Egypt. J. of Appl. Sci., 34 (11) 2019                                              373-386                                                 

ASSESSMENT OF ORGANO-CLAY COMPLEX BEHAVIOR USING INFRARED TECHNIQUE
Nabil, M.A. Bahnasawy
Soil Chem. and Phys. Dep., DRC, Egypt
Key Words: kaolinite, bentonite, humic acid, fulvic acid, IR and active groups 
ABSTRACT
The present study aims to assessment of organo- clay complex behavior using infrared (IR) technique. The results showed that the kaolinite clay mineral saturated with organic acids (humic and fulvic acids), Ca, Mg and Na cations, after the IR examination; the dominant active groups were: OH, N-H, C-H2, C=O, C=C, Al-O, Si-OH, C-C, Al-OH and C-H and the wavenumber were (3500-3670), (3400-3445), (2900-3000), (1620-1660), 1610, 1350, 1080, (950-1070), 850 and 810 cm-1 respectively.
Also the results  showed that the bentonite clay mineral impregnated with the same treatments and after examined by IR radiation that the active groups were:  OH, N-H, C=O, C=C, C-H, Si-O, C-C and Al-OH-Al and the wavenumber were (3250-3632), 3300, (950-1634), (1525-1631), (830-1068), (1048-1068), 1070 and (915-918) cm-1 respectively.
At the least, the arrangements of active groups on kaolinite mineral were as follows: OH > CH > Al-OH = C=O> N-H = C-C = CH2>C=C = Al-O = Si-OH. While the arrangements of active groups on bentonite mineral were as follows: OH > C=O = C=C>NH = CH>Si-O>Al-OH-Al>C-C. 
INTRODUCTION
The organic complexes of clay play an effective role in determining the characteristics of physical and chemical of soil and its relationship to plant nutrition, therefore their study is important to determines the impact of these characteristics and their roles in improving soil properties (FAO, 2000). The present study clearly demonstrates that FT-IR spectroscopy is an efficient method for the characterisation of the purified bentonite in an organic mixture. This is technique remains economical, rapid, and specific (Eisazadeh, et al. 2012) .Therefore, the identification of the adhesion of clay minerals to the predominant organic acids (humic and fulvic) and the difference between them increases the importance of these acids in influencing soil properties and the binding of the necessary cations (Balcke, et al. 2002).

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The possibility of using pure clay minerals (kaolinite and bentonite) with organic acids (humic-fulvic), Ca, Mg and Na cations identify the effect of the type of cation or the type of organic acid on clay minerals and the group to which he belongs (Aderonke, et al. 2017).
Using infrared radiation is leach to reach the most suitable installation that improves soil properties and their agricultural development potentials (Balcke, et al. 2002). The chemical structure of humic and fulvic acids are shown in Fig. (1).

 (Humic acid)                                                        (Fulvic acid)
         mical formula C187H186O89N9S1                      Chemical formula C135H182O95N5S2
(Sam Boggs, et al. 1985).
Fig. (1) Chemical structure of humic and fulvic acids
Humic substances are considered as the most important constituents of soils (Sara et al. 2010). They form the largest fraction of soil organic matter and play the dominant role in improving soil productivity (Jayaganesh and Senthurpandian, 2010). Humic substances are formed by the decomposition of plant and animal residues by microorganisms (Rao et al. 2019). Organic matter is intrinsic essential components of all soils (FAO 2017). The humic and fulvic substances play an important role in increasing productivity and soil fertility (Khan et al. 2018). 
The plant growth, improvement of tuber nutritive value and quality and improvement in tuber size and weight in potato plants co-inoculated with Bacillus strains and humic acid, a significant improvement was observed in potato production. Despite a low total tuber yield (28.3 t ha-1) in control treatment, combined inoculation increased potato production by around 140% which was with in the high range of the increase expected for both Bacillus strains and humic acid mono treatments. The stability and increased consistency of the potato plant response to bacterial inoculation in the presence of humic acid indicated a promising biotechnological tool to improve growth and adaptation of potatoes to field conditions  Ekin, (2019).

