Phytochemical Studies and Antioxidant Activities of Artocarpus scortechinii King

Supporting Information Rec. Nat. Prod. 10:3 (20) 299-303 Phytochemical Studies and Antioxidant Activities of Artocarpus scortechinii King Norzafneza Mohd Arriffin 1, Shajarahtunnur Jamil* 1, Norazah Basar 1, Shamsul Khamis 2, Siti Awanis Abdullah 1 and Siti Mariam Abdul Lathiff 1 1 Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 1310 Johor Bahru, Johor, Malaysia 2 Department of Landscape Architecture, Faculty of Design and Architecture, Universiti Putra Malaysia, 300 UPM Serdang, Selangor Darul Ehsan, Malaysia Table of Contents Page Experimental Details 3 Antioxidant Activities 3 DPPH Assay 3 ABTS Assay 3 FRAP Assay 3 Statistical Analysis of Data S1: IR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy--γ,γdimethylallylflavone (1) S2: 1 H NMR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy- -γ,γ-dimethylallylflavone (1) S3: 1 H NMR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy- -γ,γ-dimethylallylflavone (1) (Expansion) S: 13 C NMR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy- -γ,γ-dimethylallylflavone (1) S5: EIMS spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy- - γ,γ-dimethylallylflavone (1) 5 5 1

S: IR spectrum of cudraflavone C (2) S: 1 H NMR spectrum of cudraflavone C (2) S: 1 H NMR spectrum of cudraflavone C (2) (Expansion) S9: 13 C NMR spectrum of cudraflavone C (2) S10: EIMS spectrum of cudraflavone C (2) S11: IR spectrum of artocarpin (3) S12: 1 H NMR spectrum of artocarpin (3) S13: 1 H NMR spectrum of artocarpin (3) (Expansion) S1: 13 C NMR and DEPT spectrum of artocarpin (3) S15: EIMS spectrum of artocarpin (3) S1: IR spectrum of cycloartobiloxanthone () S: 1 H NMR spectrum of cycloartobiloxanthone () S1: 1 H NMR spectrum of cycloartobiloxanthone () (Expansion) S19: 13 C and DEPT spectra of cycloartobiloxanthone () S20: EIMS spectrum of cycloartobiloxanthone () S21: IR spectrum of artonin E (5) S22: 1 H NMR spectrum of artonin E (5) S23: 13 C NMR spectrum of artonin E (5) S2: EIMS spectrum of artonin E (5) S25: IR spectrum of oxyresveratrol () S2: 1 H NMR of oxyresveratrol () S2: 13 C NMR spectrum of oxyresveratrol () S2: EIMS spectrum of oxyresveratrol () S29: IR spectrum of macakurzin C () S30: 1 H NMR of macakurzin C () S31: 13 C NMR of macakurzin C () S32: EIMS spectrum of macakurzin C () S33: IR spectrum of flemichapparin A () S3: 1 H NMR of flemichapparin A () S35: 1 H NMR of flemichapparin A () (Expansion) S3: 13 C NMR spectrum of flemichapparin A () S3: EIMS spectrum of flemichapparin A () S3: IR spectrum of luteolin (9) 9 9 10 11 11 12 12 13 1 1 15 15 1 1 19 20 20 21 22 23 23 2 25 2 2 2 2 2 2

