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Chemical differences between two populations of Abies sachalinensis
Phytochemistry, Vol. 32, No. 2, pp. 331-334,1993 PrintedIIIGreatBritain.
003l-9422/93 $6.00+ 0.00 PergamonPressLtd
CHEMICAL DIFFERENCES BETWEEN TWO POPULATIONS
OF
ABIES SACHALINENSIS KENZO KAWAI, CHIKA TAKAHASHI, TAMIE MIYAMOTO, ATSUSHI NUMATA,*
HISAKATSU IWABUCHI~
and
MASAHIROYOSHIKURA~ Osaka University of Pharmaceutical Sciences, 2-10-65 Kawai, Matsubara, Osaka 580, Japan; tSan-Ei Chemical Industries, Ltd, l-l11 Sanwa-cho, Toyonaka, Osaka 561, Japan (Received 1 May 1992)
Key Word Index-Abies
sachalinensis; Pinaceae; wood; needle; juvabione
analogue; sesquiterpenoid.
Abstract-Juvabione analogues in the wood of the nine Abies sachalinensis trees growing natively in Hokkaido, Japan were investigated. The analytical results divided the population into a juvabione group that contained only juvabionetype sesquiterpenoids with the 4R,l’R_stereownfiguration and a epijuvabione group that contained only epijuvabione-type compounds with the 4R,l’&stereoconfiguration.
INTRODUCnON
Previously we investigated the wood extracts of two Abies Japan. Only juvabione-type (4R, 1’R) sesquiterpenoids were isolated from one of the trees [ 11, whereas only epijuvabione-type (4R, 1’S) ones were obtained from the other. This evidence suggested that there is a tree-to-tree variation in constituents of A. sachalinensis wood. However, a precise site location of where these trees had grown unfortunately could not be determined. The intention of the present study was to confirm the above results. This time we examined the juvabione analogues, leaf oil compositions and cone characteristics of nine A. sachalinensis trees growing natively in Hokkaido. sachalinensis trees sampled in Hokkaido,
RESULTS AND DISCUSSION
It is known that the three species of Japanese fir, A. mayriuna Miyabe et Kudo (Japanese name, aotodomatsu), A. suchalinensis (Fr. S&m.) Mast. (Japanese name, akatodomatsu) and A. wilsonii Miyabe et Kudo (Japanese name, karafutoshirabiso), grow in Hokkaido. Cone characteristics provide a basis for a morphological differentiation between them [2, 31. Japanese fir trees are divided into types A to H ranked in order of descending length of a part of bract scale which projects from the inside of a seminiferous scale due to a difference of lengths between both scales. It is shown that types A to D having cones with the longest to middle projecting part of bract scales are A. mayriana, types E to G A. sachalinensis and type H, having cones with a shorter bract scale than a seminiferous scale, A. wilsonii [2, 31. All the nine trees collected in Hokkaido this time were identified as type E
*Author to whom correspondence should be addressed.
or F, namely A. sachalinensis, on the basis of their cones and other taxonomic characteristics (Table 1). Acetone extracts of the debarked whole wood of these trees were separately purified by Sephadex LH-20 and silica gel column chromatographies followed by HPLC to afford juvabione analogues as listed in Table 2. All of the isolates from trees nos 1,3-7 and 9 were juvabione-type compounds with the 4R,l’R-stereoconfiguration, whereas all of the components from trees nos 2 and 8 were epijuvabione-type compounds with the 4R,l’Sconfiguration. The question of whether this chemical feature of the wood also exists in the needles led to investigation of the leaf oil components. The volatile components in the needles of the nine trees were examined by GC-MS. Component concentrations were calculated from GC peak areas without correction factors. The compositions of the essential oils from the needles are given in Table 3; components are arranged in order of GC elution. The leaf oils of all the trees examined contained the same components, but in different amounts. Trees nos 1, 3-6, 8 and 9 showed a similar terpenoid pattern, character&d by a high content of cc-pinene, camphene and bornyl acetate, whereas in the case of trees nos 2 and 7 the content of camphene was lower than that in the other trees and instead the content of limonene was markedly higher. This feature of the composition of the leaf essential oils did not correlate with the chemical features of the wood. The content of each volatile component in the leaf oils can be influenced by temperature, rainfall and other growing conditions. On the other hand, it seems that a tree-to-tree variation of stereoisomers in the wood constituents is caused by a dilTerence in enzymes which are under genetic control. It is, therefore, considered that the stereochemical difference in constituents of the wood can provide key chemotaxonomic information rather than 331
