Determination of the absolute configuration of quercivorol, (1S,4R)-p-menth-2-en-1-ol, an aggregation pheromone of the ambrosia beetle Platypus quercivorus (Coleoptera: Platypodidae)

Article abstract of DOI:10.1271/bbb.60211

A pair of enantiomers of trans-p-menth-2-en-1-ol, an aggregation pheromone of Platypus quercivorus, was synthesized from (S)- and (R)-limonene. The retention time of the aggregation pheromone from the insect coincided with that of (1S,4R)-p-menth-2-en-1-ol synthesized from (S)-limonene from GC analyses with a chiral column, enabling the absolute configuration of the aggregation pheromone to be determined as (1S,4R).

Full text of DOI:10.1271/bbb.60211

Biosci. Biotechnol. Biochem., 70 (10), 2544–2546, 2006
Note
Determination of the Absolute Configuration of Quercivorol,
(1S,4R)-p-Menth-2-en-1-ol, an Aggregation Pheromone
of the Ambrosia Beetle Platypus quercivorus (Coleoptera: Platypodidae)
y
2
3
3;
Takehiro KASHIWAGI,1; Tadakazu NAKASHIMA, Shin-ich TEBAYASHI, and Chul-Sa KIM
*
1Department of Biomechanical System, Faculty of Agriculture, Ehime University,
3-5-7 Tarumi, Matsuyama 790-8566, Japan
2Department of Forest Entomology, Forestry and Forest Products Research Institute,
Matsunosato 1, Tsukuba 305-8687, Japan
3Department of Bioresources Science, Faculty of Agriculture, Kochi University,
B200 Monobe, Nankoku 783-8502, Japan
Received April 12, 2006; Accepted May 9, 2006; Online Publication, October 7, 2006
[doi:10.1271/bbb.60211]
A pair of enantiomers of trans-p-menth-2-en-1-ol, an
aggregation pheromone of Platypus quercivorus, was
synthesized from (S)- and (R)-limonene. The retention
time of the aggregation pheromone from the insect
coincided with that of (1S,4R)-p-menth-2-en-1-ol syn-
thesized from (S)-limonene from GC analyses with a
chiral column, enabling the absolute configuration of the
aggregation pheromone to be determined as (1S,4R).
mixture identified the aggregation pheromone of the
beetle as trans-p-menth-2-en-1-ol, although its the
stereochemistry has not yet been clarified.6) To clarify
the absolute configuration of the aggregation pheromone
of the beetle, (1S,4R)- and (1R,4S)-p-menth-2-en-1-ol
were synthesized (shown in the Scheme 1), and the
retention times of these synthetic products and that of
the aggregation pheromone from crushed male volatiles
were compared.
Key words: (1S,4R)-p-menth-2-en-1-ol; quercivorol; ag-
gregation pheromone; Platypus quercivorus
(S)-limonene (1) was converted into (S)-carvomen-
thene (2) by Raney nickel W4-catalyzed reduction, and
the carbon-carbon double bond at C1 of 2 was oxidized
by m-CPBA to give cis- and trans-epoxy-(S)-carvomen-
thane (3). The epoxide ring of 3 was opened to form a
new carbon-carbon double bond at C2 by sodium 2-
heptanoxide to be converted into (1S,4R)- and (1R,4R)-
p-menth-2-en-1-ol (4 and 5). Each isomer was identified
by its retention time from a GC analysis with the same
column7–9) and by comparing its GC–MS data with that
in the literature.9) The total yields of 4 and 5 were
respectively 11.2% and 10.2% based on (S)-limonene.
(1S,4S)- and (1R,4S)-p-menth-2-en-1-ol (6 and 7) were
synthesized from (R)-limonene in the same way.
The retention times of the four diastereoisomers of p-
menth-2-en-1-ol synthesized from (R)- and (S)-limonene
and the aggregation pheromone of P. quercivorus from
GC analyses with a chiral column are shown in Table 1.