S39: 1 H NMR of luteolin (9) 29 S0: 1 H NMR of luteolin (9) (Expansion) 29 S1: 13 C NMR of luteolin (9) 30 S2: EIMS spectrum of luteolin (9) 30 S3: IR spectrum of apigenin (10) 31 S: 1 H NMR of apigenin (10) 32 S5: 13 C NMR of apigenin (10) 32 S: EIMS spectrum of luteolin (10) 33 References 33 Experimental Details Antioxidant Activities 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay: The DPPH assay was conducted according to method by Fu et al. with minor modification [1]. 100 µl of DPPH stock solution (0.1 mm) was mixed with sample 100 µl and incubated for 30 minutes in the dark at room temperature. The absorbance was measured using EPCH microplate reader at 5 nm. The sample was replaced with methanol for blank sample. Percentage inhibition was calculated using the following formula: Percentage inhibition = [(Absorbance blank DPPH Absorbance sample)/ Absorbance blank DPPH] 100% 2,2'-Azino-bis(3-ethylbenzthiazoline--sulphonic acid) (ABTS) assay: The ABTS assay was carried out based on method described by Zou et al. with minor modification [2]. ABTS and potassium persulfate were dissolved in distilled water to obtain concentration mm and.9 mm respectively. Equal amount of these two solutions were mixed and let stand for 12 to 1 hours at room temperature before use. The ABTS radical was added with distilled water to absorbance of 0. at 3 nm. 10 µl of sample was added to 9-well plates together with 190 µl of ABTS solutions. The absorbance was recorded after 30 minutes incubation in the dark at room temperature. The percentage of antioxidant activity was calculated using the following formula: Scavenging concentration = [Abs (ABTS) Abs (ABTS+Sample)/Abs (ABTS)] 100% Ferric reducing antioxidant potential (FRAP) assay: Experiment was carried out according to Channarong et al. with minor modification [3]. FRAP reagent was freshly prepared, consist of stock solution with ratio 10:1:1 of 300 mm acetate buffer, 10 mm TPTZ in 0 mm HCl and 20 mm FeCl 3.H 2 solution. 5 µl of sample, 15 µl of methanol and 150 µl of FRAP reagent were added to the 9-well plates. The absorbance at 53 nm was read after 10 minutes of incubation at 3 C. FeS.H 2 solution (0.1 mm 1.0 mm) was used to build up calibration curves of standard antioxidants. 3

Statistical Analysis of Data: Three replicates of each sample were used for statistical analysis with values reported as mean ± SD. Standard curves were generated and calculation of the 50% inhibitory concentration (IC 50 ) values was performed using GraphPad Prism for Windows (Version 5.02) software. The test was carried out using SPSS (version 1) software to study the comparison between treatment of samples and untreated control. A value of p < 0.05 was considered significantly different.,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy--γ,γ-dimethylallylflavone (1) : Pale yellow solid (. mg, 1.%). R f 0.3 (n-hex:etac = 1:1); IR (ATR) υ max cm -1 : 3393 (), 293 (sp 3 CH), 19 (C=), 122 and 151 (C=C aromatic), and 120 (C-); 1 H NMR (CD 3 CCD 3, 00 MHz) ppm: δ 13. (1H, s, 5-),.3 (1H, d, J =. Hz, H-'),. (1H, s, H-3),.3 (1H, dd,, J = 1. Hz and.2 Hz, H-10),.5 (1H, dd, J =. Hz and 2. Hz, H-5'),.0 (1H, d, J = 1. Hz, H-9),.5 (1H, d, J = 2. Hz, H-),.2 (1H, d, J = 9.2 Hz, H-1), 5. (1H, d, J = 9.2 Hz, H-15),.00 (3H, s, CH 3 ), 2.5 ( 1H, m, H-11), 1.9 (3H, s, H-), 1.0 (3H, s, H-1), 1.09 ( H, s, H-12 and H-13); 13 C NMR (CD 3 CCD 3, 100 MHz) ppm: δ.5 (C-), 13.2 (C-), 12. (C-2'), 15.9 (C-a), 155. (C-2), 155.3 (C-5), 11. (C- 10), 13.0 (C-1), 125. (C-'), 121.2 (C-15), 115. (C-9), 109.9 (C-5'), 109.5 (C-), 109.2 (C-), 10.5 (C-1'), 105.1 (C-a), 10.0 (C-), 90.2 (C-3), 9.5 (C-1), 55. (-CH 3 ), 33.1 (C-11), 2.9 (C-1), 22.2 (C-12 and C-13),. (C-); EIMS m/z (% rel. int.): 3 [M] +, C 2 H 2. sp 3 CH 12 13 1 15 1 1 11 9 10 a 5 a 1' 2' 2 3 ' 5' CH 3 S1: IR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy--γ,γ-dimethylallylflavone (1)