K. KAWAI et al.
332
R’ 1MtH 3H 5 Me 6 Me 7 H 8 H
Rz
R3 0
2Me 4H
Me0 H a-OH H B-OH Me a-OH Me B-OH
R’ 9Mc 10 H
H Me
R* H Me
Table 1. Source of Abies sachaRnensis wood studied Tree number
Type*
Place of collectiont
Elevation (m)
Hidaka-cho Shikaoi-cho Rubcshibe-cho Aibetsu-cho Shimokawa-cho Teshio-cho Esashi-cho Kucchan-cho Shimokawa-cho
1000 700 520 200 10 20 20 200
900
Age (yr)
Date of collection 1989
53 93 32 48 38 66 81 88 60
5 Sept. 5 Sept. 31 Aug. 6 Sept. 7 Sept. 7 Sept. 11 Oct. 6 Oct. I Sept.
*Based on cone charactenstics. tSite location in Hokkaido, Japan.
Table 2. Occurence of juvabione analogues in A. sachalinensis woods Tree number Compound
Configuration
1
2*
Juvabione (I) 4’Dehydrojuvabione (2) Epijuvabione (3) 4’-Dehydroepijuvabione (4) Juvabiol (5) Isojuvabiol (6) Epijuvabiol(7) Isoepijuvabiol (8) 3’-Dehydrojuvabi-S-01(9) 3’-Dehydroepijuvabi-S-01 (10)
(4R, 1’R) (4R. 1’R)
+ + _
-+ + + +-----+-
(4R, (4R, (4R, (4R, (4R, (4R, (4R, (4R,
l’s) l’s)
l’R, 3’S) l’R, 3’R) 1’S, 3’S) 1’S, 3’R) 1’R) 1’S)
3
4
5
6
7
8
9
+ + -
+ + -
+ + -
+ + -
-+
+ + _-.
n
+ + -+ _ + +-_--++f-
-+ _
+ + --
+ + -._ -.
+ + -
+ + .
+
n -
+
-
-
_-
-
_
-
--
+
n
+
*Previously reported [4, 51. + Compound present in wood. n: Compound not looked for due to small amounts. - Not present in detectable amounts.
the content of volatile components in the leaf case. Thus, the results of the analysis of the analogues in the A. sachalinensis wood (Table the population into the juvabione group that
oil in this
only juvabione-type
juvabione 2) divided contained
stereoconfiguration and the epijuvabione group that contained only eijuvabione-type compounds with the 4R,l’S-stereoconfiguration.
sesquiterpenoids
with the 4R,l’R-
333
Two populations of Abies sachalinensis Table 3. Volatile oil composition of A. sachalinensis needles* Tree number 1
Component Santene Tricyclenc a-Pinene Camphere B-Pinene Sabinene b-3-Carene Myrcene a-Phellandrene a-Terpinene Limonene fi-Phellandrene y-Terpinene flymene a-Terpinolene Hexanol (Z)-3-Hexenol Acetic acid Fenchyl acetate Linalool Linalyl acetate Bornyl acetate B-Caryophyllene Nonanol a-Humulene y-Selinene a-Terpinyl acetate Borne.01 &Cadinene Unknown ( [Ml’ 204) (E, E)-a-Famesene p-Cymen-S-01 Nonadecane Nerolidol Heneicosane Xadinol Docosane Unknown ([Ml’ 222) Tricosane
2
0.12 1.90 17.94 16.79 8.17 t t 3.20 t 0.11 3.37 3.05 0.08 t 0.66 t t t 1.13 t 0.69 17.01 4.23 t 2.17 0.50 0.37 8.19 0.74 0.60 0.19 t t 0.67 t 0.34 t 5.01 t
0.11 1.11 14.02 8.77 3.62 0.07 t 5.08 t t 29.60 5.69 0.08 t 0.52 t t t 0.36 0.46 0.75 11.53 1.27 0.19 0.69 0.62 0.05 3.91 0.34 0.72 0.09 t t 0.21 0.11 0.09 t 6.37 0.22
3 0.19 2.02 25.94 14.68 274 t t 4.62 0.24 0.12 4.43 1.88 0.08 0.08 0.73 t t 0.15 0.63 1.15 0.41 7.82 4.36 0.15 234 0.50 0.64 13.96 0.19 0.45 0.24 0.08 t 0.40 t 0.08 t 3.93 0.16
4
5
0.21 1.90 20.67 19.17 5.03 t t 1.47 0.43 0.18 4.59 7.09 0.09 t 0.64 0.16 0.58 0.67 0.90 18.67 2.59 t 1.43 t 0.71 10.07 0.24 0.09 t t t 0.37 0.16 t t t
0.22 2.01 22.13 19.44 4.24 t t 3.87 t t 3.48 4.37 0.08 t 0.51 t t 0.82 0.27 0.82 23.39 1.56 0.13 0.79 0.30 0.49 5.48 0.10 0.30 0.08 0.09 t 0.18 t t t 3.07 0.15