Egypt. J. of Appl. Sci., 34 (11) 2019                                                      375                                                 

Foliar application of FA has a significant beneficial influence on general quality of FCV tobacco of the SBCS region. Specifically, FA resulted in increased leaf K and reduced starch contents. Molecular analysis revealed that FA activated expression of high-affinity K transporters (enhanced K uptake), degraded stored starch, and energized plant metabolism (efflux of trios- PO4 for sucrose synthesis). We believe this is the first report on FCV tobacco grown on black soils that provides encouraging and consistent results that comprehensively highlight inclusive benefits of FA both from agronomic and molecular dimensions. Priya et al. (2014).
The present review was aimed to the technical procedural methods used by FT-IR for clay surface characterization because, FT-IR remain is an economical, rapid and common technique. A spectrum can be obtained in a few minutes and the instruments are sufficiently inexpensive as to be available in many laboratories according to Djomgoue and Njopwouo (2013). 
Although there are different studies on the clay and organic matter complexes, they did not address the effective role of both fulvic and humic alone and their association with the cation structure on the surface of pure clay minerals to guide the importance of the type of clay minerals and their reciprocal cations in the reclaimed soil.
MATERIALS AND METHODS
Homoionic Clays
      Kaolinite and bentonite clays were purchased from Sigma-Aldrich Comp. Each of 10 g of clays was saturated with, Ca2+, Mg2+,and Na+ by five repeated treatments with 50 ml of 1M concentration of metal chloride. The cation-exchanged clays were thoroughly washed with water, and centrifuged for 10 min at 5000 rpm. The washing and centrifugation procedures were repeated until chloride ion was no longer detected by the AgNO3 test by FAO, (2008).
Extraction of humic and fulvic substances
Humic (HA) and fulvic (FA) acids were extracted from sheep manure according to the standard method described by Sánchez-Monedero et al. (2002). The manure was mixed with deionized water at a rate of 1:5 (w/v) and then treated with 0.5 N NaOH solution to extract the humic substances. Humic acid was separated from humus extract by acidification with 0.1 M HCl to reach a pH of 2.0, after being left over-night. The precipitate was collected and the supernatant was again adjusted to pH between 4.5 and 5 using NaOH solution. The fulvic acid fraction got was settled after 24 h. Both the humic substances and fulvic acid fractions were washed with distilled water to remove impurities. Finally, black coloured fine crystals of humic acid and brown coloured crystals of fulvic acid fractions were obtained.

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Preparation of organic clay complex
Organo clay complex will prepared by using two homoiconic clay for kaolinite and bentonite with (Ca, Mg, and Na) separately with humic and fulvic acids. Organic clay complex were prepared at ratio 1:100 following the procedure of Li et al., (2003) and Wang and Xing (2005). In brief, 0.5 g of HA and  FA were dissolved in a minimum volume of 0.5 M NaOH separately and stand overnight then up to 1000 ml to obtain final concentration, 500 mgl -1. The organic clay complex suspensions were intermittent shaking for one week. The sedimented particles were washed repeatedly with distilled water until no color, was seem in the supernatant. The washed organic clay complex was dried at 40 ᴼC, grounded and finally, passed through (170) mesh and sealed in the glass tube for use.
Infrared spectroscopy
The analysis of the infrared absorption of the pure clay minerals and  organic clay complex was carried out in KBr pellets using  Jasco FTIR-4100 from wavenumber 4000 to 400 cm-1 (Ma, et al. 2010). 
RESULTS AND DISCUSSIONS
Infrared spectra of humic and fulvic acids are shown in Fig. (2) and Table (1). Humic acid show broad absorption centered around at 3360, 1406, 1233 and 1060 cm-1 regions (Edward et al., 2014). The phenolic (OH), amide, methyl, free-NH-bond, carboxylic and carbonyl group are presented in the humic and fulvic acids (Sharmah and Bordoloi, 1993). The absorption are around 1715 cm-1 region and due to the C=O stretching, (Haworth, 1971). The absorption are around 1630 cm-1 region that due to the C=C stretching vibrations of double bonds (Dorado et al., 2010). The 1200 cm-1 band is assigned to C-O stretching of phenols, carboxylic acids, ethers and OH- deformation of –COOH (Stevenson, 1994). The relative intensity of major IR absorption bands of humic and fulvic acids are shown in Table (1).