1.03 1.025 1.0 1.0 1.031 1.052 1.00 1.0.29.50.219.22.51.5.55.5.05.0.3.2.5..95.00.13.1.0.5.9 5. 5.9 5.2 5.99 5.503 5.50 1.000 1.021 1.03 1.00 1.052 1.025 1.0 1.0 1.031 1.0 3.050 1.090 3.01 3.02.02 13. 1.9 1.90 2.0 2.23 2.0 2.55 2.5 2.2 2. 2.9.009 5. 5.9 5.2 5.99 5.503 5.50.219.22.51.5.55.5.05.0.3.2.5..95.00.13.1.0.5.9.29.50 9.01 H-12/13 H- 2'-CH 3 H-11 H-1 5- - 1 13 12 11 10 9 5 3 2 1 ppm S2: 1 H NMR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy--γ,γdimethylallylflavone (1) H-10 H-5' H-9 H-' H-3 H- H-1 H-15.0....2.0....2.0 5. 5. 5. ppm S3: 1 H NMR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy--γ,γdimethylallylflavone (1) (Expansion) 5

. 155.33 155.5 15.20 15.993 12.1 13.13 13.01 11.1 10.019 105.109 10.510 109.20 109.5 109.9 115.955 121.22 125.350 90.19 9.505 55.5 33.051.5 22.1 2.950 12 13 1 15 1 1 11 9 10 a 5 a 1' 2' 2 3 ' 5' CH 3 10 3 1 CH 3 210 200 190 10 0 10 150 10 130 120 110 100 90 0 0 0 50 0 30 20 10 ppm S: 13 C NMR spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy--γ-γ-dimethylallflavone (1) 12 13 1 15 1 1 11 9 10 a 5 a 1' 2' 2 3 ' 5' CH 3 m/z 3 [M] + C 2 H 2 S5: EIMS spectrum of,5-dihydroxy-,-(2,2-dimethylpyrano)-2'-methoxy--γ-γ-dimethylallflavone (1)

Cudraflavone A (2) : Yellow needles (5. mg, 0.03%). R f 0.5 (n-hexane: EtAc = :1), ); IR (ATR) υ max cm -1 : 303 (), 29 (sp 3 CH), 19 (C=), 122 and 151 (C=C aromatic), and 120 (C-); 1 H NMR (CD 3 CCD 3, 00 MHz) δ ppm: 13.29 (s, 5-),. (1H, d, J =. Hz, H-5'),. (1H, d, J = 10.0 Hz, H-1),. (1H, dd, J =. Hz and 2. Hz, H-),. (1H, s, H-),. (1H, d, J = 2. Hz, H- 2'),.21 (1H, d, J = 9.2 Hz, H-10), 5.9 (1H, d, J = 10.0 Hz, H-15), 5.9 (1H, d, J = 9.2 Hz, H-9), 1.9 (3H, s, H-), 1.0 (3H, s, H-1), 1. (H, s, H-12 and H-13) ; 13 C-NMR (CD 3 CCD 3, 100 MHz) δ ppm:.3 (C-), 13. (C-), 159.0 (C-5), 15.2 (C-2), 15. (C-a), 15.3 (C-), 155. (C-'), 13.9 (C-11), 12.5 (C-15), 125. (C-5'), 121.2 (C-9), 11.9 (C-1), 110.1 (C-'), 109.1 (C-), 10. (C-3), 105.2 (C-), 105.2 (C-a), 10.0 (C-2'), 9.9 (C-),. (C-1), 9. (C-10), 2. (C-12 and C-13), 2.9 (C-1),. (C-); EIMS m/z (% rel. int.): 1 ([M] +, C 25 H 22. sp 3 CH 1 1 15 1 a 5 a 2' 1' 2 ' 3 9 10 11 5' 13 Chelated C= 12 S: IR spectrum of cudraflavone A (2)