6 0.11 1.89 20.14 13.73 5.97 t t 6.12 t t 4.55 10.71 0.10 t 0.62 t t t 1.17 0.15 1.21 16.63 2.85 t 1.42 0.58 0.69 1.75 0.36 0.54 0.21 t t 0.22 0.17 0.14 t 5.96 0.33
7 0.07 0.58 16.57 5.89 10.21 0.11 0.45 1.95 t 0.08 33.12 11.19 0.10 0.04 0.39 t 0.08 t 0.37 0.18 0.93 7.56 1.34 t 0.67 0.21 0.20 1.12 0.28 0.22 0.43 t 0.10 0.22 0.22 0.13 0.09 1.84 0.55
8 0.14 2.16 19.08 21.24 4.13 0.06 2.41 t t 4.29 2.63 0.13 t 0.80 t 0.12 t 0.72 t 0.42 30.26 1.36 t 0.66 0.46 0.46 1.38 0.27 0.42 t 0.28 t t 0.08 0.07 t 4.67 0.24
9 0.19 2.40 26.62 19.95 9.50 t t 0.51 t t 5.59 3.39 0.08 t 0.53 t t t 0.68 0.65 0.25 18.46 0.93 0.28 0.50 0.18 0.53 4.77 0.18 0.20 t t t 0.10 t 0.06 t 1.31 0.07
*Uncorrected peak area % values from FID signal. t: Traces (
EXPERIMENTAL Plant material. The A. sachalinensis trees, collected in
Holckaido, Japan, were identified by Dr G. Murata (Department of Botany, Faculty of Science, Kyoto University) and Drs J. Samejima and K. Takahashi (Hokkaido Research Center, Forestry and Forest Products Research Institute). Voucher specimens are deposited at the Herbarium of the Department of Botany, Kyoto University. Isolation of juvabione analoguesfromwood. A Me,CO extract
from each of the debarked
whole wood samples
was chromatographed on a Sephadex LH-20 column with Me&O-CH,Cl, (1: l), subsequently on a silica gel column with a gradient of EtOAc-C,H, and then subjected to HPLC [ODS (80% MeOH)] according to the method described elsewhere [4, 51. The juvabione analogues obtained from each wood were identified by spectral comparison with authentic samples. Analyses of leafoil components. An Et,0 extract from the needles of each tree was analysed by GC and GC-MS. Analytical GC was carried out on a J 8z W DB-WAX column (0.32 mm i.d. x 60 m), operating with inj. and det.
334
K. KAWAI
(FID) temps of 250” with He as carrier at a flow rate of 1 ml min- l. After 5 min, the column temp. was raised from 60” to 220” at 3”min- ‘. Split ratio and inj. vol. for all samples was l/100 and 0.3 pl, respectively. GC-MS analyses were performed using the gas chromatograph described above in EI mode (20 eV). Acknowledgements-The authors are very grateful to Dr G. Murata, (Kyoto University) and Drs J. Samejima and K. Takahashi (Forestry and Forest Products Research Institute) for plant identification. For gifts of A. sachalinensis trees we also thank Drs K. Takahashi and E. Kameshita (Forestry and Forest Products Research Institute) and all the heads and staff of local forestry offices
et al.
in Hidaka, Shimizu, Rubeshibe, Asahikawa, Shimokawa, Teshio, Kucchan and Hakodate, in Hokkaido. REFERENCES
1. Numata, A., Hokimoto, K., Takemura, T., Matsunaga, S. and Morita, R. (1983) Chem. Pharm. Bull. 31,436. 2. Samejima, J. (1968) Hayashi 4, 30. 3. Uehara, K. (1959) Atlas oj Trees (Jumokudaizusetsu) Vol. 1, p. 219. Ariakeshobo, Tokyo. 4. Numata, A., Kawai, K. and Takahashi, C. (1990) Chem. Pharm. Bull. 38, 2570. 5. Numata, A., Kawai, K., Takahashi, C. and Miyamoto, T. (1992) Phytochemistry 31 (in press).
