Regio- and stereochemical study of sex pheromone of pine sawfly; Diprion nipponica

Article abstract of DOI:10.1246/bcsj.75.111

Regio- and stereoisomers of 1,2,ω-trimethyldecyl propionate (ω = 5-9) were prepared from stereochemically pure chiral building blocks as sex pheromone candidates of a pine sawfly; Diprion nipponica. Among the synthesized candidates, (1S,2R,8S)-1,2,8-trimethyldecyl propionate was found to be the sex pheromone of D. nipponica, based on compatibility of its GC-MS data with that of the extract of females, and its significantly high pheromone activity in a field bioassay. The field bioassay of the synthesized compounds also revealed that (1S,2R,SR)-1,2,8-trimethyldecyl propionate, (1S,2R,7S)-1,2,7-trimethyldecyl propionate, and (1S,2R,6S)-1,2,6-trimethyldecyl propionate could attract male sawflies to some extent as pheromone mimics.

Full text of DOI:10.1246/bcsj.75.111

Bull. Chem.
Soc. Jpn.
75
1
© 2002 The Chemical Society of Japan
Bull. Chem. Soc. Jpn., 75, 111–121 (2002)
111
The Chemical
Society of Ja-
pan
The Chemical
Society of Ja-
Regio- and Stereochemical Study of Sex Pheromone of Pine Sawfly;
Diprion nipponica
pan
OB
1
Sex Pheromone of Pine Sawfly; D. nipponica
A. Tai et al.
Akira Tai, Emi Syouno, Kazuki Tanaka, Morifumi Fujita, Takashi Sugimura,* Yasutomo Higashiura,†
Masashi Kakizaki,†† Hideho Hara,††† and Tikahiko Naito††††
15
2002
111
121
BCSJA8
0009-2673
01223
7
9
Faculty of Science, Himeji Institute of Technology, Kohto, Kamigori, Ako-gun, Hyogo 678-1297
†School of Life Science, Tokyo University of Pharmacy and Life Science, Horinouchi, Hachiouji, Tokyo 192-0392
††Hokkaido Ornamental Plants and Vegetable Research Center, Higashi-Takigawa 753, Takigawa, Hokkaido 073-0026
†††Hokkaido Forestry Research Institute, Koshunai, Bibai, Hokkaido, 079-0198
2001
9
††††Faculty of Agriculture, Kobe University, Rokkoudai, Nada, Kobe, 657-0013
6
2001
(Received July 9, 2001)
4.9.1
Regio- and stereoisomers of 1,2,ω-trimethyldecyl propionate (ω = 5–9) were prepared from stereochemically pure
chiral building blocks as sex pheromone candidates of a pine sawfly; Diprion nipponica. Among the synthesized candi-
dates, (1S,2R,8S)-1,2,8-trimethyldecyl propionate was found to be the sex pheromone of D. nipponica, based on compat-
ibility of its GC-MS data with that of the extract of females, and its significantly high pheromone activity in a field bioas-
say. The field bioassay of the synthesized compounds also revealed that (1S,2R,8R)-1,2,8-trimethyldecyl propionate,
(1S,2R,7S)-1,2,7-trimethyldecyl propionate, and (1S,2R,6S)-1,2,6-trimethyldecyl propionate could attract male sawflies
to some extent as pheromone mimics.
Pine sawflies (Hymenopetra: Diprionidae) consisting of
more than 120 spices are common insects in conifer forests.
Several of their species are considered to be severe pests of
pine trees. In 1976, Jewtt and co-workers1 found that 1,2,6-tri-
methyltetradecyl acetate (1A) and its propionate analogue (1P)
are sex pheromones of Neodiprion lecontei (North American
species) and Diprion similis (European species introduced to
North America), respectively, and hypothesized a variation of
pheromones among species resides in the stereochemistry of
alcohol moieties. Subsequent stereochemical studies from
1978 to 1990 have revealed that all Neodiprion species so far
studied in North America, Japan, China, and Europe used
(1S,2S,6S)-1A or 1P as the main pheromone and (1S,2R,6R)-
1A or 1P as a synergist or inhibitor,2 while Diprion similis
used (1S,2R,6R)-1P as the main pheromone.3,4 Thereafter,
(1S,2R,6R)-1,2,6-trimethydodecyl propionate (2P)5 and one of
the stereoisomers of 1,2,6,10-tetramethyldodecyl propionate
(3P)6 were found to be pheromones of Diprion pini and Mi-
crodiprion pallipes, respectively, by a Swedish group.
Diprion nipponica is a common sawfly in northern Japan.
This work began in 1991 on the occasion of heavy defoliation
in pine forests of eastern Hidaka, Hokkaido caused by mass
outbreaks of D. nipponica.7 After exploring suitable field-test
stations where traps fitted with caged verging females could at-
tract sufficient numbers of males, field attraction tests with
synthetic pheromones were carried out. However, the results
of those studies from 1992 to 1994 were disappointing. None
of the known pheromone components or related compounds
could attract males of D. nipponica. During this period, an in-
door breeding technique for the insect was established, and
reared virgin females and males became available in 1996.
With these insects, the analysis of natural pheromone was car-
ried out by using GC equipped with combined TCD and elec-
troanntenanographic detector (EAD), and by GC-MS.8 The re-
tention time of an EAD-active compound in an extract of vir-
gin females was found to be much shorter than those of 2P and
1P. Mass spectra corresponding to the EAD-active fraction in-
dicated that an ester of propionic acid and a saturated alcohol
having 13 carbon atoms (C13 alcohol) is the pheromone.
Among several compounds synthesized based on the MS data
and on the structural relation with known pheromones,
(1S,2R,6RS)-1,2,6-trimethyldecy propionate (4P), a 1 to1 mix-
ture of (1S,2R,6R)- and (1S,2R,6S)-4P was found to attract a
significant number of males of D. nipponica in the field attrac-
tion test. Being a shorter homologue of the pheromones of D.
similis and D. pini, either (1S,2R,6S)-4P or (1S,2R,6R)-4P was
proposed to be a promising pheromone candidate at this stage.8
However, a careful analytical study conducted thereafter with
synthesized sterochemically pure (1S,2R,6R)-4P and
(1S,2R,6S)-4P showed that their GC and MS data were slightly
different from those of the natural product, and suggested that
the true pheromone is not either epimer of 4P, but should be its
regioisomers.
This paper deals with our chemical approach for determin-
ing the sex pheromone of D. nipponica by synthesizing all
pheromone candidates of possible regio- and stereoisomers of
4P.
Results and Discussion
Synthetic Studies. The common structural feature of all