1.22 2.021 0.2 1.203 1.13 1.1 1.25 1.25.1.3.3.9.1.3..59.5.3..203.22 5.5 5.90 5.3 5. 5.0 5.9 5.500 5.503 1.000 1.22 1.25 1.25 1.1 1.203 1.13 2.021 0.2.02 3.29 3. 13.29 5.3 5. 5.0 5.9 5.500 5.503 5.5 5.90.203.22.3.9.1.3..59.5.3..1.3 1. 1.99 1.02 1.90 1.93 H- H-1 5- H-12/13 13 12 11 10 9 5 3 2 1 ppm S: 1 H NMR spectrum of cudraflavone A (2) H-5' H- H- H-2' H-10 H-15 H-1 H-9.3.2.1.0.9....5..3.2.1.0.9....5..3.2.1.0 5.9 5. 5. 5. 5.5 ppm S: 1 H NMR spectrum of cudraflavone A (2) (Expansion)

1 1 15 1 a 5 a 2' 1' 2 ' 3 9 10 11 5' 13 12 11 9 1 10 S9: 13 C NMR spectrum of cudraflavone A (2) m/z 1 [M] + C 25 H 22 S10: EIMS spectrum of cudraflavone A (2) 9

Artocarpin (3): range powder (1.2 mg, 0.23%); R f 0.3 (n-hexane: EtAc = 3:2), IR (ATR) υ max cm -1 : 335 (), 295 (sp 3 CH), 1 (C=), 115 and 1 (C=C aromatic) and 1203 (C-); 1 H-NMR (CD 3 CCD 3, 00 MHz) δ ppm: 13.9 (1H, s, 5-),. (2H, s, 2'-/-),.2 (1H, d, J =. Hz, H-'),. (1H, dd, J = 1. Hz and.2 Hz, H-15),.2 (2H, d, J = 1. Hz, H-1),.5 (1H, d, J = 2. Hz, H-),.5 (1H, s, H-),.5 (1H, dd, J =. Hz and 2. Hz, H-5'), 5.13 (1H, t, J =.2 Hz, H-10), 3.9 (3H, s, -CH 3 ), 3.1 (1H, d, J =.2 Hz, H-9), 2.5 (1H, m, H-1), 1.59 (3H, s, H-13), 1.5 (3H, s, H-12), 1.10 (H, d, J =. Hz, H- and H-1); 13 C-NMR (CD 3 CCD 3, 100 MHz) δ ppm: 12. (C-), 12.9 (C-), 11. (C-2), 10. (C-5), 15.9 (C-), 15. (C-a), 15.3 (C-2'), 11.3 (C-15), 131. (C- '), 121.1 (C-10), 121.0 (C-3), 11.1 (C-1), 112.0 (C-1'), 10.9 (C-), 10.2 (C-5'), 10. (C-a), 102.9 (C-), 9. (C-), 55. (-CH 3 ), 33.1 (C-1), 2.9 (C-12), 23. (C-9), 22.2 (C- and C-1), 1. (C- 13); EIMS m/z(% rel. int.): 3 (100) ([M] +, C 2 H 2. H 3 C 1 1 15 a 5 a H 1' 2 ' 3 2' 10 sp 3 CH 9 11 5' 13 Chelated C= 1 12 S11: IR spectrum of artocarpin (3) 10

H 3 C 1 1 15 1 a 5 a H 1' 2 ' 3 2' 10 9 12 11 5' 13 S12: 1 H NMR spectrum of artocarpin (3) H-3 H- H-' H- H-1 H-15 H-5' H-1 S13: 1 H NMR spectrum of artocarpin (3) (Expansion) 11

15 10 -CH 3 1 S1: 13 C NMR and DEPT spectrum of artocarpin (3) H 3 C 1 1 15 a 5 a H 1' 2 ' 3 2' 10 9 11 5' 13 m/z 3 [M] + C 2 H 2 1 12 S15: EIMS spectrum of artocarpin (3) 12

