EXPERIMENTAL RESULTS

Transcription

1 EXPERIMENTAL RESULTS The results obtained from various assays, characterization techniques and evaluation methods are furnished. The data and information s derived from these results are presented in the form of figures, plates and tables 4.1. GREEN SYNTHESIS OF SILVER NANOPARTICLES Preliminary screening tests of greengram sprout Phytochemicals present in greengram sprout extract, qualitatively detected using standard screening tests are presented in Table 4.1 and the results of the tests are shown in Plate GC-MS analysis of phytochemicals in greengram sprout The chromatogram of the greengram sprout is shown in Fig Using NIST library 24 compounds were detected, out of which 22 were identified. The peak report derived from the chromatogram is presented in Table 4.2. Among these identified phytochemicals, 11 compounds with peak area (%) greater than 1 are reported in Table 4.3. The chemical formula, molecular weight, compound type and bioactivity are also furnished against each compound.

2 67! " #!! $!!! % & % ' ( % ( % ( ) * + *, * - # S. No. Phytochemical Result (Qualitative) 1 Tannin Saponin Flavonoids ++ 4 Steroids Terpenoids ++ 6 Alkaloids Amino acids Polyphenol Glycoside Protein ++

3 68

4 . / 69

5 Peak# R. Time Area Area% Name Butanamine, 2-methyl-N-(2-methylbutylidene) Cyclopentane, 1-acetyl-1,2-epoxy H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl Deoxy-d-arabitol ETHYLHEXYL PENTENOATE Heptanol, 2-propyl (4-AMINOBUTYL)PIPERIDINE Unknown H-Pyrrole, 2-(2,4,6-cycloheptatrienyl) Ethyl.alpha.-d-glucopyranoside MOME INOSITOL Caffeine H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL Hexadecanoic acid, methyl ester n-hexadecanoic acid Unknown Octadecanoic acid OCTADECENOIC ACID (Z) TRIMETHYLSILYL ESTER OF TETRACOSANOIC ACID PREGNANE, SILANE DERIV Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester Cholest-5-en-3-ol (3.beta.)-, carbonochloridate Stigmasterol STIGMAST-5-EN-3-OL, (3.BETA.)

6 H F5 G G M5 IL 8 M8 K L H 7 C J E I5 7 5 B 8 F D E B A? = > < ; : 9 : Compound name Nature/Type Uses Mol. Wt. Formula Area (%) Peak# R. Time C6H8O H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl Flavonoid fraction Antimicrobial, Anti-inflammatory C5H12O Deoxy-d-arabital Unknown C8H16O6 208 Ethyl-alpha-d-glucopuranoside Sugar moiety Preservative Antialopecic, Lipotropic, antineuropathic, Sweetner anticirrhotic, Cheolesterolytic C7H14O6 194 MOME INOSITOL Inositol Both, pro and and antioxidant anticarcenogenic, bronchodialator C8H10N4O2 194 Caffeine Alkaloid Both pro and antioxidant vasodilator, diuretic C7H8N4O H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL Xanthine alkaloid Antioxidant, nematicide pesticide, flavor antiandrogenic C16H32O2 256 n-hexadecanoic acid Palmitic acid C18H36O2 284 Octadecanoic acid Fatty acid (Stearic acid) Skin care Antioxidant, nematicide, pesticide, hypocholesterolemic, hemolytic C18H36O2 284 Hexadecanoic acid ethyl ester Palmitic acid ethyl ester Antimicrobial, antioxidant antiinflammatory, antiartheritic, diuretic antiashma, anti HIV reserve transcriptase C29H48O 412 Stigmasterol Steroid Antimicrobial, antioxidant antiinflammatory, antiartheritic, diuretic antiashma, anti HIV reserve transcriptase C29H50O 414 STIGMAST-5-EN-3-OL, (3, BETA) Steroid 71

7 / 2 2 N O P Q Q R 72

8 73

9 Visual and UV-Vis monitoring of Ag NPs formation The greengram plant, seeds, germinated seeds with sprout and the various stages of silver nanoparticle formation are shown in Plate 4.2(a-d). It can be observed that significant colour change appeared after 30 min and the final brownish red colour was obtained after 2 h. The final colour change confirmed the completion of the synthesis process. The UV-Vis spectrum of the reaction medium recorded in the range 300 to 600 nm is shown in Figure 4.2. The UV-Vis spectrum showed an intense absorption band with a maximum at 445 nm is attributed to the surface plasmon resonance (SPR) which is characteristic of the silver nanoparticles (Umashankari et al., 2012). The slight broadening of the peak indicated that the particles are slightly poly-dispersed XRD pattern of Ag NPs The XRD pattern of the synthesized Ag NPs is illustrated in Fig Since thin film technique had been used to record the pattern, the amorphous characteristics of the glass were also present in the XRD pattern overlapping the Ag NPs pattern. The XRD spectrum confirmed the crystalline nature of the nanoparticles with peaks appearing at 2 values of 32, 38, 46 corresponding to (111), (200), (220) and (311) Bragg reflections, respectively. The average crystallite size (D) of silver nanoparticles, calculated using the Debye-Scherrer equation (Singh et al., 2010) was 5.6 nm. The XRD parameters are provided in Table 4.4.

10 75 (a) (b) (c) (d) 0 N R S T N R 2 T N R 2 2 U 2 N 2 R 3 V W

11 76 X Y Z [ \ [ ] [ ^ _ ` _ Y a b c a d e f g Y d h a f i j g e k l d m n Z o p a a q h g r i a Y s i t k a Y h Z Z e i i h Z e b l a f e d k g i u g e b j g X Y Z [ \ [ v [ w x y f b g g i e h d m n Z o p a a q h g r i a Y s i t k a Y h Z Z e i i h Z e b l a f e d k g i u g e b j g

12 77 z b c { i \ [ \ [ w x y f b e b l i g i e a d m n Z o p a Peak No. Position ( 2 ) FWHM, ( ) Particle size, D (nm) Mean = FE-SEM morphology of Ag NPs The FE-SEM micrograph revealing the morphology of the green synthesized Ag NPs is shown in Fig FT-IR spectra of greengram sprout and Ag NPs In order to identify the biomolecules present in the greengram sprout which are responsible for the synthesis of Ag NPs, the FT-IR spectrum was recorded and is shown in Fig. 4.5a. To study the biomolecules of greengram sprout extract bound to the surface of Ag NPs, FT-IR spectrum of the synthesized Ag NPs was recorded and is shown in Fig. 4.5b. The position and intensity of the absorption bands in the spectrum provide information about the various chemical groups present and their concentration. The vibrational frequency assignments for the prominent bands in the FT-IR spectrum of the sprout sample are provided in Table 4.5. It can be observed that the amide-i linkages in protein, amino acids, glycoside linkages and polyphenols are responsible for the reduction and stabilization processes in the synthesis of Ag NPs.

