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3.2. Flavonoids pattern in Pistacia species 20 Flavonoids-enriched red and/or green leaf and stem extracts of six species of Pistacia (that were harvested in autumn and spring) were subjected to HPLC analysis. HPLC chromatograms of these extracts revealed up to seven major characteristic peaks, which were annotated 1; 2; 3; 4; 5; 6 and 7, that appeared in the corresponding retention times (minutes): 2, 7, 8, 10, 11, 13, 15 shown schematically in Fig. 4. Additionally, HPLC analysis revealed about 30 minor peaks. Fig. 4. A schematic pattern of a representative HPLC chromatogram of Pistacia flavonoidsenriched extracts showing the seven major characteristic peaks 3.3. Identification and quantification of characteristic peaks 3.3.1. HPLC identification Cyanidin, cyanidin-3-O-glucoside, quercetin and quercetin-3-O-glucoside were used to identify characteristic peaks (Appendix, Figs. 1 to 4). Based on HPLC peaks retention time, MS and NMR analyses the following compounds were identified: cyanidin-3-O- glucoside corresponds to peak #2, quercetin-3-O-glucoside corresponds to peak #4,
21 cyanidin corresponds to peak #6 and quercetin to peak #7. The flavonoids content of peaks' #2, 4, 6 and 7 (expressed as mg/gFW) was determined by using their appropriate calibration curves (Appendix, Figs. 1 to 4). For the sake of comparison, in the case of the unidentified peaks (#1, 3 and 5), their contents are expressed in cyanidin-3-O- glucoside equivalents. The flavonoids content of all the characteristic peaks, under all experimental conditions, are given in Tables 3 to 9. 3.3.2. MS identification Three characteristic peaks (peaks #2, 4 and 6) were purified by HPLC and subjected to Mass Spectrometry analysis (Appendix, Figs. 5 to 8). MS fragmentation pattern of characteristic peaks are given in Appendix, Table 3. 3.3.3. NMR identification Three purified characteristic peaks (peaks #4 and 6) were analyzed by 1H NMR (Appendix, Figs. 9 to 11). Table 3 in the Appendix shows specific chemical shifts (δ, ppm) of the compounds. 3.4. Flavonoids content and pattern in leaves of various Pistacia species 3.4.1. P. chinensis 3.4.1.1. Spring Chromatograms of flavonoid-enriched green and red leaf extracts of P. chinensis that were harvested in spring revealed six and five major peaks, respectively (Fig. 5A, Appendix, Tables 1a, 1b). Green leaves: the highest flavonoid content was that of cyanidin, 12.66 ± 0.26 mg/gFW. The flavonoid contents of peaks #1, 3 and 5 were 0.75 ± 0.15; 5.09 ± 0.43 and
Page 1 and 2: Ben-Gurion University of the Negev
Page 3 and 4: Abstract I Anthocyanins in Pistacia
Page 5 and 6: Acknowledgement III I would like to
Page 7 and 8: V 1.7. Hypothesis and objectives ..
Page 9 and 10: VII 3.6.1.2. Spring and autumn tota
Page 11 and 12: List of Figures IX Fig. 1. Anthocya
Page 13 and 14: Table 10. Chlorophyll and carotenoi
Page 15 and 16: List of Appendix Tables XIII Table
Page 17 and 18: 1. Introduction 1.1. Red coloration
Page 19 and 20: Fig. 1. Anthocyanidin chemical stru
Page 21 and 22: 5 simple phenolics are also potenti
Page 23 and 24: 7 photosystem II (PSII) efficiency
Page 25 and 26: 9 Harborne, 1976). Yet, although mo
Page 27 and 28: 1.5. Leaf senescence 11 Leaf senesc
Page 29 and 30: 1.7. Hypothesis and objectives Hypo
Page 31 and 32: 2.3. Anthocyanin extraction 15 Leaf
Page 33 and 34: 2.6. NMR analysis 17 NMR spectra of
Page 35: A C E A 19 C D D E F F Fig. 3. Pist
Page 39 and 40: 23 4.20 ± 0.63 mg/gFW, respectivel
Page 41 and 42: 25 Table 4. Flavonoids content and
Page 43 and 44: A 27 A B B Fig. 6. Comparison of P.
Page 45 and 46: 29 The flavonoid contents of peaks
Page 47 and 48: 3.4.5. P.vera 3.4.5.1. Spring 31 Le
Page 49 and 50: 33 Fig. 8. Comparison of P. lentisc
Page 51 and 52: 55 The ratios of the combined relat
Page 53 and 54: 77 Table 9. Flavonoids content and
Page 55 and 56: 3.6.1.1. Chlorophyll a/b ratio: 39
Page 57 and 58: 41 3.6.2.2. Spring and autumn total
Page 59 and 60: 3.6.4.2.1. Chlorophyll a/b ratio: 3
Page 61 and 62: 4. Discussion 45 The Pistacia germp
Page 63 and 64: 47 red. Most probably this is relat
Page 65 and 66: 49 The amount of a separated compou
Page 67 and 68: 51 apparatus. Also the decrease in
Page 69 and 70: 53 Pistacia species (Table 12), rev
Page 71 and 72: 6. References 55 Alexieva, V., Serg
Page 73 and 74: 77 Feild, T.S., Lee, D.W. and Holbr
Page 75 and 76: 99 King, J., 1997, Reaching for the
Page 77 and 78: 61 Nagata, T., Todoriki, S., Masumi
Page 79 and 80: 63 Woodall, G.S., Stewart, G.R., 19
Page 81 and 82: Peak area (mAU, x10 6 ) 65 Fig. 3.
Page 83 and 84: B A 67 Fig. 7. MS spectrum of peak
Page 85 and 86: 7.2. Tables 69 Table 1a. Identified
Page 87 and 88:
71 Table 2. Ratios of the combined
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