The retention time for the aggregation pheromone of
P. quercivorus was 16.72 min, almost the same as that
of 4 synthesized from (S)-limonene. On the other hand,
the retention time of 7 synthesized from (R)-limonene
was 16.96 min. We concluded from these results that the
aggregation pheromone of P. quercivorus was (1S,4R)-
p-menth-2-en-1-ol, for which the name quercivorol is
Mass attack of by the ambrosia beetle, Platypus
quercivorus (Murayama) (Coleoptera: Platypodidae),
on oak trees, especially Quercus crispula Blume, has
resulted in the widely spread and continuous heavy
mortality of oak in deciduous broadleaf forests in Japan
since the late 1980s.1) A phytopathogenic fungus,
hypothesized by Hijii et al.,1) was isolated from the
gallery and mycangia of the ambrosia beetle and named
as Raffaelea quercivora.2) Dysfunction of the water flow
in attacked tree trunks was caused by long gallery
digging in sapwood by P. quercivorus and by the
invasion of R. quercivora through the gallery walls.3)
This is the first case of the ambrosia beetle being
demonstrated to kill a living host tree with a vectoring
pathogen.
Although sound communication is important as a cue
for the mating behaviour of the beetles,4) field experi-
ments have indicated that chemical communication by
using an aggregation pheromone was responsible for the
mass attack on oaks by the beetles.5) The results of GC–
EAD and GC–MS analyses of the volatiles from boring
dust and field trap experiments using a synthetic racemic
y
To whom correspondence should be addressed. Fax: +81-888-64-5186; E-mail: cs-kim@cc.kochi-u.ac.jp
Present address: Department of Bioresources Science, Faculty of Agriculture, Kochi University, B200 Monobe, Nankoku 783-8502, Japan
*

Determination of the Absolute Configuration of Quercivorol
2545
H2/Ni
m-CPBA
O
CH2Cl2
1
2
3
HO
HO
Na 2-Heptanoxide
CH3-(OCH2CH2)2-OH
4
5
Scheme 1. Synthesis of (1S,4R)- and (1R,4R)-p-Menth-2-en-1-ol, Quercivorol from (S)-Limonene.
Table 1. Retention Times of Stereoisomers of p-Menth-2-en-1-ol
from GC Analyses with a Chiral Capillary Column
measured with a JEOL JNM-AL 400 (400 MHz) instru-
ment. TMS was used as the internal standard. Letters
(br.)s, d, t, q, and m represent (broad)singlet, doublet,
triplet, quartet, and multiplet, respectively, and coupling
constants are given in Hz. The solvent used was CDCl3.
Specific rotation values were recorded with a Horiba
SEPA-200 high-sensitivity polarimeter.
Chemicals. All reagents were purchased from Nacalai
Tesque Ltd. and Wako Chemical Ltd. in Japan.
Syntheses.
Retention time
(min)
p-Menth-2-en-1-ol
trans-p-Menth-2-en-1-ol from Platypus quercivorus
cis-p-Menth-2-en-1-ol from P. quercivorus
(1S,4R)-p-Menth-2-en-1-ol (4)
16.72
16.06
16.71
16.05
16.08
16.96
(1R,4R)-p-Menth-2-en-1-ol (5)
(1S,4S)-p-Menth-2-en-1-ol (6)
(1R,4S)-p-Menth-2-en-1-ol (7)
(S)-Carvomenthene (2). A 13.6 g amount of (S)-
limonene (100 mmol) and 1.4 g of Raney nickel W4
were stirred, and a hydrogen gas stream was then fed
into the solution under atmospheric pressure at room
temperature until 3.4 liters of the hydrogen gas had been
absorbed. After purging with N2 gas, the mixture was
filtered and then distilled under atmospheric pressure to
produce 12.3 g of pure (S)-carovomenthene as a color-
less oil, bp 168–172 ꢁC at 760 mmHg in a 89.4% yield.
proposed. The precise biological assay results, in which
quercivorol attracted the beetles in field-trap experi-
ments and evoked an EAD response in antenna of the
beetle, will be reported elsewhere.
Experimental
½ꢀꢂD þ78:5ꢁ (c ¼ 2:0, CHCl3). EIMS m=z (%): 138
22
Collection of volatiles from the males. Emerged male
adults of P. quercivorus were collected from logs of
Q. crispula which had been killed by the R. quercivora
and P. quercivorus complex and harvested from dam-
aged forests in Kushibiki Town, Yamagata Prefecture
and Mikawa Village, Niigata Prefecture. The insects
were crushed in 1.5-ml vials by a tiny glass rod. The
headspace volatile components containing the aggrega-
tion pheromone of the beetle were collected by SPME
(Supelco, USA) from crushed males in the vials.