13 78 ƒ ƒ ƒ ƒ ƒ ƒ ˆ ƒ X Y Z [ \ [ \ [ X ` } ~ l Y j e d Z e b f r d m Z e i i h a q h g r i a Y s i t n Z o p a

14 X Y Z [ \ [ [ X z ` Š x a f i j g e b d m b Œ Z e i i h Z e b l a f e d k g i u g e b j g b h t c Œ a q h g r i a Y s i t n Z o p a 79

15 80 z b c { i \ [ [ n c a d e f g Y d h c b h t b a a Y Z h l i h g m d e g r i X z ` Š x a f i j g e k l d m Z e i i h Z e b l a f e d k g a b l f { i Absorption band (cm 1 ) Vibrational group and mode Chemical compound References Vanaja et al. (2013) 3398* O H stretch (hydroxyl) Phenols Awwad et al. (2013) Rajathi and Sridhar (2013) N H stretch Amino group Awwad et al. (2013) 2929 C H stretch Alkanes Vanaja et al. (2013) Rajathi and Sridhar (2013) Aldehydes Umashankari et al. (2012) O H stretch Carboxylic acid Vanaja et al. (2013) 2349 N H stretch Amino acids Umashankari et al. (2012) 1641 C=O (carbonyl) Polyphenols Amide-I in protein links Rajathi and Sridhar (2013) Jayaseelan et al. (2013) Prakash et al. (2013) N H bend Amino acids Theivasanthi and Alagar (2013) 1408 C O stretch Amino acids Theivasanthi and Alagar (2013) C C stretch Aromatics Mallikarjuna et al. (2012) 1249 C O C stretch Glycoside linkages Raghavandra et al. (2013) 1081 C N stretch (carbonyl) Amide-I in proteins Awwad et al. (2013) Aliphatic amines Vanaja et al. (2013) C O stretch Carboxylic acid 669 C H bend Phenyl ring substituents Ž š œ ž Ÿ Mallikarjuna et al. (2012) Jayaseelan et al. (2013)

16 Antibacterial assays The antibacterial activity of Ag NPs against the studied bacterial strains are shown in Plate 4.3. The values of zone of inhibition obtained from the assay are presented in Table 4.6. All Gram-negative bacteria had shown good sensitivity towards the green synthesized Ag NPs for the concentrations 10 and 15 g ml 1, while the Gram-positive bacteria showed almost equal sensitivity as that of the positive control (Imipenem) Antifungal assays Regarding the antifungal activity, all four fungal strains used in this study are found to be sensitive to the green synthesized Ag NPs as well as to the commercially available antifungal drug Itraconozole. The antifungal activities of Ag NPs are shown in Plate 4.4 and the zone of inhibition values are presented in Table 4.7. The encouraging aspect of this study is that all the fungal species are relatively more sensitive to the Ag NPs compared to the positive control. This may be due to the individual organisms response and their genotypic characters which differs in their sensitivity pattern towards the single testing agent.

17 p { b g i \ [ v [ n h g Y c b j g i e Y b { b j g Y Y g q d m n Z o p a b Z b Y h a g b Œ ª «± ² ³ µ ¹ º ²» ¼ ± ½ ¹ ¾ À Á Á  ± ² µ ³ µ º à Á Ä Å Å Æ Ç Æ À û È À» Æ À Ã Ç Ã Ä È À É Ê Ë Ì Ë È É Ä Ã Ä Í Æ» È À Ã Ç È Î 82

18 83 Ï Î Æ Ð Ê Ñ Ê Ò À à Ļ Ã Æ Ç Ä Î È Å Ó Ç Æ Æ À É Ô À Ã Õ Æ É Ä Ö Æ Á Ò Ó Ë É Ã Á Ä Å Å Æ Ç Æ À û È À» Æ À Ã Ç Ã Ä È À É Ó Ä À É Ã Ø Ã Õ È Ó Æ À Ä»» Ã Æ Ç Ä Î É Ø Æ» Ä Æ É *Zone of inhibition (mm) Label Bacteria Concentrations ( g ml 1 ) Ag NPs P Gram-negative a K. pneumoniae b P. aeruginosa c E. coli Gram-positive d S. aureus Ù Ú Û Ü Ý Ü Þ ß à Ú á Ý â Ú ã ä Ù å æ Ü ã ã Ü æ ß Ý ß â ä æ æ å æ Ü à â Ú ç â è æ é æ Ü ã ã Ü ã Ü Ý ß â ä ç æ ê ë ì í î ï Ú á ß Ú ð Ü á ñ Ü ò Ü Ý Ü Ú á Þ è ã ó ß Û è â ß ß ô õ â ß Û Û ß ö è Û æ ß è á Ú ð Ý â Ü õ ã Ü à è Ý ß æ ß è Û ó â ß æ ß á Ý Û Û Ý è á ö è â ö ö ß Þ Ü è Ý Ü Ú á í

19 Ë Î Ã Æ Ð Ê Ð Ê Ò À Ã Ä Å À Ó Î» Ã Ä Í Ä Ã Ô È Å Ò Ó Ë É Ó Ä À É Ã ø ± ù ¾ ² ú µ º ø ± ³» ø ± ù µ û ² ü µ º À Á Á ý ± þ ÿ º ³ µ û à Á Ä Å Å Æ Ç Æ À û È À» Æ À Ã Ç Ã Ä È À É Ê Ë Ì Ë È É Ä Ã Ä Í Æ» È À Ã Ç È Î 84

20 85 Ï Î Æ Ð Ê Ê Ò À Ã Ä Å À Ó Î È Å Ó Ç Æ Æ À É Ô À Ã Õ Æ É Ä Ö Æ Á Ò Ó Ë É Ã Á Ä Å Å Æ Ç Æ À û È À» Æ À Ã Ç Ã Ä È À É Ó Ä À É Ã Ø Ã Õ È Ó Æ À Ä» Å À Ó Î É Ø Æ» Ä Æ É *Zone of inhibition (mm) Label Fungi Concentrations ( g ml 1 ) Ag NPs P a A. flavus b A. niger c A. fumigatus d M. gypseum Ù Ú Û Ü Ý Ü Þ ß à Ú á Ý â Ú ã ä Ù å æ Ü ã ã Ü æ ß Ý ß â ä æ æ å æ Ü à â Ú ç â è æ é æ Ü ã ã Ü ã Ü Ý ß â ä ç æ ê ë ì í î ï Ú á ß Ú ð Ü á ñ Ü ò Ü Ý Ü Ú á Þ è ã ó ß Û è â ß ß ô õ â ß Û Û ß ö è Û æ ß è á Ú ð Ý â Ü õ ã Ü à è Ý ß æ ß è Û ó â ß æ ß á Ý Û Û Ý è á ö è â ö ö ß Þ Ü è Ý Ü Ú á í

