The doublet peak at 8.8 ppm was assigned to a proton agreement with the calculated molecular mass of the H(5) at the ortho position to N(1) atom in the pyridine compound within the precision limit of ± 0.02. moiety. The doublet signal observed was attributed to the coupling effect from proton H(8) which is at the 258.0 100 2.70+006 meta position. The doublet of triplet peak at 8.30 ppm 90 was assigned to the proton H(7) which is adjacent to 80 N(1) atom. Also, the proton in between the two aromatic 70 group H(9) was assigned to a doublet peak at 7.39 ppm. 60 The doublet peak observed is as a result meta coupling 50 effect from H(10). Meanwhile, H(10) was assigned to 40 doublet of a doublet at 6.42 ppm, while a doublet at 30 259.0 6.36 ppm was assigned to H(11). The proton at 6.42 ppm 20 experienced meta coupling effect from H(9) and ortho 177.9 10 137.8 179.0 200.0 339.1 537.2 610.6 684.4 223.1 298.9 391.2 568.0 629.0 coupling effect from H(11) and therefore resonated as 0 m/z 100.0 200.0 300.0 400.0 500.0 600.0 700,0 doublet of doublet. Also the doublet signal at 6.36 ppm assigned to H(11) resulted from the ortho coupling 1 Fig. 1. ESI mass spectrum for H3L . effect from proton H(10) at 6.42 ppm.

9 1 1 1 H H NMR spectrum of H3L . The H NMR spectrum 3 of the hydrazone (Fig. 2) showed a sharp singlet peak 7 HO OH H O which integrated as one hydrogen at d = 12.16 ppm. 8 H N It was assigned to an imine proton NH(1) (Pavia et al., N H H 10 2008). Apart from the fact that the proton is attached 1 H H H N H 6 11 to a highly electronegative element, the effect of neighbo- 5 4 ring atoms contributed to a high decrease in electron density around the imine proton (Sankar et al., 2010; Jursic et al., 2002). The infrared spectrum confirmed the involvement of the imine proton in interhydrogen bonding formation with hydroxyl group (NH OH) during tautomerism and thereby contributing to decrease in electron density around the proton. Thus, the proton 12 11 10 9 8 7 6 resonated downfield (Mustafa et al., 2009). The singlet peak observed was as a result of the absence of protons 1.08 2.34 1.00 0.99 1.20 on its neighbouring atoms and thus there was no coupling 1.18 1.13 1.13 1.10 1.14 interaction. The two other singlet peaks in the downfield region of the spectrum at 11.36 ppm and 10.06 ppm 1 1 Fig. 2. HNMR spectrum of H3L recorded in were assigned to the two aromatic protons OH(2) and DMSO-d6 at 300 MHz. OH(3), respectively (Al-Shaalan, 2011; Patel and Patel, 2011). The high d ppm values could be attributed to 1 their attachment to high electronegative element and COSY spectrum of H3L . The COSY experiment was thereby making each of the peaks to resonate as a single used to confirm the protons assignment above. The COSY peak downfield. Also in the pyridine moiety, H(4) resona- spectrum shown in Fig. 3 revealed the 1H- 1H coupling ted as a single peak with a slight shoulder at 9.10 ppm. interactions in the molecule (Berger and Sicker, 2009; Antitubercular Activity of Acylhydrazone 67

13 1 13 Silverstein and Webster, 2002). The absence of off C NMR spectrum of H3L . The C NMR spectrum diagonal peak at 12.10, 11.36, 10.04, 9.10 and 8.54 ppm (Fig. 5) displayed the presence of 13 magnetically different confirmed their assignment to H(1), ArOH(2), ArOH(3), carbon atoms in the molecule which corresponded to H(4) and H(6), respectively. This confirmed that absence the number of carbon atoms in the molecule. The assign- of coupling interaction around the protons in the mole- ments were done on the basis of proton decoupled 13C cule. However, COSY spectrum confirmed that the spectrum. The two carbon atoms attached to hydroxyl multiplet peak at 7.60 ppm, assigned to H(8), experienced group in the resorcinol ring [C(1) and C(2)] resonated coupling interaction with a doublet of triplet peak at at 162.12 ppm and 160.95 ppm downfield, respectively 8.30 ppm and at the same time correlated with a doublet due to the effect of the attached electronegative element peak at 8.80 ppm. The two peaks were assigned to H(7) coupled with the fact that both carbons are sp2 carbon. and H(5), respectively. COSY spectrum also confirmed Thus, a high decrease in electron density around the that the proton at 6.42 ppm correlated with H(9) at two carbon atoms is expected. Also, the peak at 159.56 7.39 ppm and and H(11) at 6.36 ppm, respectively. The ppm was assigned to the carbonyl carbon, C(3), which resonances at 6.42 ppm H(10) and 6.36 ppm H(11) appeared downfield due to the conjugative effect of confirmed that the two protons were attached to neighbo- N(2)-N(3) core in the hydrazone. The two adjacent carbon ring carbon atoms. The schematic contour plot of 1H- atoms to N(1) appeared downfield at 152.31 ppm and 1 1 H COSY experiment for H3L , (Fig. 3), along with the 149.65 ppm, respectively, because both of them were observed coupling interaction is given in (Fig. 4). affected by electronegative effect of N(1). The lower field of carbon atom, 148.56 ppm, C(6) is due to exten- sive p electron delocalization of N(2)-N(3)=C(6) bond chain. The remaining two carbons in the pyridine moiety, ppm C(7) and C(10), were assigned to the peak at 135.37 ppm and 123.62 ppm, respectively. The resonances assigned to resorcinol carbons are C(8), 131.32; C(12), 107.84; C(13), 102.71 ppm. The non-protonated carbons, C(11) and C(9), resonated downfield due to the conjugative effect of C(6)=N(3) on C(11) and electronic effect exerted by adjacent carbonyl on C(9).

8 HO OH O 1 2 7 3 9 2 N ppm 10 3 11 12 N 6 13 H 1 1 1 5 4 Fig. 3. H- H COSY spectrum of H3L at 300 MHz. N 1

9 H 2 3 160 155 150 145 140 135 130 125 120 115 110 105 ppm 7 HO OH H O 8 Fig. 5. 13C NMR spectrum of H L1 in DMSO-d6 H N 3 at 75 MHz. N H H 10 1 H H 6 11 H N H 1 13 1 1 13 5 4 H- C HSQC spectrum of H3L . H- C correlation spectrum (Fig. 6) was used to confirm both the proton Fig. 4. 1H-1H COSY spectrum schematic structure and carbon assignments. HSQC spectrum showed all 1 of H3L . the protonated carbon in the molecule. Only 8 carbon 68 Kehinde Olurotimi Ogunniran et al.

(C, H, N) obtained are in good agreement with the calculated values. The magnetic susceptibility values recorded at room temperature for Mn(II) and Ni(II) complexes compete favourably with the calculated spin only magnetic values. The conductivity ability of the metal complexes is very low, thus suggesting non- electrolytic character of the synthesized complexes.

9 H 2 3 HO OH 7 H O 8 1 13 1 H N Fig. 6. H- C COSY spectrum of H3L at 75 MHz. N H H 10 1 atoms appeared in the correlated spectrum and this H H 6 13 H N H 11 matches the structure of the hydrazone. C-NMR peaks 5 4 at 161.12, 160.95, 159.55, 128.85 and 110.49 ppm did not make any correlation in the HSQC spectrum and Fig. 7. Schematic diagram of 1H- 13C COSY for therefore suggested that these carbons are non-protonated 1 H3L . as assigned. However, all the protonated carbons showed correlation with the attached proton at the assigned position. Based on this, the schematic diagram for the Infrared spectra. The spectra interpretation was done hydrazone 1H-13C relation is as shown in Fig. 7. by comparing the spectra of each of the metal complexes 1 1 Analytical data of H3L metal complexes. All the metal with the spectrum of the ligand, H3L . The tentative 1 1 complexes of H3L synthesized possess characteristics infrared spectra assignments of H3L and its metal colour as reported in Table 1. The metal complexes were complexes are presented in Table 2 while some of the found to possess high melting point (300 °C) due to the spectra are shown in Fig. 8. The significant changes in coordination bonds. They were found to be sparingly the vibration bands of the metal complexes when com- soluble in methanol, DMF and THF but are soluble in pared to that of the ligand were observed by Davidson DMSO and pyridine. The results of elemental analyses (2010). A strong peak at 3432.27 cm-1 attributed to OH

1 Table 1. Analytical data of H3L metal complexes

Complex Colour M. pt. Found (Calc.), % LM meff (% Yield) (°C) M C H N S cm2/mol mB

1 [Mn(H3L )(CH3COO)2].H2O (1) Brown >300 12.52 41.52 4.21 9.22 32 5.82 (62.5) (12.95) (45.55) (4.27) (9.37) 1 [Fe(H3L )Cl2] (2) Black >300 14.12 40.59 3.22 10.72 56 5.3 (51.2) (14.58) (40.77) (2.63) (10.97) 1 [Pt(H3L )C1].C1 (3) Green >300 36.89 29.51 2.04 7.53 28 - (65.0) (37.28) (29.84) (2.12) (8.03) 1 [Zn(H3L )(CH3COO)2H2O] (4) Yellow >300 14.17 41.73 4.27 9.56 30 - (78.8) (14.26) (41.51) (4.17) (9.16) 1 [Pd(H3L )C1] (5) Brown >300 26.53 39.03 2.53 10.56 26 - (65.06) (26.66) (39.12) (2.78) (10.53)

1 L = C13H8N3O3 Antitubercular Activity of Acylhydrazone 69

1 Table 2. Infrared spectra assignments for metal complexes of H3L Ligand/metal u(OH) u(C=O) u(C=N) u(N-N) d(C-O)df. d(C-N)df. complex cm-1 cm-1 cm-1 cm-1 cm-1 cm-1

1 H3L 3432.27 vs 1639.5 s,b 1508.62 m 1466.91 m 1353.0 s 1165.23 s 1 [Mn(H3L )(CH3COO)2H2O] (1) 3402.5 s,b 1660.01 m 1574.32 ms 1476.99 w 1354.03 m 1181.03 m 1 [Fe(H3L )Cl2] (2) 3396.89 s,b 1593.74 m 1539.29 m 1438.24 w 1347.51 m 1136.67 s 1 [Pt(H3L )Cl].Cl (3) 3413.3 s,b 1609.39 m 1535.93 m 1437.95 w 1212.35 m 1129.2 m 1 [Zn(H3L )(CH3COO)2H2O] (4) 3414.47 s,b 1604.31 m 1524.69 w 1491.26 w 1374.39 m 1190.56 s 1 [Pd(H3L )Cl] (5) 3384.43 b 1606.25 m 1532.06 m 1435.82 w 1211.94 m 1129.63 s

stretching vibration in the spectrum of the ligand was a medium peak at the lower wavelength in all of the observed to have shifted to lower wavelength with complexes except in Mn(II) complex where it appeared reduction in intensity, in Mn(II), Pt(II) and Zn(II) at 1660.01 cm-1. The observation could be attributed to complexes. The observation could be as a result of the coordination of CO to the central metal in the com- coordination of the OH vibration group to the metal plexes. ions in the complexes. However, the peak was observed The azomethine band, which appeared in the spectrum at higher wavelength in Fe(II) and Pd(II) complexes 1 of H L at 1508.62 cm-1 as a medium band, appeared (3396.89 and 3384.43 cm-1, respectively). The higher 3 in all of the metal complexes at a higher wavelength. wavelength observed, coupled with broadness of the The increase in u(C=N) in the spectra of metal com- peak, is as a result of the coordination to the central plexes is due to the increase in the double bond character metal through in enol form. off-setting the loss of electron density via donation to A strong and broad band at 3384.43 cm-1 attributed to the central metal and thus further confirmed the coordi- the CO stretching vibration in the spectrum of H L1 1 3 nation of H3L through the azomethine atom. The also appeared in the spectral of the metal complexes conjugate effect of the coordination through u(C=N) in with significant changes in wavelength and intensity the complexes was noticed on u(N-N) bands which due to effect of the coordination. The peak appeared as appeared as weak band in the metal complexes. UV/Visible spectra. The electronic absorption data of

Sample: Mn L1 File: D:\TGA\KVSN\CDRI\L1.001 the ligand and the metal complexes in methanol at Size: 10.0510 mg Operator: AK Method: Ramp TGA Run Date: 27-Nov-2012 13.54 ambient temperature are recorded in Table 3. The Comment: Mn L1 Instrument: TGA Q500 V20.8 Build 34 electronic transitions were interpreted by comparing

100 43.98 OC 0.8 the spectrum of the ligand to the spectra of the corres- 291.04 OC 5 O O d 80.00 C O 361.85 C ponding metal complexes. Mn(II) is of configuration 325.60 C 375.45 OC which is high spin complex with doubly forbidden 0.6 80 399.13 OC transition. Tunabe-Sugano diagram shows that the O 5 6 380.91 C 390.55 OC O Russell-Saunders term for d high spin system is S 6 0.4 with A1g as the ground term symbol. This term gives

Weight (%) Weight rise to 4G, 4D and 4P excited states. Thus, there is no 60 422.00 OC 539.57 OC sextet spin multiplicity for octahedral complexes, the 0.2 weight (%/ C) Deriv. O 336.55 C 631.64 OC transitions are Laporte forbidden and spin forbidden. 587.74 OC In the spectrum of the Mn(II) complexes, five notable 40 0.0 transition were observed. The high intense transitions 0 600 100 200 300 400 500 700 -1 Universal V4. 5A TA Instruments at ca. 44248, 33333 and 30864 cm were due to transition Temperature (°C) of ligand chromophoric groups in the complex. The transition had undergone bathochromic shift toward 1 visible region due to coordination effect. However, two Fig. 8. TGA/DTA spectrum of [Mn(H3L )(CH3 COO)2 ]. H2O (1). low intense forbidden transitions were observed in 70 Kehinde Olurotimi Ogunniran et al.

Mn(II) complex at ca 24390 and 19920 cm-1. They were Magnetic moment data of Pt(II) complex indicate that 4 6 4 6 d 8 assigned to T1g (G) ¬ A1g and T1g (G) ¬ A1g, the complex is a diamagnetic with configuration respectively which is the characteristics of octahedral which favours square-planar geometry (Brik et al., geometry but the presence of a weak shoulder at ca 2011). This is also supported by a failed attempt to get 24390 cm-1 suggested a distorted tetrahedral pyramidal the esr data for the complex. The electronic spectrum complex. This could be pictured as octahedral complex of the complex showed transition bands at ca. 33003, with one corner occupied by a lone pair of electrons 28818 and 16891 cm-1. The strong band at ca. 33003 (Brik et al., 2011). cm-1 was attributed to the n®p* transitions in the coordinated ligand while the bands at ca. 28818 and The absorption spectrum of Fe(II) complex adapted 16891 cm-1 were assigned to 1Eg ¬ 1A and 1A ¬ greatly to the literatures (Munde et al., 2012). Only one 1g 2g 1A transitions. band with low intensity was observed in the visible 1g region at ca. 22124 cm-1. This showed that Fe(II) is of The electronic spectrum of Zn(II) complex showed a 6 d configuration with high spin system and ground shoulder band at ca. 362232 cm-1 which was assigned 5 term symbol of D. The d-d transition observed was to the n®p* transition within the ligand moiety. Also 2 2 assigned to Al2 ¬ B2. However, two absorptions at the spectrum showed a strong band at ca. 30211cm-1. ca. 33113 and ca. 27700 cm-1 were attributed to n®p* This was assigned to the n®p* transition which could -1 transitions in the ligand. The band at ca. 40816 cm be as a result of cumulative effect of transitions of was assigned to n®p*transition in the ligand had azomethine and amide chromophoric groups of the completely disappeared while bathochromic shift was ligand. The bathochromic shift observed was attributed 10 observed in n®p*transition. The observation was to the coordination effect. Zn(II) ion is of d configuration, attributed to the coordination of the ligand to the metal therefore no d-d transition is expected, but metal-ligand ion in the complexes. charge transfer band was observed at ca. 23256 cm-1 due to yellow colour of the complex. No appreciable band was observed below ca. 20,000 cm-1 which is in 1 10 Table 3. UV/Visible data of H3L and its metal com- accordance with d configuration of Zn(II) ion. plexes 10 The Pd(II) ion has a d configuration with a term Compound Transition Ground Transition 1 d d-d cm-1 term symbol, S. Due to the filled state of the - orbital, symbol transition was not observed for Pd(II) complex as expected. However, intra-ligand bands were observed H L1 40816 - p®p* ( C=C) ar 3 with hypsochromic shift for n®p* transition and 33113 n®p* (C=N ) bathcromic shift for n®p* transitions. Thus, support 29940 n®p*( C=O) 1 6 the coordination of the ligand to the central metal ion Mn(H3L )(CH3 44248 S p®p* ( C=C) ar in the complex. COO)2]H2O (1) 33333 n®p* (C=N ) 30864 n®p*( C=O) 4 6 TGA/DTA analysis. The thermal behaviour of 24390 T1g (G) ¬ A1g 4 6 compounds (1), (2) and (5) was investigated by using 19920 T2g (G) ¬ A1g 1 5 a non-isothermal thermo gravimetric, TG, and differential [Fe(H L )Cl ] (2) 33113 D n®p* (C=N ) 3 2 TG. The samples were heated at a rate of 10 °C min-1 27700 n®p*( C=O) 2 2 under N atmosphere from 0-700 °C. The selected 22124 A12¬ B2 2 1 3 thermograms obtained are as represented in Fig. 8. The Pt(H3L )2.Cl2 (3) 33003 D n®p* 1 1 28818 Eg¬ A1g TG/DTG curve of compound (1) showed 4% weight 1 1 16891 A2g ¬ A1g loss between 43 and 160 °C. This corresponds to loss 1 1 Zn(HL )2)(CH3 36232 S n®p* (C=N ) of water molecule which is outside the coordination COO)2 (4) 30211 n®p*( C=O) sphere of the complex (Artur et al., 2013). However, 23256 MLCT the thermogram shows three major decomposition steps. Pd(HL1) .Cl (5) 42373 1S p®p* ( C=C) ar 2 2 The first step occurred between 291 and 352 °C (3% 31646 n®p* (C=N ) weight loss). This corresponds to loss of uncoordinated 26316 n®p*( C=O) hydroxyl group in the complex. The major weight loss Antitubercular Activity of Acylhydrazone 71 occurred in the second step is attributed to a loss of did not decompose fully to the residue within the

2CH3CO between 361-475 °C. The remaining part of temperature range of study. the ligand decomposed to metal oxide in the third step

(475-676 °C) of the decomposition. Sample: Pd L1 File: D:\TGA\KVSN\CDRI\Pd1.001 Size: 6.9180 mg Operator: AK The TGA/DTA curves for compound (2) shows one Method: Ramp TGA Run Date: 29-Nov-2012 09.35 Comment: Pd L1 Instrument: TGA Q500 V20.8 Build 34 major decomposition in Fig. 9. The stability range

120 O extended from ambient temperature to 225 °C. The 298.72 C 0.25 decomposition of the complex occurred in three stages 0.20 as indicated by DTA peaks at 367, 557 and 595 °C. O 100 86.98 C O The first decomposition started at 225 °C and ended at 426.89 C O O O 475 °C. This corresponded to 57% (Calcd. 58%) of the 161.11 C 278.89 C 0.15 1 80 weight loss for the compound which is mainly the H3L eight (%/ C)

O O 327.77 C w Weight (%) Weight 0.10 ligand. The second and the third steps were attributed O 401.65 C 102.93 C to the loss of the coordinated chloride atoms. This Deriv. 60 O occurred between 530 and 575 °C and 575 and 625 °C 465.07 C 0.5 with a weight loss of 8.7% (Calcd. 9%) in each step. 40 0.00 The % weight of the residue, which was assumed to be 0 100 200 300 400 500 600 700 the metal oxide, is 23% (Calcd. of 24%). Universal V4.TA Instruments Temperature ( OC)

1 Sample: Fe L1 File: D:\TGA\KVSN\22-10-12\Fe L1.001 Fig. 10. TGA/DTA spectrum of [Pd(H L )Cl]H O Size: 6.3700 mg Operator: AK 3 2 Method: Ramp TGA Run Date: 31-Oct-2012 10:00 (5). Comment: Fe L1 Instrument: TGA Q500 V20.8 Build 34

