4.3. Extraction and Isolation
The dried powder (3.0 kg) was extracted by using ultrasound for three times (each for 30 minutes) with chloroform methanol (CHCl3/CH3OH) (6 L). The crude extract (160.0 g) was obtained by solvent evaporation under reduced pressure. Then silica gel column chromatography (160–200 mesh,
10
.
0
×
3
3
cm, 1000 g) was used on fractionation. Chloroform methanol solvent system (v/v ratio of chloroform, 50 : 1, 30 : 1, 10 : 1, and methanol) was used to obtain 50 fractions. Fr.42 (0.70 g) was purified by silica gel column from CHCl3/CH3OH (8 : 1) to give crystalline compound 6 (32 mg) and compound 5 (13 mg). Silica gel column chromatography (160–200 mesh,
2
.
0
×
35
cm, 100 g) of Fr.43 (1.26 g) eluting CHCl3/CH3OH (30 : 1) gave thirty subfractions (43.1–43.30). Fr.43-5 was subjected to silica gel column (160–200 mesh,
1.5
×
3
0
cm, 48 g) eluted with CHCl3/CH3OH (8 : 1) to afford compound 4 (5.5 mg) and compound 1 (4.5 mg). Fr.43-12 was subjected to silica gel column (160–200 mesh,
1.5
×
3
0
cm, 48 g) eluted with CHCl3/CH3OH (20 : 1) to afford compound 7 (5 mg) and CHCl3/CH3OH (10 : 1) to obtain compound 8 (8.3 mg). Fr.43-14 was separated by silica gel column (160–200 mesh,
1.5
×
3
0
cm, 48 g) eluted with CHCl3/CH3OH (10 : 1) to afford compound 2 (5 mg). Fr.43-15 was subjected to silica gel column (160–200 mesh,
1.5
×
3
0
cm, 48 g) eluted with CHCl3/CH3OH (15 : 1) to obtain subfraction Fr.43-15-1 to afford compound 9 (6.0 mg) and eluted with CHCl3/CH3OH (10 : 1) to obtain subfraction Fr.43-15-4 purified with CHCl3/CH3OH (8 : 1), yielding compound 3 (5 mg).
10-O-Methylhernovine (
1
). Light brown powder, soluble in chloroform, is with Dragendorff’s test positive. 1H-NMR (500 MHz, CDCl3) δ ppm: 6.74 (1H, s, H-3), 3.07 (2H, d,
J
=
12.5
Hz, H-4), 2.86 (2H, d,
J
=
12.5
Hz, H-5), 3.29 (1H, m, H-6a), 2.60 (1H, m, H-7), 3.17 (1H, m, H-7), 6.83 (1H, d,
J
=
7.9
Hz, H-8), 6.86 (1H, d,
J
=
7.9
Hz, H-9), 3.92 (3H, s, 1-OCH3), 8.86 (1H, br, s, 2-OH), 3.74 (1H, s, 10-OCH3), 3.92 (1H, s, 11-OCH3). 13C-NMR (125 MHz, CDCl3) δ ppm: 142.6 (C-1), 126.0 (C-1a), 119.8 (C-1b), 152.2 (C-2), 111.3 (C-3), 130.9 (C-3a), 128.8 (C-3b), 27.4 (C-4), 41.8 (C-5), 53.6 (C-6a), 36.5 (C-7), 128.2 (C-7a), 119.4 (C-8), 111.5 (C-9), 149.8 (C-10), 144.3 (C-11), 56.0 (1-OCH3), 62.2 (10-OCH3), 56.2 (11-OCH3). Its NMR spectral data were in accord with the reported data [27].