21 GREEN SYNTHESIS OF ZINC OXIDE NANOPARTICLES Photographs of green tea plant, dried leaves and synthesized ZnO NPs The photograph of the green tea plant, leaves in dried form and synthesized ZnO NPs is shown in Plate 4.5. The pale-white colour of the ZnO NPs arise due to capping action of biomolecules on the surface of the nanoparticles Preliminary screening test of green tea Phytochemicals present in green tea extracts, qualitatively detected using standard screening tests are presented in Table 4.8. The results of the tests are shown in Plate 4.6. Intense colour changes reveal the presence of terpenoids, polyphenols, proteins and flavonoids as major constituents in the green tea sample GC-MS analysis of green tea One of the six green tea samples (MOON tea) studied for the antioxidant potentials (Section 4.3.1) was used to green synthesize ZnO nanoparticles. The chromatogram obtained from GC-MS analysis, the peak report and the major phytochemicals identified for this green tea samples are provided in Figs and Tables respectively.

22

23 88! " # $! " " % " " " & ' & ( ) & ) & ) * +, $ S. No. Phytochemical Result (Qualitative) 1 Tannin Saponin Steroids + 4 Terpenoids Alkaloids ++ 6 Amino acids Glycoside ++ 8 Polyphenols Protein Flavonoids +++ /

24 UV-Vis spectrum of ZnO nanoparticle Confirmation of the synthesized ZnO product in nano-scale was done by recording UV-Vis spectrum. The highly blue-shifted absorption maximum occurring around 325 nm is an indication of nano-sized ZnO particles. For bulk ZnO the absorption maximum is around 385 nm approximately. The UV-Vis spectrum of ZnO particles is shown in Fig XRD analysis of ZnO NPs The XRD spectra of the as prepared and calcined at 100 C ZnO NPs are shown in Fig Calcination at 100 C is essential for complete removal of water and to obtain higher crystallinity. The prominent peaks corresponding to the diffraction planes (100), (002), (101), (102), (110), (103) and (112) agree well with the JCPDS Card No , confirming the hexagonal Wurtzite structure of the ZnO NPs. The average particle size (D) of synthesized nanoparticles was calculated using the well known Scherrer formula [24] is nearly 16 nm. The XRD parameters are given in Table FE-SEM morphology of ZnO NPs The morphology of the green synthesized ZnO NPs is illustrated in Fig Nanoparticles are spherical in shape and uniformly distributed.

25 90 0 / : ; < = 4 >? > 6 A B C C D > 7 8 9

26 91 Peak No. Position ( 2 ) FWHM, ( ) Particle size, D (nm) Mean 16

27 0 0 E 3 F E G

28 FT-IR spectra of-green tea extract and ZnO NPs The FT-IR spectrum of the green tea extract and that of the synthesized ZnO NPs are shown in Fig In the IR spectrum of green tea, the band at 3394 cm -1 is due to stretching vibrations of O H groups in water, alcohol and phenols and N H stretching in amines. The C H stretch in alkanes and O H stretch in carboxylic acid appear at 2926 and 2864 cm -1 respectively. The strong band at 1627 cm -1 is attributed to the C=C stretch in aromatic ring and C=O stretch in polyphenols. The C N stretch of amide-i in protein gives the band at 1396 cm -1. The C O C stretching in polysaccharides gives a band at 1741 cm -1 and C O stretching in amino acid causes a band at 1037 cm -1. Finally the weak band at 819 cm -1 is the result of C H out of plane bending. Thus from the IR spectrum it can be observed that green tea sample is rich in polyphenols, carboxylic acid, polysaccharide, amino acid and proteins.

29 0 H 0 3 I < 4 > 6 >

30 Antibacterial assays The antibacterial activity of ZnO NPs against the studied pathogenic strains are shown in Plate 4.7. The values of zone of inhibition obtained from the assay are presented in Table All Gram-negative bacteria had shown good sensitivity towards the green synthesized ZnO NPs for the concentration 20 g ml 1. It is quite interesting to note that all bacterial species tested in this study showed resistance to the synthetic antibiotic drug which in turn indicates the better antibacterial activity of the ZnO NPs than the commercially available synthetic drug Antifungal assay Regarding the antifungal activity, all four fungal strains used in this study are found to be sensitive to the green synthesized ZnO NPs as well as to the commercially available antifungal drug Itraconozole. The antifungal activities of ZnO NPs are shown in Plate 4.8 and the zone of inhibition values are presented in Table The fungal species Aspergillus flavus had shown medium sensitivity to ZnO NPs with a concentration of 20 g ml 1, whereas the remaining three fungal species showed good sensitivity to the ZnO NPs concentration of 20 g ml 1.

31 : J K > L M N O P Q R S P T U O V W X Y Z [ \ ] ^ _ ` a b c [ V d X e Z f b g ` h i j j X k Z [ ^ ] \ ^ c h l j m n n o p o i l d q i d o i l p h l m q i r s t u t q r m l m v o d q i l p q w x y u y o z h l m v o d q i l p q w 96

32 97 { h W w o s } ~ s i l m W h d l o p m h w h d l m v m l q n z p o o i r i l o r m o j ƒ i y t r h l j m n n o p o i l d q i d o i l p h l m q i r h z h m i r l h l q z o i m d W h d l o p m h w r o d m o r q n d w m i m d h w r q p d o r Zone of inhibition (mm) Label Bacteria Concentration of ZnO NPs ( g ml 1 ) P Gram-negative a K. pneumoniae 10 b P. aeruginosa 3 c E. coli Gram-positive d S. aureus 2 5 ˆ Š Œ Ž Œ Ž Ž š œ ž Ÿ

33 t w h l o s s i l m n i z h w h d l m v m l q n ƒ i y t r h z h m i r l h X Z ^ ` _ [ ^ c V W X Y \ a ` f ` g g ` ^ r s V d X Z g [ ^ c h i j j X Z a ` _ \ ] h l j m n n o p o i l d q i d o i l p h l m q i r s t u t q r m l m v o d q i l p q w y u y o z h l m v o d q i l p q w 98

34 99 { h W w o s } } s i l m n i z h w h d l m v m l q n z p o o i r i l o r m o j ƒ i y t r h l j m n n o p o i l d q i d o i l p h l m q i r h z h m i r l h l q z o i m d W h d l o p m h w r o d m o r q n d w m i m d h w r q p d o r Zone of inhibition (mm) Label Fungi Concentration of ZnO NPs ( g ml 1 ) P a A. fumigatus 5 3 b Penicillium sp. 7 2 c A. flavus 3 3 d A. niger 3 2 ˆ Š Œ Ž Œ Œ Ž š œ ž Ÿ 4.3. PHYTOCHEMICAL ANALYSIS OF INDIAN GREEN TEAS GC-MS analysis of green teas The chromatograms of the six Indian green teas are presented in Figs through The corresponding peak reports representing the detected phytochemicals in the green tea samples are provided in Tables 4.12 through From these Tables the major phytochemicals identified with peak area percentage >1 are furnished in Tables 4.18 through Finally, the important phytochemicals with health beneficial potentials are short listed and presented in Table 4.24.