100 O 2.0 356.20 C Electron paramagnetic resonance study. The EPR O 367.59 C spectra of Mn(II) and Fe(II) complexes were recorded 80 1.5 in liquid nitrogen at 77 K. Mn(II) complexes show wide O range of geometry. Reports by Charles and Horacia1 (999); Hamed and Neilands (1994) and Pilbrow (1990) 60 1.0 379.11 OC have reported that EPR have been used successfully to Weight (%) Weight 540.02 OC affirm the geometries of Mn(II) complexes. The EPR 40 weight (%/ C) Deriv. 1 O 0.5 spectrum of [Mn(H3L ) (CH3COO)2] H2O in frozen 565.14 C O 589.28 C O 595.90 C O DMSO at 77 K exhibited 2 g values at g^ = 1.7445 and O 613.97 C 557.31 C g|| = 2.1240 with no hyperfine splitting (Fig. 11). The 20 0.0 400 0 100 200 300 500 600 700 gav isotropic value was found to be 2.0569. This value Universal V4. 5A TA Instruments is very close to the free electron spin value of 2.0023. Temperature (°C) This is consistent with the typical manganese (II) ion

1 and could be responsible for the absence of spin orbital Fig. 9. TGA/DTA spectrum of [Fe(H3L ) Cl2] (2). 6 coupling in the ground state A1 without any sextet term of higher energy (Charles and Horacia,1999; Pilbrow, 1990). The spectrum is broad spectrum which is probably The TGA/DTA curves for compound (5) (Fig. 10) shows due to polar interactions and enhanced spin lattice relaxa- an endothermic peak at 102 °C which indicates the tion in the complex. The observed g|| > g > 2 suggested a presence of water molecules outside the coordinated ^ monomeric complex with tetrahedral pyramidal geometry. sphere of the complex. The 8% (Calcd. 7.7%) weight 1 loss occurred between 88 and 161 °C is accounted for The spectrum of [Fe(H3L )Cl2] in frozen DMSO at 77 the loss of the water molecule. Decompositions observed K shows hyperfine splitting with giso = 2.0204, g^ = between 220 and 340 °C; and 340 and 465 °C are 1.9937 and g|| = 1.9865 (Fig. 12). The spectrum hyperfine attributed to the partial loss of the coordinated ligand splitting constants, A|| and A^ were found to be 14 mT in the complex. As thermogram shows, the complex and 8 mT, respectively. The splitting pattern suggested 72 Kehinde Olurotimi Ogunniran et al. pentagonal coordinated environment for d6 Fe(II) in monochromator is shown in Fig. 13. Table 4 shows the the complexes (Huang and Haight Jun, 1969). However, selected diffraction data obtained which were indexed since g^ > g|| and A|| > A^, which is the characteristic (Hesse, 1948). Mn(II) complex was scanned between of tetrahedral base complex, the complex was considered 5° and 120° at a wavelength of 1.543 Å. The diffrac- to be tetrahedral base pyramidal. tograms and associated data depict the 2q value for each peak, the relative intensity and inter-planar spacing (d-values). The X-ray diffraction patterns of the complex

1500 with respect to major peaks of relative intensity greater than 10% were indexed using a computer programme 1000 (Kozakov et al., 2011; Harikumaran and Thankamani, 2009). The selected indexed data yielded the Miller 500 indices (hkl), the unit cell parameters and the unit cell volume. The unit cell of Mn(II) complex yielded values 0 of lattice constants: a = 12.441 Å, b = 14.582 Å and c Intensity = 7.842 Å, and a unit cell volume V = 1227.2353 Å3. In -500 concurrence with these cell parameters, conditions such as a ¹ b ¹ c and a = g = 90° ¹ b required for a monoclinic -1000 sample were tested and found to be satisfactory. Hence, it can be concluded that [Mn(H L1) (CH COO) ]H O -1500 3 3 2 2 200 250 300 350 400 450 complex has a monoclinic crystal system. B (mT) Antimycobacterial results. Based on the results of the biological studies, it was found that the complexes Fig. 11. EPR spectrum of [Mn(H L1) (CH COO) ] 3 3 2 (PtL1) (MIC = 0.56 µg/mL), (ZnL1) (MIC = 0.61 H O. 2 µg/mL), (MnL1) (MIC = 0.71 mg/mL) and (FeL1) (MIC = 0.82 mg/mL), exhibited a significant activity when 3000 compared with first line drugs such as isoniazid (INH) (MIC = 0.9 mg/mL). These results suggest that they 2000 may be selectively targeted to M. tuberculosis growth. They are therefore regarded as potential drug candidates 1000 but the results of the toxicity study indicated that the complexes are more toxic than the isoniazid and the 1 0 ligand. However, Fe(II) complex of H3L displayed the Intensity

-1000

400 -2000

300 -3000 1 200 250 300 350 400 450 10 2 5 89 B (mT) 200 6 Lin (Counts) 4 3 11 7 12 1 Fig. 12. EPR spectrum of [Fe(H3L )Cl2] in DMSO 100 at 77 K.

0 5 10 20 30 40 50 60 70 80 90 100 110 120 Powder X-ray diffraction study. The powder XRD 2-Theta 1 pattern for [Mn(H3L )(CH3COO)2]H2O recorded on a 1 Bruker AXS D8 advance diffractometer operating in Fig. 13. X-ray diffraction pattern of [Mn(H3L ) the q:q mode, equipped with a secondary beam graphite (CH3COO)2]H2O (1). Antitubercular Activity of Acylhydrazone 73

lowest toxicity with the IC50 value of 0.92 µM which were found to be non significant different (P >0.05) to 1 that of the ligand (H3L ) (IC50 = 3.01mg/mL) and isoniazid (INH) (IC50 = 4.72 mM). The results obtained were displayed in bar chart (Fig. 15).

1 Table 4. X-ray powder diffraction data for [Mn(H3L ) (CH3COO)2] H2O (1) 2q d (Å) Count Intensity h k l count (%)

1 [Mn(H L )(CH COO) ]H O (1) 8.076 10.9383 212 10.9 1 0 0 3 3 2 2 9.393 5.5773 105 5.57 4 9 8 13.865 3.8474 115 3.84 5 4 5 16.800 3.1415 162 42.2 1 0 0 18.724 2.6809 166 43.2 1 0 0 19.790 2.5411 155 40.3 1 1 0 23.098 1.3748 95.8 24.9 2 0 0 25.738 1.3420 92.7 24.1 2 1 0 25.993 1.15342 93.2 24.2 2 2 0 32.067 1.04703 86 22.3 3 0 0 48.277 0.92321 89.4 23.2 3 1 1

1 [Fe(H L )(Cl )] (2) 3 2 Table 5. MIC (µg/mL) and IC50 (µM) values of the 1 metal complexes of H3L

Entry MIC IC50 (µg/mL) (µM) MnL1 0.71 0.85 FeL1 0.82 0.92 PtL1 0.56 1.03 ZnL1 0.61 1.17 1 H3L 1.02 3.01 INH 0.92 4.72

1 6 Pt(H3L )2Cl2 (3) 5 **

4 M)

g/mL) ** m (

m 3 50 IC

MIC ( 2 ** 1 ** ** **

0 MnL1 FeL1 PtL1 ZnL1 H3L1 INH **(p > 0.05)

1 MIC values IC50 values Zn(H3L )2(CH3OO)2 (4)

Fig. 15. Comparison of MIC/IC50 values of the metal Fig. 14. The proposed structures for the complexes. complexes with the ligand and isoniazid drug. 74 Kehinde Olurotimi Ogunniran et al.

Conclusion optical spectroscopy of six-coordinated Mn4+ ion 1 in the Y Ti O and Y Sn O pyrochlores. Optical In conclusion, H L was successfully synthesized at 2 2 7 2 2 7 3 Materials 20 °C from nicotinic hydrazide by stirring in the presence , 33: 1671-1676. Handbook of Electron of 2,4-dihydroxybenzaldehyde. The hydrazone is a Charles, P.P., Horacia, A.F. 1999. Spin Resonance. derivative of isoniazid and therefore was expected to 123 pp. Springer-Verlag Inc. New possess antitubercular activity. The structure of the York, USA. hydrazone was successfully elucidated by using mass Cui, Y., Dong, X., Li, Y., Li, Z., Chen, W. 2012. spectrum, CHN analysis, 1H, 13C and 2D NMR. All the Synthesis, structures and urease inhibition studies efforts made to grow single crystals of the metal com- of Schiff base metal complexes derived from 3, 5- . European Journal of plexes did not yield positive results. Hence, the physical dibromosalicylaldehyde Medicinal Chemistry, properties and structures of the metal complexes were 58: 323-331. elucidated using different spectroscopic analysis. The Davidson, G. 2010. Spectroscopic properties of inorganic Royal Society of information obtained was supported with the use of and organometallic compounds. Chemistry powder X-ray analysis which leads to the model , 39: 212-226. structures of the complexes. The in vitro antitubercular De Backer, A.I., Mortelé, K.J., De Keulenaer, B.L., study confirmed the efficacies of the compounds. The Parizel, P.M. 2006. Tuberculosis: epidemiology, metal complexes were found to be more active than manifestations, and the value of medical imaging JBR-BTR-Journal, Belge de Radiologie- isoniazid however, they were found to be more toxic in diagnosis. Belgisch Tijdschrift voor Radiologi, against the vero cell than isoniazid. 89: 243-250. Hamed, M.Y., Neilands, J.B. 1994. An electron spin Acknowledgement resonance study of the Mn(II) and Cu(II) complexes of the Fur repressor protein. Journal of Inorganic The authors express their gratitude to CISR and TWAS Biochemistry, 53: 235-248. for financial support through TWAS-CSIR fellowship Harper, C. 2007. Tuberculosis, a neglected opportunity? granted to carry out this work. We thank all the staff of Nature Medicine, 13: 309-312. SAIF Central Drug Research Institute, Lucknow, Indian Hesse, R. 1948. Indexing powder photographs of Institute of Technology, Roorkee, and Indian Institute tetragonal, hexagonal and orthorhombic. Acta of Technology, Bombay, India for their technical support. Crystallographica, 1: 200-207. Huang, T., Haight Jun, J.P.J. 1969. Electron spin References resonance studies of monomer–dimer equilibria Aboul-Fadl, T., Abdel-Aziz, H.A., Abdel-Hamid, M.K., involving molybdenum(V) complexes with cysteine Elsaman, T., Jane Thanassi, J., Pucci, M.J. 2011. and glutathione. Journal of the Chemical Society Schiff bases of indoline-2,3-dione: potential novel D: 985-986. DOI: 10.1039/C29690000985. inhibitors of Mycobacterium tuberculosis (Mtb) Jenkins, A.O., Cadmus, S.I.B., Venter, E.H., Pourcel, DNA gyrase. Molecules, 16: 7864-7879. C., Hauk, Y., Vergnaud, G. 2011. Molecular Al-Shaalan, N.H. 2011. Synthesis, characterization and epidemiology of human and animal tuberculosis biological activities of Cu(II), Co(II), Mn(II), Fe(II), in Ibadan, South Western Nigeria. Veterinary Micro- biology and UO2(VI) Complexes with a new schiff base , 151: 139-147. hydrazone: O-hydroxyacetophenone-7-chloro-4- Jia, L., Tomaszewski, J.E., Hanrahan, C., Coward, L., quinoline hydrazone. Molecules, 16: 8629-8645. Noker, P., Gorman, G., Nikonenko, B., Protopopova, Belskaya, N.P., Dehaen, W., Bakuleva, V.A. 2010. M. 2005. Pharmacodynamics and pharmacokinetics Synthesis and properties of hydrazones bearing of SQ109, a new diamine-based antitubercular drug. amide, thioamide and amidine functions. Journal British Journal of Pharmacology, 144: 80-87. of Organic Chemistry 2010: 275-332. Jindani, A., Nunn, A.J., Enarson, D.A. 2004. Two Berger, S., Sicker, D. (eds.), 2009. Classics in Spectro- 8-month regimens of chemotherapy for treatment scopy, Isolation and Structure Elucidation of Natural of newly diagnosed pulmonary tuberculosis: Inter- Products, vol. 72: 117 pp., Wiley-VCH, Germany. national multicentre randomized trial. Lancet, 364: Brik, M.G., Srivastava, A.M., Avram, N.M. 2011. 1244-1251. Comparative analysis of crystal field effects and Jursic, B.S., Douelle, F., Bowdy, K., Stevens, E.D. Antitubercular Activity of Acylhydrazone 75

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Quantification of Cr(VI)-Thymoquinone Complex Using Cyclic Voltammetry Farah Kishwara*, Khalid Mohammed Khanb, Rubina Perweena, Anila Anwara and Nasir Akhtara aDepartment of Chemistry, Federal Urdu University of Arts, Science and Technology, Gulshan-e-Iqbal Campus, Karachi-75300, Pakistan bH.E.J. Research Institute of Chemistry, International Centre for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan

(received December 23, 2014; revised August 10, 2015; accepted August 24, 2015)

Abstract. Quantitative studies of Cr(VI)- thymoquinone complex have been performed by cyclic voltammetry. For this purpose glassy carbon, platinum and saturated calomel electrodes were used as working, auxiliary and reference electrodes, respectively. The effects of concentrations and metal-ligand ratios on Cr(VI)- thymoquinone complex were investigated. Effect of concentration was found to follow Randles-Sevcik equation. Calibration curve method with linear regression line confirms that cyclic voltammetry can be used for quantification of Cr(VI)-thymoquinone complex for pharmaceutical assay. Complete complex formation seems to occur at metal ligand ratio 1:1. Results indicate quasi-reversible electron transfer mechanism. E° and diffusion coefficient of complex at different concentrations and metal ligand ratios were also calculated and found to be 0.244±0.01 V and 3.45´10-5 cm2s-1, respectively. The values of transfer coefficients, a and b, were found to be 0.716±0.02-1.231±0.01 and 0.814±0.01-0.906±0.01, respectively. Keywords: Cr(VI)-thymoquinone complex, quantitative studies, cyclic voltammetry

Introduction of different oxidation states etc. Furthermore, this tech- Electrochemical methods are now emerging as powerful nique helps to give information about the presence of et al and versatile analytical techniques and have found vast intermediates in various redox reactions (Skoog ., applications in many important fields including pharma- 1998; Braun, 1983). Hence, this technique is very popular ceutical industry, and biological and environmental and reliable electrochemical technique and widely used applications (Farghaly et al., 2014; Tsai et al., 2011; nowadays (Beitollahi and Mostafavi, 2014; Molaakbari et al et al Sivasubramanian and Sangaranarayanan, 2011; Beitollahi ., 2014; Baghbamidi ., 2012; Beitollahi and et al et al., 2008). The use of electro analysis is increasing Sheikhshoaie, 2011; Beitollahi ., 2012). day by day due to high sensitivity, reduction in solvent Thymoquinone (2-methyl-5-isopropyl-1,4-benzoquinone) and sample consumption, high-speed and low operating is a phytochemical compound (Fig. 1). It is found as cost (Almeida et al., 2013; Halls et al., 2013; Yuzhi Li an active component in the plant of Nigella sativa (Ali et al., 2013; Cheng et al., 2012; Aaboubi and Housni, and Blunden, 2003). It possesses several biological 2012; Farghaly et al., 2005; Farghaly and Ghandour, activities (Gali-Muhtasib et al., 2008; Syed, 2008; 2005). Especially voltammetric techniques have proved Badary et al., 2007; El-Mahdy et al., 2005). Most of to be more suitable to investigate the redox properties its pharmacological properties are due to its antioxidant of drugs and biological analytes (Baghbamidi et al., property (Mansour et al., 2002). In addition, it has the 2012; Karaaslan and Suzen, 2011). ability to form complexes. Previously its complexation Cyclic voltammetry is a potentiodynamic electrochemical with some redox active metals using potentiometry has et al technique. The primary function of this technique is to been investigated by Kishwar . (2012). The comple- give qualitative information regarding various electro- xation of thymoquinone with iron by cyclic voltammetry chemical processes, although it is equally beneficial for has also been examined qualitatively and quantitatively quantitative analysis. It is useful in finding out the by Kishwar and Haq (2013). mechanisms and kinetics of different electrochemical Chromium (VI) has been reported as a highly toxic reactions, rates of oxidation/reduction processes, stability element. It is a strong irritant; as a result it may cause *Author for correspondence; E-mail: [email protected] different types of allergic reactions. Its inhalation can

76 Cr(VI)-Thymoquinone Complex 77

O CH3 methanol was also used in their preparation in addition to 0.1 M NaCl. Cyclic voltammetric studies. At first CV of supporting electrolyte was run to get base-line and then 15.0 mL CH3 of 5´10-3 M solution of analyte and equimolar solution of metal were run to get overlay. The scan rate and current -4 sensitivity were 0.1 V/s and 1´10 A/V, respectively. H 3 C The potential range was set from -0.40 V to +0.80 V and then reversed back to -0.40 V. In order to observe O effect of several parameters, the complexation was studied by varying metal ligand ratio and concentration. Fig. 1. Structure of thymoquinone. Complex solutions having different metal-ligand ratio from 1:0.5 to 1:5 and concentrations from 0.02´10-3 M cause irritation and damage to nose, lungs, stomach and to 1.2´10-3 M were prepared in order to investigate intestine and ingestion could result in stomach mal- effect of these changes on complex formation. functioning and ulcers, convulsions, damage to kidney and liver and even death. In case of chronic exposure Results and Discussion it might cause pulmonary fibrosis and lung cancer. Effect of concentration on voltammograms of Cr(VI)- However, certain reducing substances in the food could thymoquinone complex. Calibration curve method reduce Cr(VI) to Cr(III) (Stoecker, 1999; Lukaski, was used to judge effect of concentration on Cr(VI)- 1999). Seeds of Nigella sativa are commonly used in thymoquinone complex. For this Randles-Sevcik equa- our food and thymoquinone in these seeds can also tion (Greef et al., 1985) was used which is as follows: perform the same function. Cr(VI) forms a vast variety ½ Ip = 0.4463 nFACo* (nF n D°/RT) of complexes (Sharpe, 1996) including complex with thymoquinone (Kishwar et al., 2012). Hence, it could where: be helpful in case of Cr(VI) toxicity. In this view quan- Ip = peak current (A); n = scan rate (V/s); n = number titative study of Cr(VI)-thymoquinone complex was of electron transferred; F = Faraday’s constant; A = area performed in order to get useful information regarding of electrode (cm2); Co* = concentration of Cr(VI)- complexation of Cr(VI) and thymoquinone. thymoquinone complex (moles/cm3); D° = diffusion coefficient of Cr(VI)-thymoquinone complex (cm2 s -1); Materials and Methods T = 25 ± 2 °C and R = rate constant. Instrumentation. Cyclic voltammeter. CHI–760 D The cyclic voltammograms at different concentration Electrochemical work station was used. Three electrodes showed dependence of Ip on concentration of the com- were used, a glassy carbon working electrode (Model plex. At low concentrations, 0.02´10-3 M and 0.1´10-3 M, number = CHI 104, area of the electrode = 0.07065 cm2), no oxidative wave of forward scan was observed whereas saturated calomel reference electrode and a platinum the reduction peak could be seen clearly during the wire auxiliary electrode. The working electrode was re- reverse scan. It was observed that one small peak (1) polished using alumina, the particles’ size of which was is also appearing in addition to a bigger peak (3) in both 0.3 micron. forward as well as in reverse scan (Fig. 2a). It may be Chemicals. Thymoquinone was purchased from MP due to complications in the reaction. It is possible that Biomedicals, LLC, whereas sodium chloride and the metal needs low concentration of the ligand for potassium dichromate from E. Merck. complex formation and excess ligand may be responsible for giving bigger peaks, as these peaks (i.e., peak 3 and Sample preparation. Supporting electrolyte. 0.1 M 4) were observed within the potential range similar to solution of NaCl. that of thymoquinone (Table 1-2). Direct increase of Analyte solution. The 5´10-3 M solution of thymo- current with concentration (0.02´10-3 M to 1.2´10-3 M) quinone (TQ) and equimolar solution of K2Cr2O7 were shows that effect of concentration follows Randles - prepared as measuring solution. TQ was analyte. 10% Sevcik equation (Fig. 2b). Calibration curve along with 78 Farah Kishwar et al. least square fit line showed no major deviation from Table 1. Electrochemical parameters of cyclic voltam- zero, which points out towards the possibility that no mograms of thymoquinone, Cr(VI), and Cr(VI)-thymo- adsorption has occurred on electrode surface. These quinone complex results indicate that calibration curve method can be Ipa Ipc Epa Epc used for quantification of Cr(VI)-thymoquinone complex (×10-5 A) (×10-5 A) (V) (V) -3 -3 within a wide range i.e. (0.2´ 10 M to 1.2´10 M). TQ 1.017 2.717 -0.242 -0.326 Effect of metal-ligand ratio. For this purpose cyclic ±0.01 ±0.01 ±0.01 ±0.01 Cr(VI) - - - - voltammograms of complex solutions having metal- Cr(VI)-TQ complex 0.246 1.142 0.179 -0.210 ligand ratios 1:0.5 -1:5 were studied. The overlay reveals ±0.01 ±0.01 ±0.01 ±0.01

0.6 10 0.4 3 1 0.2 8 0

-0.2 2 A)

-5 6 R =0.9581 -0.4 2 -0.6

Current/le.lA 4 -0.8 Ip (in 10 2 -1.0 R =0.942 4 2 -1.2 -0.4 -0.3 -0.2 -0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0 Potential/V vs SCE 0 0.5 1 1.5 Conc. x 10-3 M Fig. 2a. Cyclic- voltammograms of Cr(VI)-thy- Ip (A) moquinone complex showing effect of a Ipc (A) concentration (concentrations of complex solutions = 0.02×10-3 M, 0.1×10-3 M, Fig. 2b. Plot of anodic and cathodic peak current 0.2×10-3 M, 0.4×10-3 M, 0.6×10-3 M, against concentration of Cr(VI)-thymo- 0.8×10-3 M, 1×10-3 M, 1.2×10-3 M). quinone complex.

