A Suitable Base Material for Composite Resin Restorations

Food and Chemical Toxicology 45 (2007) 16501661 www. elsevier. com/locate/foodchemtox A par of chemical, antioxidant and antimicrobial studies of cinnamon bark bark interchange and mouth volatilizable anele colours, oleoresins and their scamstituents q Gurdip Singh b a,* , Sumitra Maurya a,1 , M. P. deLampasona b, Cesar A. N. Catalan b a alchemy Department, DDU Gorakhpur University, Gorakhpur 273 009, India Instituto de Quimica Organica, Universidad Nacional de Tucuman, Ayacucho 471, S. M. de Tucuman 4000, Argentina Received 31 August 2005 accepted 22 February 2007Abstract The antioxidant, antimycotic agent and antibacterial potentials of vapourific crude covers and oleoresin of Cinnamomum zeylanicum Blume ( undulate and bark) were investigated in the present study. The oleoresins confirm shown excellent activity for the forbidding of primary and tributary oxidization products in mustard inunct added at the concentration of 0. 02% which were evaluated utilize peroxide, thiobarbituric pane of glass, p-anisidine and carbonyl cling tos. Moreover, it was further supported by former(a) complementary antioxidant assays such as ferric thiocyanate method acting in linoleic acerb system, minify power, chelating and scavenging e? cts on 1,1 0 -diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl thoroughs. In antimicrobial investigations, utilise inverted petriplate and food poison techniques, the alternate and bark volatilizable oils has been gear up to be highly e? ective against all the footraceed kingdom Fungi except Aspergillus ochraceus. However, leaf oleoresin has shown inhibition only for Penicillium citrinum whereas bark oleoresin has turn in gotd drop mycelial govern inhibition for Aspergillus ? avus and A. ochraceus on with Aspergillus niger, Aspergillus terreus, P. citrinum and Penicillium viridicatum at 6 lL. Using agar well di? sion method, leaf volatile oil and oleoresin keep shown better results in comparison with bar k volatile oil, oleoresin and mercantile bactericide, i. e. , ampicillin. Gas chromatographicmass spectroscopy studies on leaf volatile oil and oleoresin resulted in the identi? cation of 19 and 25 components, which accounts for the 99. 4% and 97. 1%, respectively of the numerate amount and the major component was eugenol with 87. 3% and 87. 2%, respectively. The analysis of cinnamon bark volatile oil showed the presence of 13 components invoice for snow% of the tally amount. E)-cinnamaldehyde was build as the major component along with d-cadinene (0. 9%), whereas its bark oleoresin showed the presence of 17 components accounting for 92. 3% of the total amount. The major components were (E)-cinnamaldehyde (49. 9%), along with some(prenominal) other components. O 2007 Elsevier Ltd. All rights reserved. Keywords Cinnamomum zeylanicum Blume Eugenol Cinnamaldehyde Antioxidant assay 1. Introduction Free antecedent reactions occur in human body and food systems. Free radicals, in the form of reactive oxygen and Part 57.Corresponding author. Tel. +91 551 2200745 (R)/2202856 (O) fax +91 551 2340459. netmail address emailprotected com (G. Singh). 1 Present address Agarkar Research Institute, Pune 411 004, India. * q nitrogen species, atomic number 18 an integral part of normal physiology. An over production of these reactive species can occur, ascribable to aerophilous stress brought ab bulge by the imbalance of bodily antioxidant defence system and assuage radical formation. These reactive species can react with biomolecules, ca development cellular injury and death.This may lead to the development of chronic diseases such as cancers and those that involve the cardio- and cerebrovascular systems. The consumption of fruits and vegs (Peschel et al. , 2006) containing antioxidants has been found to o? er protection 0278-6915/$ identify front matter O 2007 Elsevier Ltd. All rights reserved. doi10. 1016/j. fct. 2007. 02. 031 G. Singh et al. / Food and Chem ical Toxicology 45 (2007) 16501661 1651 against these diseases. Dietary antioxidants can augment cellular defences and athletic supporter to prevent oxidative equipment casualty to cellular components (Halliwell, 1989).Besides playing an important role in physiological systems, antioxidants have been used in food industry to prolong the shelf liveliness of foods, specially those rich in polyunsaturated fats. These components in food ar take onily oxidized by molecular oxygen and are major cause of oxidative deterioration, nutritional losses, o? ?avour development and discoloration. The addition of semisynthetic antioxidants, such as propyl gallate, only ifylated hydroxylanisole (BHA), butylated hydroxyltoluene (BHT) and tertiary butylhydroquinone has been widely used industrially to control lipide oxidation in foods.However, the use of these synthetic antioxidants has been questioned due to their potential health risks and toxicity (Kahl and Kappus, 1993). The search for ant ioxidants from natural sources has stock much attention and e? orts have been put in to identify compounds that can act as suitable antioxidants to replace synthetic ones. In addition, these naturally occurring antioxidants can be formulated as functional foods and nutraceuticals that can help to prevent oxidative damage from occurring in the body.Plants contain a variety of substances called Phytochemicals (Pratt, 1992), that owe to naturally occurring components present in installs (Caragay, 1992). The phytochemical preparations with bivalent functionalities in preventing lipid oxidation and antimicrobial properties have tremendous potential for extending shelf life of food products. Several research groups around the world have succeeded in ? nding and identifying natural antioxidants from herbs and spices using di? erent elanl systems.The antioxidant activity of Labiatae herbs such as rosemary, sage, summer savory and borage are also well authenticated (Bandoniene et al. , 2 002 Djarmati et al. , 1991 Ho et al. , 2000 Aruoma et al. , 1996 Cuvelier et al. , 1994 Wong et al. , 1995 Chang et al. , 1997 Madsen et al. , 1996 Gordon and Weng, 1992 Takacsova et al. , 1995). However, the aromatic spicy and medicinal grounds from Laureceae family are less extensively studied. Cinnamon (Cinnamomum zeylanicum Blume, syn C. verum, family Laureceae) is a widely used spice and have many applications in perfumery, ? voring and pharmaceutical industries. Although, the chemical constituents of leaf and bark essential oils of cinnamon have been studied (Raina et al. , 2001 ? Simic et al. , 2004 Jayaprakash et al. , 1997), the potential antioxidant properties have yet not been studied and it seems that investigation on oleoresins are scarce. Hence, in the present work, attempt has been made to explore the possible antioxidant and antimicrobial properties by di? erent methods which can give more comprehensive information especially when the e? ectiveness of multi componen t natural oleoresins is investigated.The objective of present investigation is to compare the chemical com lay out of leaf and bark essential oils and oleoresins as well as demonstrate the possibility of protecting the stored food materials against micro-organism and antioxidative behaviour on mustard oil using as additive by various methods. 