35 p q h l q z p h q n ª y l o h 100

36 101 Peak# R.Time Area Area% Name Azabicyclo[3.2.1]octane (N,N-DIETHYL)AMINOPROPYNE ,5-ANHYDRO-6-DEOXYHEXO-2,3-DIULOSE (HYDROXYMETHYL)-2-METHYL-1-PYRROLIDINECA Decane, 3,7-dimethyl Decane, 3,7-dimethyl Undecanol ,2,3-BENZENETRIOL Nonane, 5-(2-methylpropyl) PHENOL, 2,6-BIS(1,1-DIMETHYLETHYL)-4-METHYL-, M Benzoic acid, 4-ethoxy-, ethyl ester Nonane, 5-(2-methylpropyl) Nonane, 5-(2-methylpropyl) Tridecene ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI Dodecane, 2,6,11-trimethyl ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI Dodecane, 2,6,11-trimethyl DECANOIC ACID Cyclotetradecane Oxabicyclo[4.1.0]heptane, 1,5-dimethyl ANTHRACENOL, 1,4,4A,5,8,8A,9,9A,10,10A-DECAHYD ,7,11,15-Tetramethyl-2-hexadecen-1-ol Caffeine H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL Hexadecanoic acid, methyl ester n-hexadecanoic acid ,10-Hexadecadienoic acid, methyl ester ,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z) Phytol HEXADECANOIC ACID, METHYL ESTER ,12,15-Octadecatrienoic acid, (Z,Z,Z) Octadecanoic acid Cyclohexanol, 4-[(trimethylsilyl)oxy]-, cis PREGNANE, SILANE DERIV Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester ,2-BENZENEDICARBOXYLIC ACID Olean-12-ene-3,28-diol, (3.beta.) Cyclopropane, 1,1-dichloro-2,2,3,3-tetramethyl ETHYL (9Z,12Z)-9,12-OCTADECADIENOATE # ,3,5-Trisilacyclohexane TRICYCLO[ E7,16]TRIACONTAN, 1(22),7(16)-DIE Squalene Vitamin E Stigmasterol

37 p q h l q z p h q n { y l o h 102

38 103 { h W w o s } «s t o h p o q p l q n { y l o h d q q i j r Peak# R.Time Area Area% Name METHYL-5,6-DIHYDRO-2(1H)-PYRIDINONE ,2,5,6-Tetrahydropyridin-2-one, 5-methyl Azetidin-1-yl-propionic acid, methyl ester ,3-DIHYDRO-3,5-DIHYDROXY-6-METHYL-4H-PYRAN (HYDROXYMETHYL)-2-METHYL-1-PYRROLIDINECA Butane, 2,2-dimethyl Decane, 3,7-dimethyl Pentafluoropropionic acid, octyl ester ,2,3-BENZENETRIOL Nonane, 5-(2-methylpropyl) CHLOROHEXADECANE Benzoic acid, 4-ethoxy-, ethyl ester Nonane, 5-(2-methylpropyl) Tridecene ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI ,2,4-BUTANTRIOL, 4-O-OCTYL Nonane, 5-(2-methylpropyl) Dodecane, 2,6,11-trimethyl Cyclotetradecane Caffeine H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL Hexadecanoic acid, methyl ester n-hexadecanoic acid n-heptadecanol ,12-Octadecadienoic acid (Z,Z) ,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z) HEXADECEN-1-OL, 3,7,11,15-TETRAMETHYL-, [R-[R* HEXADECANOIC ACID, METHYL ESTER OCTADECENOIC ACID (Z) Nonadecane Ethyl-3-trimethylsilyloxydecane Oxalic acid, 3,5-difluorophenyl tetradecyl ester Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester ,2-BENZENEDICARBOXYLIC ACID, DIISOOCTYL EST Heptadecane, 2,6,10,15-tetramethyl Cyclohexanol, 4-[(trimethylsilyl)oxy]-, cis Squalene Vitamin E Trihydroxycholanic acid, (3.alpha., 7.beta., 12.alpha.)

39 ± ² ³ ² ± µ ³ ² 104

40 105 ² ¹ º» ¼ ½» ¼ ¾ ² À ³ µ ³ ² Á ± À Â Ã Ä Å Peak# R.Time Area Area% Name METHYL-5,6-DIHYDRO-2(1H)-PYRIDINONE (N,N-DIETHYL)AMINOPROPYNE (1H)-Pyrimidinone, 6-hydroxy Butane, 2,2-dimethyl Decane, 3,7-dimethyl Tridecene ,2,3-BENZENETRIOL Nonane, 5-(2-methylpropyl) Benzoic acid, 4-ethoxy-, ethyl ester Tridecene ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI Nonane, 5-(2-methylpropyl) Dodecane, 4,6-dimethyl ,3,7-TRIMETHYL-3,7-DIHYDRO-1H-PURINE-2,6-DIONE H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL n-hexadecanoic acid ,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z) Phytol ,12-Octadecadienoic acid (Z,Z) ,12,15-Octadecatrienoic acid, (Z,Z,Z) Octadecanoic acid Butanedioic acid, 2-hydroxy-2-methyl-, dimethyl ester Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester ,2-BENZENEDICARBOXYLIC ACID, DIISOOCTYL EST (Z)6,(Z)9-Pentadecadien-1-ol Octadecanoic acid, 2,3-dihydroxypropyl ester