Table 2. The values of Ep, Ep/2, Ep-Ep/2, Epa-Epc,, Ip, ana, bnb and diffusion coefficients from cyclic voltammograms of Cr(VI)-thymoquinone complex with different concentrations

2 -1 Concen- Epa Epa/2 Epa-Epa/2 Ipa Ipa/Ipc bnb D (cm s ) -5 tration (V) (V) (V) ×10 (A) =0.048/Epa- (10-3 M) Epa/2 0.02 -0.236 ± 0.01 -0.29 ± 0.01 0.054 ± 0.011 1.431 ± 0.01 1.15 0.889 ± 0.01 - 0.4 -0.238 ± 0.01 -0.292 ± 0.01 0.054 ± 0.011 1.507 ± 0.01 0.88 0.889 ± 0.01 1.90 × 10-5 0.6 -0.24 ± 0.01 -0.294 ± 0.01 0.054 ± 0.011 2.143 ± 0.01 0.566 0.889 ± 0.01 1.7 × 10-5 0.8 -0.247 ± 0.02 -0.3 ± 0.02 0.053 ± 0.012 3.419 ± 0.02 0.65 0.906 ± 0.02 2.44 × 10-5 1.0 -0.248 ± 0.02 -0.301 ± 0.02 0.053 ± 0.011 4.099 ± 0.02 0.61 0.906 ± 0.01 2.24 × 10-5 1.2 -0.248 ± 0.02 -0.301 ± 0.02 0.053± 0.012 4.252 ± 0.02 0.508 0.906 ± 0.02 1.68 × 10-5

2 -1 Concen- Epc Epc/2 Epc-Epc/2 Epa-Epc Ipc ana D (cm s ) -5 tration (V) (V) (V) (V) ×10 (A) =0.048/Epc- -3 (10 M) Epc/2 0.02 -0.377 ± 0.02 -0.333 ± 0.02 -0.041 ± 0.011 0.141 ± 0.01 1.248 ± 0.02 1.2 ± 0.02 - 0.4 -0.375 ± 0.01 -0.33 ± 0.01 -0.045 ± 0.012 0.137 ± 0.01 1.706 ± 0.01 1.1 ± 0.01 2.43 × 10-5 0.6 -0.353 ± 0.01 -0.312 ± 0.01 -0.041 ± 0.011 0.113 ± 0.01 3.785 ± 0.01 1.20 ± 0.01 5.31 × 10-5 0.8 -0.348 ± 0.01 -0.295 ± 0.01 -0.053 ± 0.011 0.101 ± 0.01 5.248 ± 0.01 0.906 ± 0.01 5.75 × 10-5 1.0 -0.342 ± 0.02 -0.294 ± 0.02 -0.048 ± 0.012 0.094 ± 0.01 6.716 ± 0.02 1.0 ± 0.01 6.02 × 10-5 1.2 -0.35 ± 0.01 -0.301 ± 0.01 -0.049 ± 0.012 0.102 ± 0.01 8.37 ± 0.01 0.980 ± 0.01 6.50 × 10-5 Cr(VI)-Thymoquinone Complex 79 that complete complexation occurred at metal ligand prominent. At higher metal-ligand ratios i.e., 1:3-1:5 ratio 1:1 because here anodic and cathodic peaks (1 and distortion in the anodic peaks was observed which may 2) were observed in a potential range entirely different be due to the superimposition of two peaks lying very from that of thymoquinone (Table 3, Fig. 3). By further close to each other. These two peaks may be of complex increasing metal-ligand ratio, sudden change in the peak and thymoquinone. potential was observed. These observations suggest that In case of effect of metal-ligand ratio the cyclic voltam- probably thymoquinone suppressed the metal at high mograms seem to fulfill the criteria for quasi-reversible concentrations and its own peaks (3 and 4) became reactions (Bard and Faulkner, 2001). As Ipa/Ipc was not equal to one and DEp was found to be greater than 59/n 2.0 3 1 mV, it seems that quasi-reversible behaviour is favoured 1.0 (Table 3). For each case a and b were also calculated 0 -1.0 using the relation 0.048/Ep-Ep/2 which were found in 2

A -2.0 the range of 0.716 ± 0.02 to 1.231 ± 0.01 and 0.814 ± e-5 -3.0 0.01to 0.906 ± 0.01, respectively. Effect of metal-ligand -4.0 ratio on peak potential (Epa and Epc) gave a straight line -5.0 2

Current/1 -6.0 4 with very good R value (Fig. 4), showing independence -7.0 of peak potential on change in metal ligand ratio. -8.0 -0.4 -0.3 -0.2 -0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 An increase in Ipa and Ipc was noted with the increase Potential/V vs SCE in metal-ligand ratio but the peak current became nearly constant at metal-ligand ratio 1:3, showing nearly Fig. 3. Cyclic- voltammograms of Cr(VI)-thymo- maximum complexation below this ratio (Fig. 5). quinone complex showing effect of metal - ligand ratio (Metal-ligand Ratio = 1:0.5, Analysis of diffusion coefficient for Cr(VI)-thymo- 1:1,1:2,1:3, 1:4, 1:5, concentration of quinone complex. Diffusion coefficient of different TQ = 5×10-3 M, concentration of Cr(VI) complexes and compounds is an important constant and solution = 5×10-3 M). it could be easily and accurately measured by cyclic

Table 3. The values of Ep, Ep/2, Epa-Epc, Ip and ana, bnb and diffusion coefficients from cyclic voltammograms of Cr(VI)-thymoquinone complex with different metal-ligand ratios

2 -1 Ratio L/M Epa Epa/2 Epa-Epa/2 Ipa Ipa/Ipc bnb D (cm s ) (V) (V) (V) ×10-5(A) =0.048/Epa- Epa/2 0.5 0.149 ± 0.01 0.09 ± 0.01 0.059 ± 0.01 0.2 ± 0.01 0.256 ± 0.01 0.814 ± 0.01 - 1 0.179 ± 0.01 0.12 ± 0.01 0.059 ± 0.01 0.246 ± 0.01 0.215 ± 0.01 0.814 ± 0.01 - 2 -0.232 ± 0.01 -0.29 ± 0.01 0.058 ± 0.01 0.932 ± 0.01 0.46 ± 0.01 0.828 ± 0.01 4.64 × 10-6 3 -0.236 ± 0.02 -0.291 ± 0.01 0.055 ± 0.02 1.431 ± 0.01 0.434 ± 0.01 0.873 ± 0.02 1.09 × 10-5 4 -0.239 ± 0.01 -0.292 ± 0.01 0.053 ± 0.01 1.411 ± 0.01 0.352 ± 0.01 0.906 ± 0.01 1.06 × 10-5 5 a - - - - -

2 -1 Ratio L/M Epc Epc/2 Epc-Epc/2 Epa - Epc Ipc ana D (cm s ) -5 (V) (V) (V) (V) ×10 (A) =0.048/Epc- Epc/2 0.5 -0.17 ± 0.01 -0.103 ± 0.02 -0.067 ± 0.02 0.021 ± 0.02 0.78 ± 0.01 0.716 ± 0.02 - 1 -0.21 ± 0.01 -0.15 ± 0.01 -0.06 ± 0.01 0.031 ± 0.02 1.142 ± 0.01 0.8 ± 0.01 - 2 -0.338 ± 0.01 -0.299 ± 0.01 -0.039 ± 0.01 0.106 ± 0.01 2.028 ± 0.01 1.231 ± 0.01 2.2 × 10-5 3 -0.346 ± 0.01 -0.302 ± 0.01 -0.044 ± 0.01 0.11 ± 0.01 3.3 ± 0.01 1.091 ± 0.01 5.82 × 10-5 4 -0.37 ± 0.01 -0.31 ± 0.01 -0.06 ± 0.02 0.131 ± 0.02 4.011 ± 0.01 0.8 ± 0.01 8.59 × 10-5 5 ------a = peak distorted. 80 Farah Kishwar et al.

0 -1E.15 0 0 -0.05 1 2 3 4 1 2 3 4 -0.1 -0.1 -0.2 -0.15 R 2 =0.9356 -0.2 R 2 =0.9932 -0.3 pa pc E (V) E (V) -0.25 -0.4 R 2 =0.9231 -0.3 R 2 = 1 -0.5 -0.35

-0.4 M:L Ratio -0.6 M:L Ratio

Epa Epa/2 Epc Epc/2

Fig. 4. Variation of anodic and cathodic peak potentials with change of metal-ligand ratio in cyclic voltammo- grams of Cr(VI)-thymoquinone complex.

5 Table 4. Half wave potential (E°= E1/2) for Cr(VI)- thymoquinone complex at different concentrations and 4 metal-ligand ratios

-5 3 Concentration (E°)a Ratio (E°)a (10-3 M) (V) L/M (V) 2 p I (in 10 A) I (in 10 0.02 0.264 ± 0.01 0.5 0.12 ± 0.02 1 0.4 0.265 ± 0.01 1 0.15 ± 0.02 0 0.6 0.267 ± 0.01 2 0.261 ± 0.02 0 1 2 3 4 0.8 0.274 ± 0.02 3 0.264 ± 0.01 1.0 0.275 ± 0.02 4 0.266 ± 0.01 M:L Ratio -5 -5 1.2 0.275 ± 0.02 5 - Ipa x10 Ipa x10

Fig. 5. Variation of anodic and cathodic peak currents with change of metal-ligand ratio that in case of metal- ligand ratio 1:0.5 and 1:1anodic in cyclic voltammograms of Cr(VI)-thymo- and cathodic peaks (1 and 2 in Fig. 3) were observed quinone complex. in a potential range entirely different than rest of the metal-ligand ratios. Hence, it is suggested that change voltammetry (Anwer, 2006). Diffusion coefficient of in peak potential, both in Epa and Epc (Table 3), resulted the complex was determined using Randles- Sevcik in changed values of E°. et al (Greef ., 1985) equation by varying concentrations Briefly, quantitative studies of Cr(VI)-thymoquinone and metal-ligand ratios (Table 2-3). No reasonable complex were performed at glassy carbon electrode effect of varying concentration or metal-ligand ratio on against saturated calomel electrode which include diffusion coefficient was observed and its value remained determination of E°, D, a, and b. Effects of different nearly the same under all above mentioned conditions. parameters, i.e., concentration and metal ligand ratio, 2 Area of electrode (A) was 0.07065 cm whereas number on complexation were observed by varying these of electron transfer (n) was supposed to be 3 (Sharpe, parameters. Horizontal base line indicates the purity of 1996). the base electrolyte. The Cr(VI)-thymoquinone complex Analysis of E°, a characteristic property. For Cr(VI)- seems to be stable at lower concentrations. It was also thymoquinone complex values of E° were determined suggested that here best complex formation occurred at different concentrations and metal-ligand ratios and at metal-ligand ratio 1:1. At higher metal- ligand ratios it was found to be approximately constant at all concen- (i.e., 1:2 to 1:5) distortion in the anodic peak was observed tration and ratios (Table 4), except in first two cases of which may be due to the presence of two peaks very metal- ligand ratio. 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Effect of Processing on Physicochemical Properties and Fatty Acid Composition of Fluted Pumpkin (Telfairia occidentalis) Seed Oil Jacob Olabode Alademeyin and Jacob Olalekan Arawande* Department of Science Laboratory Technology, Rufus Giwa Polytechnic, Owo, Ondo State, Nigeria

(received February 17, 2015; revised August 18, 2015; accepted August 19, 2015)

Abstract. This paper reports the physicochemical properties and fatty acid composition of the seed oil extracted from fluted pumpkin (Telfairia occidentalis). The extracted oil was degummed, neutralised and bleached. The oil yield was 42.26±0.20%. The specific gravity (at 25 °C) of the oil was 0.923±0.003 and the refractive index (at 25 °C) was 1.475±0.002. Processing of the crude oil resulted in progressive decrease in turbidity, colour, free fatty acid, acid value, peroxide value and saponification value. However, there was increase in smoke point (243.00±0.03 to 253.00±0.03 °C), flash point (285.00±1.20 to 304.00±1.10 °C) and fire point (345.00±1.10 to 358.00±1.55 °C) as well as iodine value (113.00 to 121.50 g/100 g) and fatty acid composition during the processing of the oil. The fatty acids detected in the oil samples were myristic, palmitic, stearic, oleic, arachidic, behenic, linoleic and linolenic acids. The predominant fatty acid was oleic acid (47.40-47.90%) followed by linoleic acid (26.36-30.44%) while the least fatty acid was linolenic acid (0.01-0.05%). Keywords: fluted pumpkin, seed oil, degumming, neutralisation, bleaching, fatty acid composition

Introduction tially valuable as a high protein oil seed for human and et al Cucurbitaceae is one of the largest floras which consist animal food (Nkang ., 2003). The seeds of fluted of nearly 100 genera and 750 species. The plant family pumpkin are valuable both as an oil seed (54%) and is known for its great genetic diversity and wide spread also as a protein source (27%) with a fairly well balanced et al adaptation which include tropical and sub-tropical amino acid composition (Hamed ., 2008; Akwaowo et al regions, arid desert and temperate locations. Cucurbits ., 2000). Unfortunately, 78-91% of the fruits wasted et al are known for their high protein and oil content (Giwa annually (Fagbemi ., 2005). et al., 2010). There are many studies on the proximate composition T. occidentalis The seed and seed oil characteristic of some members and nutritive value of leaves and seeds of et al et al of the cucurbits are documented in literature and they (Effiong ., 2009; Hamed ., 2008; Akwaowo et al are generally referred to as melons. Telfairia occidentalis ., 2000; Giami and Isichei, 1999; Giami and Bekebian, is a member of the cucurbitaceae family, commonly 1992; Asiegbu, 1987), scanty information is available called fluted pumpkin. Its common names include, on the physicochemical properties of the seed oil and et al Fluted guard, Colestillada (Spanish), Krobanko (Ghana), fatty acid profile (Bello ., 2011), but little or no Gonugbe (Sierra Leone) and Ugu (South-East, Nigeria) information on physicochemical properties and fatty (Chukwuonso et al., 2010). The crop is grown mainly acid composition of the processed (refined) seed oil. for the leaves, which constitute an important component Oil seeds tend to contain a much larger proportion of of the diet in many West African countries (Gill, 1992). solid material associated with the requiring careful Fluted pumpkin (T. occidentialis Hook F.) is one of the reduction in size and usually some heat treatment before food crops with considerable value of energy and protein being processed or solvent extracted to recover the oil that is grown in Nigeria. It is a vine with large lobed (Gunstone and Norris, 1983). The chemical composition leaves and long twisting tendrils (Chukwuonso et al., of oil extract consequently gives a qualitative identifi- 2010; Okoli and Mgbeogu, 1983). Although pumpkin cation of oil, and is an important area in the selective seeds are rich in oil storage reserves, it presently has application guide in the commercialization and utility very low commercial value as an oil seed but is poten- of oil products (Salunkhe et al., 1992). A number of *Author for correspondence; E-mail: [email protected] seed oils have been characterized for the identification

83 84 Jacob Olalekan Arawande et al. of several fatty acids of nutritional and nutraceutical Materials and Methods importance but the vast majority have not been adequa- Collection and sample preparation . Fluted pumpkin tely evaluated. fruits were bought from a local market in Owo, Ondo Like all vegetable oils, fluted pumpkin (T. occidentalis) State, Nigeria. The fruits were sliced, and the seeds seed oil is composed of triglycerides (98-99%) and were removed. The seeds were cleaned and freed from other substances in unsaponifiable fractions (non- unwanted materials before they were shelled manually. glycerides) which are also known as the ‘minor com- The seeds of good quality were sliced into pieces, sun ponent’ (NRC, 2001). Crude T. occidentalis seed oil dried and smoothly milled into powdery form. The produced by solvent extraction contains both oil-soluble flour was then stored into tight container prior to oil and oil-insoluble substances that need to be removed extraction. (with minimum loss of oil, and damage to the nature Oil extraction. The powdered fluted pumpkin sample of the oil ‘glycerides’ and tocopherol). The oil-insoluble was then subjected to soxhlet extraction using hexane materials may be removed through filtration. However, as solvent. the soluble material must be removed by several methods which lead to production of edible oil. And such methods Ground seeds (50 g) were placed into a cellulose paper include degumming, neutralisation, bleaching and deodo- cone and extracted, using, hexane (B.P 65 °C) in a rization (Gunstone and Norris, 1983). Crude T. occidentalis Soxhlet extractor for 8 h. The oil was then recovered seed oil contains phosphatides (hydratable and non- by simple distillation and residual solvent was removed hydratable) as one of the soluble materials, hydratable by drying in a hot air oven at 45 °C for 2 h (Erickson et al one can be removed with 2-3% hot water while the ., 1980; Bligh and Dyer, 1959). The extracted oil non-hydratable ones are removed with addition of was stored in an air-tight bottle for further analysis. phosphoric or citric acid (Lusas, 2002; Gunstone and Refining process. The crude T. occidentalis seeds oil Norris, 1983). These phosphatides are referred to as extracted was then subjected to degumming, neutrali- gum and removal of this gum is called degumming. sation and bleaching processes. The next stage is neutralisation which is the process Degumming process. 400 cm3 of the crude oil was that removes free fatty acids from the degummed oil. heated to temperature of 70 °C followed by addition of This is done by mixing a calculated volume of a specific 0.80 cm3 of 50% phosphoric acid and the mixture was concentration of caustic soda (sodium hydroxide) with then vigorously stirred for 10 min. Thereafter 10 cm3 the oil at a definite temperature (60-80 °C) and atmos- of water heated to 80 °C was added and whole mixture pheric pressure (1.01325×104 N/M2), for a definite time agitated for another 10 min. The agitation was stopped and with prescribed agitation conditions. The alkali and the mixture was allowed to stand undisturbed for treatment is designed to remove the undesirable crude 1 h so that the mixture was separated into two layers oil impurities without saponifying any degummed oil i.e. oil and gum. The gum was drained off while the oil which would increase refining loss (Erickson et al., obtained was termed as degummed oil (Salunkhe et al., 1980). Vegetable oils generally contain colour pigments 1992, Carlson, 1991; Erickson et al., 1980). The degum- which are predominantly yellow, red and green. The med oil was further subjected to alkali neutralisation. yellow, orange and red pigments are known as Neutralisation process. Degummed oil (200 cm3) sample ‘carotenoids’. Other colours include chlorophyll, steroid, was heated up to 70 °C with constant stirring in a beaker, tocopherols and gozypoll (Bernardini, 1973). Colour then 3.3 cm3 of 3.59 M (20 Baume) sodium hydroxide pigment must be removed so as to produce oil of brighter solution was added to the oil with vigorous stirring and colour acceptable to consumers and these pigments are the temperature rose to 90 °C. Thereafter, 10 cm3 of removed from the oil with bleaching earth or fullers’ saturated solution of sodium chloride (an electrolyte) earth (John, 1990) through a process known as bleaching. was added and the resulting mixture was stirred The present work was aimed to assess the physico- vigorously at 90 °C for 30 min. Then left undisturbed chemical properties and fatty acid composition of crude, in a separating funnel for 6 h resulting into separated degummed, neutralised and bleached Telfairia occidentalis two layers, the lower layer which is known as ‘soap seed oil with a view of establishing the effects of these stock’ was then heated to 90 °C and washed with processes on the parameters. water then heated to 95 °C. The washing was done six Fluted Pumpkin Seed Oil Analysis 85 consecutive times to remove any excess caustic soda 250 °C and 300 °C, respectively. The peaks were identified and water soluble gum remaining in the oil (Salunkhe by comparison with standard fatty acid methyl esters et al., 1992, Erickson et al., 1980). The resulting neutral (ASTM, 1985). oil was then dried in a hot air oven, and later cooled in the desiccators. The dried oil was further bleached. Results and Discussion Bleaching processes. Neutralised (100 cm3) oil was Table 1 shows physicochemical parameter of crude, heated to 75 °C with constant agitation. Then 1 g of the degummed, neutralised and bleached oils obtained from bleaching earth was added and the mixture was heated fluted pumpkin seeds. Fluted pumpkin seeds contain to 110 °C with constant stirring for 45 min (Salunkhe 42.26±0.20% crude oil and the oil is light yellow with et al., 1992, Erickson et al., 1980). It was then filtered colour unit of 20 in half inch cell. This high yield value and the resulting oil was termed as bleached oil. indicates that the processing of the oil for industrial or edible purpose would be economical. The oil yield is Physicochemical characterisation of the oil samples. lesser than 54% reported for T. occidentalis (Akwaowa The crude, degummed, neutralised and bleached oil et al., 2000). There was no moisture in the crude sample samples were analysed for physicochemical properties. analysed and this implied that the oil may have prolong The moisture content and specific gravity were deter- shelf life. The colour of degummed, neutralised and mined according to AOAC (1990), while the refractive bleached oil samples were 10.0, 8.0 and 5.0 lovibond index was measured using Abbey Refractometer coupled unit, respectively, in one inch cell. This was calculated with thermometer (ASTM, 1985). The colour was based on the expression (5R+Y-B): determined using Lovibond Tintometer (Model 520).The colour of crude oil was determined in half (½") inch where: cell while that of degummed, neutralised and bleached R = the red pigment, Y = yellow pigment; B = blue oils were determined in 1” inch cell. The flash and fire pigment. points were measured using GallenKamp Authomatic The progressive decrease in colour from crude oil to Pensky-Martens flash point and fire point tester with bleached oil was as a result of phosphoric acid used for thermometer while the smoke point was determined degumming and bleaching earth specifically used to using Cleveland Open Cup apparatus (Lawson, 1995; remove colour pigments during bleaching (Abitogun ASTM, 1985). The temperature at which turbidity was and Oshodi, 2010; Bernardini, 1973). There was no first detectable also measured using Palm Test turbidity remarkable difference in the values of both specific tube (ASTM, 1985). The free fatty acid, acid value, gravity (0.923-0.924) and refractive index (1.474-1.475) saponification value and peroxide value were determined of crude, degummed, neutralised and bleached oil using methods described by AOAC (1990), while iodine samples. The specific gravities of the oil sample were value was determined by method described by Morris slightly higher than the specific gravity (0.913) of (1999) and Pearson (1976). pumpkin seed kernel oil reported by Mohammed (2004). Fatty acids identification. The oil samples were con- The refractive indices of the oil samples were slightly verted to fatty acid methyl esters (FAMEs) using the higher than 1.4721 reported for Adenopus breviflorus method described by Oshodi (1996) and Hall (1982). Benth seed oil (Akintayo and Bayer, 2002). The smoke The fatty acid methyl esters were analysed using an HP point (°C) for crude, degummed, neutralised and bleached 6890 gas chromatograph fitted with flame ionization oils were 243.00±0.03, 248.00±0.01, 250.00±0.02 and detector and powered with HP chemistation Rev.09.01 253.00±0.03, respectively, while that of flash point (°C) [206] software. The career gas was helium at pressure were 285.00±1.20, 289.00±1.30, 290.00±1.00 and of 19 psi.The FAMEs sample (1.5 µL) was injected and 304.00±1.10 and that of fire points (°C) were 345.00± the separation was carried out on an HP capillary column 1.10, 350.00±1.25, 353.00±1.20 and 358.00±1.55, (HP-INNowax; cross-linked PEG); 30.0 m length, respectively. The progressive increase in values of 0.32 mm i.d., and 0.50 µm film thickness. The oven smoke, flash and fire points from crude oil to bleached temperature was held initially at 60 °C for 2 min, incre- oil might be as a result of removal of impurities such ased from 180 °C at 12 °C/min to 320 °C at 14 °C/min as volatile organic material and the residual extraction and then maintained at 320 °C for 5.0 min. The tempera- solvent during the oil processing (Erickson et al., 1980). ture of the injection port and the detector were set at The high smoke, flash and fire points of the oil suggest 86 Jacob Olalekan Arawande et al.