Instruments. GC–MS data was recorded with a JEOL
JMS600 mass spectrometer with a capillary column
(HP-5, crosslink 5% PH ME Siloxane, 30 m ꢀ 0:32 mm
i.d., 0.25 mm thickness, He carried gas:, temperature
program from 80 ꢁC with a 1-min hold to 150 ꢁC at
4 ꢁC/min). To elucidate the absolute structure, a sample
was analyzed in a chiral capillary column (Chirasil-Dex
CB, 25 m ꢀ 0:25 mm i.d., 0.25 mm thickness, Varian,
USA) installed in a Hewlett-Packard (HP) 6890 GC
instrument equipped with an HP 5973 mass spectrom-
eter, using He as the carrier gas. The oven temperature
was held at 60 ꢁC for 1 min, and then programmed to
150 ꢁC at 5 ꢁC/min. 1H- and 13C-NMR data were
(11), 96 (19), 95 (99), 94 (10), 82 (22), 81 (38), 79 (17),
77 (13), 69 (30), 68 (100), 67 (22), 55 (61), 53 (34), 51
(10). 1H-NMR (CDCl3) ꢁ: 5.36 (1H, br.s, H-2), 2.00
(1H, m, H-6b), 1.96 (1H, m, H-3b), 1.93 (1H, m, H-6a),
1.75 (1H, m, H-5b), 1.72 (1H, m, H-3a), 1.63 (3H, br.s,
H-7), 1.46 (1H, octet, H-8), 1.25 (1H, m, H-4), 1.20 (1H,
m H-5a), 0.89 (3H, d, J ¼ 6:4, H-9), 0.87 (3H, d, J ¼
6:4, H10). 13C-NMR (CDCl3) ꢁ: 133.7 (s, C-1), 121.0 (d,
C-2), 40.1 (d, C-4), 32.4 (d, C-8), 30.9 (t, C-6), 29.0 (t,
C-3), 26.6 (t, C-5), 23.5 (q, C-7), 20.1 (q, C-9), 19.7 (q,
C-10).
22
(R)-Carvomenthene. ½ꢀꢂD ꢃ78:2ꢁ (c ¼ 2:0, CHCl3).
(1S,4R)- and (1R,4R)-p-menth-2-en-1-ol (4 and 5).
(S)-Carvomenthene (8.28 g, 60 mmol) was dissolved in
300 ml of CH2Cl2 with 180 ml of 0.5 N-aqueous sodium
bicarbonate. m-CPBA (7.74 mg, 60 mmol in 200 ml of
CH2Cl2) was slowly added over 1 h into the solution,
and the mixture stirred at room temperature under N2
gas for 9 h. The CH2Cl2 layer was successively washed
twice with 1 N-aqueous NaOH and saturated-aqueous
NaCl. After drying over anhydrous Na2SO4, the solvent
was removed to give 7.96 g of mixture of the two

2546
T. KASHIWAGI et al.
diastereoisomers, cis- and trans-epoxy-(S)-carvomen-
thane, as a colorless oil.
1:0, CHCl3).
22
(1R,4S)-p-menth-2-en-1-ol (7). ½ꢀꢂD þ25:2ꢁ (c ¼
After displacing the air with N2 gas, small pieces of
sodium (1.15 g, 50 mmol) were slowly added over 1 h
to 2-heptanol (13.0 g, 100 mmol) in the flask, and the
mixture stirred at 90 ꢁC until the sodium had completely
reacted. After 100 ml of diethylenglycol monomethyl
ether had been mixed with the solution, the mixture
was added dropwise to the epoxides (7.70 g in 100 ml of
diethylenglycol monomethyl ether) at 90 ꢁC under N2
gas. The reaction mixture was then stirred and heated at
140 ꢁC for 14 h. After cooling to room temperature, the
reactant was diluted with 100 ml of water and extracted
with hexane (110 ml ꢀ 3). The hexane layer was
successively washed with water, 1 N-HCl and saturat-
ed-aqueous NaCl, and dried over anhydrous Na2SO4.
