SUPPLEMENTARY MATERIAL
Experimental data of compounds 4-7, 9 and 11
-sitosteryl glucopyranoside (4): 1H NMR (pyridine-d5, 500 MHz)  3.95 (1H, m, H-3),
2.72 (1H, brd, J = 13.0 Hz, H-4a), 2.47 (1H, brt, J = 13.0, H-4b), 5.34 (1H, dd, J = 4.5, 2.0
Hz, H-6), 0.66 (3H, s, H3-18), 0.93 (3H, s, H3-19), 0.98 (3H, d, J = 6.5 Hz, H3-21), 0.88
(3H, d, J = 7.0 Hz, H3-26), 0.85 (3H, d, J = 7.0 Hz, H3-27), 0.89 (3H, t, J = 7.0 Hz, H3-29),
5.05 (1H, d, J = 7.5 Hz, H-1), 4.05 (1H, t, J = 8.0 Hz, H-2), 4.29 (1H, t, J = 8.5 Hz, H-3),
4.27 (1H, t, J = 8.5 Hz, H-4), 3.98 (1H, m, H-5); 13C NMR (pyridine-d5, 125 MHz) 
37.5(C-1), 30.3 (C-2), 78.6 (C-3), 39.4 (C-4), 140.9 (C-5), 121.9 (C-6), 32.2 (C-7), 32.1 (C8), 50.4 (C-9), 36.9 (C-10), 21.3 (C-11), 40.0 (C-12), 42.5 (C-13), 56.8 (C-14), 23.4 (C-15),
28.5 (C-16), 56.3 (C-17), 12.0 (C-18), 19.9 (C-19), 36.4 (C-20), 19.4 (C-21), 34.2 (C-22),
26.4 (C-23), 46.1 (C-24), 29.5 (C-25), 19.2 (C-26), 19.0 (C-27), 24.5 (C-28), 12.2 (C-29),
102.6 (C-1), 75.3 (C-2), 78.6 (C-3), 71.7 (C-4), 78.5 (C-5), 62.9 (C-6).
p-Hydroxyphenethyl trans-ferulate (5): 1H NMR (CD3OD, 500 MHz)  7.10 (1H, d, J = 1.5
Hz, H-2), 6.78 (1H, d, J = 8.0 Hz, H-5), 7.01 (1H, dd, J = 8.5, 2.0 Hz, H-6), 7.42 (1H, d, J
= 15.5 Hz, H-7), 6.39 (1H, d, J = 15.5 Hz, H-8), 7.04 (2H, dd, J = 8.5, 2.0 Hz, H-2'. H-6'),
6.71 (2H, dd, J = 8.5, 2.0 Hz, H-3', 5'), 3.45 (2H, t, J = 7.5 Hz, H-1''), 2.74 (2H, t, J = 7.5
Hz, H-2''), 3.87 (3H, s, OCH3); 13C NMR (CD3OD, 125 MHz)  128.8 (C-1), 111.6 (C-2),
149.3 (C-3), 149.8 (C-4), 116.5 (C-5), 123.2 (C-6), 142.0 (C-7), 118.8 (C-8), 169.2 (C-9),
131.3 (C-1'), 130.7 (C-2'), 116.3 (C-3'), 156.9 (C-4'), 116.3 (C-5'), 130.7 (C-6'), 42.5 (C-1''),
35.8 (C-2''), 56.4 ( OCH3).