Table 1. Physicochemical parameters of crude, degummed, neutralised and bleached oils obtained from fluted pumpkin seeds Parameters Crude oil Degummed oil Neutralised oil Bleached oil Specific gravity (at 25 °C) 0.924±0.002 0.923±0.003 0.923±0.003 0.923±0.002 Refractive index (at 25 °C) 1.475±0.002 1.474±0.004 1.474±0.003 1.474±0.010 Moisture content (%) 0.00 ± 0.00 0.00 ± 0.00 0.00 0.00±0.00 Turbidity point (JTU) 9.00±0.20 5.00±0.25 5.00±0.15 4.00±0.10 Smoke point (°C) 243.00±0.03 248.00±0.01 250.00±0.02 253.00±0.03 Flash point (°C) 285.00±1.20 289.00±1.30 290.00±1.00 304.00±1.10 Fire point (°C) 345.00±1.10 350.00± 1.25 353.00± 1.20 358.00±1.55 Colour (unit) 20.0 10.0 8.0 5.0 Free fatty acid (%) as Oleic 1.83±0.10 1.08±0.20 0.48±0.10 0.60±0.12 Acid value (mg/KOH/g) 3.64±0.29 2.14±0.20 0.94±0.25 1.18±0.14 Iodine value (g/100 g) 113.00± 0.17 115.70±0.12 116.20±0.18 121.50±0.20 Peroxide value (meq peroxide/kg) 1.80±0.28 0.90±0.12 0.50±0.07 0.30± 0.19 Saponification value (mg/KOH/g oil) 198.20±1.82 194.30±1.09 186.60±1.40 180.00± 0.06 Yield (%) = 42.26±0.20

Mean ± standard deviation of triplicate determination. that it can be suitable for deep frying purpose (Bello peroxide values (meq/peroxide/kg) of crude, degummed, et al., 2011; Akintayo and Bayer, 2002). Free fatty acid neutralised and bleached oil are 1.80±0.28, 0.90±0.12, (FFA) and acid values are among the characteristics 0.50±0.07 and 0.30±0.19, respectively. The low value features that are necessary for the confirmation of the was an indication that the oil has a high resistance to identity and edibility of oil. FFA can stimulate hydrolytic peroxidation and low rate of spoilage (Abayeh et al., deterioration of oils to form off- flavour components. 1998). This value is low as compared to the maximum The free fatty acid (% oleic acid) for the crude, degummed, acceptable value of 10 meq/KOH/g set by the Codex neutralised and bleached oils were 1.83±0.10, 1.08±0.20, Alimentarius Commission for edible oils (Abayeh 0.48±0.10 and 0.60±0.12, respectively while acid values et al., 1998). The oil is thus stable and will not easily (mgKOH/g) were 3.64±0.29, 2.14±0.15, 0.94±0.25 and go rancid. The iodine value (g/100 g) of the crude, 1.18±0.14, respectively. These values are relatively low degummed, neutralised and bleached oils are 113.00 that suggests application of the oil as good edible oil ±0.17, 115.70±0.12, 116.20±0.18 and 121.50± 0.20, (Akintayo and Bayer, 2002). respectively. The iodine value of the oil samples classi- It is noted that oil samples containing low FFA give fied the oil among the semi drying oil (Fernando and high smoke, flash and fire points and this quality will Akujobi, 1987). In addition, the high iodine value of enhance the suitability of the oil for deep fry cooking the oils indicates that the oil contains more unsaturated (Akintayo and Bayer, 2002). The FFA and acid value fatty acid than saturated fatty acid (Nielsen, 1994) since decreased from crude oil to neutralised oil but sudden iodine value is a measure of the extent of unsaturation increase is observed in bleached oil. The decrease in of fatty acid present in fats and oils (Nielsen, 1994). FFA and acid value is due to the effective use of caustic The iodine values are comparable to iodine value of alkali in neutralisation of the oil sample which led to 112.10 g/100 g for Adenopus Benth seed oil (Das et al., reduction in the free fatty acids, acid values and other 2002) and 121.03 g/100 g for African pea, Dacryodes impurities while the increase in FFA and acid values of edulis (Ajiwe et al., 1997). On the other hand, the values bleached oil is a result of acidic nature of bleaching obtained are higher than 83.50 g/100 g reported as earth used for colour removal (Salunkhe et al., 1992; iodine value of Moringa oleifera seed oil (Ogbunugafor Bernardini, 1973). The acid values of the samples were et al., 2011). Moreover, the iodine value of the oil incre- lower than the minimum acceptable value of 4.0% (for ases progressively at each stage of the processing owing crude oil) recommended by the Codex Alimentarius to gradual removal of some impurities present in it. The Commission for oil seed (Abayeh et al., 1998). The saponification values (mg/KOH/g oil) of the crude, Fluted Pumpkin Seed Oil Analysis 87 degummed, neutralised and bleached oil samples are acid has the highest composition and next to it is linoleic 198.20±1.82, 194.30±1.09, 186.60±1.40 and 180.00±0.06, acid. Palmitoleic acid was not detected in the crude oil respectively. The saponification values are relatively but was detected in degummed, neutralised and bleached lower as compared to palm kernel oil (253.781 mg/ oil samples. It is observed that fatty acid value increased KOH/g) (Arawande, 2013) and this indicates that the as the processing progressed from one stage to another. oil will not be good for soap making. The predominant fatty acid was oleic acid (47.40- 47.90%) and this high content suggest the oil to have Table 2 depicts the fatty acid composition of crude, ability to reduce incidence of coronary heart disease degummed, neutralised and bleached oils obtained from (CHD) because oleic acid decreases total cholesterol T. occidentalis fluted pumpkin ( ) seeds. The fatty acids (10%) and LDL cholesterol (Dennys et al., 2006). Also detected in the crude, degummed, neutralised and bleached the high level of unsaturated fatty acids content of the oil samples are myristic, palmitic, stearic, arachidic, oil in conjunction with its low free fatty acid and acid behenic, palmitoleic, oleic, linoleic and linolenic acids. values confirm the reasons for its edibility. The amount (%) of fatty acids in the crude oil are: myristic, 0.12; palmitic acid, 10.80; stearic acid, 0.24; Table 3 gives the summary of the total fatty composi- arachidic acid, 1.17; behenic acid, 0.42; oleic acid, tion of the oil. The values for saturated fatty acid in 47.40; linoleic acid, 26.36 and linolenic acid, 0.01. Also, crude, degummed, neutralised and bleached oils are the values (%) of these fatty acids in degummed oil are 12.75, 16.52, 17.59 and 19.15%, respectively, that of myristic acid (0.30), palmitic acid (12.40), stearic acid mono-unsaturated fatty acids are 47.40, 48.03, 48.15 (1.65), arachidic acid (1.70), behenic acid (0.46), and 49.32%, respectively, while that of polyunsatu- palmitoleic acid (0.17), oleic acid (47.86), linoleic acid rated fatty acids are 26.37, 27.37, 28.21 and 30.49%, (27.36) and linolenic acid (0.01), respectively. In respectively. The total fatty acid in crude, degummed, addition, the following fatty acids are present in the neutralised and bleached oils are 86.52, 91.92, 93.95 neutralised oil sample, myristic acid (0.45%), palmitic and 98.96%, respectively. There is progressive increase acid (13.00%), stearic acid (1.80%), arachidic acid in the saturated, monounsaturated, polyunsaturated (1.83%), behenic acid (0.51%), palmitoleic acid (0.25%), and total fatty acid of fluted pumpkin seed oil during oleic acid (47.90%), linoleic acid (28.20%) and linolenic processing from crude oil to bleached oil. In addition, acid (0.01%). Moreover the amount (%) of fatty acids these results show that the proportion of unsaturated present in the bleached oil is reported as follows: myristic fatty acids was much higher i.e., 73.77%-79.81%, and acid is 0.63, palmitic acid is 13.57, stearic acid is 2.33, that of saturated fatty acids was less than 25%. This arachidic acid is 2.00, behenic acid is 0.62, palmitoleic is in accordance with the values reported for other acid is 0.30, oleic acid is 49.02, linoleic acid is 30.44 vegetable oils (Salman and Tanver, 2005; Raie et al., and linolenic acid is 0.05. In all the oil samples, oleic 1992) which is characteristics for these oils.

Table 2. Fatty acid composition of crude, degummed, neutralised and bleached oils obtained from fluted pumpkin seeds Fatty acid Fatty acids Carbon number Crude oil Degummed oil Neutralised oil Bleached oil methylester (%) (%) (%) (%) Myristate Myristic 14:0 0.12 0.30 0.45 0.63 Palmitate Palmitic 16:0 10.80 12.40 13.00 13.57 Stearate Stearic 18:0 0.24 1.65 1.80 2.33 Arachidate Arachidic 20:0 1.17 1.70 1.83 2.00 Behenoate Behenic 22.0 0.42 0.46 0.51 0.62 Palmitoleate Palmitoleic 16:1 N.D 0.17 0.25 0.30 Oleate Oleic 18:1 47.40 47.86 47.90 49.02 Linoleate Linoleic 18:2 26.36 27.36 28.20 30.44 Linolenate Linolenic 18:3 0.01 0.01 0.01 0.05 ND = not detected. 88 Jacob Olalekan Arawande et al.

Table 3. Summary of fatty acid composition of crude, and antinutrients in fluted pumpkin (Telfairia degummed, neutralised and bleached oils obtained from occidentalis Hook). Food Chemistry, 70: 235-240. fluted pumpkin seeds Alfawaz, M.A. 2004. Chemical composition and oil Oil sample Saturated Mono- Poly- Total characteristics of Pumpkin (Cucurbita maxima) fatty acid unsaturated unsaturated (%) seed kernels. Food Science and Agriculture Research (%) fatty acid fatty acid Centre, King Saud University, Research Bulletin (%) (%) No. 129, 5-18. Crude 12.75 47.40 26.37 86.52 AOAC, 1990. Official Methods of Analysis, pp. 1250- Degummed 16.52 48.03 27.37 91.92 1255, Association of Official Analytical Chemist, Neutralised 17.59 48.15 28.21 93.95 Washington D.C., USA. Bleached 19.15 49.32 30.49 98.96 Arawande, J.O. 2013. Antioxidative Activities of Extract of Fruit Peels and Vegetables on Edible Oils. Ph.D. Thesis, 61 pp., Federal University of Technology, Conclusion Akure, Ondo State, Nigeria. Processing of fluted pumpkin seed oil from crude to Asiegbu, I.E. 1987. Some biochemical evaluation of Journal of Science, Food and bleached oil increases iodine value, smoke point, flash fluted pumpkin seed. Agriculture point, fire point, saturated, monounsaturated, polyunsatu- , 40: 151-155. rated and total fatty acid composition of the oil. Whereas, ASTM, 1985. American Society for Testing Materials. ASTM Publication processing of the oil from crude oil to bleached oil , pp. 31-36, 40-48. resulted in decrease in values of saponification, peroxide, Bello, M.O., Akindele, T.L., Adeoye, D.O., Oladimeji, colour and turbidity. It is obvious that if the bleached A.O. 2011. Physicochemical properties and fatty acids profile of seed oil of Telfairia occidentalis oil is deodourized, it will supply essential fatty acid Hook, F. International Journal of Basic and Applied needed in the body and the final refined oil will be of Sciences, 11: 9-14. high quality in terms of physicochemical properties and Bernardini, E. 1973. Oil and Fat Technology, pp. 709- this will enhance its edibility. It is further suggested 719. Technologie Publishing House, S.R.L. Rome, that the bleached oil should be deodourised and analysed Italy. for physicochemical properties and fatty acid compo- Bligh, E.G., Dyer, W.J. 1959. A rapid method of total sition. lipid extraction and purification. Canadian Journal of Biochemistry and Physiology, 37: 911-917. References Carlson, K.F. 1991. Fats and oils processing. INFORM, Abayeh, O.J., Aina, E.A., Okonghae, C.O. 1998. Oil 2: 1046-1060. content and oil quality characteristics of some Chukwuonso, E.C.C.E., Paschal, C.U., Lawrence, U.S.E. Nigerian oil seeds. Journal of Pure and Applied 2010. Dietary incorporation of boiled fluted Sciences, 1: 17-23. pumpkin (Telfairia occidetalis Hook F.) seeds 1: Abitogun, A.S., Oshodi, A.A. 2010. Effects of degum- Growth and toxicity in rats. Research Journal of ming and bleaching on physicochemical properties Biological Sciences, 5: 140-145. of crude sunflower oil seeds. Journal of Chemical Das, M., Das, S.K., Suthar, S.H. 2002. Composition of Society of Nigeria, 35: 57-61. seeds and characteristics of oil from karingda Ajiwe, V.I.E., Okeke, C.A., Nnabuike, B., Ogunleye, [Citrullus lanatus (Thumb) Mansf]. International G.A., Elebo, E. 1997. Application of oil extracted Journal of Food Science & Technology, 37: 893- from African star apple (Chrysophyllum africanum), 896. horse eye bean (Mucuna sloanei) and African pear Dennys, E.C.C., Andre, G.V.C., Maria do, C.G.P., (Dacryodes edulis). Bioresource Technology, 59: SergioMatta, D.S., Marco, T.C.S., Neuza, M.B.C. 259-261. 2006. Lipid profile of rats fed high-fat diets based Akintayo, E.T., Bayer, E. 2002. Charaterisation and on flaxseed, peanut, trout, or chicken skin. Nutrition, some possible uses of Plukenetia conophora and 22: 197-205. Adenopus breviflorus seeds and seed oils. Boire- Effiong, G.S., Ogban, P.I., Ibia, T.O., Adam, A.A. 2009. source Technology, 85: 95-97. Evaluation of nutrient-supplying potential of fluted Akwaowo, E.U., Ndon, B.A., Etuke, E.U. 2000. Minerals pumpkin (Telfairia occidetalis Hook F.) and Okra Fluted Pumpkin Seed Oil Analysis 89

(Abelmoschus esculentus) (L.) Moench. Academic Lawson, H. 1995. Food Oils and Fats-Technology, Journal of Plant Sciences, 2: 209-214. Utilization and Nutrition, pp. 65-69, Springer, USA. Erickson, D.R., Pryde, E.H., Brekks, O.L.,Mount, T.L., Lusas, E.W. 2002. Oil seeds and oil bearing materials. Falb, R.A. 1980. Handbook of Soy Oil Processing In: Handbook of Cereal Science and Technology, and Utilization, pp. 65-68, American Soybean K. Kulp and J. G. Porte, Jr. (eds), pp. 297-362, Marcel Association St. Louis and the American Oil Dekker Inc., New York, USA. Chemists Society, Champaign, Illinois, USA. Morris, B., Jacobs. 1999. The Chemical Analysis of Fagbemi, T.N., Oshodi, A.A., Ipinmoroti, K.O. 2005. Food and Food Product, pp. 357-390, 3rd edition, Processing effects on some antinutritional factors CBS Publisher, New Dehli, India. and in vitro multienzyme protein digestibility (IVPD) Nielsen, S.S. 1994. Introduction to the Chemical Analysis of three tropical seeds: Breadnut (Artocarpus altilis), of Foods, pp. 257-390, Champan & Hall, New Cashewnut (Anacardium occidentale) and Fluted York, NY, USA. pumpkin (Telfairia occidentalis). Pakistan Journal Nkang, A., Omokaro, D., Egbe, A., Amanke, G. 2003. of Nutrition, 4: 250-256. Variation in fatty acid proportions during desiccation Fernando, C.E.C., Akujobi, E.O. 1987. Chemical analysis of Telfairia occidentalis seeds harvested at physio- of selected vegetable oils and fats of Sokoto State logical and agronomic maturity. African Journal of Nigeria. Journal of Basic and Applied Sciences, of Biotechnology, 2: 33-39. 1: 11-15. NRC, 2001. Nutrient Requirement of Dairy Cattle, 1333 Giami, S.Y., Bekebian, D.A. 1992. Proximate composi- pp., 7th revised edition, National Academies Press, tion and functional properties of raw and processed National Research Council, Washington DC., USA. full fat fluted pumpkin (Telfairia occidentalis) seed Ogbunugafor, H.A., Eneh, F.U., Ozumba, A.N., Igwo- flour. Journal of Science, Food and Agriculture, Ezikpe, M.N., Okpuzor, J.,Igwilo, I.O., Adenekan, 59: 321-325. S.O., Onyekwelu, O.A. 2011. Physicochemical and Giami, S.Y., Isichei, I. 1999. Preparation and properties antioxidant properties of Moringa oleifera seed of flours and protein concentrated from raw, oil. Pakistan Journal of Nutrition, 10: 409-414. fermented and germinated fluted pumpkin (Telfairia Okoli, B.E., Mgbeogu, C.M. 1983. Fluted pumpkin occidetalis Hook) seeds. Plant Foods for Human (Telfairia occidentalis): West African vegetable Nutrition, 54: 67-77. crop. Economic Botany, 37: 145-149. Gill, L.S. 1992. Ethnomedical Uses of Plants in Nigeria, Oshodi, A.A. 1996. Amino acid and fatty acid compo- pp. 165-248, Universty of Benin City, Nigeria. sition of Adenopus breveflorus Benth seed. Inter- Giwa, S., Abdullah, L.C., Adam, N.M. 2010. Egnsi national Journal of Food Sciences and Nutrition, (Citrullus colocynthis L.) seed oil as potential bio- 47: 295-298. diesel feedstock. Energies, 3: 607-618. Pearson, D. 1976. The Chemical Analysis of Foods. Gunstone, F.D., Norris, F.A. 1983. Lipids in Food: pp. 6-14, 200-227, 7th edition, Churchill Living Chemistry, Biochemistry and Technology, pp. 58-68, Stone, London, UK. Pergamon Press, New York, USA. Raie, M.Y., Ijaz, A., Akhtar, M.W. 1992. Distribution Hall, G.M. 1982. Silage from tropical fish. Lipid of fatty acids in triglycerides of Carum capticum. behavior. Journal of Food Technology, 21: 45-54. Pakistan Academic Science, 26: 199-206. Hamed, S.Y., El-Hassan, N.M., Hassan, A.B., Eltayeb, Salman, R., Tanver, A.C. 2005. Intramolecular fatty M.M., Babiker, E.E. 2008. Nutritional evaluation acids distribution in the triglyceride of Hordeum and physicochemical properties of processed vulgare. Pakistan Journal of Scientific and Indus- pumpkin (Telfairia occidentalis Hook) seed flour. trial Research, 48: 389-392. Pakistan Journal of Nutrition, 7: 330-334. Salunkhe, D.K., Chavan, R.W., Adsule, Kadam, S.S. John, M.D. 1990. Principles of Food Chemistry, 2nd 1992. World Oil Seed, Chemistry Technology and edition, pp. 57-70, Van Nostrand Reinhold, New Utilization, pp. 148-192, An AVI Book Published York, USA. by Van Nostrand Reinhold, New York, USA. Pak. j. sci. ind. res. Ser. A: phys. sci. 2016 59(2) 90-95