Cycloartobiloxanthone (): range powder ( mg, 0.0%) : R f 0.5 (n-hexane: EtAc = 3:2), IR (ATR) υ max cm -1 : 30 (), 295 (sp 3 CH), 122 (C=); 1 H-NMR (CD 3 CCD 3, 00 MHz) δ ppm: 13.1 (s, 5- ),.95 (1H, d, J = 10.0 Hz, H-1),.3 (1H, s, H-),.1 (1H, s, H-), 5.0 (1H, d, J = 10.0 Hz, H- 15), 3. (1H, dd, J =15.2 and. Hz, H-9b), 3.23 (1H, dd, J = 15.2 and. Hz, H-10), 2.0 (1H, t, J = 15.2 Hz, H-9a), 1. (3H, s, H-13), 1. (H, s, H-/1), 1.3 (3H, s, H-12); 13 C-NMR (CD 3 CCD 3, 100 MHz) δ ppm: 10.5 (C-), 11. (C-), 10. (C-2), 15. (C-5), 151.2 (C-a), 150. (C-2'), 1.2 (C-), 13.0 (C-5'), 132. (C-'), 12.9 (C-15), 115.0 (C-1), 111. (C-3), 10.5 (C-1'), 103.9 (C-a), 100.9 (C-), 99.0 (C-), 92. (C-11),. (C-1),. (C-10), 31.3 (C-12), 2.5 (C- and C-1), 22. (C-13), 19.5 (C-9); EIMS m/z (% rel. int.): 3 [M] +, C 25 H 22. sp 3 CH 1 15 2' 1 1 a 5 a H 2 3 1' ' 10 11 9 5' 13 12 Chelated C= S1 : IR spectrum of cycloartobiloxanthone () 13

1.1 1.220 1.119 1.1.002.03.933.95.155 5. 5.0 1.000 1.1 1.220 1.119 1.1 0.9 0.993 1.31 0.9.1.09.9 13.09.933.95.002.03.155 5. 5.0 1.303 1.3 1.33 1. 1.900 1.90 1.909 1.91 2.32 2.20 3.20 3.221 3.21 3.259 3.1 3.32 3.53 3.0 1 15 2' 1 1 a H 2 1' ' 5' H-13 a 5 3 10 11 9 13 12 H-/1 5- H-9a H-9b H-10 H-12 13 12 11 10 9 5 3 2 1 ppm S: 1 H NMR spectrum of cycloartobiloxanthone () H-15 H-1 H- H-.5..3.2.1.0.9....5..3.2.1.0 5.9 5. 5. 5. 5.5 5. 5.3 ppm S1: 1 H NMR spectrum of cycloartobiloxanthone () (Expansion) 1

10.535 15.33 10.19 11.2 1.1 150.1 151.5 15 13.059 132.9 12.99 12.00 12.00 12.00 111.05 115.0 115.051 115.0 115.0 99.0 100.9 103.9 10.519 10. 10. 10. 99.05 99.01 99.0 92.10.33.3.0.2.3 19.5 21.9 22.3 2.33 2.35 2.553 31.39 31.3 19.5 21.950 2.33 2.3 2.553 19. 21.93 22.30 13.10 13.3 13.39 11 1 10 9 210 200 190 10 0 10 150 10 130 120 110 100 90 0 0 0 50 0 30 20 10 ppm 210 200 190 10 0 10 150 10 130 120 110 100 90 0 0 0 50 0 30 20 10 ppm 210 200 190 10 0 10 150 10 130 120 110 100 90 0 0 0 50 0 30 20 10 ppm 210 200 190 10 0 10 150 10 130 120 110 100 90 0 0 0 50 0 30 20 10 ppm S19: 13 C NMR and DEPT spectrum of cycloartobiloxanthone () m/z 3 [M] + C 25 H 22 S20: EIMS spectrum of cycloartobiloxanthone () 15