2. Materials and methods 2. 1. Chemicals Thiobarbituric point, pure components eugenol and cinnamaldehyde were received form Merck, Germany. Diphenylpicrylhydrazyl (DPPH), carbendazim were procured from Sigma (SigmaAldrich GmbH, Sternheim, Germany) and linoleic acid from Across (New Jersey, USA).BHT, BHA, and 2,4-dinitrophenylhydrazine were purchased from s. d ? ne-chem Ltd, Mumbai, India. Ampicillin was purchased from Ranbaxy Fine chemicals Ltd. , New Delhi, India. Crude mustard oil was purchased from local oil mill, Gorakhpur, India. All resultants used were of analytical grade. 2. 2. Sample extraction Cinnamon leaves and barks were purchas ed from local market of Gorakhpur, Uttar Pradesh, during January 2004 and voucher specimens were kept at the Herbarium of the Science faculty, DDU Gorakhpur University, Gorakhpur.Cinnamon leaves (250 g) and barks (50 mesh particle size) were hydrodistilled using Clevengers apparatus to ante up essential oils (3. 1% and 2. 5%, respectively). Oleoresins were obtained by extracting 25 g of powdered spice with 250 mL of acetone for 2 h in a Soxhlet extractor. The solvent was evaporated by placing the judge in a vacuum drier under reduced pressure. The viscous oleoresins for leaves and barks, with yield 6. 9% and 9. 7%, respectively, were obtained. Both essential oils and oleoresins were stored in cold condition and until further use. 2. 3. Chemical characterization 2. . 1. Gas chromatography (GC) A Hewlett Packard 6890 ( analytic Technologies SA, Buenos Aires, Argentina) gas chromatograph equipped with column HP-5 (5% phenyl methylsiloxane, length 30 m inner diameter 0. 25 mm ? lm t hickness 0. 25 lm) was used for the analysis whose injector and detector temperatures were maintained at 240 and 250 C, respectively. The amount of the samples injected was 0. 1 lL in split mode (801). Carrier gas used was helium with a ? ow rate of 1. 0 mL minA1. The oven temperature for essential oils were programmed linearly as follows 60 C (1 min), 60 185 C (1. C minA1), 185 C (1 min), 185275 C (9 C minA1 ), 275 C (5 min) whereas for oleoresins it was as follows 70 C (1 min), 70170 C (1. 5 C minA1), 170 C (1 min), 170180 C (9 C minA1), 280 C (5 min). 2. 3. 2. Gas chromatographymass spectrometry (GCMS) Analysis of volatile oils and oleoresins were run on a Hewlett Packard (6890) GCMS system (Analytical Technologies SA, Buenos Aires, Argentina) coupled to a quadrupole mass spectrometer (model HP 5973) with a capillary column of HP-5MS (5% phenyl methylsiloxane, length = 30 m, inner diameter = 0. 25 mm and ? lm thickness = 0. 5 lm). The injector, GCMS interface, ion source and sele ctive mass detector temperatures were maintained at 280, 280, 230 and 150 C respectively. The oven temperature programmed for the volatile oils were akin as provided for GC whereas for oleoresins, it was programmed linearly as follows 60 185 C (1. 5 C minA1), 185 C (1 min), 185275 C (9 C minA1), 275 C (2 min). The extract was held at 70 C (5 min), 70220 C (3 C minA1), 220280 C (5 C minA1) and held at 280 C for 5 min. 2. 3. 3. Components identi? cation The components of essential oil and oleoresins were identi? d on the basis of comparison of their retention indices and mass spectra with published data (Adams, 2001 Massda, 1976) and computer matching with WILEY 275 and National Institute of Standards and applied science (NIST 3. 0) libraries provided with computer controlling the GCMS system. The results were also con? rmed by the comparison of the compounds elution order with their relative retention indices on non-polar phase 1652 G. Singh et al. / Food and Chemical Toxicology 45 (2007) 16501661 2. 4. 2. DPPH and hydroxyl radical scavenging e? ects The DPPH assay was carried out as described by Brand-Williams and his co-workers (1995). , 10, 15, 20, 25 lL of the sample were added to 5 mL of 0. 004% methanol beginning of DPPH. later on a 30 min incubation point in time at room temperature, the absorbance was read against a blank at 515 nm. The assay was carried out in triplicate and analyses of all samples were run in duplicate and results are averaged. This try out was adopted from a method described by Halliwell et al. (1987). Solutions of the reagents were always prepared freshly. The reaction variety contained in a ? nal volume of 1. 0 mL, one hundred lL of 2-deoxy-2ribose (28 mM in KH2PO4K2HPO4 bu? er, pH 7. ), 500 lL of various concentrations of the tested oils or the pure compounds in bu? er, 200 lL of 1. 04 mM EDTA and 200 lM FeCl3 (11 v/v), century lL of 1. 0 mM H2O2 and one C lL of 1. 0 mM ascorbic acid. Test samples were kept at 37 C for 1 h. The free radical damage imposed on the substrate, deoxyribose, was thrifty using the thiobarbituric acid test (Ohkawa et al. , 1979 Shimada et al. , 1992). 1. 0 mL of TBA (1%), and 1. 0 mL tricholoroacetic acid (2. 8%) were added to the test tubes and were incubated at ampere-second C for 20 min. After cooling, absorbance was measured at 532 nm against a blank containing deoxyribose and bu? r. Reactions were carried out in triplicate. Inhibition (I) of deoxyribose degradation in percent was mensural in the following way I? %? ? degree CelsiusX ? A0 A A1 =A0 ? where A0 is the absorbance of the control reaction, and A1 is the absorbance of the test compound. 2. 4. 