Liberation Studies of Padhrar Coal By Using Fractionation Method, XRD Analysis and Megascopic and Microscopic Techniques Muhammad Shahzad*, Zulfiqar Ali, Yasir Majeed, Zaka Emad, Muhammad Aaqib and Bilal Adeel Mining Engineering Department, University of Engineering & Technology, Lahore, Pakistan

(received October 2, 2014; revised September 14, 2015; accepted December 31, 2015)

Abstract. This research study aims to establish liberation characteristics of Padhrar coal by using various methods including fractionation method, megascopic and microscopic analysis and X-ray diffraction (XRD) technique. Sieve analysis revealed that more than 83% of the coal lied in the medium particle size range of -26.670+6.680 mm. The results of fractionation analysis indicated that most of the sulphur was found in the smaller sized fractions having particle size less than 6.680 mm while most of the ash was found to be associated at larger particle size (+26.670 mm) and at relatively smaller particle size (-6.680 mm). It was found that Padhrar coal consisted of three major minerals namely; quartz, pyrite and kaolinite. These minerals were found to be associated with organic matter at different particle size levels, thus making the nature of the Padhrar coal more complex for its cleaning. Keywords: liberation study, Padhrar coal, fractionation method, XRD of coal, megascopic analysis, microscopy of coal

Introduction located in the Eastern and the Central Salt Range and in the Makerwal area of Surghar range. In the recent Coal is a complex heterogeneous mixture of organic past, the coal resources of Punjab have been defined to and inorganic constituents (Ural, 2007). These are be located in the seven zones, Padhrar coal zone is one usually present in the form of solid, liquid, and gaseous of them containing coal resource of 63.83 million tonnes phases intimately mixed with each other (Liu et al., (Snowden, 2010). 2005; Vassilev and Vassileva, 1996). Although, the nature of coal (rank and type) and the utilization of coal Until now, Padhrar coal could not find its applications in different processes are primarily dependent upon its in electricity production and cement industry due to its organic components (Liu et al., 2005; Vassilev and low quality. It contains large amounts of mineral matter Tascon, 2003), but the amount and type of inorganic and sulphur. Few attempts were made to upgrade the matter also play a key role in defining the end use. quality of Padhrar coal through physical cleaning methods (Shahzad et al., 2015). But these were not The inorganic matter of coal is generally classified into proved successful due to the lack of or poor knowledge three major classes namely; dissolved salts, discrete of liberation characteristics of Padhrar coal. particles, and inorganic elements mingled within organic constituents (Vessilev and Vassileva, 1996; Ward, 2002). Most of the difficulties associated with coal utilization Dissolved salts are usually present in the pore water of arise from inorganic mineral matter rather than maceral coal. Discrete inorganic particles which may be composition. Mineral matter act as a diluent in coal, crystalline or non-crystalline represents true mineral displacing combustible material with non-combustible components of coal (Ward, 2002). The crystalline matter (Ward, 2002). mineral matter typically comprised of oxides-hydroxides, The form and quantity of inorganic components have sulphides-sulphosalts, sulphates, silicates, carbonates, profound effect on the behaviour of coal during phosphates, vanadates, tungstates, chlorides, native combustion. Quartz particles give rise to erosion in the elements, and other mineral classes (Ural, 2007; Vassilev grinding mills and on exposed surfaces of the furnace. and Vassileva, 1996). Sulphur in various forms have long been recognized as a cause of corrosion of furnace and boiler pipes as well Punjab area of Pakistan has a coal resource potential as the major source of environmental pollution during of 596 million tonnes. Coal reserves in Punjab are burning of coal. Slagging in furnace is mostly associated *Author for correspondance; E-mail: [email protected] with iron sulphides, siderite or calcite (Creelman and 90 Liberation Studies of Padhrar Coal 91

Ward, 1996). These problems demand the removal of was divided into four equal parts by drawing two undesirable elements from coal before its final use in perpendicular lines on its top surface and the centre of the industry. Since the nature and distribution of mineral each quarter was selected as one of the four sampling matter present in the coal, have fundamental effect on points. Four increments each having an amount of 5 kg the coal cleaning technologies, the characterization and were taken from each point by using a shovel. The liberation studies of the mineral matter are critically number of increments collected in this way were totaled important (López and Ward, 2008; Ward, 2002). to be 48 and the total amount of the gross sample collected from three coal stockpiles was found Several methods are available to determine the type approximately equal to 240 kg. The gross sample was and amount of mineral matter and to assess their preserved into two plastic drums already lined with associations with coal macerals. Fractionation method plastic sheets of 0.4 mm size along their inner walls. was used by several researchers (Cloke et al., 2002; Spears and Booth, 2002) to study associations of mineral The gross sample was crushed by using Denver components with organic material of coal in different laboratory jaw crusher set at ¾ inches. The gross sample size fractions. Microscopic (petrographic) study of coal was further divided into four sub samples by the standard has long been used to study the kinds and amounts of procedure of coning and quartering. Sieve analysis was macerals and minerals. It was also utilized to determine performed on one sub-sample. The individual sieve the degree of coalification by measuring the percentage fractions were weighed and subsequent weights were of reflectance in reflected light (Valentim et al., 2006). recorded. After that, all the size fractions were further The powder X-ray diffraction (XRD) is probably the ground to -0.177 mm by using laboratory disc mill. most widely employed technique both for qualitative These ground fractions were analyzed for their ash and and quantitative analysis of minerals in coal (Ritz and sulphur contents following ASTM standard test methods Klika, 2010; Saikia et al., 2007; Ward et al., 2001; (ASTM D-3174, ASTM E-775). All these tests were Wertz and Collins, 1998). conducted on air-dried basis. This study aims to determine the quality of coal from ASTM standards (D-3173, 3174 and 3175) were used Padhrar area of district Khushab, Punjab, Pakistan. to perform a proximate analysis on another coal sub Another objective of the present research is to establish sample obtained from bulk sample which was first the liberation characteristics of this coal. Moreover, this ground to -0.177 mm. The percentage of total sulphur paper also describes the nature of associations of mineral was also determined for this sub-sample by using matter with coal matrix. standard test method (ASTM E-775). Bomb calorimeter was utilized to find out the calorific value of coal sample. Materials and Methods Megascopic pictures of randomly selected coal pieces Coal sample in bulk amount was collected from Punjab were captured on the site while microscopic photographs Mineral Development Corporation (PUNJMIN) coal were taken by using MM6C-AF-2 microscope (Fig. 1) mine located in Padhrar coalfield of Punjab, Pakistan. in the reflected light. A very fine size coal sample Since primitive methods of mining are being used in (-0.150 mm) was prepared and subjected to X-ray Punjab coal mines, it was not possible to take sample diffraction analysis using computer controlled Philips by using mechanical means. XPERT PRO diffractometer system with Cu Ka The sample was collected from a coal stock pile in radiation having a wavelength of 1.54 Å. The scan accordance to ASTM D-2234 and ASTM D-6883. The range was kept starting from 4.990° and ending at production of the mine was about 80-90 metric tonnes 120.000° with step size of 0.035°. The total measuring per day. There were three piles each containing time was observed to be 8.15 min. The data thus obtained approximately 20 metric tonnes of coal (the amount of was stored in a digital format. JCPDS Powder Diffraction coal that was loaded into a single truck). These stockpiles File was used to identify the minerals from diffractogram. contained fresh coal that was mined at that day. The height of each coal pile was measured to be less than Results and Discussion 1.5 meter. The top surface of each pile was levelled. Sieve analysis. The results of sieve analysis are given For the collection of gross sample, four points were in Table 1. The graph between cumulative mass selected in each coal pile in such a way that the pile percentage and average aperture size of each fraction 92 Muhammad Shahzad et al.

Table 2. Summary of results of proximate analysis Characteristic Value Moisture content (%) 3.13 Ash content (%) 32.40 Volatile matter (%) 34.07 Fixed carbon (%) 30.40 Total sulphur (%) 5.60 Calorific value (Kcal/kg) 4753

Padhrar coal to sub-bituminous C type class (ASTM D388). It also contains higher amounts of ash and sulphur which restrict its use in power generation and Fig. 1. MM6C-AF-2 microscope. cement manufacturing. is shown in Fig. 2. It may be noted that about 83.48% Fractionation method. The results of ash and sulphur of the total mass lies in the size range of -26.67 determination tests performed on all the seven fractions mm+6.68mm. are given in Table 3. It can be seen that vast variations exist in the values of ash and sulphur at various coal Proximate analysis. The results of proximate analysis particle sizes. It is interesting to note that the largest- and combustion analysis (sulphur and calorific value) sized fraction (+26.670 mm) has the highest amount of of the Padhrar coal samples are given in Table 2. The ash contents while the lowest values of total sulphur results obtained from the proximate analysis categorize percentage are also found in the same size class. The ash contents were the lowest for the medium-sized Table 1. Masses of the fractions retained at sieves of fraction i.e., for the size range of -13.330+6.680 mm. different mesh size The sulphur contents were the highest in the size range Fraction size Mass retained Cumulative mass of -6.680+3.327 mm. The ash contents in other size (mm) (%) Passing (%) Retained (%) fractions vary from 30.53 to 35.33 %. These abrupt variations in the fractionation analysis of Padhrar coal +38.00 0.00 100.00 0.00 may be attributed from the complex associations of -38.00+26.67 1.57 98.43 1.57 mineral matter with the organic materials. It may be -26.67+18.86 22.50 75.93 24.07 safely concluded that Padhrar coal contains various -18.86+13.33 28.34 47.59 52.41 -13.33+6.68 32.64 14.95 85.05 minerals in different forms which have associations at -6.68+3.327 8.47 6.48 93.52 different levels of particle size and liberate accordingly. -3.327+1.680 2.90 3.58 96.42 The highest ash contents in the largest-sized fraction - 1.680 3.58 0.00 100.00 may be due to the presence of mineral matter in the form of nodules or bands at larger sizes. When these

120 Table 3. Results of ash and sulphur determination tests 100 performed on different size fractions of Padhrar coal 80 Aperture size Average ash content Total sulphur 60 (mm) (%) (%) Passing 40 Retaining +26.670 48.96 3.430

Cumulative mass % 20 -26.670+18.860 30.63 5.280 -18.860+13.330 34.33 5.490 0 0 5 10 15 20 25 30 35 40 -13.330+6.680 25.30 4.935 Aperture size (mm) -6.680+3.327 35.33 7.050 -3.327+1.680 32.40 6.556 Fig. 2. Graph between aperture size and cumulative -1.680 30.53 6.934 mass percentage. Liberation Studies of Padhrar Coal 93 large-sized particles are broken, these nodules are fragmented into very fine particles which go down into the smaller-sized fractions leaving more organic rich coal particles in the medium-sized fractions. X-ray diffraction analysis. Diffractogram of Padhrar coal sample is shown in Fig. 3 while Table 4 presents the major minerals identified by the peak pattern. These minerals are kaolinite, quartz and pyrite. Megascopic and microscopic analysis. Megascopic and microscopic studies were also performed on Padhrar coal samples. Figure 4 shows bands and nodules of quartz packed in the coal. It may be noted that quartz is present in the form of amalgamates of loosely adhered fine particles. The size of these amalgamates varies approximately from 5 mm to greater than 25 mm. These masses of quartz are found to be the major reason of higher ash contents at larger coal particle sizes. Since these amalgamates consist of fine particles loosely bound together, when the lumps of coal are broken, they are separated from the coal and disintegrate into individual fine grains. These disintegrated fine grains Fig. 4. (A) Lump of coal containing quartz ultimately go into the finer fractions during sieve particles in the form of concentrated masses analysis, leaving the organic rich coal particles in the and bands, (B) Quartz and pyrite nodules middle fractions. along with coal particles resulted by the breakage of a coal lump.

800 Figure 5 shows that pyrite is present in various forms in the Padhrar coal. It is found both in thick and thin 600 bands as well as in the form of nodules. These bands or nodules are relatively smaller than the quartz masses, 400 having greater adherence among the particles. When Counts the coal lumps are broken, these nodules and bands

200 separate from the organic materials and pass down into the medium sized coal fractions due to their relatively larger size. This results an increased total sulphur 0 10 20 30 40 50 60 70 80 90 100 110 percentage in the medium sized (-6.680+3.271 mm) Position (2°q) coal particles. In Fig. 5(B) a clay rich coal band can be seen at the left side just before the pyrite nodule. It Fig. 3. X-ray diffractogram of Padhrar coal sample represents that clayey material is intimately associated (K=Kaolinit; Q=Quartz; P=Pyrite). with organic matter and does not liberate at coarser sizes, thus causing higher ash values in lower size classes. Table 4. Pattern list of Padhrar Coal Figure 6 presents the microscopic photographs of Ref. Code Compound Chemical Score selected Padhrar coal pieces taken in reflected light by name formula using MM6C-AF-2 microscope. A very thin band of quartz (white) can be seen in the top left corner of the 00-001-0527 Kaolinite Al Si O (OH) 12 2 2 5 4 picture. Pyrite particles (golden colour) are unevenly 00-005-0490 Quartz SiO 25 2 distributed in the coal while clayey particles (dirty 00-042-1340 Pyrite FeS2 16 white) are associated with coal matrix at finer sizes. 94 Muhammad Shahzad et al.

Fig. 5. (A) Pyrite and quartz bands of medium thickness in the coal, (B) Pyrite nodules and thin bands in the coal lump.

Fig. 6. Microscopic views of coal particles.

All the above results confirm that the Padhrar coal Conclusion exhibits both epigenetic and syngenetic nature. Mineral This research work was conducted to assess the quality matter is present in the form of large nodules, bands, and liberation characteristics of Padhrar coal. The results and amalgamates of fine materials as well as in the of proximate analysis categorized the Padhrar coal into form of finer particles intimately associated with coal sub-bituminous C type class. It was also showed that matrix at very small size. Padhrar coal contained high amount of sulphur and Liberation Studies of Padhrar Coal 95 mineral matter. Sieve analysis revealed that more than López, I.C., Ward, C.R. 2008. Composition and mode 83% of the coal lied in the medium particle size range of occurrence of mineral matter in some Colombian of -26.670+6.680 mm. The results of fractionation analysis coals. International Journal of Coal Geology, 73: indicated that most of the sulphur was found in the smaller 3-18. sized fractions having particle size less than 6.680 mm Ritz, M., Klika, Z. 2010. Determination of minerals in while most of the ash was found to be associated at larger coal by methods based on the recalculation of the particle size (+26.670 mm) and at relatively smaller bulk chemical analyses. Acta Geodynamica et particle size (-6.680 mm). X-ray diffraction (XRD) studies Geomaterialia, 7: 453-460. showed the presence of three major minerals namely; Saikia, B.K., Boruah, R.K., Gogoi, P.K. 2007. FT-IR kaolinite, pyrite and quartz. The megascopic and and XRD analysis of coal from Makum coalfield Journal of Earth System Science microscopic analysis confirmed their presence. 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Modeling the Land Suitability using GIS and AHP for Cotton Cultivation in Punjab, Pakistan Nabila Naza and Haroon Rasheedb* aDepartment of Computer and Software Engineering, Bahria University, Karachi Campus 13, National Stadium Road, Karachi-75260, Pakistan bDepartment of Electrical Engineering, Bahria University, Karachi Campus 13, National Stadium Road, Karachi-75260, Pakistan

(received October 29, 2014; revised September 10, 2015; accepted October 19, 2015)

Abstract. The main goal of this research was to establish a spatial model for identification of suitable land for cotton in Punjab, Pakistan through evaluation of multidisciplinary variables by applying geographic information system (GIS) and analytical hierarchy process (AHP) approach. In this model, rivers were used as constraint and seven factors were temperature, soil physical and chemical properties, soil pH, aridity classes, agro-ecological zones, and river command area. On the basis of these parameters suitability maps were generated. By pair-wise comparison matrix (PWCM) of AHP, weights were extracted by means of principal Eigen vector by Saaty’s method, with accepted consistency ratio of 0.09. Multi-criteria evaluation (MCE) employing weighted linear combination aggregates all suitability maps to generate final suitability map. It was found that more potential sites exist along with existing cotton practiced area. The result provided important information for farmers to establish linkage between policy decisions and regulatory actions and to improve agricultural land management. Keywords: cotton, multi criteria evaluation, analytic hierarchy process, land suitability, geographic information systems, pair-wise comparison matrix

Introduction the best location for agricultural production, necessitate Cotton is Pakistan’s main cash crop and is planted on the use of geo-spatial domain which accelerates the rate 15% of its total arable land during the Kharif season. of adoption of sustainable agriculture and benefit the This production is concentrated in two provinces with farmers and decision makers in agriculture planning et al Punjab accounting for nearly 75% and Sindh nearly (Joshua ., 2013). 25% of this arable land. An estimated 1.6 million farmers One of the most important applications of GIS is the grow cotton in Pakistan. The textile sector is the largest display and analysis of data for environmental decision- industrial sector in Pakistan and accounts for about 40% making (Eastman, 1999). GIS-based MCDA combines of the industrial labour force (Shafiq-ur-Rehman, 2015). and transforms spatial and aspatial data (input) into a Pakistan is the 4th largest producer, 7th largest consumer, resultant decision (output) (Malczewski, 2004). The 7th largest importer, and 12th largest exporter of cotton hierarchical model i.e., analytical hierarchy process (AHP) in the world (FAS-USDA, 2015). consists of objectives, criteria and sub-criteria. To evaluate As a semi-industrialized country, Pakistan is heavily the criteria, scoring was made with preference scale, dependent on the agricultural crops such as cotton. As and a pair-wise comparison matrix (PWCM) was created, a result, the policymakers are facing a great challenge for which consistency should be below 0.10 (Akinci et al to carry out agricultural reform for sustainability and ., 2013). Multi-criteria evaluation (MCE) model optimization of resources. To meet the increasing demand finds solutions to decision-making problems character- of agri-products, it is not feasible to bring more area ized by multiple factors, and weighted linear combination under cultivation due to limited resources. To tackle this, (WLC) aggregate them into a final land suitability index focus has shifted from extensive to intensive farming (Khoi and Murayama, 2010). for precision farming and sustainable agriculture (Perveen Using GIS based MCE model, cotton land suitability et al., 2013). To deal with the complexities for selecting in Sindh, Pakistan was evaluated by agro-informatics *Author for correspondence; data of soil, ground water availability, irrigation methods, E-mail: [email protected] climate, land use, cropping patterns and agro-ecological

