SUPPLEMENTARY MATERIAL

A new hemiterpene glycoside from the ripe tomatoes

Masateru Onoa,*, Shin Yasudaa, Yuki Shionoa, Chisato Furusawaa, Shinya

Inabaa, Takayuki Tanakaa, Tsuyoshi Ikedab and Toshihiro Noharab

aSchool of Agriculture, Tokai University, Minamiaso 5435, Aso, Kumamoto 869-

1404, Japan; bFaculty of Pharmaceutical Sciences, Sojo University, 4-22-1 Ikeda,

Nishi-ku, Kumamoto 860-0082, Japan

*Corresponding author: E-mail: mono@agri.u-tokai.ac.jp

Abstract

A new hemiterpene glycoside (1) was isolated from ripe tomatoes (the fruit of Lycopersicon

esculentum, Solanaceae) along with eight known compounds. The chemical structure of 1 was

determined to be 2-methylbutan-1-ol -D-glucopyranosyl-(1→6)--D-glucopyranoside, based

on spectroscopic data as well as chemical evidence. In addition, the radical-scavenging

activities of the isolated compounds on the free radical of 1,1-diphenyl-2-picrylhydrazyl were

examined. Among the tested compounds, tryptophan, 4-O--D-glucopyranosyl caffeic acid,

and dihydro-p-coumaryl alcohol -O--D-glucopyranoside demonstrated 42.0%, 50.1%, and

76.0% scavenging activities, respectively, at a concentration of 0.5 mM.

Table S1. 1H- and 13C-NMR spectral data for 1 (in pyridine-d5)

H C

Agl-1a 4.08* 75.0Agl-1b 3.99 dd(6.5, 9.5)Agl-2 1.72 m 35.4Agl-3a 1.51 m 26.4Agl-3b 1.11 mAgl-4 0.80 t(7.5) 11.4Agl-5 0.94 d(7.0) 16.8Glc-1 4.76 d(8.0) 104.9Glc-2 4.00 dd(8.0, 8.5) 75.1Glc-3 4.21 dd(8.5, 8.5) 78.5Glc-4 4.16 dd(8.5, 8.5) 71.8Glc-5 4.11 m 77.3Glc-6a 4.87 dd(1.5, 11.5) 70.2Glc-6b 4.36 dd(6.5, 11.5)Glc'-1 5.15 d(7.5) 105.5Glc'-2 4.06* 75.3Glc'-3 4.26* 78.5Glc'-4 ca. 4.27 71.7Glc'-5 3.95 m 78.5Glc'-6a 4.54 dd(1.5, 12.0) 62.8Glc'-6b 4.39 dd(4.5, 12.0)

in ppm from TMS (coupling constants (J) in Hz are given in parentheses). Agl, aglycone moiety. Glc, inner glucosyl group. Glc', terminal glucosyl group. *, Signals were overlapping. 1H-NMR spectral data (500 MHz). 13C-NMR spectral data (125 MHz).

Table S2. 1H- and 13C-NMR spectral data for 8 (in pyridine-d5)

H C H C

Agl-1a 1.48* 37.2 Gal-1 4.90 d(7.5) 102.4Agl-1b 0.79* Gal-2 4.41 dd(7.5, 9.5) 73.2Agl-2a 2.06 m 29.8 Gal-3 4.18* 75.4Agl-2b 1.61* Gal-4 4.61 d(3.5) 79.9Agl-3 3.94* 78.7 Gal-5 4.05* 75.6Agl-4a 1.81* 34.8 Gal-6a 4.64 dd(8.5, 11.0) 60.6Agl-4b 1.40* Gal-6b 4.23*

Agl-5 0.90* 44.6 Glc-1 5.15 d(8.0) 105.1Agl-6a 1.14* 28.8 Glc-2 4.34 dd(8.0, 8.5) 81.3Agl-6b 1.09* Glc-3 4.11 dd(8.5, 8.5) 86.8Agl-7a 1.50* 32.3 Glc-4 3.78 dd(8.5, 8.5) 70.5Agl-7b 0.81* Glc-5 3.85* 77.3Agl-8 1.27* 34.9 Glc-6a 4.49 dd(1.5, 10.0) 63.0Agl-9 0.51 ddd(2.5, 8.5, 8.5) 54.5 Glc-6b 4.03*

Agl-10 35.8 Glc'-1 5.54 d(7.5) 104.8Agl-11a 1.37* 20.7 Glc'-2 4.07 dd(7.5, 8.5) 76.2Agl-11b 1.11* Glc'-3 4.09* 77.6Agl-12a 1.65* 38.2 Glc'-4 4.17* 71.1Agl-12b 1.00 m Glc'-5 3.93* 77.8Agl-13 41.8 Glc'-6a 4.52 dd(1.5, 10.5) 62.5Agl-14 0.92* 54.5 Glc'-6b 4.37*

Agl-15a 2.17 ddd(6.5, 7.5, 14.0) 33.2 Xyl-1 5.19 d(8.0) 104.9Agl-15b 1.39* Xyl-2 3.95 dd(8.0, 8.5) 75.1Agl-16 4.99 ddd(4.5, 7.5, 7.5) 82.7 Xyl-3 4.18* 78.6Agl-17 1.84 br d(7.5) 59.0 Xyl-4 4.12* 70.7Agl-18 0.70 s 13.8 Xyl-5a 4.23* 67.3Agl-19 0.62 s 12.2 Xyl-5b 3.68 dd (10.5, 10.5)Agl-20 2.70 q(7.5) 36.3Agl-21 1.31 d(7.5) 17.9Agl-22 181.1

in ppm from TMS (coupling constants (J) in Hz are given in parentheses). Agl, aglycone moiety. Gal, galactosyl group. Glc, inner glucosyl group. Glc', terminal glucosyl group. Xyl, Xylosyl group. *, Signals were overlapping. 1H-NMR spectral data (500 MHz). 13C-NMR spectral data (125 MHz).

Figure S1. Connectivities elucidated by the 1H-1H COSY spectrum (bold lines) and 1H-13C long-range correlations (arrows) observed in the HMBC spectrum of 1 (in pyridine-d5, 500 MHz)

Figure S2. DPPH radical-scavenging effects of 3, 4, 6, and Trolox. Data shown represent mean ± S.D. derived from four determinations. The final concentration of each sample tested was 0.5 mM.

Figure S3. 1H-NMR spectrum of 1 (in pyridine-d5, 500 MHz)

Figure S4. 13C-NMR spectrum of 1 (in pyridine-d5, 125 MHz)

Figure S5. 1H-1H COSY spectrum of 1 (in pyridine-d5, 500 MHz)

Figure S6. HMQC spectrum of 1 (in pyridine-d5, 500 MHz)

Figure S7. HMBC spectrum of 1 (in pyridine-d5, 500 MHz)

Figure S8. Positive FAB-MS of 1

Figure S9. Positive FAB-MS of 1

Figure S10. 1H-NMR spectrum of 8 (in pyridine-d5, 500 MHz)

Figure S11. 13C-NMR spectrum of 8 (in pyridine-d5, 125 MHz)

Figure S12. 1H-1H COSY spectrum of 8 (in pyridine-d5, 500 MHz)

Figure S13. HMQC spectrum of 8 (in pyridine-d5, 500 MHz)

Figure S14. HMBC spectrum of 8 (in pyridine-d5, 500 MHz)