41 ± ² ³ ² ± µ Æ ³ ² 106

42 107 ² ¹ º» ¼ ½ Ç ¼ ¾ ² À ³ µ Æ ³ ² Á ± À Â Ã Ä Å Peak# R.Time Area Area% Name ,3-DIHYDRO-3,5-DIHYDROXY-6-METHYL-4H-PYRAN Butane, 2,2-dimethyl NONANE, 3,7-DIMETHYL Decane, 3,7-dimethyl DL-Proline, 5-oxo-, methyl ester Tetradecane ,2,3-BENZENETRIOL PENTADECANE Benzoic acid, 4-ethoxy-, ethyl ester Nonane, 5-(2-methylpropyl) Undecanol Tetradecane ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI Undecene, 5-methyl Nonane, 5-(2-methylpropyl) Nonane, 5-(2-methylpropyl) ISOPROPENYL-2-METHYL-7-OXA-BICYCLO[4.1.0]HE ,3,7-TRIMETHYL-3,7-DIHYDRO-1H-PURINE-2,6-DIONE H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL HEXADECANOIC ACID, METHYL ESTER n-hexadecanoic acid ,12-Octadecadienoic acid (Z,Z) ,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z) HEXADECEN-1-OL, 3,7,11,15-TETRAMETHYL-, [R-[R* HEXADECANOIC ACID, METHYL ESTER cis,cis,cis-7,10,13-hexadecatrienal Octadecanoic acid Octadecynoic acid trans-9-octadecenoic acid, trimethylsilyl ester Floxuridine Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester ,2-BENZENEDICARBOXYLIC ACID, DIISOOCTYL EST

43 ± ² ³ ² ± µ È É Ê ³ ² 108

44 109 ² ¹ º» ¼ ½ Ë ¼ ¾ ² À ³ µ È É Ê ³ ² Á ± À Â Ã Ä Å Peak# R.Time Area Area% Name Azabicyclo[3.2.1]octane (N,N-DIETHYL)AMINOPROPYNE (2-Hydroxyethyl)-1,2,4-triazole ,5-ANHYDRO-6-DEOXYHEXO-2,3-DIULOSE (1H)-Pyrimidinone, 6-hydroxy Hexane, 3,3-dimethyl Decane, 3,7-dimethyl Tridecene ,2,3-BENZENETRIOL ,6,10-DODECATRIENE, 7,11-DIMETHYL-3-METHYLEN Nonane, 5-(2-methylpropyl) Benzoic acid, 4-ethoxy-, ethyl ester Nonane, 5-(2-methylpropyl) Sulfurous acid, hexyl octyl ester HEXANE, 3,3,4-TRIMETHYL Undecanol ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI Hexadecane Dodecane, 2,6,11-trimethyl Dodecane, 4,6-dimethyl Heptadecene, 1-chloro ,3,7-TRIMETHYL-3,7-DIHYDRO-1H-PURINE-2,6-DIONE H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL HEXADECANOIC ACID, METHYL ESTER n-hexadecanoic acid ,12-Octadecadienoic acid (Z,Z) ,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z) HEXADECANOIC ACID, METHYL ESTER cis,cis,cis-7,10,13-hexadecatrienal OCTADECENOIC ACID (Z) Ethyl-3-trimethylsilyloxydecane Tridecanoic acid, 3-hydroxy-, ethyl ester Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester Cyclohexanol, 4-[(trimethylsilyl)oxy]-, cis

45 ± ² ³ ² ± µ Ì Í Í ³ ² 110

46 111 ² ¹ º» ¼ ½ Î ¼ ¾ ² À ³ µ Ì Í Í ³ ² Á ± À Â Ã Ä Å Peak# R. Time Area Area% Name METHYL-5,6-DIHYDRO-2(1H)-PYRIDINONE Cyclopentane, 1-acetyl-1,2-epoxy (N,N-DIETHYL)AMINOPROPYNE H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl (HYDROXYMETHYL)-2-METHYL-1-PYRROLIDINECA ,2,3-Propanetriol, monoacetate Tridecene Nonane, 5-(2-methylpropyl) Nonane, 5-butyl ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI Dodecane, 2,6,11-trimethyl ,3,4,5-TETRAHYDROXY-CYCLOHEXANECARBOXYLI (4H)-BENZOFURANONE, 5,6,7,7A-TETRAHYDRO-6-HY ,3,7-TRIMETHYL-3,7-DIHYDRO-1H-PURINE-2,6-DIONE H-PURINE-2,6-DIONE, 3,7-DIHYDRO-3,7-DIMETHYL n-hexadecanoic acid (Z)6,(Z)9-Pentadecadien-1-ol Octadecenoic acid (Z)-, methyl ester Phytol ,12-Octadecadienoic acid (Z,Z) ,12,15-Octadecatrienoic acid, (Z,Z,Z) Octadecanoic acid Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester Octadecanoic acid, 2,3-dihydroxypropyl ester dl-.alpha.-tocopherol Stigmasterol

47 112 ² ¹ º» ¼ ½ Ï ¼ Ð ² Ñ À Ò ³ Á ± Ó Á ² º Å Ô ² ² À Á à ³ ² Õ ½ Ö µ Ð É É ³ ² Peak R.T. Area (%) Molecular formula M. wt. Name of the compound Type Therapeutic use C6H6O ,2,3-Benzenetriol Pyrogallol Antioxidant Antiseptic fungicide, Antidermatitic insecticide, candidicide C7H12O ,3,4,5-Tetrahydroxycyclohexanecarbonyl Quinic acid Antimicrobial, anti-inflammatory induces antioxidant C8H10N4O Caffeine Alkaloid Both pro and antioxidant C7H8N4O H-Purine-2,6-dione, 3,7-Dihydro-3,7-Dimethyl (Theobromine) Xanthine alkaloide Vasodilaton both pro-and antioxidant natural diuretic diuretic C16H32O n-hexadecanoic acid Palmitic acid Antioxidant Nematicide pesticide hemolytic anti-inflammatory hemolytic C20H48O Phytol Diterpene Antioxidant Antimicrobial, anticancer, diuretic C18H30O ,12,15-Octadecatrienoic acid (z,z,z) -linolenic acid Antiarthritic, antihistaminic, anticoroanary, antiandrogenis, antinematicide, anticancer, antibacterial

48 113 ² ¹ º» ¼ ½ Ø ¼ Ð ² Ñ À Ò ³ Á ± Ó Á ² º Å Ô ² ² À Á à ³ ² Õ ½ Ö µ Ì ³ ² Peak R.T. Area (%) Molecular formula M. wt. Compound name Type/nature Therapeutic use * * 1,2,3-Benzenetriol Pyrogallol * * * 1,3,4,5-Tetrahydroxycyclohexanecarbonyl Quinic acid * * * Caffeine Alkaloid * * * 1H-Purine-2,6-dione, 3,7-Dihydro-3,7-Dimethyl (Theobromine) Xanthine, Alkaloid * * * n-hexadecanoic acid Palmitic acid * C18H36O ,12,15-Octadecatrienoic acid (z,z,z) Palmitic acid, ethyl ester Antioxidant, nematicide pesticide, hypocholesterolemic, hemolytic Ù Ú Û Ü Ý Þ ß à á â ã ä ß å æ å