20-Hydroxyecdysone (6): 1H NMR (CD3OD, 400 MHz)  1.78 (1H, m, H-1a), 1.42 (1H, t, J
= 14.4 Hz, H-1b), 3.82 (1H, ddd, J = 12.0, 4.0, 2.8 Hz, H-2), 3.93 (1H, brq, J = 2.8 Hz, H3), 1.76 (1H, m, H-4a), 1.67 (1H, m, H-4b), 2.37 (1H, dd, J = 12.8, 4.8 Hz, H-5), 5.80 (1H,
d, J = 2.4 Hz, H-7), 3.14 (1H, ddd, J = 10.4, 7.2, 2.4 Hz, H-9), 1.74 (1H, m, H-11a), 1.66
(1H, m, H-11b), 2.13 (1H, td, J = 13.2, 5.2 Hz, H-12a), 1.87 (1H, brd, J = 13.2 Hz, H-12b),
1.95 (1H, m, H-15a), 1.58 (1H, brt, J = 10.8 Hz, H-15b), 1.98 (1H, brt, J = 11.2 Hz, H-16a),
1
1.81 (1H, m, H-16b), 2.39 (1H, m, H-17), 0.88 (3H, s, H3-18), 0.96 (3H, s, H3-19), 1.18 (3H,
s, H3-21); 3.32 (1H, dd, J = 10.4, 1.6 Hz, H-22), 1.68 (1H, m, H-23a), 1.28 (1H, m, H-23b),
1.79 (1H, m, H-24a), 1.42 (1H, t, J = 11.6 Hz, H-24b), 1.18 (3H, s, H3-26), 1.19 (3H, s, H327); 13C NMR (CD3OD, 100 MHz)  37.4(C-1), 68.7 (C-2), 68.5 (C-3), 32.9 (C-4), 51.8 (C5), 206.4 (C-6), 122.1 (C-7), 168.0 (C-8), 35.1 (C-9), 39.3 (C-10), 21.5 (C-11), 32.5 (C-12),
48.5 (C-13), 85.2 (C-14), 31.8 (C-15), 21.5 (C-16), 50.3 (C-17), 18.1 (C-18), 24.4 (C-19),
77.9 (C-20), 21.1 (C-21), 78.4 (C-22), 27.4 (C-23), 42.4 (C-24), 71.3 (C-25), 29.3 (C-26),
29.0 (C-27).
20-Hydroxy-24-hydroxymethylecdysone (7): 1H NMR (CD3OD, 500 MHz)  1.77 (1H, m,
H-1a), 1.45 (1H, m, H-1b), 3.83 (1H, dt, J = 12.0, 4.0 Hz, H-2), 3.94 (1H, brs, H-3), 1.72
(1H, m, H-4a), 1.67 (1H, m, H-4b), 2.37 (1H, dd, J = 12.0, 4.5 Hz, H-5), 5.80 (1H, d, J =
1.5 Hz, H-7), 3.15 (1H, m, H-9), 1.77 (2H, m, H2-11), 2.13 (1H, td, J = 13.0, 5.0 Hz, H12a), 1.87 (1H, brd, J = 13.0 Hz, H-12b), 1.96 (1H, m, H-15a), 1.58 (1H, m, H-15b), 1.96
(1H, m, H-16a), 1.77 (1H, m, H-16b), 2.38 (1H, t, J = 9.5 Hz, H-17), 0.88 (3H, s, H3-18),
0.96 (3H, s, H3-19), 1.10 (3H, s, H3-21); 3.33 (1H, m, H-22), 1.66 (1H, m, H-23a), 1.28 (1H,
m, H-23b), 1.90 (1H, m, H-24), 1.18 (3H, s, H3-26), 1.19 (3H, s, H3-27), 3.57 (1H, dd, J =
11.0, 6.0 Hz, H-28a), 3.50 (1H, dd, J = 11.0, 5.0 Hz, H-28b); 13C NMR (CD3OD, 125 MHz)
 37.4(C-1), 68.7 (C-2), 68.5 (C-3), 32.9 (C-4), 51.8 (C-5), 206.5 (C-6), 122.1 (C-7), 167.9
(C-8), 35.1 (C-9), 39.3 (C-10), 21.5 (C-11), 32.5 (C-12), 48.5 (C-13), 85.2 (C-14), 31.8 (C15), 21.5 (C-16), 50.5 (C-17), 18.0 (C-18), 24.4 (C-19), 79.9 (C-20), 21.1 (C-21), 78.4 (C22), 27.4 (C-23), under CD3OD signal (C-24), 71.3 (C-25), 29.7 (C-26), 28.9 (C-27), 64.4
(C-28);
C NMR (DMSO d6, 100 MHz)  36.6 (C-1), 66.5 (C-2), 66.7 (C-3), 31.5 (C-4),
13
48.6 (C-5), 202.6 (C-6), 120.4 (C-7), 165.2 (C-8), 33.1 (C-9), 37.6 (C-10), 20.2 (C-11),
30.8 (C-12), 46.8 (C-13), 82.9 (C-14), 30.3 (C-15), 20.0 (C-16), 50.1 (C-17), 17.1 (C-18),
23.9 (C-19), 75.7 (C-20), 21.0 (C-21), 76.2 (C-22), 26.1 (C-23), 48.5 (C-24), 68.7 (C-25),
30.0 (C-26), 29.0 (C-27), 63.0 (C-28).