3. Chelating e? ect and reducing power Chelating e? ect was determined accord to the method of Shimada et al. (1992). To 2 mL of the mixture, consisting of 30 mM hexamine, 30 mM potassium chloride and 9 mM ferrous sulphate were added to 5, 10, 15, 20, 25 lL of essential oil or oleoresin in methanol (5 mL) and 200 lL of 1 mM tetramethyl murexide.After 3 min at room temperature, the absorbance of the mixture was determined at 485 nm. A abase absorbance indicates a higher chelating power. EDTA was used as a positive control. The reducing power was carried out as described before (Oyaizu, 1986). Various amount (5, 10,15, 20 lL) of essential oil or oleoresin (dissolved in 2. 5 mL of methanol) mixed with 2. 5 mL of 200 mM phosphate bu? er (pH = 6. 6) and 2. 5 mL of 1% potassium ferricyanide, and the mixture was incubated at 50 C for 20 min. After adding 2. 5 mL of 10% trichloroacetic acid, the mixture was centrifuged at 200 g for 10 min in Sigma 3K30 model centrifuger.The organic layer (5 mL) was mixed with 5 mL of deionised water and 1 mL of 0. 1% ferric chloride and the absorbance read at 700 nm in a UVvisible spectrophotometer. reported in the literature (Adams, 2001). The retention indices were calculated for all volatile constituents using a homologous series of n-alkanes C8C16. 2. 3. 4. Antio xidative assays in mustard oil Oxidative deterioration was monitored under modi? ed Shaal Oven test (Economou et al. , 1991). click and bark essential oils and oleoresins along with synthetic antioxidants and major components were added individually to unre? ned mustard oil at levels of 0. 2% (v/v). The initial PV value of oil is 1. 7 meq of O2/kg. Oxidative deterioration was periodically assessed by measuring the antioxidant parameters such as peroxide (PV), thiobarbituric acid (TBA), p-anisidine (p-An) and total carbonyl (TC) values. 2. 3. 5. PV and TBA values The rate of oil oxidation was monitored by the increase of peroxide values. or so 3 g of each(prenominal) oil sample was weighed and subjected to iodimetric determination (AOCS, 1990). TBA values were evaluated according to the methods previously give tongue to by some authors (Sidwell et al. , 1954) with elfin changes. To 10 g of oil sample, 0. 7% aq. thiobarbituric acid (20 mL) and benzine (25 mL) solution were added . This mixture was shaken continuously for 2 h using mechanical shaker. After 2 h, supernatant was taken and placed in boiling water-bath for 1 h. After cooling, absorbance of supernatant was measured at 540 nm with Hitachi-U-2000 spectrophotometer. 2. 3. 6. p-Anisidine value The test was performed according to the methods (AOCS, 1998,) previously stated by earlier workers (Ottolenghi, 1959 Kikuzaki and Nakatani, 1993). In a 50 mL volumetric ? ask, 0. 6 g of oil sample was taken and volume was made using isoctane solution.From this solution, 5 mL was treated with 1 mL of 0. 25% of p-anisidine reagent and kept in bleached for 10 min and absorbance was measured at 350 nm using a UVVIS spectrophotometer. 2. 3. 7. Total carbonyl value Carbonyl value was evaluated according to the methods as reported earlier (Frankel, 1998). About 4 g of sample was taken in a 50 mL volumetric ? ask and the volume was made up using carbonyl free benzene. Out of this, 5 mL was pippeted out and mixed with 3 mL of 4. 3% trichloroacetic acid and 5 mL of 2,4-dinitrophenyl hydrazine (0. 05% in benzene) in 50 mL volumetric ? asks.The mixture was incubated at 60 C for half an hour to convert free carbonyls into hydrazones. After cooling, 10 mL of KOH solution (4% in ethanol) was added and the volume was made with ethanol. After 10 min, absorbance was measured at 480 nm using UVVIS spectrophotometer. Blank was prepared in the same manner substituting 5 mL of benzene instead of sample. A standard curve was drawn using valeraldehyde (50250 lg) in 5 mL of benzene instead of sample. The total carbonyl was calculated with the help of the standard curve and expressed as mg of valeraldehyde per nose candy g of sample. 2. 5. Antimicrobial activity 2. 5. . fungicide investigations In order to determine the antifungal e? cacy of the volatile oil and its oleoresin, the morbific fungus Aspergillus niger, Aspergillus ? avus, Aspergillus ochraceus, Aspergillus terreus, Fusarium moniliforme, Fusarium graminearum, Penicillium citrinum and Penicillium viridicatum were undertaken. These fungus kingdom were isolated from food materials such as onion, vegetable waste, wheat straw, fruits of Musa species, sweet potato, decaying vegetation and vegetable, respectively and were procured from microbic Type Culture Collection (MTCC), Institute of Microbial Technology, Chandigarh, India.The MTCC code No. of these strains are 2479, 1884, 1810, 3374, 1893, 2088, 2553 and 2007, respectively. Cultures of each of the fungi were maintained on Czapek (DOX) agar media with adjusting pH 6. 06. 5 and slants were stored at 4 C. The antifungal activity of the volatile oil and oleoresin against fungi were undertaken using inverted petriplate (Ramdas et al. , 1998) and poison food techniques (Amvam Zolla et al. , 1998). In inverted petriplate method, the required dose (2, 4 and 6 lL) of undiluted sample were soaked on a small piece (diameter 12 mm) of Whatmann No. 1 ? ter paper and it was kept on the lid of petriplate which is in inverted position whereas in poison food 2. 4. complementary antioxidant assays 2. 4. 1. Antioxidant activity in linoleic acid system Antioxidant activity was carried out using the method proposed by Osawa and Namaki (1983) with small changes. Samples (1 mL) in ethanol were mixed with 2. 5% linoleic acid in ethanol (4. 1 mL), 0. 05 M phosphate bu? er (pH = 7, 8 mL) and distilled water (3. 9 mL) and kept in screw cap containers under dark condition at 40 C. This solution (0. 1 mL) was added to the solution of 9. 7 mL of 75% ethanol and 0. mL of 30% ammonium thiocyanate. After 3 min, 0. 1 mL of 0. 02 M ferrous chloride in 3. 5% hydrochloric acid was added to the reaction mixture, the absorbance of red color was measured at 500 nm in the spectrophotometer, for every two days. The control and standards were subjected to the same procedure except for the control, where there was no addition of sample and for the standard 1 mL of sample was replaced with 1 mg of BHA and BHT. G. Singh et al. / Food and Chemical Toxicology 45 (2007) 16501661 technique, the required dose (2, 4 and 6 lL) of the undiluted sample were mixed with the 20 mL of culture medium.Each test was replicated for 3 times and fungi toxicity was measured after 6 days in terms of percent mycelial zone inhibition. 2. 5. 