Artonin E (5): range powder (5.3 mg, 0.0%) R f 0.3 (n-hex:etac = 3:2); IR (ATR) υ max cm -1 : 3393 (), 293 (sp 3 CH), 19 (C=), 122 and 151 (C=C aromatic), and 120 (C-), (CD 3 CCD 3, 00 MHz) δ ppm: 13.2 (s, 5-),.9 (1H, s, H-'),.3 (1H, d, J = 10.0 Hz, H-1),.1 (1H, s, H-),. (1H, s, H-), 5.9 (1H, d, J = 10.0 Hz, H-15), 5.1 (1H, t, J =.2 Hz, H-10), 3.15 (2H, d, J =.2 Hz H- 9), 1.59 (3H, s, H-13), 1. (3H, s, H-12), 1. (H, s, H- and 1); 13 C-NMR (CD 3 CCD 3, 100 MHz) δ ppm: 12. (C-), 11. (C-5), 11.1 (C-2), 159.0 (C-), 152.3 (C-a), 1. (C-), 13.1 (C-5'), 131. (C-11), 12.1 (C-15), 121.5 (C-10), 120. (C-3), 11.1 (C-ʹ), 11.5 (C-1), 110.5 (C-1'), 10. (C-a), 103. (C-), 100. (C-), 9. (C-),. (C-1), 2.3 (C- and C-1), 2.9 (C-12), 23. (C-9), 1. (C-13). EIMS m/z (% rel. int.): 3 [M] +, C 25 H 2. sp 3 CH Chelated C= 1 15 1 5 1 a a H 1' 5' 2 ' 3 2' 10 9 11 12 13 S21 : IR spectrum of artonin E (5) 1

12.00 159.0 11.9 1.01 1.9 13.229 9.09 100.2 103.0 10. 11.55 11.19 120.9 121.02 12.121 131.1. 23. 2.91 1. 1.000 1.2 1.13 0.25 1.22 1.50 1.01 2.91.03 5.11 3.1 13.2 3.15 3. 5.12 5.132 5.135 5.12 5.1 5.19 5.153 5.15 5.13 5. 5.0 5. 5.92.15.1.09.12.13.3.3.9 1.0 1. 1.590 1.593 5-1 15 1 5 1 a a H 1' 5' 2 ' 3 2' 10 9 13 11 12 H-1 H- H- H-13 H-12 H-' H-15 H-10 H-9 H-/1 13 12 11 10 9 5 3 2 1 ppm S22 : 1 H NMR spectrum of artonin E (5) 5' ' 1 13 1 15 1 5 1 a a H 1' 5' 2 ' 3 2' 10 9 11 12 13 210 200 190 10 0 10 150 10 130 120 110 100 90 0 0 0 50 0 30 20 10 0 ppm S23 : 13 C NMR spectrum of artonin E (5)

m/z 3 [M] + C 25 H 2 S2: EIMS spectrum of artonin E (5) 1

xyresveratrol () Yellow solid; (2. mg, 0.5%). R f 0.3 (n-hexane:et 2 = 2:3); IR (ATR) υ max cm -1 : 3209 (), 2921 (sp 3 CH), 159 (C=C aromatic); 1 H NMR (CD 3 CCD 3, 00 MHz) δ ppm :.3 (1H, d, J =. Hz, H-),.35 (1H, d, J =1. Hz, H-β),.91 (1H, d, J = 1. Hz, H-α ),.53 (1H, d, J = 2.0 Hz, H-2' and H- '),.5 (1H, d, J = 2. Hz, H-3),.39 (1H, dd, J =., 2. Hz, H-5),.2 (1H, t, J = 2.0 Hz, H-); 13 C-NMR (CD 3 CCD 3, 100 MHz) δ ppm : 15. (C-,5'), 15.2 (C-2), 15.0 (C-), 10. (C-1'), 12. (C-), 125.5 (C-α), 123.5 (C-β), 11. (C1), 10. (C-5), 10. (C-2' and C-'), 102. (C-3), 101. (C-). EIMS m/z (% rel. int.): 2 [M] +, C 1 H 12. sp 3 CH H ' 5' α 1' 2' 5 1 β 2 3 S25: IR spectrum of oxyresveratrol () 19

15.021 15.229 15.2 10.9 123. 125. 12.359 11.25 10.55 101.35 102.5 10.0 1.000 1.022 1.005 2.150 0.95 1.090 0.93.15.3.31.330.92.5.533.53.33.39.0.10.50.5.23.29.25 H ' 5' α 1' 2' 5 1 β 2 3 H-2', H-' H-3 H- H- H-β H-α H-5.5..3.2.1.0.9....5..3 ppm S2: 1 H NMR spectrum of oxyresveratrol () H ' 5' α 1' 2' 5 1 β 2 3 2ˈ/ˈ 1' 1 5 0 15 10 155 150 15 10 135 130 125 120 115 110 105 100 95 ppm S2: 13 C NMR spectrum of oxyresveratrol () 20

