Bioscience, Biotechnology, and Biochemistry
ISSN: 0916-8451 (Print) 1347-6947 (Online) Journal homepage: http://www.tandfonline.com/loi/tbbb20
Isolation and Structural Elucidation of the Major Genuine Soybean Saponin Shigemitsu Kudou, Masahide Tonomura, Chigen Tsukamoto, Makoto Shimoyamada, Teiji Uchida & Kazuyoshi Okubo To cite this article: Shigemitsu Kudou, Masahide Tonomura, Chigen Tsukamoto, Makoto Shimoyamada, Teiji Uchida & Kazuyoshi Okubo (1992) Isolation and Structural Elucidation of the Major Genuine Soybean Saponin, Bioscience, Biotechnology, and Biochemistry, 56:1, 142-143, DOI: 10.1271/bbb.56.142 To link to this article: http://dx.doi.org/10.1271/bbb.56.142
Published online: 12 Jun 2014.
Submit your article to this journal
Article views: 165
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Citing articles: 46 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=tbbb20 Download by: [University of Lethbridge]
Date: 06 May 2016, At: 04:23
Biosci. Biotech. Biochem., 56 (1), 142-143, 1992
Note
Isolation and Structural Elucidation of the Major Genuine Soybean Saponin Shigemitsu KUDOU, Masahide TONoMuRA, Chigen TSUKAMOTO, Makoto SHIMOYAMADA, Teiji UCHIDA, and Kazuyoshi OKUBO Kanesa Co., Ltd., 202 Hamada, Tamagawa, Aomori 030, Japan * Faculty of Agriculture, Tohoku University, 1-1 Tsutumitori Amamiya-machi, Aoba-ku, Sendai 981, Japan
Downloaded by [University of Lethbridge] at 04:23 06 May 2016
Received June 14, 1991
Many· kinds of saponin have been isolated from soybean seeds, and their structures have been determined. 1- 10 ) In our previous paper,10) we divided soybean saponin into three groups, namely "group A saponin," "group B saponin," and "group E saponin," on the basis of their aglycone structure (soyasapogenols A, B, and E, Fig. 1). It was also reported that Bd and Be in the "group E saponin" were very unstable and readily converted into Ba and Bb, respectively.11) Further investigations showed that Bd and Be were mixtures containing BdA and BeA, respectively. This paper describes the isolation and chemical structure of BeA. While the extraction of soybean saponins has been routinely conducted with heating, 4 - 11) we carried it out at room temperature. After 0.5 g of soybean hypocotyls had been soaked in 10 ml of 70% aqueous ethanol for 30 min at room temperature or extracted for 5 hr at 80°C, the saponin composition of both supernatants was examined by HPLC (Fig. 2). The major compounds in the solution extracted at room temperature were BdA and BeA, whereas those found in the hot extract were Ba and Bb. Saponins BdA and BeA showed a maximum absorption at 292 nm, whereas Ba and
Bb showed no absorption above 220 nm. These results suggest that BdA and BeA were the genuine saponins in soybean seeds, while Ba and Bb were artifacts produced by heating during the extraction or concentration process. Such an investigation led us to isolate BdA and BeA, and to determine their structures. One kilogram of ground soybean hypocotyls (kindly supplied by Perikan Co., Ltd.) was shaken with 4 liters of 50% aqueous methanol for 30 min, and the solution filtered through filter paper. The filtrate was applied to an ODS column (YMC Co., Ltd., ODS-A 60-60/30,5 x 74cm) equilibrated with 50% aqueous methanol, eluted with 80% methanol and then fractionated (l5g/tube). The BeA fractions (No. 121-140), which were eluted after "group A saponin" and "group B saponin," were combined, evaporated under reduced pressure below 40°C to remove the methanol, and lyophylized to provide 260 mg of the BeA fraction. A sample (100 mg) Table I.
13C_ and 1H-NMR Spectral Assignments for Bb and BeA. Bb
~
'I'DHh99' 0 H DH
12
HD
22
3
,
CH2DH
.
RIO
soYa~sapog.nOI A .'
,
I
CH2DH
o#soyasapogenOI 8,.
V
O
o :
2
RIO
CH3
I
,
OH
5
0
CH2DH
RIO,. C
Soyasapogenol E
H20H
SeA
Aglycone I II III
Ba Bb Bd
B B E
N V
Be BeA
E
Fig. 1.
f3 -D-glc(1 ~2)- f3 -D--gal (1->2)- f3 -D-glcUA a -L-rha(1 ~2)- f3 -D-ga I (l ~2)- f3 -D-glclJA f3 -D-glc(1 ~2)- f3 -D-gal (l ~2)- f3 -D-glcUA a -L-rha(1-->2)- f3 -D-gal (1->2)- {3 -D-g]cUA a -L-rhaO ~2)- f3 -D-gal 0 ~2)- f3 -D-glcUA
Structures of Ba, Bb, Bd, Be, BeA, and Soyasapogenols.