Egypt. J. of Appl. Sci., 34 (11) 2019                                                      377                                                 

              (Humic acid)                                        (Fulvic acid)     
Fig. (2) Infrared spectra of humic and fulvic acids
Table (1): Relative intensity of major IR absorption bands of humic and   fulvic acids
Acids	Frequency
(cm-1)	Assignment	Bond nature	

Humic acid	3360	H-bonded OH, or free		Broad	
	2930	 C-H2 stretching	Weak	
	1715	C=O stretching 	Weak	
	1630	C=C stretching vibration of double bonds 	Shoulder	
	1406	Aliphatic C-H deformation	Medium	
	1233	 C-O stretching of phenols, carboxylic acids, ethers and  COOH groups  	Medium	
	1059	Si-O stretching	Weak	

Fulvic
acid	3445	H-bonded OH, Free or bonded 	Broad	
	1631	C=C stretching vibration 	Strong	
	1404	 C-H 	Weak	
	1068	Si-O stretching	Medium	
 Jayaganesh and Senthurpandian (2010)

Kaolinite mineral
Figs. (3 and 4) and Table (2) shows that Kaolinite Clay- organic complexes can be attributed to kaolinite by IR absorption spectra. Their IR spectra demonstrate fully-represented and well-resolved OH, Al-O and Al-OH bands of the mineral lattice around 3650, 1350 and 850 cm-1 respectively. Kaolinite-Mg clearly-seen absorption bands of OH, Si-OH and Al-OH around 3500, 1080 and 850 cm-1 respectively. As the infrared absorption spectra showed, the kaolinite mineral adsorbed with humic acid is that the active groups are O-H stretching (H bonded OH group), N-H stretching, C-H2 stretching, C=O stretching of amide group and CH out of plane bending around wavenumber 3400, 2900, 1660 and 810 cm-1 respectively.  As for the kaolinite-humic-Mg, the dominant active groups were: OH, CH2 , C=C aromatic stretching and /or asymmetric –COO stretching, C-C stretching of aliphatic groups, C-O stretching of polysaccharides or polysaccharide-like, CH out of plane bending around the wavenumber  3680, 3000, 1620, 1070, 950, 810 and 775 cm-1 respectively.

378                                                      Egypt. J. of Appl. Sci., 34 (11) 2019                                                      

Fig. (3): Infrared spectra of Kaolinite Clay-Organic Complexes

Fig.(4) Kaolinite structure, showing the interlayer hydrogen bonds (Blanca, 2015)

Egypt. J. of Appl. Sci., 34 (11) 2019                                                      379                                                 

Table (2): Infrared spectra of Kaolinite clay–Organic complexes
W.N.
(cm-1)	Kaolinite	W.N.
(cm-1)	Kaolinite- Mg	*W.N.
(cm-1)	Kaolinite -humic	W.N.
(cm-1	Kaolinite –humic-Mg	W.N.
(cm-1	Kaolinite fulvic-Na	W.N.
(cm-1	Kaolinite fulvic-Ca	W.N.
(cm-1	Kaolinite fulvic -Mg	
3650
	OH
Weak	3500	OH
Medium	3400	N-H stretching
Medium	3670	OH
Weak	3670	OH Stretching
Weak	3670	OH Stretching
Weak	3670	OH Stretching
Weak	
1350
	Al-O
Medium	1080	Si –OH Stretching
Strong	2900	C-H2 stretching
Weak	3000	C-H2
Broad	3445	N-H stretching
Weak	1620	C=O
Weak	1631	C=O stretching
Strong	
850
	Al-OH
Strong	850	Al-OH
Medium	1660	C=O stretching
Weak	1620	C=C stretching
Strong	810	CH
Weak	810	Weak	850
	Al-OH
Weak	