20-Hydroxyecdysone 20,22-monoacetonide (9): White amorphous powder;
1
H NMR
(CDCl3, 500 MHz)  1.84 (1H, m, H-1a), 1.69 (1H, m, H-1b), 3.85 (1H, dt, J = 11.5, 3.5 Hz,
H-2), 4.04 (1H, brq, J = 2.5 Hz, H-3), 1.82 (1H, m, H-4a), 1.43 (1H, m, H-4b), 2.42 (1H, dd,
J = 13.0, 4.0 Hz, H-5), 5.85 (1H, d, J = 2.0 Hz, H-7), 2.98 (1H, brt, J = 9.5, Hz, H3-9), 1.77
2
(H, m, H-11a), 1.63 (1H, m, H-11b), 2.04 (1H, m, H-12a), 1.85 (1H, m, H-12b), 2.00 (1H,
m, H-15a), 1.50 (1H, m, H-15b), 2.03 (1H, m, H-16a), 1.80 (1H, m, H-16b), 2.18 (1H, t, J =
8.0 Hz, H-17), 0.80 (3H, s, H3-18), 0.98 (3H, s, H3-19), 1.16 (3H, s, H3-21); 3.63 (1H, dd, J
= 9.5, 3.0 Hz, H-22), 1.53 (2H, m, H2-23), 1.75 (2H, m, H2-24), 1.23 (3H, s, H3-26), 1.28
(3H, s, H3-27), 1.33 (3H, s, H3-2'), 1.41 (3H, s, H3-3'); 13C NMR (CDCl3, 125 MHz) 
36.7(C-1), 67.8 (C-2), 67.4 (C-3), 31.7 (C-4), 49.0 (C-5), 204.0 (C-6), 121.6 (C-7), 164.6
(C-8), 33.8 (C-9), 38.1 (C-10), 20.4 (C-11), 31.5 (C-12), 47.2 (C-13), 85.1 (C-14), 31.0 (C15), 21.1 (C-16), 50.0 (C-17), 17.1 (C-18), 23.9 (C-19), 84.3 (C-20), 21.9 (C-21), 82.0 (C22), 23.7 (C-23), 41.3 (C-24), 70.6 (C-25), 28.7 (C-26), 31.1 (C-27), 106.9 (C-1'), 28.9 (C2'), 26.9 (C-3'); 13C NMR (CD3OD, 100 MHz)  37.4(C-1), 68.7 (C-2), 68.5 (C-3), 32.9 (C4), 51.7 (C-5), 206.5 (C-6), 122.1 (C-7), 167.7 (C-8), 35.1 (C-9), 39.2 (C-10), 21.5 (C-11),
31.7 (C-12), 48.5 (C-13), 85.3 (C-14), 32.4 (C-15), 22.4 (C-16), 50.5 (C-17), 17.5 (C-18),
24.5 (C-19), 85.8 (C-20), 22.6 (C-21), 83.3 (C-22), 24.5 (C-23), 42.2 (C-24), 71.1 (C-25),
28.9 (C-26), 29.4 (C-27), 108.0 (C-1'), 27.9 (C-2'), 27.3 (C-3').