2. Antibacterial investigations Six pathogenic bacteria Bacillus genus Cereus (430), Bacillus subtilis (1790), Staphylococcus aureus (3103) (gram-positive), Escherichia coli (1672), Salmonella typhi (733), Pseudomonas aeruginosa (1942) (gram-negative) were selected for present study. All the bacterial strains were procured from Microbial Type Culture Collection (MTCC), Institute of Microbial Technology, Chandigarh, India. They were sub cultured on nutrient agar broth (Hi-media) and stored at 4 C.Active cultures for experiments were prepared by transferring one loopful of cells from stock cultures to ? ask of nutrient agar broth, which were incubated without agi tation for 24 h at 37 C. In order to determine the antibacterial activity of the essential oils and oleoresins, agar well di? usion method was followed. 0. 1 mL of 101 time diluted bacterial strain in ringers solution were ? ood inoculated on to the surface of well settled sterilized culture medium. The swell (10 mm diameter) were cut from agar, and 0. 2 mL of sample (2, 4 and 6 lL of essential oil or oleoresin diluted in 1 mL of DMSO) was delivered into them.For standard, 0. 2 mL of aqueous solution of ampicillin (1 mg mLA1) was used. After incubation for 24 h at 37 C, all plates were examined for any zones of harvest-tide inhibition according to method developed by Davidson and Parish (1989). All the plates were replicated twice and the results were averaged. 2. 5. 3. Statistical analysis For the oil or oleoresin, three samples were prepared for each experiment. The data were presented as mean standard deviation of three determinations (data were not shown). The quantitative da ta of major components of oil and oleoresin were statistically examined by analysis of variance (Sokal, 1973) and signi? ant di? erences among some(prenominal) groups of data were examined by Ducans multiple range test. A probability value of p 0. 05 was considered signi? cant. Table 1 Chemical composition of cinnamon leaf volatile oil and oleoresin Compound Volatile oil MS % a-Thujene a -Pinene b-Pinene Myrcene a-Phellandrene p-Mentha-1(7),8-diene p-Cymene 1,8-Cineole Terpinolene a-Terpineol a-Cubebene Eugenol b-Caryophyllene Aromadendrene a-Amorphene Germacrene-D Bicyclogermacrene d-Cadinene Spathulenol Sabinene c-Terpinene Terpinen-4-ol d-Elemene Viridi? orol Methoxy-eugenol Isospathulenol Neophytadiene Docosane Nonacosane Vitamin-E Total 0. 1 0. tr tr 1. 9 tr 0. 7 0. 7 tr tr tr 87. 3 1. 9 1. 1 tr 0. 6 3. 6 0. 4 0. 5 99. 4% a 1653 Oleoresin KI 931 941 980 993 1007 1011 1026 1033 1088 1191 1350 1358 1420 1441 1490 1490 1496 1527 1576 MSa % 0. 3 tr tr 87. 2 1. 4 0. 8 0. 4 0. 2 1. 7 0. 6 1. 7 tr tr tr 1. 0 0. 3 0. 1 0. 3 0. 3 0. 1 0. 1 0. 2 97. 1% KI 1007 1026 1191 1358 1420 1441 1490 1490 1496 1527 1576 975 1064 1177 1340 1594 3. Results and discussion 3. 1. Chemical analysis GC and GCMS analysis of cinnamon leaf volatile oil showed the presence of 19 components accounting for 99. % of the total amount (Table 1). The major component was eugenol (87. 3%) followed by bicyclogermacrene (3. 6%), a-phellanderene (1. 9%), b-carryophyllene (1. 9%), aromadendrene (1. 1%), p-cymene (0. 7%) and 1,8-cineole (0. 7%). Moreover, its oleoresin showed the presence of 25 components accounting for 97. 1% of the total amount (Table 1). The major components accounting were eugenol (87. 2%), spathulenol (1. 7%), bicyclogermacrene (1. 7%), b-caryophyllene (1. 4%) and d-elemene (1. 0%). The analysis of cinnamon bark volatile oil showed the presence of 13 components accounting for 100% of the total amount (Table 2). E)-cinnamaldehyde was found as the major component along with d-cadinene (0. 9%), a-copaene (0. 8%) and a-amorphene (0. 5%), whereas its bark oleoresin showed the presence of 17 components accounting for 92. 3% of the total amount (Table 2). The major components were (E)-cinnamaldehyde (49. 9%), coumarin (16. 6%), d-cadinene (7. 8%), a-copaene (4. 6%), (Z)-cinnamaldehyde (1. 5%), ortho-methoxy cinnamaldehyde (1. 5%) and b-bisabolene (1. 4%) along with several other compo- pctages are the mean of three runs and were obtained from electronic integration measurements using selective mass detector tr 0. 1. a nents. Recently, Raina et al. (2001) reported eugenol (76. 6%), linalool (8. 5%) and pipertone (3. 31%) as major components from its leaf oil grown in small Andman whereas the steam distilled volatile oil of cinnamon fruit ? grown at Karnataka and Kerala consists (Simic et al. , 2004 Jayaprakash et al. , 1997) of hydrocarbons (32. 8% and 20. 8%) and oxygenated compounds (63. 7% and 73. 4%) and trans-c innamyl acetate rayon and b-caryophyllene were found to be major component. 3. 2. Antioxidative assays in mustard oil The changes of PV in mustard oil of all investigated samples are presented in Fig. 1.The rate of oxidative reactions in mustard oil with additives was almost similar to that of the blank sample. The stability of the mustard oil samples to the formation of peroxides can be ranked in the following locomote order Leaf oleoresin BHT PG % eugenol scramble oleoresin % BHA Leafoil cinnamaldehyde bark oil 1654 G. Singh et al. / Food and Chemical Toxicology 45 (2007) 16501661 Table 2 Chemical composition of cinnamon bark volatile oil and extract Compound Volatile oil MS % a-Pinene Camphene Sabinene b-Pinene Limonene 1,8-Cineole Camphor Z-cinnamaldhyde E-cinnamaldhyde a-Copaene a-Amorphene -Cadinene Terpinen-4-ol b-Caryophyllene Coumarin a-Muurolene b-Bisabolene Cadina-1(2), 4-diene Ortho-methoxy cinnamadehyde Cubenol 1-Heptadecene 1-Nonadecene Tetracosane Octacosane Nonacosane Total a a Oleoresin KI 941 953 975 980 1031 1035 1144 1225 1279 1379 1490 1527 MSa % 1. 5 50. 0 4. 6 7. 8 0. 1 1. 0 16. 6 4. 4 1. 4 1. 8 1. 5 0. 5 0. 2 0. 4 0. 1 0. 1 0. 2 92. 3% KI 1225 1279 1379 1527 1177 1420 1436 1500 1506 1530 1532 tr tr tr tr tr tr tr tr 97. 7 0. 8 0. 5 0. 9 100% ays. The e? ects of volatile oils and oleoresins on malonaldehyde formation for mustard oil in terms of incubation time versus TBA value at 60 C are shown in Fig. 2. The malondehyde formation of all the additives increases with storage time. The oil showed a bind inhibition at 0. 