3-(3,4-Dihydroxyphenyl)-2-Propenoic Acid Synthesis

3-(3,4-Dihydroxyphenyl)-2-Propenoic Acid SynthesisAntimicrobic bodily function of Co (II) complex of 3-(3,4-Dihydroxyphenyl)-2-propenoic acid and sonochemical synthesis of nanoscale mixed ligand EDA coordination for preparation of CoCl2.6H2O anoparticleAbstract3-(3,4-Dihydroxyphenyl)-2-propenoic acid abbreviated as EDA was synthesized and characterized. Co (II) metal complex of this ligand prepared by response of chloride salt with EDA in dry acetonitrile. Phenolic compounds (a group of secondary metabolites) are wide distributed in plants and break shown to possess antimicrobial properties. Anti bacteriuml activity was studied for ligand and its metal complex. This complex were foot raceed for their antibacterial activity against Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes and Escherichia coli relatively with that of free ligand. Differential response to phenolic compounds was observed among bacteria. Also this complex was synthesized in Nano-scale an d was characterized by SEM, XDR (X- Ray powder diffraction). Antibacterial activity of complex and nano- literal were studied and compared with each other.Keywords 3, 4-dihydroxy benzene acrylic acid, Cu (II) and Co (II) complexes, Antibacterial activity, nano- scaleIntroductionAcrylic acid or hydroxycinnamic acid compounds are widely distributed in the plants. They usually exist number of them exist as free acids in nature 4, 5, 6.Much work has been realized by bioin ingrained as well as medicinal chemists to launch the relationship between the metal ions and their complexes as antimicrobial agents 7-8. Phenolic compounds are secondary plant metabolites and innate(p)ly present in almost all plant materials, including food products of plant origin. These compounds are thought to be an integral part of both human and animal diets 13. The chemical structure of phenolic acids shows that they are simple phenols. Hydroxycinnamic acid is the major subgroup of phenolic compounds 11,12. Hydroxycinnamates are phenylpropanoid metabolites and occur widely in plants4,and plant products2. Hydroxycinnamates and their derivatives are bioactive plant food ingredients. The other natural ligand from plants such as alkaloids compound also can be employ in synthesis of metal complex 1.Nanophasic and nanostructured materials are attracting a great piss it off of attention because of their potential for achieving specific processes and selectivity, especially in biological and pharmaceutical applications 3. Nanoparticles are made of natural or artificial polymers ranging 10.In particular, those conjugated with biological moieties have enormous potential in drug delivery and therapeutic applications. In fact, much progress has been achieved in the past ten years based on inorganic nanomaterials 9.In this context we have undertaken the antimicrobial evaluation of Co (II) complex of 3-(3,4-Dihydroxyphenyl)-2-propenoic acid. For this purpose the in vitro expertness of tow gram positive bacteria (Staphylococcus aureus, Streptococcus pyogenes) and tow gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli) to the synthesized compounds was investigated.Materials and MethodsSynthesis of the metal complex General Method3-(3,4-Dihydroxyphenyl)-2-propenoic acid, cobalt chloride was Merck chemicals and was used without further purification. Organic solvents were reagent grade. electronic spectra were recorded by Camspec UVVisible spectrophotometer model Perkin Elmer Lambda 25. The IR spectra were recorded using FT-IR Bruker Tensor 27 spectrometer. 