m/z 2 [M] + C 1 H 12 S2: EIMS spectrum of oxyresveratrol () 21

Macakurzin C (): Yellow needle (3.3 mg, 0.0%); R f 0.3 (n-hexane:etac = 1:1); IR (ATR) υ max cm - 1 : 3319 (), 29 and 2 (sp 3 CH), 152 (C=) and 15 (C=C aromatic); 1 H NMR (CDCl 3, 00 MHz) δ ppm: 12.21 (1H, s, 5-),.3 (1H, s, 3-),.32 (2H, d, J =. Hz, H-2' and H-'),.5 (2H, t, J =. Hz, H- and H-5'),.55 (1H, d, J =.5 Hz, H-),.9 (1H, d, J = 10.0 Hz, H-9),.23 (1H, s, H-), 5.0 (1H, d, J = 10.0 Hz, H-10), 1.50 (H, s, H-12 and H-13); 13 C NMR (CDCl 3, 100 MHz) δ ppm: 5. (C-), 10.5 (C-), 10.0 (C-9), 151.2 (C5), 1. (C-2), 13. (C-3), 130.9 (C-1'), 130.3 (C- ), 12.9 (C-'), 12. (C- and C-5'), 12.5 (C-2' and C-'), 11. (C-'), 103. (C-), 101. (C-10), 99. (C-),.3 (C-9'), 29. (C-10'), 2.2 (C-11'); EIMS m/z (% rel. int.): 33 [M] + C 20 H 1 5 ). 12 13 9 a 5 a 2 1' 3 sp 3 CH S29: IR spectrum of macakurzin C () 22

5- S30: 1 H NMR spectrum of macakurzin C () H- H-2'/' H-10 3- H-/5' H- H-9 S31: 1 H NMR spectrum of macakurzin C () (Expansion) 23

/5' 2'/' 11' 10' 5 2 S32: 13 C NMR spectrum of macakurzin C () m/z 33 [M] + C 20 H 1 5 2

S33: EIMS spectrum of macakurzin C () Flemichapparin A (): red solid (.5 mg, 0. %); R f 0.5 (n-hexane:etac = :1); IR (ATR) υ max cm -1 : 322 (), 29 (sp 3 CH), 159, 15 (C=C aromatic); 1 H-NMR (CD 3 CCD 3, 00 MHz) δ ppm : 1.13 (1H, s, 5-),.13 (1H, d, J = 1.0 Hz, H-β),.9 (1H, d, J=1.0 Hz, H-α),.1-. (5H, m, H-2'-H- '), δ.1 (1H, d, J=10.0 Hz, H-1"),.00 (1H, s, H-), 5.52 (1H, d, J=10.0 Hz, H-2"), 1.5 (H, s, H- "/5"); 13 C NMR (CD 3 CCD 3, 100 MHz) δ ppm: 192.9 (C-1), 1. (C-3), 15.3 (C-), 15. (C-5), 12.3 (C-α), 135. (C-1'), 130.9 (C-1'), 12. (C-,C- and C-5'), 12.5 (C-2' and C-'), 12.5 (C-β), 12. (C-2"), 10. (C-1"), 102. (C-), 9. (C-),.2 (C-3"), 2.1 (C-5"), 2.0 (C-"); EIMS m/z (% rel. int.): 322 [M] +, C 20 H 1. sp 3 CH ' 5'' ' 2'' 1'' 5 3 2 1 2' 1' ' 5' S33: IR spectrum of flemichapparin A () 25

5- S3: 1 H NMR spectrum of flemichapparin A () H-β H-α H-2'/' H-//5' H-2" H-1" H- S35: 1 H NMR spectrum of flemichapparin A () (Expansion) 2

1 5 α 3" 5" " S3: 13 C NMR spectrum of flemichapparin A () m/z 322 [M] + C 20 H 1 S3: EIMS spectrum of flemichapparin A () 2