02% concentration, and was comparable to BHA and PG but much lower than BHT. These results were well correlated with p-anisidine and total carbonyl values (Fig. 4). However, the sequence is slightly di? erent as compared with the one obtained during measurements of peroxide values.For instance, bark oleoresin had a little greater activity for preventing the formation of secondary oxidation products than primary ones. On contrary, volatile oils were slightly less e? ective in preventing the formation of secondary oxidation products than primary ones. From the above results, it should be said that the formation of the primary oxidation species, peroxides, were also quite similar with the secondary oxidation products, and the changes of both oxidation characteristics are in a good correlation. Hence, the inhibition activity of leaf and bark oleoresins were excellent among all the additives and there was a signi? ant di? erence between the blank and antioxidants at the P 0. 05 level. 3. 3. Antioxidant activity in linoleic acid system To evaluate the antioxidant potential of volatile oils and oleoresins of leaf and bark, their lipid inhibitory activities were compared with selected antioxidants and their major components by using ferric thiocyanate method of measuring the amounts of peroxides formed in emulsion during incubation. High absorbance is an indication of a high concentration of formed peroxides. The absorbance values of volatile oils and oleoresins of cinnamon along with synthetic antioxidants are shown in Fig. . The absorbance Percentages are the mean of three runs and were obtained from electronic integration measurements using selective mass detector tr 0. 01. Simultaneously with the measurements of peroxide value, the changes the secondary oxidation products such as malonaldehyde and 2-alkenals, which are measured by thiobarbituric (Fig. 2), p-anisidine (Fig. 3) and total carbonyl values (Fig. 4), were also determined after every 7 one hundred twenty Control BHT C. L. oil C. L. Oleoresin eugenol BHA PG C. B. Oil C. B. Oleoresin E-cinnamaldehyde 100 Peroxide value (meq/kg) 80 60 40 20 0 0 7 14 21 28Incubation time (days) Fig. 1. Inhibitory e? ect of volatile oil and oleoresin of cinnamon leaf and bark on the primary oxidation of mustard oil measured using peroxide value method. G. Singh et al. / Food and Chem ical Toxicology 45 (2007) 16501661 1655 6 5 Control BHT Leaf oil Leaf oleoresin Eugenol BHA PG verbalize oil speak oleoresin E-cinnamaldehyde TBA value (meq/g) 4 3 2 1 0 0 7 14 21 28 Incubation time (days) Fig. 2. Inhibitory e? ect of volatile oil and oleoresin of cinnamon leaf and bark on the malonaldehyde formation in mustard oil measured using TBA value method. 7 6 Control BHT C. L. Oil C. L.Oleoresin eugenol BHA PG C. B. Oil C. B. Oleoresin E-cinnamaldehyde p-anisidine value 5 4 3 2 1 0 0 7 14 21 28 Incubation time (days) Fig. 3. Inhibitory e? ect of volatile oil and oleoresin of cinnamon leaf and bark on the formation of 2-alkenals in mustard oil measured using p-anisidine method. 16 14 Carbonyl value (mg) 12 10 8 6 4 2 0 7 Control BHT C. L. Oil C. L. Oleoresin Eugenol BHA PG C. B. Oil C. B. Oleoresin E-cinnamaldehyde 14 21 28 Incubation time (days) Fig. 4. Inhibitory e? ect volatile oil and oleoresin of cinnamon leaf and bark on the total carbonyls present in mustard oil. 16 56 G. Singh et al. Food and Chemical Toxicology 45 (2007) 16501661 1. 9 1. 7 Absorbance at 500 nm 1. 5 1. 3 1. 1 0. 9 0. 7 0. 5 0 Control BHT Leaf oleoresin verbalise oleoresin Cinnamaldehyde BHA Leaf oil bark oil eugenol 25 50 75 100 125 150 175 200 Incubation time (h) Fig. 5. Inhibitory e? ect of volatile oil and oleoresin of cinnamon leaf and bark on the primary oxidation of linoleic acid system measured using ferric thiocyanate method. of linoleic acid emulsion without additive increased rapidly, and there was a signi? cant di? erence between blank and antioxidants at the P 0. 05 level. As can be seen in this ? , bark oleoresin was most e? ective among all the additives followed by leaf oleoresin. However, there are no signi? cant (p 0. 05%) di? erences between antioxidative activities of oleoresins, oils, BHA, BHT and PG. 3. 4. DPPH and hydroxyl radical scavenging e? ects Table 6 shows the DPPH and hydroxyl radical scavenging activity of leaf and bark volatile oils and oleor esins with various concentrations. As positive control, BHA and BHT were also examined. Bark oleoresin showed the best result through all concentrations for DPPH assay. The volatile oils have shown almost equal and moderate radical scavenging activity.At a concentration of 5 lL, signi? cant di? erences in DPPH scavenging activities was observed between BHA (78. 4%), BHT (81. 2%) and oleoresins of both leaf (51. 3%) and bark (75. 6%). However, as concentration increased, the di? erences in scavenging activities between BHA, BHT and oleoresins become less signi? cant. For hydroxyl radical scavenging test AOH radicals were generated by reaction of ferric-EDTA together with H2O2 and ascorbic acid to attack the substrate deoxyribose. The resulting products of the radical attack form a pink chromogen when heated with TBA in acid solution (Ohkawa et al. , 1979 Shimada et al. 1992). When the oils or oleoresins were incubated with this reaction mixture they were able to interfere with free r adical reaction and could prevent damage to the sugar. The results are shown in Table 6. At 5 lL, scavenging e? ects on hydroxyl radicals were 31. 2%, 51. 2%, 43. 6% and 57. 6% for leaf and bark volatile oils and oleoresin. However, at 25 lL BHA and BHT exhibited scavenging activities of 84. 9% and 83. 2%, respectively. There was a little change in the order of DPPH and hydroxyl radical scavenging activity of leaf oleoresin (86. 1%), bark volatile oil (79. 6%) and bark oleoresin (78. 6%).A close to linear correlation between radical scavenging activity and concentration of polyphenolic compounds in various vegetable and fruits have been reported (Pyo et al. , 2004 Robards et al. , 1999). These reports indicated that the radical scavenging activity of oleoresins tycoon be mostly a? ected by position of the phenolic hydroxyl group which is present in eugenol. Yepez et al. (2001) used eugenol as standard which removed 95% of the initial DPPH free radical. 3. 5. Chelating e? ect and re ducing power Chelating e? ects of the leaf and bark oleoresins on ferrous ions increased from 20. 5% at 5 lL to 24. % at 10 lL and maintained a tableland of 28. 235. 5% at 15 25lL (Fig. 6). The bark oleoresin showed a better chelating e? ect than those leaf oleoresin and both volatile oils. In addition, chelating e? ects of oleoresins were relatively parallel and increased from 20. 523. 6% at 5 lL to 38. 5 42% at 25 lL. However, at 5 lL, the chelating ability of EDTA was 90. 4%. Apparently, the cinnamon leaf and bark oleoresins could chelate ferrous ions but were not as e? ective chelators as EDTA. Reducing powers of leaf and bark oleoresins of cinnamon were excellent and were in the range 56. 