1H- NMR was recorded on a Bruker AVANCE DRX 500 spectrometer at 500 and 125MHz respectively. All the chemical shifts are quoted in ppm using the high-frequency positive convention 1H -NMR spectra were referenced to external SiMe4. The percent composition of elements was obtained from the Microanalytical Laboratories, Department of Chemistry, University of tarbiyatmoallem, Tehran.A solution of metal salt dissol ved in acetonitrile added a dually to a stirred acetonitrile solution of the ligand (EDA), in the molar ratio 11 (metal ligand). The reaction mixture was further stirred for 4-5h to ensure of the completing and precipitation of the formed complexes. Finally, the complexes dried in vacuum desiccators over anhydrous CaCl2.Microorganisms and culture mediaThe following microorganisms were used in this study to test antimicrobial activity of complex. Escherichia coli, Staphylococcus aureus were kindly provided by (anistitue pastor ). The strains were maintained on PD3 agar at 26oC. For long- term storage, glycerol stocks of microorganisms were prepared in the corresponding growth media with a final glycerol concentration of 12%. The bacterial glycerol stocks were quickly frozen in lucid nitrogen and stored at -80oC.In-vitro anti-bacterial activityAll methodology and steps were followed according to diffusion criminal record method. An inoculum of 0.5 McFarland standard (1.5*108 cfu/ml ) was applied on Mueller Hintonagar (a depth of 4 mm in a petridish of 100 mm diameter) 14. Maximum 6 discs were applied on each plate and they were incubated at 37 C for 24 hours. Zone of inhibition was measured including the disc diameter (6mm).Preparation of nanoparticlesCo nanocrystallites were prepared by the reaction of C9H8O4 with Co (C9H7O4) Cl2 in THF as solvent under echography power. Then the suspension was irradiated for 1h with a high- density ultrasonic probe immersed directly into the solution under various conditions. A maltiwave ultrasonic generator ( sonicator 3000 Italstructure MPD 3000). The samples were characterized with a scan electron microscope (SEM) with gold coating.Results and DiscussionsStructural description of the complexThe reaction of Co(II) salt with the ligand, EDA, results in the formation of ML for M=Co (II). complex is quite s add-in and could be stored without any appreciable change. The EDA ligand and the Co(C9H6O4)Cl2.2H2O complex have 223-2 25C and 195-198C melting point respectively also complex is insoluble in common organic solvents, such as n-hexane and dichloromethane. However, that is soluble in DMSO, ethanol and DMF. its structure was characterized by elemental analysis, 1H-NMR and IR. Their elemental analyses are in accord with their proposed formula. The spectral data of the complexes have good relationship with the literature data.Fig.1. Structure of the ligand, EDA.Analysis of Co ( C9H6O4 ) Cl2.2H2O (EDACC) Dark Blue crystals yield 86% .elemental analyses, 11 metal to ligand stoicheiometry is designate to all the chelates ( table 1).Table1 Elemental analyses data on the caffeic acid and its Co (II) complex. (31.40) (1.74) (17.15)1H-NMR ( ppm DMSO, 500MHz) 5, 97-7.21 5H, 2q, aroma 11.91 1H, s, acid) 8.98-9,362H, s, alkene. IR absorptions(cm-1 KBr) 1620 (C=C), 972 (=C-H), 1352 (C-O), 574 (Co-O) and 456 (Co-Cl).The electronic spectral data of the complex in acetonitrile are presented in table 2. there are one peak in spectrum of ligand which can be assigned to * transition. The electronic complex shows a broad band at 680 nm attributable to the 4T1g ( F ) 4 A2g ( F) and the other one at 640- 550 nm attributable to the 4T1g ( F ) 4T1g ( P ) transition for Co( II ) ion .table 2 Electronic spectra of caffeic acid and its Co(II) complex in nmRaman shift (cm1) 500 Co-O), 325 (Co-Cl), 975(C-H), 1618C=C), 1189C-O) (Fig.2.Left).Fig.2. Structure of Co (II) complex with ligand, EDA.In-vitro anti-bacterial activityThe mean diameters of microbial growth inhibited by different complexes are shown in Table 4. All complexes had antimicrobial activity. Inhibition zones larger than 5mm indicated that antimicrobial activity. The data obtained by the disk diffusion method showed that all complexes have antibacterial activity. Among the bacteria, Escherichia coli was the most sensitive bacteria both ligand and complex Co(C9H7O4)Cl2 Normal- scale had antibacterial effect on this bacteria , whereas only lig and C9H8O4 had antibacterial activity against Streptococcus pyogenes and Pseudomonas aeruginosa . Ligand