				810	CH
Medium	1070	C-C Stretching
Weak	
	
			810
	CH
Weak	
		
				950	C-O stretching
Weak							
				
		810
	CH
Weak			

*W.N= wavenumber

380                                                      Egypt. J. of Appl. Sci., 34 (11) 2019                                                      

However kaolinite mineral saturated with fulvic acid and sodium cation (kaolinite- fulvic-Na), the effective groups were OH stretching, N-H stretching and CH out of plane was bending around 3670, 3445 and 810 cm-1 respectively .While kaolinite saturated with fulvic acid and calcium cation (kaolinite-fulvic-Ca) the active groups were OH stretching, C=O, CH out of plane and CH out of plane bending around 3670, 1620, 810 and 810 cm-1 respectively. At least, kaolinite mineral saturated with fulvic acid and magnesium cation (kaolinite-fulvic-Mg), the effective groups were OH stretching, H-bending OH, free OH, intermolecular bonded OH, C=O stretching vibration of double bounds of cyclic and acyclic compounds benzene ring substitution around wavenumber 3670, 3500, 1631, 910 and 850 cm-1 respectively.     
Bentonite mineral
Figs. (5 and 6) and Table (3) shows that bentonite clay- organic complexes can be attributed to bentonite by IR absorption spectra. Their IR spectra demonstrate fully-represented and well-resolved OH stretching, H-bending OH, free OH, intermolecular bonded OH, C=O, Si-O stretching and Al-OH-Al bending   bands around wavenumber 3632, 3422, 1634, 1048 and 918  cm-1 respectively. Bentonite-Mg clearly-seen absorption bands of OH, C=O, Si-O stretching, Al-OH-Mg bending and Si-O stretching around 3250, 1634, 1048, 843 and 778 cm-1 respectfully. As the infrared absorption spectra showed, the bentonite mineral adsorbed with humic acid (bentonite-humic) that the active groups were N-H stretching, C=C aromatic stretching and /or asymmetric –COO stretching,-CH- polysaccharides, C=O stretching and CH out of plane around wavenumber 3300, 1600, 1050, 950 and 775 cm-1 respectively.  As for the bentonite-humic-Mg, the dominant active groups were: OH,  stretching, N-H stretching, C=C aromatic stretching and /or asymmetric –COO stretching, C-C stretching of aliphatic groups, C-O stretching of polysaccharides or polysaccharide-like, CH out of plane bending around the wavenumber  3600, 3300, 1620, 1070, 950, and 830 cm-1 respectively. 
 However bentonite mineral saturated with fulvic acid and sodium cation (bentonite-fulvic-Na) the effective groups were  OH stretching, N-H stretching, C=C aromatic stretching, Si-O stretching, Al-OH-Al bending and Al-OH- around 3632, 3300, 1525, 1068, 915 and 842 cm-1 respectively. While bentonite saturated with fulvic acid and calcium cation (bentonite-fulvic- Ca) the active groups were OH stretching, C=C aromatic stretching and, CH out of plane around 3632, 1620, and 830 cm-1 respectively. At least, bentonite mineral saturated with fulvic acid and magnesium cation (bentonite-fulvic-Mg) indicated the effective groups were H-bonded OH, free OH, N-H stretching, C=C aromatic stretching, CH stretching, C=O stretching of polysaccharides or polysaccharide –like substances and CH out of plane around wavenumber 3400, 3300, 1631, 1068, 950 and 775 cm-1 respectively.  

Egypt. J. of Appl. Sci., 34 (11) 2019                                                      381                                                 

Fig.(5): Infrared spectra of Bentonite clay-organic complexes

Fig. (6) Bentonite structure, showing the interlayer hydrogen bonds (Jun, et al. 2019).

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Table (3): Infrared spectra of Bentonite clay –Organic complexes
W.N.
(cm-1)	Bentonite	*W.N.
(cm-1)	Bentonite-Mg	W.N.
(cm-1)	Bentonite-Humic	W.N.
(cm-1	Bentonite-Humic-Mg	W.N.
(cm-1	Bentonite-Fulvic-Na	W.N.
(cm-1	Bentonite-Fulvic-Ca	W.N.
(cm-1	Bentonite-Fulvic-Mg	
3632	OH stretching
Weak	3250	OH
Broad	3300	N-H stretching
Medium	3600	OH Stretching
Shoulder	3632	OH stretching
Weak	3632	OH Stretching
Shoulder	3400	OH Stretching
Shoulder	
1634	C=O
Medium	1634	C=O
Strong	1600	C=C stretching
Medium	3300	N-H stretching
Broad	3300	N-H stretching
Medium	1620	C=C stretching
Weak	3300	N-H stretching
Broad	
1048
	Si-O stretching
Broad	1048	Si-O stretching
Weak	1050	-CH-
Shoulder	1620	C=C stretching
Medium	1525	C=C stretching
Medium	830	CH
Broad	1631	C=C stretching
Strong	
918
	Al-OH-Al bending
Weak	843	Al-OH- Mg bending
Medium	950	C=O stretching
Shoulder	1070	C-C Stretching
Shoulder
	1068	Si-O stretching
Weak			1068	CH stretching
Shoulder	
						950	C-O stretching
Shoulder	915	Al-OH-Al bending
Shoulder			950