Nantenine (
2
). Yellow needle crystals, soluble in chloroform, exhibited a positive Dragendorff’s test. 1H-NMR (500 MHz, CDCl3) δ ppm: 6.63 (1H, s, H-3), 4.51 (2H, s, H-7), 6.95 (1H, d,
J
=
8.0
Hz, H-8), 7.07 (1H, s, H-9), 6.92 (1H, d,
J
=
8.0
Hz, H-11), 3.88 (3H, s, 1-OCH3), 3.87 (3H, s, 2-OCH3), 2.95 (3H, s, N-CH3), 5.97 (2H, s, –OCH2–). 13C-NMR (125 MHz, CDCl3) δ ppm: 145.7 (C-1), 125.0 (C-1a), 121.5 (C-1b), 151.2 (C-2), 112.9 (C-3), 127.7 (C-3a), 128.2 (C-3b), 29.7 (C-4), 54.9 (C-5), 63.8 (C-6a), 38.8 (C-7), 130.9 (C-7a), 106.7 (C-8), 148.8 (C-9), 147.4 (C-10), 108.6 (C-11), 60.9 (1-OCH3), 55.9 (2-OCH3), 43.5 (N-CH3), 101.6 (9, 10-OCH2O–). The 1H- and 13C-NMR spectral data were consistent with the reported data [28].
Corytuberine (
3
). Colorless columnar crystals, soluble in chloroform, were positive to Dragendorff’s test. 1H-NMR (500 MHz, CDCl3) δ ppm: 6.71 (1H, s, H-3), 3.18 (1H, td,
J
=
14
, 4 Hz, H-4), 2.69 (1H, dd,
J
=
14
, 4 Hz, H-4), 3.05 (1H, m, H-5), 2.54 (1H, m, H-5), 2.99 (1H, d,
J
=
13
Hz, H-6a), 3.04 (1H, m, H-7), 2.44 (1H, d,
J
=
1
3
Hz, H-7), 7.01 (1H, d,
J
=
7.5
Hz, H-8), 6.92 (1H, d,
J
=
7.5
Hz, H-9), 3.92 (3H, s, 2-OCH3), 2.57 (3H, s, N-CH3), 3.76 (3H, s, 10-OCH3). 13C-NMR (125 MHz, CDCl3) δ ppm: 141.8 (C-1), 124.3 (C-1a), 130.9 (C-1b), 148.8 (C-2), 111.4 (C-3), 125.2 (C-3a), 118.9 (C-3b), 28.9 (C-4), 52.8 (C-5), 62.7 (C-6a), 35.2 (C-7), 127.9 (C-7a), 125.1 (C-8), 114.7 (C-9), 148.2 (C-10), 142.2 (C-11), 62.0 (1-OCH3), 44.0 (N-CH3), 62.5 (10-OCH3). The 1H- and 13C-NMR spectral data were identical with the literature data [29].
Lagesianine A (
4
). Colorless columnar crystals, soluble in chloroform, gave a positive visualization to Dragendorff’s test. 1H-NMR (500 MHz, CDCl3) δ ppm: 7.05 (1H, s, H-3), 4.59 (1H, br, s, H-4), 3.22 (1H, br, dd,
J
=
1
2
, 1.7 Hz, H-5), 2.80 (1H, dd,
J
=
1
3
, 3.1 Hz, H-5), 3.06 (1H, m, H-6a), 3.10 (1H, dd,
J
=
13
, 3.1 Hz, H-7), 2.62 (1H, m, H-7), 6.86 (1H, d,
J
=
8
Hz, H-8), 6.89 (1H, d,
J
=
8
Hz, H-9), 3.98 (3H, s, 1-OCH3), 3.76 (3H, s, 2-OCH3), 2.67 (3H, s, N-CH3), 3.94 (3H, s, 10-OCH3), 8.76 (1H, s, 11-OH). 13C-NMR (125 MHz, CDCl3) δ ppm: 144.2 (C-1), 128.3 (C-1a), 119.3 (C-1b), 152.2 (C-2), 111.8 (C-3), 131.9 (C-3a), 125.8 (C-3b), 62.1 (C-4), 60.1 (C-5), 66.4 (C-6a), 35.1 (C-7), 128.8 (C-7a), 119.9 (C-8), 111.2 (C-9), 149.7 (C-10), 143.7 (C-11), 63.1 (1-OCH3), 55.9 (2-OCH3), 43.3 (N-CH3), 56.1 (10-OCH3). The above data were in accord with the literature data [30].