96 Modeling the Land for Cotton Cultivation 97 zones (Perveen et al., 2013). For the delineation of suitable to the east, Sindh to the south, Balochistan to the soil for Zero-Till Wheat cultivation in Gujranwala, southwest, and Khyber Pakhtunkhwa to the west and Pakistan based on soil profile, ground water survey data Islamabad to the north. Temperature ranges from -2°- and satellite imagery was employed with GIS framework 45 °C. Climatically, Punjab has three major seasons using weighted overlay (Iqbal and Mehdi, 2008). To namely; hot weather (April to June); rainy season (July achieve the goal of land suitability assessment in to September); mild weather (October to March). Six Sheikhupura and Nankana Sahib Districts of Punjab, rivers of Punjab named Indus, Jhelum, Beas, Chenab, Pakistan, GIS based AHP model was employed by using Ravi and Sutlej provide heavy irrigation system by factors of soil texture and water supply along with other canals throughout the province. The province is mainly factors for production of rice (Waqar et al., 2013). a fertile region along the river valleys. The region Delineation of land suitability for agricultural production contains Thal and Cholistan deserts. Despite of its was employed by applying GIS based AHP approach tropical wet and dry climate, extensive irrigation makes in Yusufeli district of Artvin city, Turkey, using soil, it a rich agricultural region. Wheat and cotton are the land use capability class, land use capability sub-class, major crops. Cotton and rice are important cash crops soil depth, slope, aspect, elevation, and erosion degree (Saif, 2014). to identify highly, moderately, marginally and unsuitable Criteria for cotton cultivation. In the study area, cotton areas (Akinci et al., 2013). GIS and MCDA model using is sown in May or June and picking starts in September AHP technique employed for suitable agricultural land or October. The data source (Table 1) describes the in Greater Karu, Nasarawa state, Nigeria using soil, source for obtaining all the factors and constraint utilized slope, water bodies and geological maps to indicate for evaluating the cotton land suitability. The suitable highly, moderately and not suitable areas (Joshua et al., criteria (Arain et al., 2014) for cotton cultivation was 2013). For the identification of suitable areas for cropland characterized by the factors: canal command area; soil in the Tam Dao National Park region, Vietnam, a GIS- pH (6.6-8.4); soil chemical properties classes (Acid based MCE model using biophysical factors and Landsat soils: slightly acid, neutral soils: non-calcareous to slight ETM+ imagery indicated the location and extent of calcareous, mildly alkaline soils: non-calcareous to crop farming areas at different suitability levels (Khoi strongly calcareous, moderately alkaline soils: non- and Murayama, 2010). calcareous to strongly calcareous); mean maximum The main research problem was the cotton requirements annual temperature (34 °C); soil physical properties and limited cotton cultivation in the study area. Also a constant growth in Pakistani yarn exports was observed from 2009-2013 (Hussain, 2014). This research aims Table 1. Data source for the research work to identify suitable land for cotton cultivation in Punjab, Data Source Pakistan using the GIS based AHP modeling. It starts Statistical data for cotton production from geometric correction, digitization of the re-sampled for Punjab and maps to form the vector layers, then rasterization for Statistical data for area sown under all the factors and constraint was performed. After Cotton for Punjab. (Rafique, 2013) standardizing the maps for suitability, AHP was applied Digital scanned map of Punjab rivers, to weight the factors using Saaty’s PWCM for suitability Digital scanned map of soil physical analysis. Finally, the MCE technique was applied using properties, Digital scanned map of soil pH and WLC for all the factors and constraints, to decide the Digital scanned map of soil chemical land suitability for cotton cultivation in the study area. properties. (Panagos et al., 2011) Shape file of mean maximum annual Materials and Methods temperature. (ICIMOD, 2008) Site description. Punjab is the second largest province Digital map of canal command area. (Brabben, 2000) of Pakistan in terms of land area at 205,344 km2 with Shape file of Punjab administrative coordinate extending from 27° 42' 20.16'' N to 34° 00' boundary. (PCO, 2013) 59.04'' N latitude and from 69° 20' 6'' E to 75° 22' 49.44'' Digital map of aridity classes and E longitude. The province is bordered by Kashmir to Digital map of agro-ecological zones. (PARC, 2007) the north-east, the Indian states of Punjab and Rajasthan 98 Nabila Naz and Haroon Rasheed classes (River plain and terraces:non-calcareous and Table 2. Standardized criteria for the constraint and calcareous loamy soils); aridity classes (Arid (Kharif, factors used in this work Rabi) and Arid (Kharif), Hyper-arid (Rabi); agro- Layer Standardization description of factors and constraint ecological zones (irrigated plains-D.G. Khan irrigated Canal Suitable: Canal command area and irrigated plains-cotton zone). Rivers were used as command Not suitable: area 1. Rivers constraint. The detailed description of suitability (Table 2. Non-canal command area 2) for factors and constraint describes both suitable and Soil pH Suitable: 6.6 - 8.4 not suitable criteria for cotton cultivation (Arain et al., Not suitable: 1. 6.1 - 6.5 2014). These values were in agreement with those 2. > 8.4 considered in the literature. Soil Suitable: chemical 1. Acid soils: slightly acid Developing GIS based AHP model. For cotton land properties 2. Neutral soils: non to slight calcareous 3. Mildly alkaline soils: non to slight calcareous suitability, GIS-based AHP model (Fig. 1) describes all 4. Mildly alkaline soils: moderately calcareous the steps used in this study. The geometric rectification 5. Mildly alkaline soils: strongly calcareous 6. Moderately alkaline soils: non to slight calcareous of the downloaded digital scanned maps, which were 7. Moderately alkaline soils: moderately calcareous originally constructed in geographic latitude longitude 8. Moderately alkaline soils: strongly calcareous Not suitable: projection, for all the factors and constraint was done 1. Salt affected soils: saline 2. Salt affected soils: saline-sodic in Erdas Imagine 9.2. Using rectification module of 3. Salt affected soils: slight to strong saline-sodic Erdas Imagine, Linear rubber sheet map transformation 4. Miscellaneous areas was used. Coordinates were assigned by using about Mean Suitable: 34 °C maximum Not suitable: 10 easily recognizable GCPs. Maps of all the criteria annual 1. 26 °C were geo-referenced to WGS84. First polynomial order temperature 2. 28 °C 3. 30 °C and nearest neighborhood re-sampling method computes 4. 32 °C the new coordinates for output image. The projection Soil Suitable: of shape file of Punjab administrative boundary was physical 1. River plain and terraces: non calcareous loamy soil properties 2. River plain and terraces: calcareous loamy soils already in WGS84 UTM zone 42N so all the geometri- Not suitable: cally rectified factor and constraint maps were then 1. River plain and terraces: non calcareous clayey soil 2. River plain and terraces: calcareous sandy soil, dune 3. River plain and terraces: calcareous clayey soils 4. River plain and terraces: salt affected soils 5. Piedmont plains: non-calcareous loamy soils 6. Piedmont plains: non-calcareous clayey soils 7. Piedmont plains: calcareous sandy soils 8. Piedmont plains: calcareous loamy soils 9. Piedmont plains: calcareous clayey soils 10. Piedmont plains: salt affected soils 11. Loess and weathered rock plains 12. Mountains and hills 13. Seasonally flooded soils 14. Sand dunes and sandy soils Aridity Suitable: classes 1. Arid (Kharif, Rabi) 2. Arid (Kharif), Hyper-arid (Rabi) Not suitable: 1. Humid 2. Sub-humid 3. Semi-arid Agro- Suitable: ecological 1. Irrigated plains: D.G. Khan irrigated zones 2. Irrigated plains: cotton zone Not suitable: 1. Barani region: high rainfall 2. Barani region: low rainfall 3. Thal region: arid zone 4. Thal region: irrigated zone 5. Marginal land: Suleiman mountains 6. Irrigated plains: central mixed zone 7. Marginal land: Cholistan 8. Irrigated plains: rice zone River Suitable: Land Fig. 1. The research procedure used in this study. constraint Not suitable: Rivers Modeling the Land for Cotton Cultivation 99 re-projected to WGS84 UTM zone 42N using ArcGIS criteria (Table 2) describe suitability and non suitability 10.2.2. In ArcGIS 10.2.2 all factors and constraint were for cotton cultivation (Arain et al., 2014). In this study, then manually digitized using the projected shape file the standardized factors: canal command area (Fig. 10), of Punjab administrative boundary with geometrically soil pH ranges (Fig. 11), soil chemical properties classes rectified maps as base maps. In this work, the used (Fig. 12), mean maximum annual temperature (Fig. 13), factors: canal command area (Fig. 2), soil pH ranges soil physi-cal properties classes (Fig. 14), aridity classes (Fig. 3), soil chemical properties classes (Fig. 4), mean (Fig. 15), agro-ecological zones (Fig. 16), and constraint: maximum annual temperature (Fig. 5), soil physical Punjab rivers (Fig. 17), describe only the suitable and properties classes (Fig. 6), aridity classes (Fig. 7), agro- not suitable class in a particular layer for cotton ecological zones (Fig. 8), and constraint: Punjab rivers cultivation in the study area. (Fig. 9), describe all the classes in a particular layer of Analytic hierarchy process. In Idrisi Selva AHP method the study area. For generating the standardized map for was used to derive a set of weights for all the stan- each parameter and the final aggregated land suitability dardized factors by utilizing PWCM with the principal map in Idrisi Selva, all the parameters should be in Eigen vector of this matrix representing a best fit set of raster form. For this, rasterization was performed in weights by Saaty’s method. In the comparison matrix, Idrisi Selva with the resolution of 100 ´ 100 m. two factors were compared at a time in terms of their The logic of aggregation demands that all the rasterized importance on a scale from 1/9 to 9, where 1/9 indicates criteria be standardized to the same scale to make that relative to the column factor, the row factor is less comparisons possible. Ranges of values that are not of important; 9 indicates that in relation to the column interest are explicitly set to 0 while, the range of interest factor, the row factor is more important (Eastman, 2012; is set to 1 (Eastman, 2012). The re-class tool of Idrisi Selva Teknomo, 2006). After constructing comparison matrix, was used to standardize the input criteria. Standardized Eigen vectors were computed, which were the normalized

Fig. 2. Digitized map of Punjab canal command area. Fig. 3. Digitized map of Punjab soil pH. 100 Nabila Naz and Haroon Rasheed

Table 3. Pairwise Comparison Matrix of all the factors relevant to cotton crop land suitability Factors Soil Canal Soil Mean annual Soil Aridity Agro- pH command chemical maximum physical classes ecological area properties temperature properties zones Soil pH 1 ------Canal command area 1/3 1 - - - - - Soil chemical properties 1 3 1 - - - - Mean annual maximum temperature 3 1 3 1 - - - Soil physical properties 5 3 5 5 1 - - Aridity classes 7 7 3 3 1 1 - Agro-ecological zones 9 7 5 5 3 1 1

Eigen vector of the matrix obtained by normalizing Table 4. Weights of all the factors used in the study each column of the matrix. Since vectors are normal- Factors Weights ized, the elements in Eigen vector sum to 1. PWCM (Table 3) represent the relative importance of all the Canal command area 0.0397 Soil pH 0.0471 Soil chemical properties 0.0567 Mean maximum annual temperature 0.0804 Soil physical properties 0.2072 Aridity classes 0.2335 Agro ecological zones 0.3354

Fig. 4. Digitized map of Punjab soil chemical Fig. 5. Digitized map of Punjab mean maximum properties. annual temperature. Modeling the Land for Cotton Cultivation 101

Fig. 7. Digitized map of Punjab aridity classes.

(lmax- n) CI = ______...... (1) (n - 1) CI CR = ____ ...... (2) RI where:

Fig. 6. Digitized map of Punjab soil physical lmax = maximum Eigen value, CI = Consistency Index, properties. CR = Consistency Ratio, RI = Random Index and n = number of criteria in each PWCM. This step was performed in Idrisi Selva. In this work, Eigen vector factors involved in this study. The hierarchy of weights has acceptable consistency ratio of 0.09. for all the factors (Table 4) was used for generating GIS based MCE is concerned with the allocation of final land suitability map. land to suit a specific objective on the basis of variety Weights are used to derive consistency ratio (CR) which of attributes that the selected areas should possess. In indicates any inconsistency made in PWCM. If CR > MCE, each standardized factor was combined by 0.10, then some pair-wise values need to be reconsidered weighted linear combination that is, each factor was until desired value of CR < 0.10 is reached (Eastman, multiplied by a weight, with results being summed up, 2012). The formula for consistency ratio (Kihoro et al., which was then multiplied by the product of constraints. 2013) was described in equation 1 and 2: The final image was measure of aggregated suitability 102 Nabila Naz and Haroon Rasheed

Fig. 9. Digitized map of Punjab river constraint.

Fig. 8. Digitized map of Punjab agro-ecological zones. for non-constrained locations (Fig. 18). The formula for WLC (Eastman, 1999) is given in equation 3:

s = S wixi . P cj ...... (3) where:

S = the composite suitability score, wi = weight of a factor, xi = factor, cj = constraint, S = the sum of weighted factors, and P = product of constraints.

Results and Discussion The final suitability map (Fig. 19) generated by GIS based AHP modeling technique for cotton cultivation potential sites in Punjab by using decision support tool Fig. 10. Standardized factor map for Punjab canal of multi criteria. Number of hectares suitable for cotton command area. Modeling the Land for Cotton Cultivation 103

Fig. 11. Standardized factor map of Punjab soil Fig. 13. Standardized factor map of Punjab mean pH. maximum annual temperature.

Fig. 12. Standardized factor map of Punjab soil Fig. 14. Standardized factor map of Punjab soil chemical properties. physical properties. 104 Nabila Naz and Haroon Rasheed

Table 5. Summary of statistical information by GIS based AHP model for cotton land suitability in Punjab, Pakistan Statistics Area Production (Bales) Area (hectare) Percentage (%) Suitable land 6141259.4685409 31 27021541.66157996 Not suitable land 13811985.0537708 69 0 Land under cultivation (2011-2012) 2533000 13 11129000 Potential increase by suitability map 3608259.4685409 18 15892541.66157996

cultivation was 6141259.4685409 hectares with The final cotton land suitability map represents 18% production capability 27021541.66157996 bales and increase in suitable area for cotton cultivation with an area not suitable for cotton cultivation was increased potential in production of 15892541.66157996 13811985.0537708 hectares. Of the study area, 31% bales were recorded. represents suitable area and 69% represents not suitable According to Punjab Development Statistics (Rafique, area (Table 5) for cotton cultivation. 2013), highly suitable districts for cotton cultivation According to Punjab Development Statistics (Rafique, were Rahim Yar Khan, Bahawalpur, Bahawalnagar, 2013), area sown under cotton for 2012 was 2533000 Multan, Dera Ghazi Khan, Muzaffargarh, Vehari, hectares with production for the year 2011-2012 was Khanewal, Sahiwal, Lodhran, and Rajanpur. Final cotton 11129000 bales. According to this statistical report, land suitability map indicates that the districts with existing cropland under cultivation covers 13% while large extent of suitability were Bahawalnagar, Bahawalpur, area not under cultivation was 87% of the study area. Chiniot, Dera Ghazi Khan, Faisalabad, Jhang, Khanewal,

Fig. 15. Standardized factor map of Punjab aridity Fig. 16. Standardized factor map of Punjab agro- classes. ecological zones. Modeling the Land for Cotton Cultivation 105

report (Cai et al., 2010); Punjab districts cotton produc- tion (SUPARCO, 2012); major and minor cotton crop areas (USDA, 2014); cotton production regions (FAO, 2004) who concluded the sites suitable for cotton cultivation in Punjab, Pakistan.

Conclusion For decision makers to select certain crop land suit- ability is a complicated issue especially when based on environmental factors. The GIS based MCE using AHP procedure involves the utilization of geographical data, the decision maker preferences, manipulation of data, and preference according to specific decision rules. The model implemented in this study, demonstrated a rational and objective approach to make decisions in agricultural applications in Punjab. MCE method was adequate to integrate databases required for different kinds of environmental variables in a GIS context. In this study, application of GIS technique to identify Fig. 17. Standardized constraint map of Punjab suitable areas for cotton crop in Punjab, Pakistan was rivers. successful. The results obtained from this study, indicate that the use of GIS and application of MCE using AHP Lodhran, Multan, Muzaffargarh, Okara, Pakpattan, could provide a superior guide map for farmers and Rahim Yar Khan, Rajanpur, Sahiwal, Toba Tek Singh, decision makers at local level to select the appropriate and Vehari. Districts with less extent suitable area were cultivation sites, crop management and diversification Bhakkar, Chakwal, Khushab, Layyah, Mianwali, and operations to achieve better agricultural production. Sargodha. Districts not suitable for cotton cultivation The approach has been used in some studies in other were Attock, Gujranwala, Gujrat, Hafizabad, Jhelum, countries. This study clearly brought out the distribution Kasur, Lahore, Mandi Bahauddin, Nankana Sahib, of cotton derived from the evaluation of environmental Narowal, Rawalpindi, Sheikhupura, and Sialkot. variables in GIS context. The final suitability map (Fig. 19) shows that the agro Additionally, the result of this study could be useful for ecological zone, aridity classes, soil physical properties other investigators who could use these results for and temperature zones comprise the major part of the diverse studies. This study has been done considering cotton suitable land in this analysis. There are several soil physical and chemical properties, temperature, other crops grown under the cotton suitable areas that water resources, aridity, and agro-ecological zones that necessitate their land suitability evaluation in order affected the suitability of cotton cultivation. Therefore, to get better yields by optimally utilizing the present it gives primary result. resources. The areas which are not suitable for cotton cultivation may be suitable for any other crop and The common thread in mapping cotton cultivation excellent results can be obtained by better management suitability is to utilize correct environmental variables in terms of millions of bales. This will eventually give to achieve greater accuracy. To accomplish this, the boost to economy of Pakistan, which also supports the study has focused deeply on those environmental textile industrial sector. variables that are worth mentioning for cotton cultiva- The final land suitability map for cotton cultivation is tion and accurate enough to provide perfect suitability also in conformity with the findings of IWMI research map. 106 Nabila Naz and Haroon Rasheed

Fig. 18. The process of combining criteria maps for cotton land suitability in Punjab, Pakistan, using GIS based AHP model. Modeling the Land for Cotton Cultivation 107

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Quality Variation Minimizer: A New Approach for Quality Improvement in Textile Industry Muhammad Amin*, Muhammad Amanullah and Atif Akbar Department of Statistics, Bahauddin Zakariya University, Multan, Pakistan

(received March 24, 2014; revised September 4, 2015; accepted October 9, 2015)

Abstract. The main theme of this research is to introduce a new technique for quality improvement in industries and services environment. The technique is called as quality variation minimizer (QVM), which is used to test and compare product quality among multiple data groups, i.e. machines, operators, and material etc. For the significant application, QVM is applied at Card department in spinning industry to determine yarn grains quality by different machines. Then comparison of QVM is made with other already developed techniques, i.e., coefficient variation (CV), sigma level etc. to determine yarn grains quality. From the results determined by t-test and chi square test, it has been found that QVM is an effective method to determine yarn grains quality with sample average near the target/demanded value as well as minimum variation. Keywords: coefficient of variation, quality variation minimizer, sigma level, yarn grains quality

Introduction the yarn manufacturing industry in the late 1940s until et al Quality is the only key to survive any organization or 1950 (Clapp ., 2001). In 1981, Milliken & the company for a certain product or services. Quality means Company (Textile Company) applied TQM in the fitness for use or to meet the customer needs and satis- company to meet the customer demands. Currently, faction. In this situation, quality improvement activities there are 30,000 approximately textile-related companies have become a part of the business culture and a way of in the United States, of which mostly use TQM tools life (Gijo and Rao, 2005). for reducing cost and enhancing customer satisfaction (Mukhopadhyay and Ray, 2006). Mukhopadhyay and Across all types of industries, a well-known fact is that Ray (2006) applied Six Sigma to reduce yarn packing ‘process and product variation’ is the strongest factor defects and they had the techniques of Six Sigma i.e., affecting the product and process costs, process yield, control chart, sigma level, MSA, regression, etc. Feili et al product quality and customer satisfaction (Hild ., and Fekraty (2010) constructed the control charts on 2000). During the past half of the century, various tools, the basis of probability and fuzzy theory to monitor the i.e. statistical process control (SPC), quality circles yarn quality. They have found that fuzzy theory performs (QC), total quality management (TQM), benchmarking, better than probability theory for monitoring product quality management system (QMS), enter resource quality. Maros et al. (2011) applied control charts on planning (ERP) packages, Kaizen and Six Sigma have chenille yarn defect types to see the variations. been developed for quality improvements and custo- mer satisfaction (Antony et al., 2005; Dedhia, 2005; Textile industry and its markets both are so complex Montgomery, 2005). and the customer is always demanding for better quality of textile products and on time delivery with minimum Nowadays, billions of dollar are spent annually on good cost. To satisfy these challenges, companies are imple- quality products in the world. In order to remain in the menting SQC techniques for their customer satisfactions main stream of global competition, market manufacturers (Das, 2013). Acceptable quality is identified by the end have to produce good quality products in various fields, user while fabric makers are the main user of yarn i.e. health, the internet, textile, food etc. (Dhillon, 2007). (Lord, 2003). In textile spinning industry, quality is valued In 2000, World fiber production was predicted 50 million by utilizing the parameters yarn count (NEC), count et al tonnes (Clapp ., 2001). In the textile industry, to coefficient of variation (CV%) or evenness (consistency), ensure the product quality, quality control is necessary. strength, CLSP, TPI (twist per inch), uniformity (Um%), Statistical quality control (SQC) was first time used in thin, thick places and so on. Yarn quality consistency *Author for correspondence; E-mail: [email protected] is measured by variance, standard deviation or CV of