	C=O stretching
Shoulder	

		830	CH
Shoulder			
				
*W.N= wavenumber

Egypt. J. of Appl. Sci., 34 (11) 2019                                                      383                                                 

Assessment of effective groups
          Fig. (7) reveal a common of frequency function groups on kaolinite mineral, dominated by OH, CH, C=O, Al-OH, N-H, CH2 , C-C, C=C, Al-O and Si-OH in a descending order.  While, contents of C=C, Al-O and Si-OH groups the least widespread.    

 Fig.(7) Frequency of function groups on kaolinite mineral
       Fig. 8 dictate that the frequency of function groups bentonite mineral dominant dominated by OH, C=O, C-C, N-H, CH, Si-O, Al-OH-Al and C-C in a descending order. It is noticeable; contents of C-C group the least common.

Fig. (8) Frequency of function groups bentonite mineral 
         
CONCLUSION
The results showed that the difference among the effective groups within the treatments of the one clay mineral is due to the prevailing cation type (Ca, Mg and Na) and behavior on element or clay mineral type. As for the difference in the effective groups between the treatment of the one mineral is due to the type of organic acid and its effects on the surface of clay mineral and the inner layers. While the difference of the effective groups between the minerals is due to the difference in the type of clay mineral (kaolinite and bentonite) and the group to which it belongs to. 

384                                                      Egypt. J. of Appl. Sci., 34 (11) 2019                                                      

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Egypt. J. of Appl. Sci., 34 (11) 2019                                                      385                                                 

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تقييم سلوك معقد الطين العضوى  بإستخدام نظام ألأشعه تحت الحمراء
نبيل محمد عبداللطيف بهنساوى
 قسم كيمياء وطبيعة ألأراضى - مركز بحوث الصحراء- مصر
          تهدف الدراسة إلى تقييم سلوك معقد الطين العضوى  بإستخدام نظام ألأشعه تحت الحمراء. وأوضحت النتائج أن معدن طين الكاؤلينيت المشبع بالأحماض العضوية وكاتيونات الكالسيوم والماغنسيوم والصوديوم بعد الفحص بالأشعة تحت الحمراء أن  المجاميع الفعاله السائدة هى:-                                       
 OH, N-H, CH2, C=O, C=C, Al-O, Si-OH, C-C, Al-OH and CHوذات أرقام موجية هى: (3670-3500) ، (3445-3400) (3000-2900) ، (1660-1620) ، 1610، 1350، 1080، (1070-950) ، 850 و810 سم-1 على الترتيب.
كما أوضحت النتائج أن معدن طين البنتونيت المشبع بنفس المعاملات السابقة  بعد فحصها بالأشعة تحت الحمراء أن المجاميع الفعاله السائدة هى:-
 OH, N-H, and C=O, C=C, CH, Si-O C-C and Al-OH-Al وذات أرقام موجية هى:
(3632-3250) ، 3300، (1634-950) ، (1631-1525) ، (1068-830 )  (1068-1048)،1070 و (918-915 )    سم-1على الترتيب.
ومن الدراسة إتضح أن ترتيب المجاميع الفعالة على معدن طين الكاؤلينيت كالتالى:-
OH > CH > Al-OH = C=O > N-H = C-C = CH2 > C=C = Al-O = Si-OH 
بينما معدن طين البنتونيب فكان ترتيب المجاميع الفعالة هى:-
OH > C=O = C=C >NH = CH > Si-O >Al-OH-Al >C-C.