Corydine (
5
). It is colorless columnar crystals. 1H-NMR (500 MHz, CDCl3) δ ppm: 6.71 (1H, s, H-3), 3.21 (1H, td,
J
=
13.4
, 6.5 Hz, H-4), 2.71 (1H, m, H-4), 3.09 (1H, dd,
J
=
7
, 3.5 Hz, H-5), 2.57 (1H, m, H-5), 3.06 (1H, m, H-6a), 3.08 (1H, m, H-7), 2.48 (1H, t,
J
=
13
Hz, H-7), 7.11 (1H, d,
J
=
8.3
Hz, H-8), 6.90 (1H, d,
J
=
8.3
Hz, H-9), 8.73 (1H, s, 1-OH), 3.93 (3H, s, 2-OCH3), 2.58 (3H, s, N-CH3), 3.94 (3H, s, 10-OCH3), 3.76 (3H, s, 11-OCH3). 13C-NMR (125 MHz, CDCl3) δ ppm: 142.4 (C-1), 123.8 (C-1a), 130.7 (C-1b), 149.3 (C-2), 111.4 (C-3), 126.4 (C-3a), 119.3 (C-3b), 28.8 (C-4), 52.7 (C-5), 62.7 (C-6a), 35.4 (C-7), 127.7 (C-7a), 124.4 (C-8), 111.0 (C-9), 151.9 (C-10), 143.9 (C-11), 56.0 (2, 10-OCH3), 43.7 (N-CH3), 62.0 (11-OCH3). The 1H- and 13C-NMR spectral data were consistent with the literature data [31].
Isocorydine (
6
). It is colorless columnar crystals. 1H-NMR (500 MHz, CDCl3) δ ppm: 6.73 (1H, s, H-3), 3.20 (1H, m, H-4), 2.72 (2H, d,
J
=
1
7
Hz, H-4), 3.02 (1H, m, H-5), 2.49 (1H, t,
J
=
1
2
Hz, H-5), 2.89 (1H, d,
J
=
12
Hz, H-6a), 3.06 (1H, dd,
J
=
1
3
.
5
, 3.1 Hz, H-7), 2.45 (1H, q,
J
=
1
3.5
Hz, H-7), 6.86 (1H, d,
J
=
8.0
Hz, H-8), 6.88 (1H, d,
J
=
8.0
Hz, H-9), 3.72 (3H, s, 1-OCH3), 3.94 (3H, s, 2-OCH3), 2.55 (3H, s, N-CH3), 3.92 (3H, s, 10-OCH3), 8.85 (1H, s, 11-OH). 13C-NMR (125 MHz, CDCl3) δ ppm: 142.2 (C-1), 126.0 (C-1a), 120.1 (C-1b), 151.3 (C-2), 111.1 (C-3), 129.0 (C-3a), 130.0 (C-3b), 29.4 (C-4), 52.8 (C-5), 62.9 (C-6a), 35.9 (C-7), 130.0 (C-7a), 119.0 (C-8), 111.0 (C-9), 149.5 (C-10), 144.0 (C-11), 62.1 (1-OCH3), 56.1 (2-OCH3), 43.9 (N-CH3), 56.0 (10-OCH3). The 1H- and 13C-NMR spectral data were identical with published data [32].