003l-9422/93 $6.00+ 0.00 PergamonPressLtd
CHEMICAL DIFFERENCES BETWEEN TWO POPULATIONS
OF
ABIES SACHALINENSIS KENZO KAWAI, CHIKA TAKAHASHI, TAMIE MIYAMOTO, ATSUSHI NUMATA,*
HISAKATSU IWABUCHI~
and
MASAHIROYOSHIKURA~ Osaka University of Pharmaceutical Sciences, 2-10-65 Kawai, Matsubara, Osaka 580, Japan; tSan-Ei Chemical Industries, Ltd, l-l11 Sanwa-cho, Toyonaka, Osaka 561, Japan (Received 1 May 1992)
Key Word Index-Abies
sachalinensis; Pinaceae; wood; needle; juvabione
analogue; sesquiterpenoid.
Abstract-Juvabione analogues in the wood of the nine Abies sachalinensis trees growing natively in Hokkaido, Japan were investigated. The analytical results divided the population into a juvabione group that contained only juvabionetype sesquiterpenoids with the 4R,l’R_stereownfiguration and a epijuvabione group that contained only epijuvabione-type compounds with the 4R,l’&stereoconfiguration.
INTRODUCnON
Previously we investigated the wood extracts of two Abies Japan. Only juvabione-type (4R, 1’R) sesquiterpenoids were isolated from one of the trees [ 11, whereas only epijuvabione-type (4R, 1’S) ones were obtained from the other. This evidence suggested that there is a tree-to-tree variation in constituents of A. sachalinensis wood. However, a precise site location of where these trees had grown unfortunately could not be determined. The intention of the present study was to confirm the above results. This time we examined the juvabione analogues, leaf oil compositions and cone characteristics of nine A. sachalinensis trees growing natively in Hokkaido. sachalinensis trees sampled in Hokkaido,
RESULTS AND DISCUSSION
It is known that the three species of Japanese fir, A. mayriuna Miyabe et Kudo (Japanese name, aotodomatsu), A. suchalinensis (Fr. S&m.) Mast. (Japanese name, akatodomatsu) and A. wilsonii Miyabe et Kudo (Japanese name, karafutoshirabiso), grow in Hokkaido. Cone characteristics provide a basis for a morphological differentiation between them [2, 31. Japanese fir trees are divided into types A to H ranked in order of descending length of a part of bract scale which projects from the inside of a seminiferous scale due to a difference of lengths between both scales. It is shown that types A to D having cones with the longest to middle projecting part of bract scales are A. mayriana, types E to G A. sachalinensis and type H, having cones with a shorter bract scale than a seminiferous scale, A. wilsonii [2, 31. All the nine trees collected in Hokkaido this time were identified as type E
*Author to whom correspondence should be addressed.
or F, namely A. sachalinensis, on the basis of their cones and other taxonomic characteristics (Table 1). Acetone extracts of the debarked whole wood of these trees were separately purified by Sephadex LH-20 and silica gel column chromatographies followed by HPLC to afford juvabione analogues as listed in Table 2. All of the isolates from trees nos 1,3-7 and 9 were juvabione-type compounds with the 4R,l’R-stereoconfiguration, whereas all of the components from trees nos 2 and 8 were epijuvabione-type compounds with the 4R,l’Sconfiguration. The question of whether this chemical feature of the wood also exists in the needles led to investigation of the leaf oil components. The volatile components in the needles of the nine trees were examined by GC-MS. Component concentrations were calculated from GC peak areas without correction factors. The compositions of the essential oils from the needles are given in Table 3; components are arranged in order of GC elution. The leaf oils of all the trees examined contained the same components, but in different amounts. Trees nos 1, 3-6, 8 and 9 showed a similar terpenoid pattern, character&d by a high content of cc-pinene, camphene and bornyl acetate, whereas in the case of trees nos 2 and 7 the content of camphene was lower than that in the other trees and instead the content of limonene was markedly higher. This feature of the composition of the leaf essential oils did not correlate with the chemical features of the wood. The content of each volatile component in the leaf oils can be influenced by temperature, rainfall and other growing conditions. On the other hand, it seems that a tree-to-tree variation of stereoisomers in the wood constituents is caused by a dilTerence in enzymes which are under genetic control. It is, therefore, considered that the stereochemical difference in constituents of the wood can provide key chemotaxonomic information rather than 331