25-Acetoxy-20-hydroxyecdysone 3-O--ᴅ-glucopyranoside (11): White amorphous powder;
1
H NMR (CD3OD, 500 MHz)  1.86 (1H, m, H-1a), 1.47 (1H, m, H-1b), 3.85 (1H, brd, J =
12.0 Hz, H-2), 4.08 (1H, brs, H-3), 1.86 (1H, m, H-4a), 1.69 (1H, m, H-4b), 2.46 (1H, dd, J
= 13.0, 4.0 Hz, H-5), 5.82 (1H, d, J = 2.0 Hz, H-7), 3.16 (1H, brt, J = 9.5, Hz, H-9), 1.96
(2H, m, H2-11), 2.13 (1H, ddd, J = 13.0, 13.0, 4.5 Hz, H-12a), 1.86 (1H, m, H-12b), 1.95
(1H, m, H-15a), 1.58 (1H, m, H-15b), 1.86 (1H, m, H-16a), 1.69 (1H, m, H-16b), 2.37 (1H,
t, J = 8.5 Hz, H-17), 0.88 (3H, s, H3-18), 0.97 (3H, s, H3-19), 1.18 (3H, s, H3-21); 3.30 (1H,
under CD3OD signal, H-22), 1.55 (H, m, H-23a), 1.28 (1H, m, H-23b), 1.99 (H, m, H-24a),
1.85 (1H, m, H-24b), 1.40 (3H, s, H3-26), 1.47 (3H, s, H3-27), 4.34 (1H, d, J = 8.0 Hz, H1'), 3.25 (1H, dd, J = 9.0, 8.0 Hz, H-2'), 3.37 (2H, t, J = 9.0 Hz, H-3', H-4'), 3.29 (1H, m, H5'), 3.84 (1H, brd, J = 12.0 Hz, H-6'a), 3.63 (1H, dd, J = 12.0, 5.0 Hz, H-6'b), 1.95 (3H, s,
CH3COO); 13C NMR (CD3OD, 125 MHz)  38.4(C-1), 68.2 (C-2), 76.8 (C-3), 30.5 (C-4),
51.7 (C-5), 206.6 (C-6), 122.1 (C-7), 168.5 (C-8), 35.1 (C-9), 38.4 (C-10), 21.5 (C-11),
32.5 (C-12), 48.5 (C-13), 85.1 (C-14), 31.8 (C-15), 21.5 (C-16), 50.6 (C-17), 18.0 (C-18),
24.1 (C-19), 77.8 (C-20), 21.0 (C-21), 78.4 (C-22), 26.9 (C-23), 39.5 (C-24), 83.9 (C-25),
26.6 (C-26), 26.3 (C-27), 103.2 (C-1'), 77.9 (C-2'), 71.5 (C-3'), 74.2 (C-4'), 78.0 (C-5'), 62.6
3
(C-6'), 172.7 (CH3COO), 22.4 (CH3COO); 1H NMR (pyridine-d5, 400 MHz)  4.06 (1H, m,
H-2), 4.33 (1H, brs, H-3), 2.94 (1H, ddd, J = 13.2, 2.8 Hz, H-5), 6.23 (1H, d, s, H-7), 3.56
(1H, brt, J = 8.4, Hz, H-9), 2.57 (1H, ddd, J = 12.8, 12.8, 4.4 Hz, H-12a), 1.83 (1H, m, H12b), 3.00 (1H, t, J = 9.2 Hz, H-17), 1.22 (3H, s, H3-18), 0.92 (3H, s, H3-19), 1.63 (3H, s,
H3-21); 3.86 (1H, brd, J = 10.0 Hz, H-22), 1.44 (3H, s, H3-26), 1.51 (3H, s, H3-27), 4.91
(1H, d, J = 7.6 Hz, H-1'), 4.07 (1H, dd, J = 9.0, 8.0 Hz, H-2'), 3.93 (1H, t, m, H-3'),
4.21(1H, t, J = 8.8 Hz, H-4'), 4.24 (1H, , J = 8.8 Hz, H-5'), 4.54 (1H, dd, J = 11.6, 11.6 Hz,
H-6'a), 4.34 (1H, dd, J = 11.6, 5.2 Hz, H-6'b), 1.97 (3H, s, CH3COO).