109 110 Muhammad Amin et al. the yarn quality parameter according to yarn product carding, drawing (breaker and finisher), simplex, ring (Lord, 2003). One of the most important prerequisites and autocone. The card is the heart of spinning industry for a spinner is to keep the average count and count and variation arises due to material, machine, man, and variation within control. Most customers are interested the environment. Therefore, the sample of grains/yard in average count, counts CV%, average CLSP and average from the card machine wise was taken and detected single yarn strength over a specified yarn length. Most which machine produce excellent quality with minimum important parameter for measuring the yarn quality is variation and which has maximum variation. In this the yarn strength because fabric durability and structure spinning industry, 22 card machines were sets in card completely depend on yarn strength (Lord, 2003). Modern department, however, machine Card MK5-12 was not high performance machine in knitting and weaving in working condition when the samples from this industry mills require a constant yarn quality. Most of the success- of these machines were collected. ful spinning mills have to produce the demanded quality Methods. Different statistical tools were used for in close cooperation with knitter and weavers through measuring the required yarn quality. These tools include most constant and cost-efficient manner. In parliamentary mean, standard deviation, coefficient of variation (CV), law to predict yarn behaviour, it is not sufficient merely sigma level, QVM (our proposed technique), T-test, chi to use individual quality characteristic, i.e. CV% for the square test with respective p-values. The formulation valuation of a narration. Because CVs does not discri- of sigma level, process capability and QVM (proposed minate between the sources of erroneous beliefs, then technique) methods are given as under: alternative methods are required for measuring yarn quality (Lord, 2003). Sigma level. A statistical tool which is used to measure the process quality level to meet the customer require- Reliable and accurate tools invention are necessary for ment, which is also a technique of six sigma and very evaluating the product quality. Customers are always important tool for measuring the product quality. Sigma demanding excellent product quality with a process level is grounded along the voice of customer, process average near the target (Demand) with minimum average and process variation (standard deviation). variation. Now, the problem is arising how to measure the process quality to meet the customer demand with Six Sigma is implemented at textile spinning mills of minimum variation. In the present study a new technique India, reduction in delay procurement of material that Quality Variation Minimizer (QVM) has been introduced results sigma level increase from 1.80 to 2.4 and yield to measure the product quality. The principal objectives improved from 62-76% (Das, 2005). Six Sigma is of QVM are as follows: implemented in textile industry to reduce dying shade i. To predict product quality, variation and the sigma level is increased from 0.81 to et al ii. To compare different brands/companies/depart- 2.34, saving annually over $40,000 (Das ., 2007). ments, product quality, Mathematically sigma level is defined as:

iii. To detect a minimum and maximum product

ø ø

_ _ ë

_ _

ö quality variation. ö é ______USL x ö ______x LSL ö é Sigma level = process shift + min , ë s ø s ø This QVM was applied in the textile industry to compare machines quality in terms of yarn grains quality. where: _ Materials and Methods m = target or demanded average; x = the estimated process mean; s = the estimated standard deviation Data and company. The data are taken from the Mehr from the process; USL = the upper specification limit Dastgir Spinning (MDS) Mills Ltd. Multan, Pakistan. and LSL = the lower specification limit. The MDS manufacturing the following yarn products, i.e., 7/1s, 10/1s and Super 40/1s. A sample from the Generally the process shift is 1.5 from the target i.e., carding department for product 40/1s was taken which m. Because some quality standards on average are m ± is nominated fine yarn to produce various types of 3s and at one side, may be left or right, the distance of clothes. The spinning industry for yarn manufacturing target values. In other words, we can take the process contains the following departments: mixing, blow room, shift allowed variation given by the customer is on Quality Variation Minimizer for Textile Industry 111 either side from the target value is 1.5 for further detail process average near the target? Here a new technique about process shift of 1.5 (Bothe, 2002). has been introduced which has been called Quality

Variation Minimizer (QVM) to measure the product ø _ _

_ _ ö ______USL x ______x LSL ö Sigma level = 1.5 + minimum , quality. QVM mathematically is defined as: s s ø _ _ |______x m | QVM = Za_ ´ s + There is direct relationship between sigma level and 2 n customer expectations. So higher sigma level results where: better customer expectations, meaning the fewer defects Z a_ are produced in the process. Sigma level is inversely 2 = the critical value of the normal distribution at a proportional to process standard deviation. It shows specified level of significance. QVM is affected by that if process standard deviation increases, then the level of significance, the absolute distance between sigma level is decreased. Similarly, if process standard process average and the target value, process standard deviation is decreased, then the sigma level is increased. deviation and the sample size. The smaller the value of Our main objective to minimize the process variation alpha indicates more reliable QVM results as compared is the main reason to use sigma level to measure the to QVM at larger alpha. As in the literature there are process quality for comparison purposes (Breyfogle, standard quality testing tools, i.e. Cp, sigma level. This 1999). proposed method QVM has been proved practically that it performs in a better way than already developed Process capability. Another measure for determining method with simple and minimum computation. the product quality is process capability (Cp). Cp measures customer specifications in terms of acceptable spread Results and Discussion or dispersion, without reference to the target value In Table 1, studies of yarn grains/yards quality of (Ehrlich, 2002). Cp mathematically is defined as: different machines have been presented alongwith _ performance of QVM. A comparison among QVM with C p = ______USL LSL s SD, CV, Cp and sigma level is depicted here.

The difference is the tolerance of manufacturing process From Table1, it is clear that on the basis of smaller and the voice of the customer. standard deviation and CV, the best quality was given by machine Card MK5-19, while T-test says that process According to Keller (2011), Montgomery (2005) and average grains do not meet the customer needs and also Park (2003) the interpretation of Cp is as follows: chi-square test indicates that the process variation is

If Cp <1, then the process variation is greater than not acceptable. Now, considering the machine Card allowable variation and more defective items are MK5-03, Card MK5-18 and Card MK5-22, these have produced the process average near the target but their CV is so large. Similar results are given by Cp because it depends If Cp = 1, it indicates the sample variation exactly only on process standard deviation. On the other hand, equals the allowable variation (i.e. tolerance) sigma level and QVM observes both target and process

If Cp >1, then the process variation is smaller than variation. Table 1 also shows that sigma level is maxi- allowable and less defective items are produced. mum where QVM is minimum and t-test is accepted. This indicates that the process average met the target The main limitation of this technique like CV is that it or demanded value and also variation by chi-square test does not see how process mean is away from the target was minimum of machine Card MK5-16 as compared value. It just only sees the voice of the customer and to other machines and therefore, considered as the best process standard deviation. machine for producing a good quality grains. On the Quality variation minimizer. As the limitation of CV other hand bad quality was produced by Card MK5-01 and Cp techniques is discussed to assess the product as its sigma level was minimum and QVM was maxi- quality, now the question arises how these limitations mum. This machine also needed to be checked for the are swept to get reliable results regarding the product mechanical faults. There are some situations where CV, quality and also how to measure the process quality to SD and QVM are minimum while sigma level, Cp is meet the customer demand with minimum variation and maximum but it is not always true. QVM depends on 112 Muhammad Amin et al. level of significance, absolute distance between process and target value. These relationships are also shown in average and target value and process standard deviation. correlation in Table 2. The significance of this proposed As the greater value of the level of significance may technique is that it has been applied in the textile spinning provide similar results as other groups. So smaller value industry and the results show that this technique is gives more cleared results of QVM to discriminate the comparatively better than CV. So this proposed technique product quality among the groups. QVM value will may be used in other manufacturing industries to com- move in the same direction with process standard pare the product quality by different machines, operators, deviation and the absolute distance between average temperature level, humidity level etc.

Table 1. Card department yarn grains/yards quality analysis by machines

Department Card Specification limits USL 61.5 Process shift 1.5 Grains/Yard machine wise analysis Target 60 Level of significance 0.05 LSL 58.5 Target SD 0.6 Descriptive and quality measuring statistics QVM QVM QVM Testing Machines Mean SD CV Cp Sigma Sample Level of significance T-test P-value Deci- Chi- P-value Deci- level size 0.1 0.05 0.01 sion square sion Card MK5-01 59.35 1.35 2.27 0.37 2.13 20 2.6025 3.2800 4.1252 -2.15 0.0444 NO 95.841 0.0000 NO Card MK5-02 59.95 0.72 1.21 0.69 3.5 86 1.1014 1.4600 1.9192 -0.67 0.5065 Yes 123.664 0.0040 NO Card MK5-03 60 1.09 1.82 0.46 2.87 82 1.5849 2.1300 2.8173 -0.03 0.9776 Yes 267.654 0.0000 NO Card MK5-04 60.28 0.81 1.34 0.62 3.01 79 1.4546 1.8600 2.3681 3.11 0.0027 NO 141.588 0.0000 NO Card MK5-05 60.33 0.76 1.26 0.66 3.03 81 1.4367 1.8200 2.2965 3.94 0.0002 NO 128.658 0.0000 NO Card MK5-06 60.3 0.87 1.44 0.58 2.89 82 1.5547 1.9900 2.5343 3.11 0.0026 NO 169.091 0.0000 NO Card MK5-07 60.29 0.79 1.31 0.63 3.04 86 1.4308 1.8300 2.3235 3.35 0.0012 NO 147.356 0.0000 NO Card MK5-08 59.88 0.99 1.65 0.51 2.9 81 1.5531 2.0500 2.6670 -1.13 0.2616 Yes 215.92 0.0000 NO Card MK5-09 59.88 0.92 1.54 0.54 2.99 83 1.4646 1.9300 2.5095 -1.22 0.2259 Yes 194.751 0.0000 NO Card MK5-10 59.91 0.87 1.45 0.57 3.12 88 1.3499 1.7800 2.3327 -0.96 0.341 Yes 182.808 0.0000 NO Card MK5-11 60.06 0.76 1.26 0.66 3.4 85 1.1614 1.5400 2.0158 0.79 0.4305 Yes 133.412 0.0000 NO Card MK5-13 59.7 0.89 1.5 0.56 2.84 87 1.5955 2.0400 2.6041 -3.15 0.0023 NO 190.323 0.0000 NO Card MK5-14 60.37 0.95 1.57 0.53 2.69 69 1.7461 2.2200 2.8199 3.22 0.002 NO 170.576 0.0000 NO Card MK5-15 60.18 0.79 1.32 0.63 3.16 81 1.3318 1.7300 2.2286 2.05 0.0434 NO 139.974 0.0000 NO Card MK5-16 60.09 0.7 1.16 0.71 3.52 88 1.1011 1.4500 1.8917 1.16 0.2488 Yes 118.304 0.0140 NO Card MK5-17 60.04 0.99 1.65 0.51 2.98 81 1.4709 1.9700 2.5890 0.33 0.7428 Yes 217.566 0.0000 NO Card MK5-18 59.98 0.76 1.26 0.66 3.45 83 1.1188 1.5000 1.9753 -0.24 0.8132 Yes 130.863 0.0000 NO Card MK5-19 59.56 0.68 1.14 0.74 3.06 89 1.4263 1.7700 2.1926 -6.16 0.0000 NO 112.431 0.0410 NO Card MK5-20 59.69 0.94 1.58 0.53 2.76 65 1.6833 2.1600 2.7507 -2.68 0.0094 NO 158.602 0.0000 NO Card MK5-21 59.52 1.01 1.7 0.49 2.51 84 1.9423 2.4500 3.0845 -4.32 0.0000 NO 235.576 0.0000 NO Card MK5-22 59.97 1.1 1.84 0.45 2.83 81 1.6342 2.1900 2.8825 -0.26 0.7925 Yes 271.193 0.0000 NO

YES = no significant difference between process value and target value; NO = there is significant difference between process value and target value.

Table 2. Correlation matrix among quality measures

QVM Mean SD CV Cp Sigma level ABS distance QVM 1.0000 -0.5171 0.9070 0.9098 -0.8553 -0.9688 0.6576 Mean -0.5171 1.0000 -0.4419 -0.4653 0.3642 0.4490 -0.3826 SD 0.9070 -0.4419 1.0000 0.9995 -0.9768 -0.8392 0.2797 CV 0.9098 -0.4653 0.9995 1.0000 -0.9754 -0.8415 0.2862 Cp -0.8553 0.3642 -0.9768 -0.9754 1.0000 0.8224 -0.2016 Sigma level -0.9688 0.4490 -0.8392 -0.8415 0.8224 1.0000 -0.7053 ABS distance 0.6576 -0.3826 0.2797 0.2862 -0.2016 -0.7053 1.0000

ABS = absolute; QVM = quality variation minimizer; CV = coefficient of variation; Cp = process capability. Quality Variation Minimizer for Textile Industry 113

Conclusion sigma methodology to reduce lot-to-lot shade varia- Journal of Industrial Textiles Different machines of card department in textile industry tion of linen fabrics. , were studied and different quality measuring tools were 36: 227-251. used for checking the machines quality. Most of the Das, P. 2005. Reduction in delay in procurement of spinning quality analyst uses only the CV for measuring materials using six sigma philosophy. Total Quality the yarn quality. They say that the machine with minimum Management & Business Excellence, 16: 645-656. CV has better quality. From the CV formula, it was Dedhia, N.S. 2005. Six sigma basics. Total Quality found that this is the ratio of process SD to the process Management & Business Excellence, 16: 567-574. mean. CV does not show the target mean but just gives Dhillon, B.S. 2007. Applied Reliability and Quality: the ratio between process standard deviation and mean. Fundamentals, Methods and Procedures, 260 pp., st Similarly, the Cp measurement only sees the process 1 edition, Springer Series in Reliability Engine- standard deviation and not the process mean. From the ering, Springer, London, UK. present results and discussion, it has come to the point Ehrlich, B.H. 2002. Transactional Six Sigma Servicing, that smaller the value of QVM results in larger quality Leveraging Manufacturing Concepts to Achieve level of yarn product and QVM is minimum where World-Class Service, pp. 126-127, St. Lucie Press, sigma level is maximum. This indicates that better Boca Raton London, New York, USA. quality of product is produced. QVM may also be preferred Feili, H.R., Fekraty, P. 2010. Comparing fuzzy charts over CV for its computational ease. QVM technique with probability charts and using them in a textile seems to be so simple to use and can be considered as company. The Journal of Mathematics and Com- good as other techniques, i.e. regression and design of puter Science, 1: 258-272. experiments, etc., in the industry to test the quality over Gijo, E.V., Rao, T.S. 2005. Six sigma implementation- the time to get quick and reliable results. hurdles and more hurdles. Total Quality Manage- ment & Business Excellence, 16: 721-725. References Hild, C., Sanders, D., Cooper, T. 2000. Six sigma on Antony, J., Kumar, M., Madu, C.N. 2005. Six sigma continuous processes: how and why it differs?. in small and medium sized UK manufacturing Quality Engineering, 13: 1-9. enterprises: Some empirical observations. Interna- Keller, P. 2011. Six Sigma Demystified, pp. 326-327, tional Journal of Quality & Reliability Management, 2nd edition, McGraw-Hill, New York, USA. 22: 860-874. Lord, P.R. 2003. Handbook of Yarn Production: Bothe, D.R. 2002. Statistical reason for the 1.5s Shift. Technology, Science and Economics, pp. 276-300, Quality Engineering, 14: 479-487. CRC Press, Woodhead Publishing Ltd., Cambridge, Breyfogle, F.W. 1999. Implementing Six Sigma: Smarter England. Solutions Using Statistical Methods, 1229 pp., 2nd Maros, T., Viladimir, B., Caner, T.M. 2011. Monitoring edition, Wiley Interscience, New York, USA. chenille yarn defects using image processing with Clapp, T.G., Godfrey, A.B., Greeson, D., Jonson, R.H., control charts. Textile Research Journal, 81: 1344- Rich, C., Seastrunk, C. 2001. Quality Initiatives 1353. reshape the textile industry. Quality Digest, October, Montgomery, C.D. 2005. Introduction to Statistical Quality http://www.qualitydigest.com/oct01/html/textile. Control, pp. 202-203, 5th edition, John Wiley & html. Sons Inc, New York, USA. Das, A. 2013. Testing and statistical quality control in Mukhopadhyay, A.R., Ray, S. 2006. Reduction of yarn textile manufacturing. In: Process Control in Textile packing defects using Six Sigma methods: A case Manufacturing, A. Majumdar, A. Das, R. Alagirusamy study. Quality Engineering, 18: 189-206. and V. K. Kotari (eds.), pp. 41-78, Woodhead Park, S.H. 2003. Six Sigma for Quality and Produc- Publishing Series in Textiles: Number 131, New tivity Promotion, pp.18-23, Asian Productivity Dehli, India. Series 32, Asian Productivity Organization, Tokyo, Das, P., Roy, S., Antony, J. 2007. An application of six Japan. Pak. j. sci. ind. res. Ser. A: phys. sci. 2016 59(2) 114-117

Effect of Different Processing Stages on the Crystallinity % and Tensile Strength of 100% Cotton Fabric Zahid Hussaina, Muhammad Qamar Tusief b*, Sharjeel Abidc , Muhammad Tauseef Khawera, Nabeel Amind and Mudassar Abbasd aInterloop (Pvt) Limited Khurrianwala-Jaranwala Road, Faisalabad, Pakistan bDepartment of Fibre & Textile Technology, University of Agriculture, Faisalabad, Pakistan cAdvance Textile Material Engineering, National Textile University, Faisalabad, Pakistan dSchool of Textile and Design, University of Management and Technology Lahore, Pakistan

(received August 31, 2015; revised October 14, 2015; accepted October 15, 2015)

Abstract. In this study, 100 % cotton fabric was used to check the impact on fabric crystallinity and tensile strength at different processing stages. Desizing, scouring, bleaching, mercerization and resin (only resin & resin+softener) application were the processes performed on the fabric. X-Ray diffractometer and tensile strength tester were used to determine the crystallinity index (CI) and tensile strength, respectively. Results revealed that from scouring to mercerization crystallinity (CI) decreased while desizing and resin application treatments showed no significant impact on the crystallinity (CI). In case of tensile strength, a decreasing trend from desizing to resin application was observed. Keywords: cotton fabric, tensile strength, crystallinity index, X-ray diffraction

Introduction potassium hydroxide (40%) for 15 min to 50 h to check Cotton is mostly used natural cellulosic fibre in textile the impact on moisture regain tensile, mechanical, products. Cellulose has crystalline and amorphous swelling and optical properties. It was concluded that regions. In crystalline region, atoms are arranged in a anhydrous ethylamine, NaOH and KOH reduced the close packing. There exist three dimensional arrangements crystallinity significantly, while diethylamine and due to hydrogen bonding between them. On the other pyridine did not produce any significant change in hand in amorphous region, atoms are randomly arranged crystallinity of cotton (Pandey and Iyengar, 1969a). because of the absence of hydrogen bonding. Number The chemical modification of cotton fibre was also of sites (hydroxyl groups) available in amorphous region analysed by applying different concentrations of the governs the bonding or reactivity in suitable conditions chemica and their impact on different properties of for modification in performance properties of cotton cotton fibre were observed and concluded that ethylamine, fibre, while a material with greater crystallinity percentage EDA, KOH and LiOH reduced the crystallinity of cotton has greater strength and a big influence on hardness, by increasing the concentration (Pandey and Iyengar, density, transparency and diffusion of the material e.g. 1969b). Cotton fibre and fabric treated with butane- cotton fibre or fabric (Parikh, 2007). tetracarboxylic acid (BTCA) with the catalyst sodium hypophosphite at different concentrations. In the same During different chemical treatments on cotton fibre or line the cotton fibre treated with BTCA reduced its fabric, the available sites in the amorphous region react and get modified. Such modifications have impact on crystallinity and strength at different concentrations the crystallinity and strength of the cotton fibre or fabric. (Xu, 2003). Similarly, the impact of crosslinking and Much research work has been carried out to measure bleaching treatment on crystallinity of cotton fibres was the crystallinity of cotton by using X-ray diffraction determined by using formaldehyde and dimethy- method. The crystallite orientation of cotton cellulose loldihydroxy ethylene urea (DMDHEU) or citric acid was analysed by using improved method of X-ray (CA) as cross linking agents for bleached cotton fabric diffraction spectrometer (Creely et al., 1956). In an (Parikh, 2007). While in another study the cotton was other research two types of cotton were treated with treated with neutralized vegetable oil base macro- anhydrous ethylamine, diethylamine, pyridine and molecular crosslinkers to check the impact on cotton aqueous solutions of sodium hydroxide (30%) and crystallinity by using X-ray diffraction methodology *Author for correspondence; E-mail:[email protected] (Ford etal., 2010). 114 Proceeing Stages Effect on Cotton Fabric 115