Luteolin (9): yellow solid (13 mg, 0.13%) ; R f 0.55 (n-hexane:etac = 2:3); IR (ATR) υ max cm -1 : 321 (), 295, 2925 and 232 (sp 3 CH), 153 (C=), 10 (C=C aromatic); 1 H NMR (CD 3 CCD 3, 00 MHz) δ ppm: 13.05 (1H, s, 5-),.15 (1H, d, J = 2. Hz, H-2'),.9 (1H, dd, J =. Hz and 2. Hz, H-'),.01 (1H, d, J =. Hz, H-5'),.0 (1H, s, H-3),.5 (1H, d, J = 2.0 Hz, H-),.2 (1H, d, J = 2.0 Hz, H-); 13 C-NMR (CD 3 CCD 3, 100 MHz) δ ppm: 12.1 (C-), 1.3 (C-2), 1.1 (C-5), 12.5 (C-), 15.9 (C-a), 19.5 (C-), 15. (C-), 122. (C-1'), 119.2 (C-'), 115. (C-5'), 113.2 (C-2'), 10. (C- a), 103.3 (C-3), 9. (C-5), 93. (C-). EIMS m/z (% rel. int.): 2 [M] +, C 15 H 10. sp 3 CH H a 2 ' 1' a 3 5' 2' S3: IR spectrum of luteolin (9) 2

5- S39: 1 H NMR spectrum of luteolin (9) H-2' H-' H-5' H-3 H- H- S0: 1 H NMR spectrum of luteolin (9) (Expansion) 29

2 5 a 1' ' 5' 2' a 3 5 S1: 13 C spectrum of luteolin (9) m/z 2 [M] + C 15 H 10 S2: EIMS spectrum of luteolin (9) 30

Apigenin (10): yellow solid (5 mg, 0.05%); R f 0.5 (n-hexane:etac = 1:1); IR (ATR) υ max cm -1 : 32 (), 292, 2920 and 22 (sp 3 CH), 15 (C=), 10 (C=C aromatic); 1 H NMR (CD 3 CCD 3, 00 MHz) δ ppm : 13.0 (1H, s, 5-),.9 (2H, d, J =. Hz, H-2' and H-'),.0 (2H, d, J =. Hz, H- and H-5'),.5 (1H, s, H-3 ),.5 (1H, d, J = 2.0 Hz, H-),.2 (1H, d, J = 2.0 Hz, H-); 13 C-NMR (CD 3 CCD 3, 100 MHz) δ ppm : 12.2 (C-), 1.2 (C-5), 1.0 (C-2), 12.5 (C-), 11.0 (C-9), 15.9 (C-), 12. (C-2'/'), 122. (C-1'), 115.9 (C-/5'), 10.5 (C-10), 103.2 (C-3), 9. (C-), 93. (C-); EIMS m/z (% rel. int.): 20 [M] +, C 15 H 10 5. sp 3 CH H a 2 ' 1' a 3 5' 2' S3: IR spectrum of apigenin (10) 31

H-/5' H-3 5- H- H-2'/' H- S: 1 H NMR spectrum of apigenin (10) /5' 2'/' 2 5 9 1' 10 3 S5: 13 C NMR spectrum of apigenin (10) 32

m/z 20 [M] + C 15 H 10 5 S: EIMS spectrum of apigenin (10) References [1] Fu, R., Zhang, Y., Guo, Y., Liu, F., Chen, F. (201). Determination of Phenolic Contents and Antioxidant Activities of Extracts of Jatropha curcas L. Seed Shell, A By-Product, A New Source of Natural Antioxidant, Industrial Crops and Products, 5, 25-20. [2] Zuo, Y., Chang, S. K. C., Gu, Yan. And Qian, Y. (2011). Antioxidant activity and phenolic compositions of lentil (Lens culinaris var. Mortaon) extract and its fractions, J. Agric Food Chem, 59 (), 22-22. [3] Channarong, S, Jutiviboonsuk, A. and Korsanan, S. (2012). Total Reducing Antioxidant Capacity of Thai Herbal Aromatic Powder (Ya-Hom) Measured by FRAP Assay, Thai Pharmaceutical and Health Science Journal, (3), 111-11. 33