PREPARATION, CHARACTERIZATION, ANTIOXIDANT ACTIVITY OF FUNCTIONALIZED 2-(FERROCENYLMETHYLAMINO) BENZONITRILE BY ZnO NANOPARTICLE USING 3 AMINOPROPYLTRIETHOXYSILANE(APTES)

The aim of the present study was study the interaction between 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) and ZNO@APTES@2-(ferrocenylmethylamino)benzonitrile and Galic acide Then determination of binding parameters like binding constant and binding free energy The determination of binding constant and binding free energy is based upon the decrease in absorbance of the electronic absorption spectrum .The ZNs was synthesise according to chemical method. The characterizations of ZnO NPs were measured by FTIR and X-ray diffraction. (


Introduction
Nanoparticles are a piece of nanomatrials that are known as a single particles 1-100 nm in diameter from another years nanoparticles have been a common material for the development of new cutting-edge applications in communications, energy [1] On the other hand Zinc oxide nanoparticles have attracted more care due to their multilateral and promising applications in biological sciences like an antibacterial, antifungal,and antifouling agent [2] As indicated in more reports ZnO is regarded as superior functional material due to the numerous actives sites on its cover wich can be efficiently secure with other materials.[3]

Preparation ZnO@APTES
ZnO NPS was prepared according to chemical method [5].Aqueous ammonia (0.2M) was slowly dripped at a rate of 30 mL/ h in a solution of 100 mL of zinc acetate(0.1 M) to pH of 10.This mixture was kept at 60 C for 12 h and then centrifuged for20 min at a speed of 3800 rpm/min.Finally, the solid was rinsed several times with ultrapure water and ethanol by centrifugation and then dried in an oven for 1 h..The prepared ZnO (10 mg) was dissolved in 10 mL ethanol and ultrasonicated for 1h.APTS (10 mL) was added to the solution and sonicated for 1 h to complete the reaction.The amino-functionalized ZnO was separated from the solution via centrifugation, washed with absolute ethanol.

Characteristic measurement
The ultrasonic system used in this study were a VCX 505 sonics.X-ray diffraction(XRD; PROTO) This device works under a wavelength λ CuKα1 = 1.54 Å was performed at 40Kv voltage and 40mA current.Fourier transform infrared (FT-IR) spectra of the samples were recorded at room temperature (Agilent Technoogies Cary 630 FTIR) in the 300-4000cm -1 region.
UV-Vis experiments were performed using a (SP-UV300SRB) and a quartz voltammetric cell with a volumetric capacity of 5 mL.Graphs plot and calculus were carried out using OriginLab software version 2.0 (Integral Software, France).

Spectrophotometric studies of DPPH-ZnO@APTES@2(ferrocenylmethylamino)benzonitril interaction
The interaction of @APTES@2(ferrocenylmethylamino)benzonitril with DPPH was studied by UV-Vis absorption titration for getting further clues about the mode of interaction and binding strength.Has been calculated the Binding constant Depending on the Benesi-Hildebrand equation: where K is the binding constant, A 0 and A are absorbance of DPPH in the absence and in the presence of antioxidant standards, 0and are their absorption coefficients respectively, [AS] concentration of antioxidant standard.
Table 1 represents the FWHM value for every peak assigned for particle size calculation.Table 1.Parameter calculation for average size calculation for nanoparticle.

FT-IR analysis
Substance-specific vibrations of the molecules lead to the specific signals obtained by IR spectroscopy.FT-IR spectra and fuctional group involved in ZnO NPs synthesis illustrated peak in the rang of 400-550cm -1 [7] (fig 2 ) which is belonged to Zno stretching vibration.After the silane treatment (fig3)several new peak were observed, broak peak obtained at 3350cm -1 corresponded to OH stretching vibration [8] and the peak in the rang of 1020-1250cm -corresponded Si-O [9]The peak at 1610 cm -belonged to the NH bond and -NH2 deformation vibration, which proved the presence of NH2 groups [7] .The peak at 687 cm -1 can be assigned to Si-C stretching vibration.and the table 2 show the most functional groups.
Such red shift of the FTIR spectrum in ZnO@APTES@2(ferrocenylmethylamino)benzonitril composites are due to the conjugation between ZnO and2(ferrocenylmethylamino)benzonitril surface, the peak at 490cm-1 correspond Fe-c and the peak at 2870 correspond C-H aromatic stretch .The bands at 1425 and 1610 cm-1 can be attributed to the stretching of N-H groups , the peak at 1358 and 1537 correspond NO stretch .Therefore, the FT

Determination of IC 50
The addition growing of ontiaxidants to (10 -4 DPPH Dissolved in ACN ) a remarkable overall decrease in absorbance.(table3) And through the equation I=((A 0 -A 1 ))/A 0 )*100 We can calculated Percent of inhibition I where I : Percent of inhibition, A0 and A are absorbance of DPPH• in the absence and in the presence of antioxidant standards [10] Table 3. Inhibition values of DPPHof ZNO@APTES@2-(ferrocenylmethylamino)benzonitril and Galic Acide.We observe when it is IC 50 small the antioxidant effectiveness it is big and from it we conclude ZNO@APTES@2-(ferrocenylmethylamino)benzonitril has antioxidant effectiveness less than Galic Acide.

Binding constant
The binding constant and binding free energy were calculated based upon the decrease in absorbance using the equation of Benesi-Hildebrand And ZNO@APTES@2-(ferrocenylmethylamino)benzonitril and it is chemical "interaction chemical" while indicates the small value of the binding constant between G.A and free radical DPPH .That this interaction electrostatic "electrostatic interaction physical" The negative values of the binding free energy resultant from overlap between free radical DPPH .And Galic Acide ,ZNO@APTES@2-(ferrocenylmethylamino)benzonitril indicate on automatic interaction

Table 2 .
Functional group and absorption band of the ZnO after APTES treated.

Table 4 .
Results of the evaluation of antioxidant activity.

Table 5 .
Values of A and A 0 /A-A 0 For each concentration.

Table 6 .
Binding constants and binding free energy values.