4
Figure 1S. 1H NMR (500 MHz, pyridine-d5) of compound 4
Figure 2S. 13C NMR (125 MHz, pyridine-d5) of compound 4
5
Figure 3S. 1H NMR (CD3OD, 500 MHz) of compound 5
Figure 4S. 13C NMR (CD3OD, 125 MHZ) of compound 5
6
Figure 5S. HSQC of compound 5
Figure 6S. HMBC of compound 5
7
Figure 7S. DARTMS of compound 5
Figure 8S. 1H NMR (CD3OD, 400 MHz) of compound 6
8
Figure 9S. 13C NMR (CD3OD, 100 MHz) of compound 6
Figure 10S. HSQC of compound 6
9
Figure 11S. HMBC of compound 6
Figure 12S. DARTMS of compound 6
10
Figure 13S. 1H NMR (DMSO, 400 MHZ) compound 7
Figure 14S. 13C NMR compound 7 (DMSO, 100 MHz)
11
Figure 15S. 1H NMR (CD3OD, 500 MHz) of compound 7
Figure 16S. 13C NMR (CD3OD, 125 MHz) of compound 7
12
Figure 17S. HSQC compound 7
Figure 18S. HMBC compound 7
13
Figure 19S. 1H NMR (CHCl3, 500 MHz) of compound 9
Figure 20S. 13C NMR (CHCl3, 125 MHz) of compound 9
14
Figure 21S. HSQC of compound 9
Figure 22S. HMBC of compound 9
15
Figure 23S. DARTMS of compound 9
Figure 24S. 1H NMR (CD3OD, 500 MHz) of compound 11
16
Figure 25S. 13C NMR (CD3OD, 125 MHz) of compound 11
Figure 26S. HSQC of compound 11
17
Figure 27S. HMBC of compound 11
18
Table 1S. Concentration-response evaluation of acetone extracts of S. acuta and S.
rhombifolia, and of compounds 4 and 5 on yeast a-glucosidase inhibition test
Sample
Concentration (mol L1
SAAa
SRAa
Compound 4
OD 405 nm
Inhibition (%)
IC50 mol L1
)
8.49 ± 0.66a
control
2.174  0.128
1.00
1.946  0.27
10.80  4.50
1.78
1.887  0.208
27.70  8.82
3.16
1.788  0.177
51.72  10.78
5.62
1.451  0.171*
33.68  4.34*
10.00
0.932 ± 0.120** 57.48 ± 3.18**
17.78
0.200 ± 0.127**
90.82 ± 0.95**
31.62
0.047 ±0.028**
97.68 ± 1.50**
control
2.174  0.128
1.00
2.027  0.160
6.95  2.2
1.78
1.892 ± 0.161
13.16  3.28
3.16
1.669  0.102*
23.12  2.94*
5.52
1.338 ± 0.094**
38.41 ± 2.61**
10.00
0.982 ± 0.130**
55.23 ± 3.61**
17.78
0.267 ± 0.074**
87.99 ± 2.77**
31.62
0.024 ± 0.012**
98.97 ± 0.52 **
control
1.799  0.001
19
)
8.10 ± 0.34a
32.70 ± 1.35
Compound 5
Quercetin
3.16
1.692  0.022
5.86  1.21
5.62
1.571  0.016
12.57  0.85
10.00
1.460  0.015
17.78
1.246 ± 0.005**
30.64  0.29**
31.62
0.901 ± 0.015**
49.89  0.80**
56.23
0.602 ± 0.059**
66.49  3.30**
100.00
0.142 ± 0.060**
76.68  3.33**
control
2.174  0.128
3.16
1.839  0.132
15.54  1.48
5.62
1.651  0.116*
24.14  1.37*
10.00
1.409  0.107**
35.27  2.02**
17.78
1.104 ± 0.086**
49.28  1.79**
31.62
0.815 ± 0.059**
62.51  1.39**
56.23
0.571 ± 0.038**
73.76  0.57**
100.00
0.346 ± 0.032**
83.30  0.77**
control
2.019  0.092
2.95
1.883  0.080
6.71  0.40
5.26
1.774  0.087
12.14  1.29
9.34
1.530  0.125*
24.24  4.94*
16.61
0.944 ± 0.195*
53.39  8.74*
20
18.78  0.80
19.24 ± 1.73
15.61 ± 1.61
Acarboseb
82.89  3.34*
29.56
0.344 ± 0.069**
control
1.43  0.05
3.16
1.001  0.061*
29.9  2.17*
5.62
0.811  0.037*
43.14  1.64*
10.00
0.583  0.036*
56.17  1.66**
17.78
0.393 ± 0.035*
72.54  1.55*
31.62
0.243 ± 0.08*
82.93  0.10*
7.26 ± 0.34b
Concentrations and IC50 values in g mL-1. bConcentrations and IC50 values in mM. Each
a
value represents the mean of three experiments  standard error. Data were analysed using
Dunnett’s test, values of *p≤0.05 and **p≤0.01 were considered significant.