K. KAWAI et al.
332
R’ 1MtH 3H 5 Me 6 Me 7 H 8 H
Rz
R3 0
2Me 4H
Me0 H a-OH H B-OH Me a-OH Me B-OH
R’ 9Mc 10 H
H Me
R* H Me
Table 1. Source of Abies sachaRnensis wood studied Tree number
Type*
Place of collectiont
Elevation (m)
Hidaka-cho Shikaoi-cho Rubcshibe-cho Aibetsu-cho Shimokawa-cho Teshio-cho Esashi-cho Kucchan-cho Shimokawa-cho
1000 700 520 200 10 20 20 200
900
Age (yr)
Date of collection 1989
53 93 32 48 38 66 81 88 60
5 Sept. 5 Sept. 31 Aug. 6 Sept. 7 Sept. 7 Sept. 11 Oct. 6 Oct. I Sept.
*Based on cone charactenstics. tSite location in Hokkaido, Japan.
Table 2. Occurence of juvabione analogues in A. sachalinensis woods Tree number Compound
Configuration
1
2*
Juvabione (I) 4’Dehydrojuvabione (2) Epijuvabione (3) 4’-Dehydroepijuvabione (4) Juvabiol (5) Isojuvabiol (6) Epijuvabiol(7) Isoepijuvabiol (8) 3’-Dehydrojuvabi-S-01(9) 3’-Dehydroepijuvabi-S-01 (10)
(4R, 1’R) (4R. 1’R)
+ + _
-+ + + +-----+-
(4R, (4R, (4R, (4R, (4R, (4R, (4R, (4R,
l’s) l’s)
l’R, 3’S) l’R, 3’R) 1’S, 3’S) 1’S, 3’R) 1’R) 1’S)
3
4
5
6
7
8
9
+ + -
+ + -
+ + -
+ + -
-+
+ + _-.
n
+ + -+ _ + +-_--++f-
-+ _
+ + --
+ + -._ -.
+ + -
+ + .
+
n -
+
-
-
_-
-
_
-
--
+
n
+
*Previously reported [4, 51. + Compound present in wood. n: Compound not looked for due to small amounts. - Not present in detectable amounts.
the content of volatile components in the leaf case. Thus, the results of the analysis of the analogues in the A. sachalinensis wood (Table the population into the juvabione group that
oil in this
only juvabione-type
juvabione 2) divided contained
stereoconfiguration and the epijuvabione group that contained only eijuvabione-type compounds with the 4R,l’S-stereoconfiguration.
sesquiterpenoids
with the 4R,l’R-
333
Two populations of Abies sachalinensis Table 3. Volatile oil composition of A. sachalinensis needles* Tree number 1
Component Santene Tricyclenc a-Pinene Camphere B-Pinene Sabinene b-3-Carene Myrcene a-Phellandrene a-Terpinene Limonene fi-Phellandrene y-Terpinene flymene a-Terpinolene Hexanol (Z)-3-Hexenol Acetic acid Fenchyl acetate Linalool Linalyl acetate Bornyl acetate B-Caryophyllene Nonanol a-Humulene y-Selinene a-Terpinyl acetate Borne.01 &Cadinene Unknown ( [Ml’ 204) (E, E)-a-Famesene p-Cymen-S-01 Nonadecane Nerolidol Heneicosane Xadinol Docosane Unknown ([Ml’ 222) Tricosane
2
0.12 1.90 17.94 16.79 8.17 t t 3.20 t 0.11 3.37 3.05 0.08 t 0.66 t t t 1.13 t 0.69 17.01 4.23 t 2.17 0.50 0.37 8.19 0.74 0.60 0.19 t t 0.67 t 0.34 t 5.01 t
0.11 1.11 14.02 8.77 3.62 0.07 t 5.08 t t 29.60 5.69 0.08 t 0.52 t t t 0.36 0.46 0.75 11.53 1.27 0.19 0.69 0.62 0.05 3.91 0.34 0.72 0.09 t t 0.21 0.11 0.09 t 6.37 0.22
3 0.19 2.02 25.94 14.68 274 t t 4.62 0.24 0.12 4.43 1.88 0.08 0.08 0.73 t t 0.15 0.63 1.15 0.41 7.82 4.36 0.15 234 0.50 0.64 13.96 0.19 0.45 0.24 0.08 t 0.40 t 0.08 t 3.93 0.16
4
5
0.21 1.90 20.67 19.17 5.03 t t 1.47 0.43 0.18 4.59 7.09 0.09 t 0.64 0.16 0.58 0.67 0.90 18.67 2.59 t 1.43 t 0.71 10.07 0.24 0.09 t t t 0.37 0.16 t t t
0.22 2.01 22.13 19.44 4.24 t t 3.87 t t 3.48 4.37 0.08 t 0.51 t t 0.82 0.27 0.82 23.39 1.56 0.13 0.79 0.30 0.49 5.48 0.10 0.30 0.08 0.09 t 0.18 t t t 3.07 0.15