49 114 ² ¹ º» ¼ ç è ¼ Ð ² Ñ À Ò ³ Á ± Ó Á ² º Å Ô ² ² À Á à ³ ² Õ ½ Ö µ ³ ² Peak R.T. Area (%) Molecular formula M. wt. Compound name Type/nature Therapeutic use * * 1,2,3-Benzenetriol Pyrogallol * * * * * Caffine 1,3,4,5-Tetrahydroxycyclohexanecarboxyl 1,3,7-Trimethyl-3,7- dihydro-1h-purine-2,6- dione Quinic acid * Alkaloid * * * 1H-Purine-2,6-dione, 3,7-Dihydro-3,7-Dimethyl Xanthine, Alkaloid * * * n-hexadecanoic acid Palmitic acid * * * 9,12,15-Octadecatrienoic acid (z,z,z) Lainolenic acid * * * Hexadecanoic acid ethylester Palmitic acid, ethyl ester * Ù Ú Û Ü Ý Þ ß à á â ã ä ß å æ å

50 115 ² ¹ º» ¼ ç ½ ¼ Ð ² Ñ À Ò ³ Á ± Ó Á ² º Å Ô ² ² À Á à ³ ² Õ ½ Ö µ Æ ³ ² Peak R.T. Area (%) Molecular formula M. wt. Compound name Type/nature Therapeutic use * * Tetradecane Alkane * * * 1,2,3-Benzenetriol Pyrogallol * * * * * Caffine 1,3,4,5-Tetrahydroxycyclohexanecarboxyl 1,3,7-Trimethyl-3,7- dihydro-1h-purine-2,6- dione Quinic acid * Alkaloid * * * 1H-Purine-2,6-dione, 3,7-Dihydro-3,7- Dimethyl Xanthine, Alkaloid * * n-hexadecanoic acid Palmitic acid * C 6H 26O 234 Cis, cis, cis-7,10,13- Hexadecatriental Alcohol *

51 116 ² ¹ º» ¼ ç ç ¼ Ð ² Ñ À Ò ³ Á ± Ó Á ² º Å Ô ² ² À Á à ³ ² Õ ½ Ö µ È É Ê ³ ² Peak R.T. Area (%) Molecular formula M. wt. Compound name Type/nature Therapeutic use * * 1,2,3-Benzenetriol Pyrogallol * * * * * 1,3,4,5- Tetrahydroxycyclohexanecarboxyl 1,3,7-Trimethyl-3,7- dihydro-1h-purine- 2,6-dione Quinic acid * Caffeine * * * 1H-Purine-2,6-dione, 3,7-Dihydro-3,7- Dimethyl Xanthine, Alkaloid * * * n-hexadecanoic acid Palmitic acid * C 6H 26O 234 Cis, cis, cis-7,10,13- Hexadecatriental Fatty aldehydes é ê ë ì í î ï ð ï ñ ò ï ó ò ô

52 117 ² ¹ º» ¼ ç õ ¼ Ð ² Ñ À Ò ³ Á ± Ó Á ² º Å Ô ² ² À Á à ³ ² Õ ½ Ö µ Ì Í Í ³ ² Peak R.T. Area (%) Molecular formula M. wt. Compound name Type/nature Therapeutic use Methyl-5,6-dihydro- 2(1H)-pyridinone 1-(N,N- Dimethyl)aminoporpyne Unknown Unknown * * 4H-Pyran-4-one, 2,3- dihydro-3,5-dihydro-6- methyl Flavonoid fraction * * * 1,3,4,5,-tetrahydroxycyclohexanecarboxyl Quinic acid * * * 1,3,7,-trimethyl-3,7- dihydro-1h-purine-2,6- dione Coffeine alkaloid * * * 1H-Purine-26-Dione, 3,7- Dihydro-3,7-dimethyl Xanthine, Alkaloid * * * n-hexadecanoic acid Palmitic acid * * * 9,12,15-Octadecatrienoic acid (z,z,z) Linolenic acid * é ê ë ì í î ï ð ï ñ ò ï ó ò ô

53 118 ² ¹ º» ¼ ç» ¼ Ð ² Ñ ² º ³ ¹ à µ Ó Á Ó ² º À Ò ³ Á ± Ó Á ² º Å µ à ³ ² Å ² ± À º Å ² à ² º Ò ö Ä ¹ Ò Ð Í Compounds Peak area (%) of green tea samples MOON TAN GT TET KOL ASS ø ù ø Þ ß Ý ú á ß Û Ý ú á Þ ß á ß û á ß Þ Pyrogallol Quinic acid Caffeine (alkaloid) Xanthine (Alkaloid) Palmitic acid Palmitic acid Phytol ethyl ester linolenic acid Total

54 ± Ó Á ² º Å ³ Â Á ³ Â µ Å º Á ³ Ä Á ± À Â Ã Ä Å µ Ã ³ ² Å ² ± À º Å Ô ü µ ² ² Õ ½ ¼ è è Ö 119

55 120

56 FT-IR analysis of phytochemical green teas The FT-IR spectra of the six green tea samples are shown in Fig (a) through (f).

57 122

58 123 ý Ó ¼» ¼ ½ Ë ¼ ý þ ÿ Å À Á ³ ² µ Ã ³ ² Å ² ± À º Å Ô ² Ö Ð É É Ô ¹ Ö Ì Ô Á Ö Ô Ä Ö Æ Ô Ö È É Ê ² Ã Ä Ô µ Ö Ì Í Í

59 124 The vibrational band assignment for the prominent peaks and the chemical compounds identified are provided in Table ² ¹ º» ¼ ç Ç ¼ þ à µ ² Ä Ó ¹ ² ³ Ó Ã ² º ¹ ² Ã Ä Å µ à ³ ² Å ² ± À º Å Wavenumber (cm 1 ) Vibration band/group Chemical compound Reference 3270 ~ 3320 O H stretch H Bonded Phenols, alcohols Awwad et al. (2013) Umashankari et al. (2012) Vanaja et al. (2013) Theivasanthi et al. (2013) 2946 C H stretch (asym.) Alkanes Vanaja et al. (2013) O H stretch Carboxylic acid Sharmin et al. (2013) 2833 C H stretch (sym.) Alkanes Umashankari et al. (2012) Arokiyaraj et al. (2013) 1629 ~ 1663 C=O stretch (carbonyls) Flavonoids Heneczkowski et al. (2001) Polyphenols, catechins Rajathi and Sridhar (2013) C=C stretch Aromatics Kong and Yu (2007) 1449 C C stretch (in ring) Aromatics 1239 C N stretch Aliphatic amines Umashankari et al. (2012) Heneczkowski et al. (2001) Mallikarjuna et al. (2012) Heneczkowski et al. (2001) Nagajyothi et al. (2013) 1113 C O stretch Alcohols, esters, carboxylic acids Mallikarjuna et al. (2012) 1014 ~ 1019 C O stretch Alcohols, esters, carboxylic acids Rajathi and Sridhar (2013) Mallikarjuna et al. (2012) C N stretch Aliphatic amines Nagajyothi et al. (2013) C OH stretch Secondary alcohols Jayaseelan et al. (2013) The FT-IR spectra of the green tea samples clearly reveals the presence of polyphenols, flavonoids, amines as major phytochemicals. This fact enables to speculate higher antioxidant potentials of the green tea samples.