Table 2S. Concentration-response evaluation of DPPH free radical scavenging activity of
acetone and methanol extracts of S. acuta and S. rhombifolia, and of compound 5
Sample
SAA
SRA
Concentration (g mL-1)
OD 515 nm
control
0.665  0.006
56.23
0.543  0.005**
18.23  0.12**
100
0.481  0.008**
27.56  0.58**
177.83
0.381  0.011**
42.67  1.07**
316.23
0.236  0.010**
64.51  1.24**
562.34
0.066 ± 0.021**
90.14 ± 3.07**
control
0.665  0.006
56.23
0.542  0.005**
21
Inhibition (%)
IC50 g mL-1)
220.54 ± 6.47
221.50 ± 5.55
18.48  0.13**
SAM
SRM
Compound 5a
100
0.476 ± 0.004**
28.37  0.17**
177.83
0.380  0.003**
42.83  0.57**
316.23
0.244 ± 0.007**
63.29 ± 1.19**
562.34
0.088 ± 0.006**
86.81 ± 1.03**
control
0.665  0.006
56.23
0.577  0.007**
13.13  0.42**
100
0.527  0.011**
20.63  0.95**
177.83
0.447  0.013**
32.68  1.30**
316.23
0.310 ± 0.021**
53.36  2.79**
562.34
0.118 ± 0.023**
82.30  3.26**
control
0.665  0.006
56.23
0.591  0.004**
11.06  0.81**
100
0.549  0.003**
17.32  0.52**
177.83
0.470  0.003**
29.26  0.55**
316.23
0.356 ± 0.002**
46.38  0.51**
562.34
0.009 ± 0.005**
70.83  0.54**
control
0.679 ± 0.018
10
0.572  0.019**
15.86  1.35**
17.788
0.515  0.013**
24.18  0.43**
31.62
0.416  0.013**
38.74  0.47**
22
295.01 ± 8.19
346.28 ± 3.47
46.18 ± 0.83a
-tocopherola
53.26
0.293 ± 0.013**
56.89  0.83**
100.00
0.170 ± 0.008**
74.93  0.65**
177.83
0.088  0.004**
87.10  0.29**
316.23
0.054  0.002**
92.07  0.14**
control
0.741  0.018
13.34
0.609 ± 0.026**
17.96  1.74**
17.78
0.553 ± 0.027**
25.51  2.02**
23.71
0.488  0.020**
34.31  1.80**
31.62
0.400 ± 0.019**
46.13  1.98**
42.17
0.284 ± 0.017**
61.65  2.19**
56.23
0.135  0.009**
81.76  1.11**
74.13
0.106  0.007**
85.79  0.63**
13.67 ± 0.45a
Concentrations and IC50 values in mol L-1. Each value represents the mean of three
a
independent experiments  standard error. Data were analysed using Dunnett’s test, values
of *p≤0.05 and **p≤0.01 were considered significant.
23