112 Bull. Chem. Soc. Jpn., 75, No. 1 (2002)
Sex Pheromone of Pine Sawfly; D. nipponica
Fig. 1. Structures of pheromone candidates.
known pheromones of pine sawflies is a branching of methyl
groups at the 1, 2, and 6 positions. Judging from the variation
of the known pheromone structure, the functionality and their
stereochemistries at the 1 and 2 positions are rigorously recog-
nized by the pheromone receptor, and thus they must be an es-
sential unit for pheromone activity in all Diprion and Neodip-
rion species. The structure of an alkyl chain, including a num-
ber of branched methyl groups and their positions, could be di-
verse and its recognition with the receptor is rather loose. In
fact, we have experienced that (1S,2S)-1,2-dimethyltetradedyl
acetate, a compound lacking the 6-methyl group in 1A, func-
tioned as a pheromone mimic for N. sertifer in a field test.9
When GC and MS data of the EAD-active compound from the
D. nipponica extract are taken into account, the pheromone
candidates to be synthesized are concentrated to certain stere-
oisomers of six regioisomers, as listed in the Fig. 1.
Our synthetic procedure for the pheromone candidates is
summarized in Scheme 1. C8 blocks 23–28 were converted to
Grignard reagents, and were coupled with a C5 block 31 de-
rived from methyl 3-hydroxy-2-methybutanoate (29) through
three steps.10 A combination of a specified regio- and stereo-
isomer of the C8 block and a specified stereoisomer of the C5
block gave one of the stereoisomers of C13 alcohols (4H, 5H,
6H, 7H, 8H, and 9H). The propionates (4P, 5P, 6P, 7P, 8P,
and 9P) derived from the corresponding C13 alcohols, were
purified by preparative GC and employed for analytical and
field studies.
Optically pure C8 blocks were prepared from (R)- and (S)-
methyl 3-hydroxy-2-methylpropioate (10) supplied by the
courtesy of Kaneka Co. The procedures used to obtain (R)-C8
blocks are shown in Scheme 2. After protection of the hydroxy
group in (S)-10, the ester moiety was reduced with LiAlH4,
followed by tosylation of the resulting hydroxy group to give
(S)-2-methyl-3-tetrahydropyranyloxypropyl tosylate (11),
which was utilized as a common starting material for the fol-
Scheme 1.
lowing steps. The coupling of (S)-11 with Grignard reagents