6 0.11 1.89 20.14 13.73 5.97 t t 6.12 t t 4.55 10.71 0.10 t 0.62 t t t 1.17 0.15 1.21 16.63 2.85 t 1.42 0.58 0.69 1.75 0.36 0.54 0.21 t t 0.22 0.17 0.14 t 5.96 0.33
7 0.07 0.58 16.57 5.89 10.21 0.11 0.45 1.95 t 0.08 33.12 11.19 0.10 0.04 0.39 t 0.08 t 0.37 0.18 0.93 7.56 1.34 t 0.67 0.21 0.20 1.12 0.28 0.22 0.43 t 0.10 0.22 0.22 0.13 0.09 1.84 0.55
8 0.14 2.16 19.08 21.24 4.13 0.06 2.41 t t 4.29 2.63 0.13 t 0.80 t 0.12 t 0.72 t 0.42 30.26 1.36 t 0.66 0.46 0.46 1.38 0.27 0.42 t 0.28 t t 0.08 0.07 t 4.67 0.24
9 0.19 2.40 26.62 19.95 9.50 t t 0.51 t t 5.59 3.39 0.08 t 0.53 t t t 0.68 0.65 0.25 18.46 0.93 0.28 0.50 0.18 0.53 4.77 0.18 0.20 t t t 0.10 t 0.06 t 1.31 0.07
*Uncorrected peak area % values from FID signal. t: Traces (
EXPERIMENTAL Plant material. The A. sachalinensis trees, collected in
Holckaido, Japan, were identified by Dr G. Murata (Department of Botany, Faculty of Science, Kyoto University) and Drs J. Samejima and K. Takahashi (Hokkaido Research Center, Forestry and Forest Products Research Institute). Voucher specimens are deposited at the Herbarium of the Department of Botany, Kyoto University. Isolation of juvabione analoguesfromwood. A Me,CO extract
from each of the debarked
whole wood samples
was chromatographed on a Sephadex LH-20 column with Me&O-CH,Cl, (1: l), subsequently on a silica gel column with a gradient of EtOAc-C,H, and then subjected to HPLC [ODS (80% MeOH)] according to the method described elsewhere [4, 51. The juvabione analogues obtained from each wood were identified by spectral comparison with authentic samples. Analyses of leafoil components. An Et,0 extract from the needles of each tree was analysed by GC and GC-MS. Analytical GC was carried out on a J 8z W DB-WAX column (0.32 mm i.d. x 60 m), operating with inj. and det.
334
K. KAWAI
(FID) temps of 250” with He as carrier at a flow rate of 1 ml min- l. After 5 min, the column temp. was raised from 60” to 220” at 3”min- ‘. Split ratio and inj. vol. for all samples was l/100 and 0.3 pl, respectively. GC-MS analyses were performed using the gas chromatograph described above in EI mode (20 eV). Acknowledgements-The authors are very grateful to Dr G. Murata, (Kyoto University) and Drs J. Samejima and K. Takahashi (Forestry and Forest Products Research Institute) for plant identification. For gifts of A. sachalinensis trees we also thank Drs K. Takahashi and E. Kameshita (Forestry and Forest Products Research Institute) and all the heads and staff of local forestry offices
et al.
in Hidaka, Shimizu, Rubeshibe, Asahikawa, Shimokawa, Teshio, Kucchan and Hakodate, in Hokkaido. REFERENCES
1. Numata, A., Hokimoto, K., Takemura, T., Matsunaga, S. and Morita, R. (1983) Chem. Pharm. Bull. 31,436. 2. Samejima, J. (1968) Hayashi 4, 30. 3. Uehara, K. (1959) Atlas oj Trees (Jumokudaizusetsu) Vol. 1, p. 219. Ariakeshobo, Tokyo. 4. Numata, A., Kawai, K. and Takahashi, C. (1990) Chem. Pharm. Bull. 38, 2570. 5. Numata, A., Kawai, K., Takahashi, C. and Miyamoto, T. (1992) Phytochemistry 31 (in press).