058. 4, comparable with that of BHA (63. ) and BHT (65. 2) at 5 lL (Fig. 7). However, at 25 lL, the reducing power of the leaf and bark oleoresins, BHA and BHT were comparable (78. 587. 9). The reducing powers of the oleoresins might be due to the hydrogen donating abilities (Shimada et al. , 1992). 3. 6. Antimicrobial studies The results of volatile oils and oleoresins of cinnamon leaf and bark by inverted petriplate and poison food tech- G. Singh et al. / Food and Chemical Toxicology 45 (2007) 16501661 1657 100 90 Chelating effect (%) 80 70 60 50 40 30 20 10 0 0 EDTA Leaf oleoresin Bark oleoresin E-Cinnamaldehyde Leaf oil Bark oil Eugenol 10 15 20 25 30 Concentration ( L) Fig. 6. Chelating e? ect of volatile oil and oleoresin of cinnamon leaf and bark along with synthetic antioxidants. 100 Reducing power (%) 80 BHA Leaf oil Bark oil Eugenol BHT Leaf oleoresin Bark oleoresin Cinnamaldehyde 60 40 20 0 5 10 15 20 25 30 Concentration ( L) Fig. 7. Reducing power of volatile oil and oleoresin of cinnamon leaf and bark along with synthetic antioxidants. niques are reported in Tables 3 and 4, respectively. Using inverted petriplate method (Table 3), the leaf volatile oil was found to be 100% antifungal against all the tested fungi except A. chraceus and A. terreus at 6 lL. I t was interesting to note that complete inhibition against A. ?avus was obtained only at 2 lL. However, leaf oleoresin has shown complete mycelial zone inhibition only for P. citrinum. More than 75% activity was obtained for P. veridicatum, F. moniliforme and A. ?avus. Bark volatile oil has shown complete inhibition against the fungi such as F. gramenearum, F. moniliforme, P. citrinum, P. viridicatum and A. terreus at 6 lL. Using poison food technique (Table 4), leaf volatile has caused complete inhibition against all the tested fungi except P. itrinum whereas oleoresin has caused complete inhibition only against P. citrinum. Bark volatile oil has shown complete inhibition against almost all the tested fungi except for A. ?avus, A. ochraceus whereas its oleoresin has caused complete inhibition for A. ?avus and A. ochraceus along with A. niger, A. terreus, P. citrinum and P. viridicatum at 6 lL. Using agar well di? usion method (Table 5), leaf volatile oil has shown better results in comparison with oleoresin and commercial bactericide, i. e. , ampicillin. Complete mycelial zone inhibition was obtained using leaf volatile oil against P. eruginosa and B. cereus. However, it has moderate inhibitory e? ect on B. subtilis and S. aureus whereas its oleoresin has shown almost 100% activities against S. typhi and B. cereus. Bark volatile oil has been found to be better than bark oleoresin as it has caused more than 50% inhibition against all the tested fungi. There are several reports (Singh et al. , 1995 Hili et al. , 1997) stating that C. zeylanicum Blume exhibit antimicrobial activity. Their results demonstrate that the leaf oil completely inhibit the growth of E. coli, S. aureus and P. aeruginosa at the 1658 G. Singh et al. Food and Chemical Toxicology 45 (2007) 16501661 Table 3 Antifungal activity of volatile oils and oleoresins of cinnamon leaf and bark by inverted petriplate method Test Dose (lL) Percent mycelial inhibition zonea AN Leaf volatile oil 2 4 6 2 4 6 2 4 6 2 4 6 2 4 6 2 4 6 91. 5 100 100 25. 0 50. 0 58. 7 85. 3 93. 1 100 6. 3 38. 7 87. 2 62. 5 100 100 6. 3 35. 1 78. 3 AF 100 100 100 45. 6 76. 3 89. 3 100 100 100 6. 3 8. 8 13. 8 81. 2 100 100 65. 3 93. 2 100 AO 18. 7 56. 3 87. 5 46. 3 56. 3 68. 7 15. 6 52. 8 85. 3 12. 5 25. 0 37. 5 54. 3 78. 7 100 12. 5 25. 0 30. 8 FG 50. 0 52. 5 100 37. 5 50. 56. 3 36. 3 45. 8 95. 2 87. 5 87. 5 100 25. 0 50. 0 58. 7 75. 0 87. 5 100 FM 50. 0 52. 5 100 57. 5 80. 0 92. 5 31. 2 43. 2 83. 6 75. 0 87. 5 100 58. 6 79. 5 83. 3 58. 7 75. 3 83. 8 PC 37. 5 56. 3 100 67. 8 93. 3 100 25. 5 45. 8 86. 3 100 100 100 100 100 100 100 100 100 PV 37. 5 56. 3 100 38. 9 65. 5 87. 5 28. 5 47. 3 93. 7 100 100 100 76. 5 87. 5 100 85. 5 91. 5 100 AT 18. 7 36. 5 75. 0 46. 3 56. 3 68. 7 41. 3 53. 2 69. 1 37. 5 56. 3 100 87. 5 94. 1 100 56. 3 85. 6 100 Leaf oleoresin Eugenol Bark volatile oil Bark oleoresin E-cinnamaldehyde AN = Aspergillus niger AF = Aspergillus ? vus AO = Aspergillus ochraceus FG = Fusarium graminearum FM = Fusarium moniliforme PC = Penicillium citrinum PV = Penicillium viridicatum AT = Aspergillus terreus. a mediocre of three replicates. Table 4 Antifungal activity of volatile oils and oleoresins of cinnamon leaf and bark by food poisoned method Test Dose (ppm)a Percent mycelial inhibition zonea AN Leaf volatile oil 2 4 6 2 4 6 2 4 6 2 4 6 2 4 6 2 4 6 g-force 2000 3000 100 100 100 62. 5 77. 5 87. 5 100 100 100 73. 5 100 100 48. 9 65. 3 83. 6 52. 3 68. 7 72. 3 78. 2 82. 2 96. 3 AF 31. 3 87. 5 100 18. 8 50. 0 100 15. 6 63. 2 95. 6 () 51. 3 87. 5 88. 7 91. 3 100 52. 87. 6 91. 2 85. 3 91. 2 96. 2 AO 50. 0 100 100 35. 0 82. 5 97. 5 45. 6 95. 6 100 75. 0 81. 2 100 100 100 100 100 100 100 84. 2 91. 2 98. 4 FG 75. 0 100 100 62. 5 77. 5 87. 5 63. 5 82. 1 93. 8 50. 0 75. 0 87. 5 65. 3 83. 2 100 47. 2 67. 8 85. 3 90. 2 96. 3 94. 5 FM 100 100 100 38. 7 46. 3 78. 7 45. 6 53. 6 78. 3 75. 0 83. 2 100 48. 7 56. 3 78. 7 63. 2 65. 8 87. 1 97. 2 100 100 PC 50. 0 75. 0 87. 5 35. 0 62. 5 97. 5 48. 6 73. 1 82. 6 43. 7 51. 3 65. 0 100 100 100 85. 2 89. 7 91. 2 100 100 100 PV 87. 5 100 100 50. 0 65. 5 70. 0 73. 2 85. 6 93. 6 50. 0 75. 0 87. 5 60. 0 85. 3 100 55. 3 63. 1 91. 2 100 100 100 AT 18. 7 50. 0 56. () 50. 0 100 15. 5 50. 0 75. 2 32. 5 45. 0 76. 3 35. 0 76. 2 83. 7 42. 3 45. 6 89. 3 98. 5 100 100 Leaf oleoresin Eugenol Bark volatile oil Bark oleoresin E-cinnamaldehyde Carbendazimb AN = Aspergillus niger AF = Aspergillus ? avus AO = Aspergillus ochraceus FG = Fusarium graminearum FM = Fusarium moniliforme PC = Penicillium citrinum PV = Penicillium viridicatum AT = Aspergillus terreus. a Average of three replicates. b Aqueous solution was used. G. Singh et al. / Food and Chemical Toxicology 45 (2007) 16501661 Table 5 Antibacterial activity of volatile oils and oleoresins of cinnamon leaf and bark by agar well di? sion method Test Concentration (ppm) Inhibition zone (mm)a Gram (+) bacteria Bs Leaf volatile oil 1000 2000 3000 1000 2000 3000 1000 2000 3000 1000 2000 3000 1000 2000 3000 1000 2000 3000 1000 2000 3000 17. 1 0. 4 20. 0 0. 6 32. 6 1. 2 14. 6 1. 2 19. 0 0. 2 25. 4 0. 