60 ANTIOXIDANT POTENTIALS OF INDIAN GREEN TEAS Total phenolic contents-fcr method The optical density values representing the total phenolic contents (TPC) of the six green tea samples and the standard galic acid for various concentrations (15, 30, 60, 125 and 250 mg ml -1 ) measured using spectrophotometer are provided in Table The dose response curves representing TPC of the green tea are shown in Fig ² ¹ º» ¼ ç Ë ¼ É À ³ Ó Á ² º Ä Ã Å Ó ³ Ò ² º  ŠÀ Å Ã ³ Ó Ã ³ ³ ² º À à º Ó Á Á à ³ à ³ Å µ à ³ ² Å ² Ã Ä ² º Ó Á ² Á Ó Ä µ ² Ó Â Å Á à Á à ³ ² ³ Ó Ã Å Optical density values Concentration of extracts g ml -1 Gallic acid (standard) Green tea samples MOON TAN GT TET KOL ASS

61 126 E C D >8 A B 7 ; <=> :! " # $ % & & ' ( ) * + ), -., / ( ) 0 % F G H I J I K L I M N O P Q R P O S N T O P U V R W P O N X Y N Y Z [ S \ P T N [ G U U N T Y P T Y O N X H R P P T Y P Z O Z ] S [ P O

62 Total flavonoids-aluminium chloride method The optical density values of the six green tea samples and the standard quercetin for various concentrations (15, 30, 60, 125 and 250 g ml -1 ) are provided in Table The dose response curves representing TF of the green tea are shown in Fig ^ Z _ [ P J I ` L I a S Y G U Z [ b P T O G Y c N X W Z [ V P O R P S R P O P T Y G T H Y N Y Z [ X [ Z W N T N G b O N X Y \ P H R P P T Y P Z O X N R W Z R G N V O U N T U P T Y R Z Y G N T O Optical density values Concentration of extracts g ml -1 Green tea samples MOON TAN GT TET KOL ASS

63 128 E C A B ; D 7 ; <=> : d! " # $ % & & ' ( ) * + ), -., / ( ) 0 % F G H I J I K e I M N O P Q R P O S N T O P U V R W P O N X Y N Y Z [ X [ Z W N T N G b O N X H R P P T Y P Z O Z ] S [ P O

64 DPPH-RSA assay The optical absorbance percentage values of the six green tea samples and the standard quercetin for various concentrations (15, 30, 60, 125 and 250 g ml 1 ) are provided in Table The dose-response curves representing the DPPH-radical scavenging activity (%) for the six green tea sample and the standard ascorbic acid are shown in Fig The DPPH scavenging activity of all samples appeared to depend on the extract concentration up to 60 g ml 1, above which the activity approaches saturation level. This is due to the quantity of DPPH used in the test reaction. The DPPH-radical scavenging activity values expressed in ascorbic acid equivalents are furnished in Table ^ Z _ [ P J I ` e I M f f g R Z b G U Z [ O U Z W P T H G T H Z U Y G W G Y G P O N X Y \ P h T b G Z T H R P P T Y P Z O Concentration of extracts g ml -1 Absorbance (%) Ascorbic Green tea samples acid (standard) MOON TAN GT TET KOL ASS

65 oe 6? n m 130 i d j * ( - k / *. * / l! " # $ % & & d i d ' ( ) * + ), -., / ( ) 0 % F G H I J I K p I M N O P Q R P O S N T O P U V R W P O N X M f f g R Z b G U Z [ O U Z W P T H G T H Z U Y G W G Y c q r s Z T b Y \ P O Y Z T b Z R b Z O U N R _ G U Z U G b

66 FRAP assay The optical density values of the six green tea samples and the standard ascorbic acid for various concentrations (15, 30, 60, 125 and 250 g ml 1 ) are provided in Table The dose-response curves representing the ferric reducing power of the six green teas and that of the standard ascorbic acid are illustrated in Fig The FRAP activity values expressed in ascorbic acid equivalents are furnished in Table ^ Z _ [ P J I ` p I F t u f Z O O Z c W Z [ V P O N X Y \ P h T b G Z T H R P P T Y P Z O Absorbance (%) Concentration of extracts g ml -1 Ascorbic acid (standard) Green tea samples MOON TAN GT TET KOL ASS

67 132 E C A B ; D 7 ; <=> : d j * ( - k / *. * / l! " # $ % & & d v ' ( ) * + ), -., / ( ) 0 % F G H I J I ` w I M N O P Q R P O S N T O P U V R W P O N X F t u f Z O O Z c N X Y \ P H R P P T Y P Z O Z ] S [ P O Z T b Y \ P O Y Z T b Z R b Z O U N R _ G U Z U G b

68 133 ^ Z _ [ P J I x w I M f f g R Z b G U Z [ O U Z W P T H G T H Z U Y G W G Y c q t y u s z X P R R G U R P b V U G T H Z T Y G N { G b Z T Y S N P R q F t u f s z Y N Y Z [ S \ P T N [ G U U N T Y P T Y q ^ f } s Z T b Y N Y Z [ X [ Z W N T N G b O q ^ F s N X Y \ P H R P P T Y P Z O Z ] S [ P O S. No. Green tea sample DPPH-RSA (mg AE g -1 ) FRAP (mg AE g -1 ) TPC (mg QAE g -1 ) TF (mg QE g -1 ) 1 MOON * TAN GT TET KOL ASS ~ ƒ ƒ ˆ ƒ ˆ