After removing the solvent under reduced pressure, the
product was distilled under reduced pressure to give
2.32 g of mixture of (1S,4R)- and (1R,4R)-p-menth-2-
en-1-ol as a colorless oil, bp 120–137 ꢁC/23 mmHg. The
mixture (1.5 g) was chromatographed in a silica gel
(Wako C300) column eluted with 3% and 5% diethyl
ether in hexane to obtain 723 mg of (1S,4R)-p-menth-
2-en-1-ol and 663 mg of (1R,4R)-p-menth-2-en-1-ol as
colorless oils in respective yields of 12.1% and 11.4%.
1:0, CHCl3).
Acknowledgments
We thank Mr. Shoichi Saito, Forest Research and
Instruction Center of Yamagata Prefecture, and Mr.
Koichi Nunokawa, Forestry Research Institute of
Niigata Prefecture for their assistance with collecting
the logs.
References
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Zool., 36, 317–321 (2001).
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quercivorus (Murayama) (Coleoptera: Platypodidae) on
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Tokoro, M., Kashiwagi, T., Tebayashi, S., and Kim, C.-S.,
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quercivorus (Murayama) (Coleoptera: Platypodidae).
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22
(1S,4R)-p-menth-2-en-1-ol (4). ½ꢀꢂD ꢃ25:2ꢁ (c ¼
1:0, CHCl3). EIMS m=z (%): 154 (3), 139 (21), 136 (59),
134 (15), 121 (41), 119 (54), 111 (11), 105 (11), 94 (31),
93 (100), 92 (43), 91 (85), 84 (11), 83 (11), 81 (13), 80
(14), 79 (37), 78 (13), 77 (74), 71 (12), 69 (19), 67 (10),
65 (16), 55 (10), 53 (10). 1H-NMR (CDCl3) ꢁ: 5.60 (2H,
s, H-2 and 3), 1.93 (1H, dt, J ¼ 9:2 and 5.6, H-4), 1.84
(1H, ddd, J ¼ 12:4, 6.0 and 2.8, H-6a), 1.72 (3H, ddd,
J ¼ 13:2, 5.6 and 2.8, H-5a), 1.60 (1H, td, J ¼ 12:4 and
3.2, H-6b), 1.57 (1H, m, H-8), 1.36 (1H, dddd, J ¼ 13:2,
12.4, 9.2 and 3.2, H-5b), 1.21 (3H, s, H-7), 0.89 (3H, d,
J ¼ 6:8, H-9) 0.87 (3H, d, J ¼ 6:8, H-10). 13C-NMR
(CDCl3) ꢁ: 134.5 (d, C-2), 131.2 (d, C-3), 69.6 (s, C-1),
41.7 (d, C-4), 38.0 (t, C-6), 31.6 (d, C-8), 28.5 (q, C-7),
23.6 (t, C-5), 19.8 (q, C-9), 19.4 (q, C-10).
22
(1R,4R)-p-menth-2-en-1-ol (5). ½ꢀꢂD þ28:7ꢁ (c ¼
1:0, CHCl3). EIMS m=z (%): 154 (2),136 (16), 121 (11),
119 (14), 94 (20), 93 (100), 92 (17), 91 (32), 79 (23), 77
(36), 71 (11), 69 (13), 65 (11). 1H-NMR (CDCl3) ꢁ: 5.66
(2H, s, H-2 and 3), 1.79 (1H, m, H-4), 1.76 (H, m, H-6a),
1.59 (1H, m, H-8), 1.54 (1H, m, H-5a), 1.41 (1H, m,
H-6b), 1.38 (1H, m, H-5b), 1.21 (3H, s, H-7), 0.92 (3H,
d, J ¼ 6:8, H-9), 0.89 (3H, J ¼ 6:8, H-10). 13C-NMR
(CDCl3) ꢁ: 133.4 (d, C-2), 133.0 (d, C-3), 67.4 (s, C-1),
42.1 (d, C-4), 37.2 (t, C-6), 31.7 (d, C-8), 29.6 (q, C-7),
21.6 (t, C-5), 19.7 (q, C-9), 19.3 (q, C-10).
9) Adams, R. P., ‘‘Identification of Essential Oil Compo-
nents by Gas Chromatography/Quadrupole Mass Spec-
troscopy,’’ Allured Publishing Corporation, Illinois,
pp. 111–117 (2001).
(1S,4S)-p-menth-2-en-1-ol (6). ½ꢀꢂD ꢃ28:7ꢁ (c ¼
22