~N
I
BeA
:::>
BeA
~on 0
N
N
:::>
:::>
ISSN: 0916-8451 (Print) 1347-6947 (Online) Journal homepage: http://www.tandfonline.com/loi/tbbb20
Isolation and Structural Elucidation of the Major Genuine Soybean Saponin Shigemitsu Kudou, Masahide Tonomura, Chigen Tsukamoto, Makoto Shimoyamada, Teiji Uchida & Kazuyoshi Okubo To cite this article: Shigemitsu Kudou, Masahide Tonomura, Chigen Tsukamoto, Makoto Shimoyamada, Teiji Uchida & Kazuyoshi Okubo (1992) Isolation and Structural Elucidation of the Major Genuine Soybean Saponin, Bioscience, Biotechnology, and Biochemistry, 56:1, 142-143, DOI: 10.1271/bbb.56.142 To link to this article: http://dx.doi.org/10.1271/bbb.56.142
Published online: 12 Jun 2014.
Submit your article to this journal
Article views: 165
View related articles
Citing articles: 46 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=tbbb20 Download by: [University of Lethbridge]
Date: 06 May 2016, At: 04:23
Biosci. Biotech. Biochem., 56 (1), 142-143, 1992
Note
Isolation and Structural Elucidation of the Major Genuine Soybean Saponin Shigemitsu KUDOU, Masahide TONoMuRA, Chigen TSUKAMOTO, Makoto SHIMOYAMADA, Teiji UCHIDA, and Kazuyoshi OKUBO Kanesa Co., Ltd., 202 Hamada, Tamagawa, Aomori 030, Japan * Faculty of Agriculture, Tohoku University, 1-1 Tsutumitori Amamiya-machi, Aoba-ku, Sendai 981, Japan
Downloaded by [University of Lethbridge] at 04:23 06 May 2016
Received June 14, 1991
Many· kinds of saponin have been isolated from soybean seeds, and their structures have been determined. 1- 10 ) In our previous paper,10) we divided soybean saponin into three groups, namely "group A saponin," "group B saponin," and "group E saponin," on the basis of their aglycone structure (soyasapogenols A, B, and E, Fig. 1). It was also reported that Bd and Be in the "group E saponin" were very unstable and readily converted into Ba and Bb, respectively.11) Further investigations showed that Bd and Be were mixtures containing BdA and BeA, respectively. This paper describes the isolation and chemical structure of BeA. While the extraction of soybean saponins has been routinely conducted with heating, 4 - 11) we carried it out at room temperature. After 0.5 g of soybean hypocotyls had been soaked in 10 ml of 70% aqueous ethanol for 30 min at room temperature or extracted for 5 hr at 80°C, the saponin composition of both supernatants was examined by HPLC (Fig. 2). The major compounds in the solution extracted at room temperature were BdA and BeA, whereas those found in the hot extract were Ba and Bb. Saponins BdA and BeA showed a maximum absorption at 292 nm, whereas Ba and
Bb showed no absorption above 220 nm. These results suggest that BdA and BeA were the genuine saponins in soybean seeds, while Ba and Bb were artifacts produced by heating during the extraction or concentration process. Such an investigation led us to isolate BdA and BeA, and to determine their structures. One kilogram of ground soybean hypocotyls (kindly supplied by Perikan Co., Ltd.) was shaken with 4 liters of 50% aqueous methanol for 30 min, and the solution filtered through filter paper. The filtrate was applied to an ODS column (YMC Co., Ltd., ODS-A 60-60/30,5 x 74cm) equilibrated with 50% aqueous methanol, eluted with 80% methanol and then fractionated (l5g/tube). The BeA fractions (No. 121-140), which were eluted after "group A saponin" and "group B saponin," were combined, evaporated under reduced pressure below 40°C to remove the methanol, and lyophylized to provide 260 mg of the BeA fraction. A sample (100 mg) Table I.
13C_ and 1H-NMR Spectral Assignments for Bb and BeA. Bb
~
'I'DHh99' 0 H DH
12
HD
22
3
,
CH2DH
.
RIO
soYa~sapog.nOI A .'
,
I
CH2DH
o#soyasapogenOI 8,.
V
O
o :
2
RIO
CH3
I
,
OH
5
0
CH2DH
RIO,. C
Soyasapogenol E
H20H
SeA
Aglycone I II III
Ba Bb Bd
B B E
N V
Be BeA
E
Fig. 1.
f3 -D-glc(1 ~2)- f3 -D--gal (1->2)- f3 -D-glcUA a -L-rha(1 ~2)- f3 -D-ga I (l ~2)- f3 -D-glclJA f3 -D-glc(1 ~2)- f3 -D-gal (l ~2)- f3 -D-glcUA a -L-rha(1-->2)- f3 -D-gal (1->2)- {3 -D-g]cUA a -L-rhaO ~2)- f3 -D-gal 0 ~2)- f3 -D-glcUA
Structures of Ba, Bb, Bd, Be, BeA, and Soyasapogenols.
~N
I
BeA
:::>
BeA
~on 0
N
N
:::>
:::>