C9H8O4 and complex Co (C9H7O4) Cl2( Normal- scale) had non any effect against Staphylococcus aureus.Table 4. Zone of growth inhibition of the test compounds against the bacteriaComplex Co (C9H7O4)Cl2 ( Normal scale), had more antibacterial activity against Escherichia coli (14mm) . Complex Co(C9H7O4)Cl2 (Nano scale), had less activity against Staphylococcus aureus (5mm). From these results it may be concluded that there is not any accordance between normal scale and nano- scale from the antibacterial activity aspect. The antimicrobial activity of complexes demonstrated in this study can be added to the already known beneficial biological properties of these compounds to the human health.Nanoparticle studyXRD pattern of mixture of caffeic acid andCoCl2 .6H2O prepared by the ultrasonic process is given in Fig. 4. The diffraction peaks accord with the amorphous crystal system.Fig .4. The XRD pattern of the mixture of caffeic acid and 6H2O .CoCl2The SEM micrographs of nanostructure are shown in Fig. 5. The nanoparticles show a low degree of crystalline with no be peaks in the XRD pattern.Fig. 5. SEM images of complex Co (C9H7O4)Cl2The IR spectrum of Co (II) nanostructure (Fig.5) shows the absorption pesk at 574 648 are assigned to the (Co-O) modes, which confirms the formation of Co (II) nanostructure.It has been reported that the negative charge on the cadre surface of Gram-negative bacteria was higher than on Gram-positive bacteria (Chunget al., 2004). Due to a higher negative charge on cell surface, thefundamental interaction between Gram-negative bacteria and nanoparticles was definitely stronger than that of Gram-positive bacteria. Moreover, results showed that Gram-positive bacteria were more sensitive to Co (II) nanoparticles.Fig.6. IR Spectrum of complex Co (C9H7O4) Cl2AcknowledgmentsWe gratefully acknowledge the pecuniary support from the Researc h Council of Ardabil Islamic Azad University and many technical supports that provided by Ardabil University of. edical SciencesReferences1 A.H. Osman, (Synthesis and characterization of cobalt (II) and nickel (II) complexes of someSchiff bases derived from 3-hydrazino-6-methyl 1, 2, 4 triazin-5(4H) one) Transition Met. Chem, 2006, 31, 35.2 Clifford, 1999 M.N. Clifford, Chlorogenic acids and other cinnamates nature, occurrence and dietary burden, J. Sci. nutrient Agric. 79 (1999), pp. 362372.3 Brigger, I., C. Dubernet, and P. Couvreur. 2002. Nanoparticles in cancer therapy and diagnosis. Adv. Drug Delivery Rev. 546316514 Herrmann, K. Occurrence and content of hydroxycinnamic and hydroxybenzoic acid compounds in foods. Crit. Rev. Food Sci.Nutr. 1989, 28, 315-3475 Herrmann, K. Contents of principle plant phenols in fruits. Fluess. Obst 1992, 59, 66-70.6 Herrmann, K. Phenolcarboxylic acids in plant foods and their antioxidative activity. Gordian 1993, 93, 92-96.7 M.J. Seven and L. A. Johnson, Metal Binding in Medicine, Lippincott Co, Philadelphia, PA, 4ta Ed (1960).8 A. J. Crowe, Metal-Based Antitumor Drugs, Vol. 1, Freund, London, 1989, p. 1039 Gong, J., Hu, X., Wong, K., Zheng, Z., Yang, L., Lau,W., Du,R. Chitosan Nanostructures with Controllable Morphology Produced by a Nonaqueous Electrochemical Approach. good materials. 2008, 20, 21112115.10 Kreuter, J. (2001). Nanoparticulate systems for brain delivery of drugs. Advanced Drug Delivery Reviews, 47, 6581.11 C. Sanchez-Moreno, J.A. Larrauri and F. Saura-Calixto, A procedure to measure the antiradical efficiency of polyphenols, J. Sci. Food Agric. 76 (1998), pp. 270276.12 Z. Sroka and W. Cisowski, Hydrogen peroxide scavenging, antioxidant and anti-radical activity of some phenolic acids, Food Chem. Toxicol. 41 (2003), pp. 753758.13 E. Psomiadou and M. Tsimidou, Stability of virgin olive oil. 1. Autoxidation studies, J. Agric. Food Chem. 50 (2002), pp. 716721.14WHO, Geneva (Sept. 1997). Manual on antimicrobial resistance and susceptibility testing. Diversion of emerging and other communicable diseases surveillance and control. WHO antimicrobial resistance monitoringprogramme.