Dihydrosanguinarine (
7
). It is pale yellow needles. 1H-NMR (500 MHz, CDCl3) δ ppm: 7.13 (1H, s, H-1), 7.69 (1H, s, H-4), 4.22 (2H, s, H-6), 6.87 (1H, d,
J
=
8.0
Hz, H-9), 7.32 (1H, d,
J
=
8.0
Hz, H-10), 7.71 (1H, d,
J
=
8.5
Hz, H-11), 7.50 (1H, d,
J
=
8.5
Hz, H-12), 6.08 (2H, s, 2, 3-OCH2O–), 6.30 (2H, s, 7, 8-OCH2O–), 2.64 (3H, s, N-CH3). 13C-NMR (125 MHz, CDCl3) δ ppm: 104.3 (C-1), 148.1 (C-2), 147.5 (C-3), 100.7 (C-4), 126.5 (C-4a), 142.5 (C-4b), 48.5 (C-6), 113.6 (C-6a), 144.6 (C-7), 147.1 (C-8), 107.2 (C-9), 116.2 (C-10), 127.3 (C-10a), 124.4 (C-10b), 120.4 (C-11), 123.9 (C-12), 130.8 (C-12a), 101.0 (2, 3-OCH2O–), 101.3 (7, 8-OCH2O–), 41.6 (N-CH3). The 1H- and 13C-NMR spectral data were in accord with published data [33].
Protopine (
8
). It is colorless columnar crystals. 1H-NMR (500 MHz, CDCl3) δ ppm: 6.66 (1H, s, H-1), 6.93 (1H, s, H-4), 6.71 (1H, d,
J
=
7.7
Hz, H-11), 6.68 (1H, d,
J
=
7.7
Hz, H-12), 5.97 (2H, s, 2, 3-OCH2O–), 5.95 (2H, s, 9, 10-OCH2O–), 2.00 (3H, s, N-CH3). 13C-NMR (125 MHz, CDCl3) δ ppm: 110.3 (C-1), 148.0 (C-2), 146.2 (C-3), 108.0 (C-4), 135.6 (C-4a), 31.4 (C-5), 57.7 (C-6), 51.1 (C-8), 117.5 (C-8a), 146.0 (C-9), 145.9 (C-10), 106.8 (C-11), 124.9 (C-12), 128.6 (C-12a), 46.1 (C-13), 197.5 (C-14), 132.3 (C-14a), 101.2 (2, 3-OCH2O–), 100.9 (9, 10-OCH2O–), 41.6 (N-CH3). The 1H- and 13C-NMR spectral data were consistent with published data [34].
Dihydrocryptopine (
9
). It is white powder. ESI-MS m/z: 372.5 [M + H]+. 1H-NMR (500 MHz, CDCl3) δ ppm: 7.04 (1H, s, H-1), 6.63 (1H, s, H-4), 6.63 (1H, s, H-4), 3.65 (2H, m, H-5), 3.40 (1H, m, H-6), 3.33 (1H, m, H-6), 4.31 (2H, s H-8), 6.93 (1H, d,
J
=
8.6
Hz, H-11), 6.88 (1H, d,
J
=
8.6
Hz, H-11), 3.18 (2H, m, H-13), 3.05 (1H, m, H-14), 3.86 (3H, s, 2-OCH3), 3.87 (3H, s, 3-OCH3), 5.97 (2H, s, 9, 10-OCH2O–), 6.30 (1H, br, s, 14-OH), 2.62 (3H, s, N-CH3). 13C-NMR (125 MHz, CDCl3) δ ppm: 107.4 (C-1), 151.3 (C-2), 146.1 (C-3), 109.0 (C-4), 130.9 (C-4a), 63.9 (C-5), 55.5 (C-6), 54.1 (C-8), 123.8 (C-8a), 147.0 (C-9), 148.5 (C-10), 125.7 (C-11), 112.3 (C-12), 124.3 (C-12a), 55.4 (C-13), 42.0 (C-14), 126.9 (C-14a), 60.8 (2-OCH3), 55.8 (3-OCH3), 101.5 (9, 10-OCH2O–), 42.6 (N-CH3). The spectrum matched the previous report [35].