8 14. 3 0. 6 17. 0 0. 3 29. 6 1. 2 14. 2 0. 5 18. 3 0. 3 26. 7 0. 7 16. 2 1. 3 20. 2 1. 1 25. 3 0. 3 12. 3 0. 1 17. 3 0. 5 23. 7 0. 6 32. 5 1. 2 34. 3 0. 3 41. 2 0. 2 Sa 26. 1 1. 5 34. 9 1. 3 48. 7 0. 5 27. 1 0. 1 38. 9 0. 2 49. 3 2. 2 23. 1 1. 1 26. 9 1. 3 38. 7 0. 3 27. 0 0. 9 44. 6 0. 56. 7 0. 1 23. 1 0. 4 28. 7 0. 2 33. 6 0. 3 23. 0 0. 7 41. 6 0. 8 53. 7 0. 1 29. 5 0. 6 32. 6 1. 6 37. 5 0. 2 Bc 43. 3 1. 7 58. 0 0. 6 + 64. 5 0. 6 80. 4 1. 1 + 33. 3 1. 5 56. 0 0. 8 72. 3 0. 2 41. 3 1. 7 52. 6 1. 2 56. 3 0. 5 38. 6 0. 2 41. 3 0. 4 45. 6 0. 7 31. 3 1. 2 48. 6 0. 2 52. 3 0. 3 31. 4 0. 2 34. 6 0. 1 38. 2 0. 3 Gram (A) bacteria Ec 13. 0 0. 2 18. 2 1. 1 25. 8 0. 5 11. 4 0. 6 13. 1 0. 7 18. 5 1. 1 11. 3 0. 1 17. 2 1. 6 21. 8 0. 3 28. 1 0. 2 33. 2 1. 3 35. 1 0. 3 33. 4 0. 5 35. 4 0. 3 37. 1 0. 3 26. 1 0. 5 33. 1. 8 34. 1 0. 2 33. 6 0. 8 37. 8 1. 4 39. 5 0. 6 St 12. 5 0. 8 14. 6 1. 1 17. 9 0. 2 53. 6 1. 3 73. 8 0. 5 78. 1 0. 8 12. 5 0. 8 14. 6 1. 1 17. 9 0. 2 20. 6 1. 8 32. 7 2. 0 41. 3 0. 3 17. 2 0. 1 18. 6 0. 7 19. 3 0. 5 18. 6 1. 4 31. 7 1. 0 40. 3 0. 3 21. 9 0. 5 25. 6 0. 7 28. 9 1. 3 Pa 1659 25. 7 0. 6 + + 20. 5 0. 1 21. 4 0. 8 25. 8 0. 1 26. 7 0. 5 + + 50. 2 1. 2 56. 5 0. 8 60. 2 0. 3 40. 6 0. 4 45. 3 0. 8 56. 2 0. 7 30. 2 1. 1 48. 5 0. 6 59. 2 0. 1 24. 3 0. 4 26. 3 1. 5 27. 3 1. 1 Leaf oleoresin Eugenol Bark volatile oilBark oleoresin E-cinnamaldehyde Ampicillin Bs = Bacillus subtilis Sa = Staphylococcus aureus Bc = Bacillus cereus Ec = Escherichia coli St = Salmonella typhi Pa = Pseudomonas aeruginosa. (+) indicates complete inhibition. a Average of three replicates. level of 500 lg mLA1. Another report (Smith-Palmer et al. , 1998) found the MICs of C. zeylanicum against E. coli and S. aureus were 0. 05% and 0. 04%, respectively. To con? rm the relationship of the constituents in cinnamon leaf and bark and antimicrobial activity, the major components were tested for antimicrobial activity. The results are shown in Tables 35.Among both constituents, E-cinnamaldehyde possessed better activity and these ? ndings are quite similar with the results of Chang et al. (2001). However, eugenol, in spite of being phenolic compound, failed to inhibit the fungal growth by inverted petriplate method but when it was added directly to the growth media in higher concentrations, it appeared to inhibit completely the microbial growth. Nevertheless, it is worth noting that essential oils and oleoresins are very heterogeneous mixtures of a single substances, biological actions are in the first place due to these components in a very complicated concert of synergistic or antagonistic e? cts. Table 6 Comparison of scavenging e? ects of cinnamon leaf and bark volatile oils and oleoresins against DPPH and hydroxyl radicals Sample Radical scavenging activitya (%) DPPH radical 5 lL Leaf oil Leaf oleoresin Eugenol Bark oil Bark oleoresin E-cinnamaldehyde BHA BHT a Hydroxyl radical 15 lL 69. 9 74. 1 65. 2 76. 2 89. 3 72. 3 92. 1 89. 2 20 lL 72. 1 76. 7 71. 3 82. 1 91. 2 75. 1 94. 7 91. 7 25 lL 73. 9 91. 2 92. 9 83. 6 95. 3 78. 3 96. 4 94. 9 5 lL 31. 2 43. 6 39. 4 51. 2 57. 6 49. 8 71. 3 66. 2 10 lL 55. 7 57. 1 45. 1 57. 6 62. 3 53. 6 75. 1 72. 1 15 lL 63. 5 70. 4 54. 3 73. 1 68. 9 57. 1 78. 75. 3 20 lL 68. 1 73. 6 61. 5 76. 9 71. 2 65. 2 81. 7 77. 5 25 lL 72. 2 86. 1 68. 2 79. 6 78. 6 68. 3 84. 9 83. 2 10 lL 58. 7 58. 9 56. 8 73. 5 87. 5 68. 1 89. 3 85. 1 45. 2 51. 3 41. 3 71. 1 75. 6 65. 3 78. 4 81. 2 Average of three replicates. 1660 G. Singh et al. / Food and Chemical Toxicology 45 (2007) 16501661 Chang, S. T. , Chen, P. F. , Chang, S. C. , 2001. Antibacterial activity of leaf essential oils and their constituents from Cinnamon osmophloeum. ledger of Ethanopharmacology 77, 123127. Cuvelier, M. E. , Berset, H. , Rich ard, H. , 1994. Antioxidant constituents in sage (Salvia o? cinalis).Journal of factory farm and Food Chemistry 42, 665669. Davidson, P. M. , Parish, M. E. , 1989. Methods for testing the e? cacy of food antimicrobials. Food Technology 43, 148155. Djarmati, Z. , Jankov, R. M. , Schwirtlich, E. , Djulinac, B. , Djoedjevic, A. , 1991. High antioxidant activity of oleoresins obtained from sage by supercritical CO2 extraction. Journal of American Oil Chemical Society 68, 731734. Economou, K. D. , Oreopoulou, V. , Thomopoulos, 1991. Antioxidant activity of some plant oleoresins of the family Labiatae. Journal of American Oil Chemical Society 68, 109115. Frankel, E. N. 1998. Lipid Oxidation. The Oily matter, Dundee, UK, 301 pp. Gordon, M. H. , Weng, C. X. , 1992. Antioxidant properties of oleoresins from tanshen (Salvia miltiorrhiza Bunge). Food Chemistry 44, 119 122. Halliwell, B. , 1989. Protection against tissue damage in vivo by Desferrioxamine what is its mechanism of action? Free Radical Biology and Medicine 7, 645651. Halliwell, B. , Gutteridge, J. M. C. , Aruoma, O. I. , 1987. The deoxyribose method a simple test tube assay for the determination of rate constants for reaction of hydroxyl radicals. Analytical Biochemistry 165, 215 219. Hili, P. , Evans, C.S. , Veness, R. G. , 1997. Antimicrobial action of essential oils the e? ect of dimethylsulphoxide on the activity of cinnamon oil. Applied and Environmental Microbiology 24, 269275. Ho, C. T. , Huang, M. T. , Lou, Y. R. , Ma, W. , Shao, Y. , Wei, G. J. , Wang, M. , Chin, C. K. , 2000. Antioxidant and antitumor activity of rosemary leaves. In Shahidi, F. , Ho, C. T. (Eds. ), Phytochemicals and Phytopharmaceuticals. Champaign, Illinois, USA, pp. 296308. Jayaprakash, G. K. , Rao, L. J. , Sakariah, K. K. , 1997. Chemical composition of the volatiles from oil from the fruits of Cinnamon zeylinicum Blume.Flavour Fragrance Journal 12, 331. Kahl, R. , Kappus, H. , 1993. Toxicity of synthetic antioxidants BHA and BHT in comparison with natural antioxidants vitamin E. Zeitschrift fur Lebensmittel-Untersuchung und Forschung 196, 329338. Kikuzaki, H. , Nakatani, N. , 1993. Antioxidant e? ect of some ginger constituents. Journal of Food Science 58, 14071410. Madsen, H. L. , Andersen, L. , Christiansen, L. , Brockho? , P. , Bertelsen, G. , 1996. Antioxidative activity of summer savory (Satureja hortensis L. ) and rosemary (Rosmarinus o? cinalis L. ) in minced cooked pork meat. Z. Lebensm.Unters Forsch. 