69 CORRELATION STUDIES Correlation analysis on the antioxidant potentials of Indian green teas with their phenolic and flavonoid contents The relationship existing between the content of phenolic compounds and antioxidant activities can be well understood by carrying out correlation analysis. The linear and positive correlation existing between the DPPH-RSA values and the TPC and TF values are shown in Fig Similarly Fig illustrates the relationship between the antioxidant activity assayed by FRAP method and the TPC and TF values. Further the significant correlation observed between the antioxidant activities assayed by the DPPH and FRAP methods is illustrated in Fig The results of the correlation analysis are presented in the form of Pearson s correlation coefficient (R) matrix in Table 4.31 Š Œ Ž Š Œ Š š š š œ ž Ÿ Š š Š Š Œ š Š š Š Š Ÿ Š Š Ÿ Œ Variables DPPH-RSA FRAP TPC TF ª «± ² ³ µ µ «ª ª «DPPH-RSA 1 FRAP TPC TF

70 ¹ š Ž º» Œ Š š ¼ ½ Š ¾ š Š Œ Š š Š š š š œ Š ž» Š ¾ œ ž ¹ Š Š Ÿ Œ 135

71 ¹ š Ž º º» Œ Š š ¹ À Š š š š œ Š ž» Š ¾ œ ž ¹ Š Š Ÿ Œ 136

72 ¹ š Ž º» Œ Š š ¼ ½ Š ¾ ¹ À Š Š 137

73 Correlation analysis between FT-IR estimates and calorimetric data of the Indian green teas From the infrared vibrational band assignments provided in Table 4.25, three bands appear to have some relevance to the antioxidant potentials of the green tea samples. The absorbance (%) of these three vibrational bands considered for the correlation studies, are presented in bold numbers in Table Since the absorbance data corresponding to the bands around 1448 and 1113 cm -1 did not seem to contain any correlation, they were not considered for correlation studies. Š Œ Ž º ¹ Á  Š Š œ à ž Œ ¾ š Š š Š Œ Š ¾ Š Š Ÿ Œ Vibrational band (cm 1 ) Absorbance (%) MOON TAN GT TET KOL ASS 3270 ~ 3320* ~ ~ ~ ~ Ä Å Æ Ç Æ È É Ê Ë Ì Í É Î Ï Ê Ð Ñ Ò Æ Ñ Ð Î Ò Ð Í Ó Ë Ñ Ô Ë Ñ Ñ Ð Ì Æ Ç È Ë É Æ É Æ Ì Õ Ò È Ò Ö

74 139 The calorimetric data namely the TPC, TF, DPPH-RSA and FARP values and the FT-IR estimates of polyphenols, flavonoides and amines considered for the correlation studies are provided in Table Š Œ Ž ¹ Á  Š ¾ Œ š Ÿ š ¾ Š Š Š Š Ÿ Œ FT-IR data (absorbance %) Colorimetric data* Green tea samples Polyphenols Flavonoids Alcohols/ amines DPPH-RSA (mg AE g 1 ) FRAP (mg AE g 1 ) TPC (mg GAE g 1 ) TF (mg QE g 1 ) MOON TAN GT TET KOL 31.30** ASS Ä Å Æ Ç Æ Ó Ñ Ë Ï Æ Î Ç Ë Ñ Ò Ø Ù Ë Ñ Ú Ô Ë Ï Ï Î É È Ô Æ Ç Ð Í Ð Ì Ò Ð Ù Ð Ñ Ð Ö Ä Ä Û Æ Ò Ð Ë Ï È Ç Ç Ð Í È É Ô Ë Ñ Ñ Ð Ì Æ Ç È Ë É Ò Ç Î Í È Ð Ò Ö Ü Ò Ô Ë Ñ Ê È Ô Æ Ô È Í Ð Ý Î È Þ Æ Ì Ð É Ç ß Ü à á â Æ Ì Ì È Ô Æ Ô È Í Ð Ý Î È Þ Æ Ì Ð É Ç ß ã Ü à á Ý Î Ð Ñ Ô Ð Ç È É Ð Ý Î È Þ Æ Ì Ð É Ç ß ä à Ö

75 140 The data presented in Table 4.33 were subjected to bivariate correlation analysis to establish the mutual correlation among them. The Pearson s linear correlation coefficient (R) matrix thus obtained is presented in Table Š Œ Ž Ž Š Œ š Š Œ Š š š š œ ž Ÿ Š š Š Š Œ š ¹ Á  Š ¾ Œ š Ÿ š ¾ Š Š Š Š Ÿ Œ FT-IR data Colorimetric data Variables Polyphenols Flavonoids Alcohols/ amines DPPH-RSA FRAP TPC TF Polyphenols 1 FT-IR data Flavonoids 0.985** 1 Alcohols/ amines 0.953* 0.940** 1 DPPH-RSA 0.940* 0.977** 0.914* 1 Colorimetric data FRAP * 0.888* 0.925** 1 TPC 0.924* 0.953** 0.930** 0.974** 0.966** 1 TF 0.932* 0.958** 0.954** 0.958** 0.969** 0.991** 1 Ä Ä Û Ë Ñ Ñ Ð Ì Æ Ç È Ë É È Ò Ò È â É È Ó È Ô Æ É Ç Æ Ç å Ö å æ Ì Ð Þ Ð Ì Ö Ä Û Ë Ñ Ñ Ð Ì Æ Ç È Ë É È Ò Ò È â É È Ó È Ô Æ É Ç Æ Ç å Ö å ç Ì Ð Þ Ð Ì Ö

76 ANTIMICROBIAL POTENTIAL OF INDIAN GREEN TEAS The antimicrobial activities of the six green teas against four bacterial species are shown in Plate 4.9. The diameter of zone of inhibition, measured to estimate the activity are provided in Table The antifungal potential of the six green tea samples against four fungal species are shown in Plate 4.10 and the diameters of the zone of inhibition are presented in Table 4.36.

77 142 í ò ò í ò ò ð ê ñ é ê ê ë ð ê ñ é ê ê ë ì ï ì î ì ì í ë ì ï ì î ì ì í ë ð ê ñ í ò ò é ê ê ë ì ï ì ð ê ñ ì í ë î ì í ò ò ì ï ì î ì ì í ë é ê ê ë Œ Š Ž è À š Š š Š Œ Š š š š Š Š Ÿ Œ

78 143 Š Œ Ž ó À š Š š Š Œ Š š š š Š Š Ÿ Œ Green tea samples Bacillus subtilis Diameter of zone of inhibition (mm) Volume of sample (30 L) Escherichia coli Staphylococcus aureus Streptococcus pyrogenes MOON TAN GT TET KOL 9 9 NA 9 ASS ô õ ö ô ø ù ú û ü û ú ý

79 144 þ ÿ ÿ ÿ

80 145 ÿ! ÿ " ÿ Green tea samples Diameter of zone of inhibition (mm) Volume of sample (30 L) Candida albicans Aspergillus niger Aspergillus flavus Aspergillus terreus MOON TAN GT TET KOL ASS