All the above mentioned studies reported on one specific at 120 °C for 1 min and cured at 150 °C for three min. process treatment (surface modification) not on the In the second method (resin+ softener), 30 g/L Reaknit complete processing stages (from grey fabric to finish) EC (DMDHEU, CHT), 30 g/L Tubingal SMF-L to analyse the gradual decreasing or increasing trend (microemulsion of silicone, CHT) and 20 g/L magnesium of crystallinity which directly relates with the strength chloride were used at 5-6 pH at normal temperature. of cotton fabric. The aim of the present study is to Fabric was padded at 70 % pick up, dried at 120 °C for analyse the impact of desizing, scouring, bleaching, 1 min and then cured at 150 °C for 3 min. mercerization followed by the resin application in two All the above mentioned chemicals were commercial ways (one is only resin, while the other is resin along grade of CHT chemical company and sourced from a with softener) on the fabric crystallinity and tensile reputed textile mills Khurrianwala, Faisalabad, Pakistan strength. Caustic soda, magnesium chloride and acetic acid were used of analytic grade. Materials and Methods For this study, 100 % cotton fabric of 110*110/ 60*60 X-ray diffractometer (Model Xpert Pro Pan Analytical) was used and analysed in reference to crystallinity and was used to determine the crystallinity index (CI). In X-ray diffraction, X-rays beams were incident on the tensile strength by using X-ray diffractometer and tensile sample diffracted a beam whose angular dependence strength tester. Lab scale jigger machine was used to was measured by photomultiplier detector travelling perform the experimental work on fabric. Five chemical upon a goniometer in circle and provide a graph of treatments were applied from desizing to resin intensity (counts) of X-ray photons detected as a function application. For desizing, a -amylase 5% on the weight of 2q (the angle between the detector position and the of fabric BEISOL SED (CHT), 2 mL/L BEIXON Q direction of incident beam) as shown in Fig. 1. (Biodegradable sequesting agent, CHT) and 2 g/L FELOSAN FOX (universal detergent with high washing From the Fig. 2, crystallinity (CI) can be measured by efficiency, CHT) were used for desizing treatment at the formula. 90-95 °C for 30 min followed by one hot wash and then cold wash. Crystallinity %= [(Ic – Iam)/Ic]*100 In caustic scouring treatment, 20 g/L NaOH flakes where: (analytical grade), 1 g/L HEPTOL BNF (sequesting Ic = the intensity of the principal cellulose peak at 2q= agent for strong alkaline treatment, CHT) and 2 g/L 22.7° , FELOSAN FOX (universal detergent with high washing efficiency, CHT) were applied at 90-95 °C for 60 min Iam = the intensity of amorphous peak at 2q= 18°. followed by 10 min hot wash and then cold wash. After Tensile strength tester Lloyd of Ametek Company was this, 2 g/L acetic acid was used to neutralize the fabric. used to check the impact on warp wise and weft wise In bleaching process, 6 g/L hydrogen peroxide 50% strength of the cotton fabric after every process. (commercial grade by Sitara peroxide), 2 g/L NaOH flakes, 5 g/LCONTAVAN ICE (stabilizer, CHT), 1 g/L Results and Discussion HAPTOL BNF and 3 g/L FELOSAN FOX were used Grey fabric. The values related to the crystallinity and at 90 °C for 60 min followed by hot wash and then cold tensile strength (warp wise and weft wise) of the grey wash followed by neutralization with 2 g/L acetic acid. fabric (Sample 1) are mentioned in Table 1. These In mercerization process, fabric was treated with 30 % results reveal a decreasing trend in the crystallinity and solution of caustic soda was used for 60 sec under tensile strength of the fabric. tension and then hot washed. After this, 2 g/L acetic Desized fabric. The effect of desizing process on fabric acid used to neutralize the fabric followed by cold wash. in respect of its crystallinity and tensile strength was Two different methods were used for the resin application. determined. The results given in Table 1 and further In first method (only resin application), 30 g/L Reaknit illustrated in Fig. 2-3 depict non significant impact on EC ( modified DMDHEU, CHT) and 20 g/L magnesium crystallinity % of the fabric after desizing. But the chloride (catalyst) were used at 5-6 pH at normal tensile strength of the fabric reduced from 478.05 N to temperature. Fabric was padded at 70 % pick up, dried 439.03 on warp wise and from 469.7N to 433.11N on 116 Muhammad Qameer Tusif et al.

Table 1. Results of crystallinity(%) and tensile strength Bleached fabric. Significant decrease in crystallinity (warp & weft wise) and tensile strength was observed after the bleaching Sample Process steps Crystallinity Tensile Tensile process as shown in Table 1 and Fig. 2-4. During bleaching No (%) strength (N) strength (N) process, natural and colour impurities removed from warp wise weft wise the fabric. Hydrogen bonding from the crystalline region 1 Grey 68.32 478.05 469.7 breaks due to the chemical degradation of the fabric 2 Desize 68.11 439.03 433.11 which reduced the crystallinity from 67.23% to 62.47% 3 Scoured 67.23 419.03 412.4 4 Bleached 62.47 362.71 353.56 and also the tensile strength reduced significantly. 5 Mercerized 61.56 366.89 352.89 6 A Resin 64.77 247.08 226.09 Mercerized fabric. During mercerization caustic soda 6 B Resin+Softener 62.05 279.36 254.6 reacts with the cellulose and breaks down the hydrogen bonding of the atoms in crystalline region of the cotton weft wise. Size material applies on yarn level before fibre. This provides greater number of sites for reaction. weaving to provide adequate strength for the next Ultimately the pick up of the cotton fabric enhanced et al., process which helps during the weaving process. But (El Badry 2013). Due to this crystallinity of the during desizing size material removed from the fabric Cryst allinity % that affects the strength of the fabric. Both these factors 70 reduce the strength after desizing in fabric form. 68

Scoured fabric. Both crystallinity and tensile strength % 66 decreased after caustic scouring as shown in Table 1 and Fig. 2-4. During this process, natural impurities of 64 cotton removed by the reaction of caustic soda at high Crystallinity 62 temperature. Chemical degradation and the effect of 60 elevated temperature reduced the crystallinity from 0 1 2 3 4 5 6 7 8 9 68.11% to 67.23% and tensile strength about 20N. Process steps y = -1.1129x + 70.494 R² = 0.659

Crystallinity % linear (crystallinity %) X-ray tube Detector Fig. 2. Graphical explanation of crystallinity at

w different processing steps. 2q Sample Tensile strength (N) warp wise 600

500 (N) 400

strength 300

200 Tensile 10000 100

0 5000 Intensity (counts) 1 2 3 4 5 6 7 8 9 Process steps

0 y = -36.861x + 554.61 R² = 0.9009 35 40 45 50 55 Tensile strength (N) warp wise Position [ 2°q] {CuK-a} Linear (tensile strength (N) warp wise)

Fig. 1. Shows the working principle of X-Ray Fig. 3. Graphical explanation of warp wise tensile diffractometer and graphical output. strength (N) at different processing steps. Proceeing Stages Effect on Cotton Fabric 117

Tensile strength (N) weft wise Conclusion 600 It is concluded from the above study that, 100% cotton 500 fabric significantly lose its degree of crystallinity

400 (crystallinity %) during caustic scouring, bleaching and (N) mercerization due to chemical degradation and elevated 300 temperature impact, while desizing has no significant

sTrength 200 effect on crystallinity % (CI). But the resin application increases the crystallinity % age due to the crosslinking 100 Tensile and bonding in the cotton fabric structure. Tensile 0 strength decreases during all the processing stages from 1 2 3 4 5 6 7 8 9 desizing to resin finish treatment. But scouring, bleaching Process steps and resin application significantly reduces the tensile y = -39.959x + 557.27 R² = 0.9085 strength of the fabric. Tensile strength (N) weft wise Linear ( tensile strength (N) weft wise) References Fig. 4. Graphical explanation of weft wise tensile Creely, J. J., Sega, L. l., Ziifle, H. M. 1956. Determination strength (N) at different processing steps. of the degree of crystallite orientation in cotton fibers by means of the recording X-Ray diffraction fabric decreased slightly from 62.47% to 61.56% in the spectrometer. Textile Research Journal, 28: 789- mercerized sample while the warp wise strength of the 795. fabric increased due to parallelization of the fibre along El Badry, K., Saleh, S.M., Bahlood, S. O. 2013. effect vertical axis under tension as shown in Table 1 and of mercerization techniques on cotton towels Fig. 2-4. properties. Journal of Applied Sciences Research., Resin finished fabric. After the resin application 9: 2386. crystallinity % of the fabric increased significantly from Ford, E. N. J., Mendon, S. K., Thames, S. F., Rawlins, 61.56 to 64.77 due to the cross linking of resin finish J. W. 2010. X-ray diffraction of cotton treated with (DMDHEU). This helps to enhance the bonding and neutralized vegetable oil-based macromolecular close packing in the crystalline region of the fibre. On crosslinkers. Journal of Engineered Fibres and the other hand the tensile strength of the fabric reduced Fabrics, 5: 10-20. while the stiffness of the fabric enhanced by the resin Pandey, S. N., Iyengar, R. L. N. 1969a. Studies of application as shown in Table 1 and Fig. 2-4. chemically modified cotton. Part I: Effect of Resin+softener finished fabric. After the application chemical treatments for varying periods on of resin with softener, a slight increase of crystallinity crystallinity and certain other properties of cotton. from 61.56% to 62.05% was observed in the fabric. Textile Research. Journal, 39: 15-23. Because the effectiveness of resin crosslinking reduced Pandey, S. N., Iyengar, R. L. N. 1969b. Studies on by the combination of softener, crystallinity of the fabric chemically modified cotton. Part II: Effect of enhanced on less % age as compared to only resin different concentrations of chemicals on crystallinity application as shown in Table 1 and Fig. 2. Fabric and certain other properties of cotton. Textile (softner+resin) was also soft comparatively from the Research Journal, 39: 24-31. resin padded fabric. It was due to the softness induced Parikh, D.V. 2007. X-ray crystallinity of bleached and by the micro-emulsion silicon based softener. Resin crosslinked cottons.Textile Research Journal, 77: with softener finish impact on tensile strength was also 612-616. less as compared to only resin padded fabric due to the Xu, W. 2003. Effect of crosslinking treatment on the flexibility enhanced by softener and less fiber stiffness crystallinity, crystallite size, and strength of cotton as shown in Table 1 and Fig. 3-4. fibres.Textile Research Journal, 73: 433-436. Pak. j. sci. ind. res. Ser. A: phys. sci. 2016 59(2) 118-120 Short Communication Biosorption Characteristics of Water Hyacinth (Eichhornia crassipes) in the Removal of Nickel (II) Ion under Isothermal Condition Chidi Obi* and Sylvester Eigbiremonlen Physical Chemistry Unit, Department of Pure and Industrial Chemistry, University of Port Harcourt, P.M.B. 5323, Port Harcourt, Rivers State, Nigeria

(received March 30, 2015; revised August 14, 2015; accepted August 19, 2015)

Abstract. This study was taken to investigate the potentiality of water hyacinth (Eichhornia crassipes) as an alternative biosorbent for the removal of Ni (II) ion from aqueous solution. The optimum pH, contact time and concentration were found to be 6.0, 40 min and 1.0 mg/L under isothermal condition. The biosorption of Ni (II) ion was found to decrease with increasing pH, initial concentration and contact time. Results obtained were analysed with Langmuir and Freundlich biosorption models. The equilibrium data fitted well to the Langmuir biosorption model with correlation coefficient (R2) value of 0.98. The monolayer adsorption capacity was 0.29 mg/g. The removal of Ni (II) ion from aqueous solution using water hyacinth biomass followed a monolayer biosorption. Keywords: monolayer biosorption, water hyacinth, nickel removal

Contamination of toxic metals such as copper, lead, area of Rivers state, Nigeria. The water hyacinth biomass zinc, nickel and chromium in the aquatic environment was sun-dried for two days. The biosorbent was prepared is a matter of attention as studied by Kaewsarn and Yu by washing it with 0.1M HCl (to convert alignates to (2001). Heavy metal contamination may cause serious alignic acid) and then rinsed with deionized water. The health problems such as cancer and brain damage leaves were then dried further in an oven for 24 h until (Mukhopadhyay, 2008). the leaves became crisp. After drying, the leaves were ground by a manual grinder, to a constant size of The presence of nickel ions in surface water is a problem 150 mm. of increasing importance in Nigeria. The nickel stock solution (1000 mg/L) was prepared The permissible limit of nickel according to World using analytical grade of CH COOONi.4H O and test Health Organization (WHO) in drinking water is 1 mg/L 3 2 solution was prepared by dilution to the desired as reported by Nemerow (1963). Current developed concentrations. The biosorption study was carried out methods for solving water contaminated related problems by adopting a column reactor system under isothermal include filtration, ion exchange, membrane separation, condition. The column experiment was performed in a nutrient stripping and adsorption. The adsorption packed bed consisting of a cotton wool and the biomass technology (biosorption) which utilizes natural biomass with inner diameter of 30 mm and length of 500 mm. materials is very effective for the detoxification of The water hyacinth leaves powder (1 g) was used to metal-bearing industrial effluents. study the effect of pH, contact time and concentration Water hyacinth (Eichhornia crassipes) is known as one at 298 K. The supernatant obtained was analysed using of the ‘world’s worst aquatic weeds’ (Malik, 2007). On atomic absorption spectrophotometer (AAS). The amount of metal ion biosorbed per gram of the biomass the other hand, it appears to be a valuable material with q was calculated using the equation below. a remediation property. Therefore, E. crassipes biomass e was used in this study to remove Ni (II) ion from Ci-Ce 50 aqueous solution under isothermal condition and qe = ´ M 1000 isotherm model equations were used to analyse the equilibrium data. where:

The leaves of water hyacinth were collected from Choba qe = the amount of metal ion biosorbed per gram of the River, Choba community in Obio-Akpor local government biomass in mg/g

*Author for correspondence; E-mail: [email protected] Ci = the initial concentration of the metal ion in mg/L, 118 Short Communication: Water Hyacinth for Nickel (II) Removal 119

Ce = the equilibrium concentration of the metal ion in 1.2 mg/L 1 M = the mass of the biomass in g; 50 = the volume of the metal ion in milliliters; 1000 = the conversion factor 0.8 to liters. The experiment was performed in triplicate 0.6 and the mean values taken for each parameter. e q (mg/g) 0.4 The effect of pH on the biosorption capacity is shown in Fig. 1. The biosorption capacity of Ni (II) ion increased 0.2 initially and then became constant with increasing pH. A pronounced dependence of Ni (II) ion biosorption on 0 0 40 50 60 the solution pH was observed (Abdullah and Devi 10 20 30 Time (Mins) Prasad, 2010). This biosorption behaviuor at low pH showed competition between Ni (II) ion and the net Fig. 2. Effect of contact time on Ni (II) ion biosorption positive charge on the biosorbent surface thereby using water hyacinth biomass at 298 K. lowering the rate of uptake. However, as the pH increased, the rate of biosorption increased and optimum thereby reducing the rate of Ni (II) ion uptake (Turp biosorption was achieved at pH6. This could be as a et al., 2011). Therefore, the optimum biosorption time result of the unsaturation of the metal binding sites on of 40 min was obtained. the surface of the biomass indicating weak chemical The initial ion concentration serves as an important interaction between Ni (II) ion and the biomass surface driving force for overcoming mass transfer resistance charges. At pH above 6, the biosorption of Ni (II) ion of Ni (II) ion between the aqueous and solid phases decreased. At this point, precipitation of nickel (II) (Pandey et al., 2007). The effect of different initial Ni hydroxide set in leading to a decrease in the rate of Ni (II) concentrations on the water hyacinth biosorption (II) ion uptake (Wang and Xing, 2002). capacity is shown in Fig. 3. The biosorption of Ni (II) The effect of contact time on the biosorption of Ni (II) ion showed a concentration dependency. The rate of ion using water hyacinth biomass is shown in Fig. 2. biosorption increased as the concentration of Ni (II) ion The result obtained showed that there was an initial fast increased from 20 - 60 mg/L. It was observed that Ni uptake of Ni (II) ion followed by a slow and constant (II) ion uptake decreased above 60 mg/L. This could biosorption. The increase in the rate of biosorption be as a result of the concentration gradient established within 10 - 40 min could be due to unsaturation of the between Ni (II) ion and the biomass surface charges active sites of the biomass and at higher time, due to precipitation. This observation is similar to the precipitation of the nickel (II) hydroxide took place research of Tsai and Chen (2010).

1.2 1.2

1 1

0.8 0.8

0.6 0.6 e e q (mg/g) q (mg/g) 0.4 0.4

0.2 0.2

0 0 0 2 4 6 8 10 12 0 20 40 60 80 100 160 pH Ci (mg/L)

Fig. 1. Effect of pH on the biosorption of Ni (II) Fig. 3. Effect of concentration on Ni (II) ion biosorption ion using water hyacinth biomass at 298 K. using water hyacinth biomass at 298 K. 120 Chidi Obi and Sylvester Eigbiremonlen

The plot of Langmuir as the best model equation is water ways and aquatic lives can be transformed into represented in Fig. 4. However, Langmuir isotherm a useful source for pollution control. constants were determined from a plot of Ce/qe against

Ce as shown in Table 1 (Akbal and Camci, 2011). The References 2 isotherm correlation coefficient (R ) of Langmuir was Abdullah, M.A., Devi Prasad, A.G. 2010. Biosorption 0.98 indicating a physical type of biosorption with of nickel (II) from aqueous solutions and electroplating monolayer capacity of 0.29 mg/g. The result of the wastewater using tamarind (Tamarindus indica L.) biosorpion capacity obtained was greater than the work bark. Australian Journal of Basic and Applied done by Hassan et al. (2010). Sciences, 4: 3591-3601. Akbal, F., Camci, S. 2011. Copper, chromium and nickel removal from metal plating wastewater by electro- 1.2 coagulation. Desalination, 269: 214-222. y=3.445x+0.201 Hasan, S.H., Ranjan, D., Talat, M. 2010. Water hyacinth 10 2 R =0.975 biomass (WHB) for the biosorption of hexavalent 8 chromium: Optimization of process parameters. e Bio Resources, 5: 563-575. e

C /q 6 Kaewsarn, P., Yu, Q. 2001. Cadmium (II) removal from aqueous solutions by pre-treated biomass of 4 marine alga Padina Sp. Environmental Pollution, 2 112 : 209-213. Malik, A. 2007. Environmental challenge vis a vis 0 opportunity: The case of water hyacinth. Environment 0 0.5 1 1.5 2 2.5 3 3.5 International, 33: 122-138. Ce Mukhopadhyay, M. 2008. Role of surface properties Fig. 4. A plot showing Langmuir adsorption model during biosorption of copper by pretreated of Ni (II) ion using water hyacinth biomass. Aspergillus niger biomass. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 329: 95-99. Table 1. The Langmuir parameters Nemerow, N.L. 1963. Theories and Practices of Industrial Waste Treatment. 557 pp., Addison-Wesley Model equation Parameters water hyacinth Pub. Co. Inc., Reading, Massachusettes, USA.

Langmuir KL 0.06 Pandey, P.K., Choubey, S., Verma, Y., Pandey, M., Qmax (mg/g) 0.29 Kamal, S.S.K., Chandrashekhar, K. 2007. Biosorptive R2 0.98 removal of Ni (II) from wastewater and industrial effluent. International Journal of Environmental Research and Public Health, 4: 332-339. The biosorption process was dependent on the pH of Tsai, W.T., Chen, H.R. 2010. Removal of malachite the aqueous solution, contact time and concentration green from aqueous solution using low-cost of Ni (II) ion in the solution. The optimum contact time Chlorella based biomass. Journal of Hazardous Materials and pH of 40 min and 6 were obtained for the biosorption , 175: 844-849. process. The equilibrium data obtained fitted well to Turp, S.M., Eren, B., Ates, A. 2011. Prediction of Langmuir adsorption model equation with a linear adsorption efficiency for the removal of nickel (II) correlation coefficient (R2) of 0.98 indicating a ions by zeolite using artificial neural network Fresenius Environmental Bulletin monolayer type of adsorption. (ANN) approach. , 20: 3158-3165. However, the results obtained from the column reactor Wang, K., Xing, B. 2002. Adsorption and desorption system under isothermal condition have shown that of cadmium by goethite pretreated with phosphate. water hyacinth leaves which is termed as nuisance to Chemosphere, 48: 665-670.