203, 333338. Massda, Y. , 1976. Analysis of essential oils by Gas Chromatography and Mass Spectrometry. Halsted/Wiley, New York. Ohkawa, H. , Ohishi, N. , Yagi, K. , 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95, 351358. Osawa, T. , Namaki, M. , 1983. A novel type antioxidant isolated from leaf wax of Eucalyptus leaves. Agricultural and Biological Chemistry 45, 735739. Ottolenghi, A. , 1959. Interaction of ascorbic acid and mitochondri al lipids. Archives of Biochemistry and Biophysics 79, 355358. Oyaizu, M. , 1986.Studies on products of browning reactions antioxidative activities of products of browning reactions prepared from glucosamine. Japanese Journal of Nutrition 44, 307315. Peschel, W. , Sanchez-Rabaneda, F. , Dickmann, W. , Plesehen, A. , Gartiza, I. , Jimenez, D. , Lamuela-Raventos, R. , Buxaderas, S. , Codina, C. , 2006. An Industrial approach in the search of natural antioxidants from vegetables and fruit wastes. Food Chemistry 97, 137 150. Pratt, D. E. , 1992. Natural antioxidants from plant materials. In Huang, I. M. T. , Ho, C. T. , Lee, C. Y. (Eds. ), Phenolic Compounds in Food and Their E? ects on Health.American Chemical Society, New York, pp. 5472. 4. Conclusion The present study provided the potential antimicrobial and antioxidant properties of the volatile oils and oleoresins of cinnamon leaf and bark. The oleoresins have shown better e? ect on primary and secondary oxidation products in must ard oil. The radical scavenging activity and other complementary assays are also in good correlation. Moreover, the potency of the constituents such as eugenol and cinnamaldehyde could provide a chemical basis for some of the health bene? ts claimed for cinnamon and warrant further studies to assess their potential as e? ctive natural remedies. Acknowledgements We are thankful to Head, Chemistry Department, DDU Gorakhpur University, Gorakhpur for providing laboratory facilities. Prof. K. D. S. Yadav of our department is also thanked for providing spectral facility. invigoration Sciences Research Board, DRDO, New Delhi and CONICET and Consejo de Investigaciones de la Universidad Nacional de Tucuman (CIUNT) Argentina are also thanked for ? nancial assistance. References Adams, R. P. , 2001. Identi? cation of Essential Oils Compounds by Gas Chromatography/Quadrupole Mass Spectrometry. Allured Publishing Corporation, Carol Stream, IL, USA.Amvam Zolla, P. H. , Biyiti, L. , Tchoumbougnan g, F. , Menut, C. , Lamaty, G. , Bouchet, P. , 1998. Aromatic plant of tropical Central Africa. Part XXXIII, Chemical composition and antifungal activity of thirteen essential oils from aromatic plants of Cameroon. Flavour and Fragrance Journal 13, 107114. AOCS. 1990. O? cial methods and recommended practices of the American Oil Chemists Society Method cd-83 and method cd-1890, fourth ed. , American Oil Chemists Society, Champaign. AOCS. 1998. O? cial methods peroxide value. In O? cial Methods and Recommended Practices of the American Oil Chemists Society, ? fth ed. AOCS Press Illinois, USA. cd 8-53. AOCS o? cial Methods p-anisidine value, 1998. In O? cial Methods and Recommended Practices of the American Oil Chemists Society, ? fth ed. , AOCS Press Illinois, cd 18-90. Aruoma, O. I. , Spencer, J. P. E. , Rossi, R. , Aeschbach, R. , Khan, A. , Mahmood, N. , Munoz, A. , Murcia, A. , Butler, J. , Halliwell, B. , 1996. An evaluation of the antioxidant and antiviral action of oleoresins of rosemary and provencal herbs. Journal of Food and Chemical Toxicology 34, 449456. Bandoniene, D. , Venskutonis, P. R. , Gruzdiene, D. , Murkovic, M. , 2002. Antioxidant activity of Sage (Salvia o? inalis L. ), Savory (Satureja hortensis L. ) and Borage (Borago o? cinalis L. ) oleoresins in rapeseed oil. European Journal of Lipid Science and Technology 104, 286 292. Brand-Williams, W. , Cuvelier, M. E. , Berset, C. , 1995. Use of a free radical method to evaluate antioxidant activity. Lebensmittl-Wissenschaft und Technologic 28, 2530. Caragay, A. B. , 1992. Cancer preventive foods and ingredients. Food Technology 46, 6569. Chang, S. S. , Ostric-Matijasevic, Hsien O. A. L. , Huang, C. L. , 1997. Natural antioxidants from rosemary and sage. Journal of Food Science 42, 11021106. G. Singh et al. Food and Chemical Toxicology 45 (2007) 16501661 Pyo, Y. H. , Lee, T. C. , Longedra, L. , Rosen, R. T. , 2004. Antioxidant activity and phenolic compounds of Swiss chard (Beta vulgaris subspeci es cycla) oleoresins. Food Chemistry 85, 1926. Raina, V. K. , Srivastava, S. K. , Aggrawal, K. K. , Ramesh, S. , Kumar, Shushil, 2001. Essential oil composition of Cinnamon zeylinicum Blume leaves from little Andaman, India. Flavour Fragrance Journal 16, 374. Ramdas, K. , Suresh, G. , Janardhanan, N. , Masilamani, S. , 1998. Antifungal activity of 1,3-disubstituted symmetrical and unsymmetrical thioureas.Pesticidal Science 52, 145151. Robards, K. , Prenzeler, P. D. , Tucker, G. , Swatsitang, P. , Glover, W. , 1999. Phenolic compounds and their role in oxidative process in fruits. Food Chemistry 66, 401436. Shimada, K. , Fujikawa, K. , Yahara, K. , Nakamura, T. , 1992. Antioxidative properties of xanthan on autooxidation of soybean oil in cyclodextrin emulsion. Journal of Agriculture and Food Chemistry 40, 945948. Sidwell, C. G. , Salwin, H. , Benca, M. , Mitchell, J. H. , 1954. The use of thiobarbituric acid as a measure of fat oxidation. Journal of American Oil Chemical Society 31, 603. ? ? ? ? ? ? Simic, A. Sokovic, M. D. , Ristic, M. , Grujic -Jovanovic, S. , Vukojevic, J. , Marin, P. D. , 2004. The chemical composition of some Lauraceae 1661 essential oils and their antifungal activities. Phytotherapy Research 18, 713717. Singh, N. B. , Srivastava, M. , Singh, A. B. , Srivastava, A. K. , 1995. Cinnamon bark oil, a potent fungi toxic against fungi causing respiratory tract mycoses. Allergy 50, 995999. Smith-Palmer, A. , Stewart, J. , Fyfe, L. , 1998. Antimicrobial properties of plant essential oils and essences against ? ve important food-borne pathogens. Letters in Applied Microbiology 26, 118122. Sokal, R. R. 1973. Introduction to Biostatistics. WH Freeman and company, San Francisco, pp. 164185. Takacsova, M. , Pribela, A. , Faktorova, M. , 1995. Study of the antioxidative e? ects of thyme, sage, juniper and oregano. Nahrung/Food 39, 241243. Wong, J. W. , Hashimoto, K. , Shibamoto, T. , 1995. Antioxidant activities of rosemary and sage oleoresins and vita min E in a model meat system. Journal of Agriculture and Food Chemistry 45, 2707 2712. Yepez, B. , Espinosa, M. , Lopez, S. , Bolanos, G. , 2001. Producing antioxidant fractions from herbaceous matrices by supercritical ? uid extraction. Fluid Phase Equilibria 4887, 16.