A. Tai et al.
Bull. Chem. Soc. Jpn., 75, No. 1 (2002) 113
Scheme 2.
prepared from bromopropane, bromoethane, and bro-
momethane gave (R)-2-methyl-1-hexanol (12), (R)-2-methyl-
1-pentanol (13), and (R)-2-methyl-1-butanol (14), respectively.
Bromides, 15, 16, and 17 derived from the above alcohols
were converted to the Grignard reagents, which were subjected
to reactions with the corresponding counter part to yield (R)-3-
methyl-1-heptanol (18), (R)-4-methyl-1-heptanol (19), and
(R)-5-methyl-1-heptanol (20), respectively. By the same pro-
cedure as mentioned above, (S)-isomers were obtained form
(R)-10 except for (S)-19, which was prepared from commer-
cially available (S)-16. Achiral 6-methyl-1-heptanol (21) and
rac-2-methyl-1-heptanol (22) were prepared by one-step reac-
tions from commercially available compounds. C8 blocks,
23–26 and 28, were obtained by bromination of the resulting
alcohols via the tosylates. 1-Bromooctane (27) was obtained
from a commercial source.
Determination of the Pheromone by Analytical and Field
Studies with Synthetic Pheromone Candidates.
Through
an analytical study of the extract of virgin females of D. nip-
ponica with GC-EAD, the EAD-active material was found to
exist in ester fractions of the Florisil column chromatography,
and its peak on GC appeared immediately after the peak of iso-
proyl dodecanoate (32). Choosing 32 as an internal standard,
the EAD-active material was subjected to a GC-MS analysis.
The retention times (rt) of the EAD-active material and 32
were 28.27 and 28.21 min, respectively. The mass spectrum
(MS) of the EAD-active material is shown in Fig. 2A.
Based on the results mentioned above, a comparative study
with synthetic pheromone candidates and the EAD-active ma-
terial was carried out with GC-MS. The rt of each compound
calibrated with the internal standard (32) is listed in Table 1 to-
gether with the difference in the rt from the EAD-active mate-
rial. Among the compounds examined, (1S,2R,8R)- and
(1S,2R,8S)-7P were found to be identical with the EAD-active
material in their rt values. The MS of the synthetic pheromone
candidates are shown in Figs. 2B–2D. The EAD-active mate-
rial and (1S,2R,8S)- and (1S,2R,8R)-7P were found to give
very compatible spectra with each other, while the spectra of
Stereochemically pure C5 blocks, (2S,3S)- and (2R,3S)-me-
thyl 3-hydroxy-2-methylbutanoate (29), were prepared by a
method established by our group.10 The starting material, (S)-
butyl 3-hydroxybutanoate, was obtained by the enantio-differ-
entiating hydrogenation of methyl acetoacetate over asymmet-
rically modified Ni and a following optical enrichment.11

114 Bull. Chem. Soc. Jpn., 75, No. 1 (2002)
Sex Pheromone of Pine Sawfly; D. nipponica
Table 1. GC Data of Synthetic Pheromone Candidates and EAD Active Material in the Female Extract
Synthetic
compound
Modified rt * calibrated
Difference in modified rt
and observed rt of sawfly
extract (rtm − 28.27)/min
by standard (rtm)/min
1
(1S,2R,5RS)-5P
27.04
27.25
27.68
27.65
27.99
27.97
28.26
28.25
28.11
27.92
29.19
−1.23
−1.02
−0.59
−0.62
−0.28
−0.30
−0.01 ꢀ 0
−0.02 ꢀ 0
−0.16
2
3
4
5
6
7
8
9
(1S,2R,6S)-4P
(1S,2R,6R)-4P
(1S,2R,7S)-6P
(1S,2R,7R)-6P
(1S,2R,8S)-7P
(1S,2R,8R)-7P
(1S,2S,8S)-7P
(1S,2R)-8P
−0.35
+0.92
10 (1S,2R)-9P
*Calibration was carried out by following equation; A × (28.21/B), where A and B are the
observed rt of each compound and that of internal standard receptively, listed in Table 4. The fig-
ure, 28.21 is the rt of internal standard when EAD active compound appeared at rt 28.27.
(1S,2R,6S)-4P and (1S,2R,7S)-6P are slightly different from
those of the EAD-active material. Compounds 4P and 6P dis-
play an additional peak at m/z 139–140. This difference was
diagnostic to specify the pheromone structure in this study.
The peaks m/z 182 (M+ − C2H5CO2) and 153 (a fragment
caused by fission at the C2–C3 bond) which existed in the
EAD-active material were important information at an early
stage of this study to postulate the structure at the 1 and 2 posi-
tions of the pheromone.
The results of a field bioassay of the synthesized pheromone
candidates are summarized in Table 2. Four compounds
((1S,2R,6S)-4P,
(1S,2R,7S)-6P,
(1S,2R,8S)-7P,
and
(1S,2R,8R)-7P were found to attract males to a trap baited with
10 µg of aliquot. Among them, a trap with (1S,2R,8S)-7P
caught the largest number of males, and had the ability to at-
tract males even with the low amount of bait (2 µg). The activ-
ity of (1S,2R,8S)-7P was also proved to be significantly higher
than that of the other three compounds by a statistical analysis.
Thus, the field study revealed that (1S,2R,8S)-7P was only eli-
gible to be the true pheromone, and that the others were phero-
mone mimics.
The analytical studies specified that the true pheromone
should be either (1S,2R,8S)- or (1S,2R,8R)-7P. Out of these
two epimers, only (1S,2R,8S)-7P fulfilled the biological re-
quirement, and thus (1S,2R,8S)-1,2,8-trimethyldecyl propi-
onate was determined to be the true sex pheromone of D. nip-
ponica.
The S configuration at the 8 position is very acceptable as a
natural product, since the S configuration of ethyl-methyl-
branching of an alkyl chain end was common in natural prod-
ucts: e.g. (2S,3S)-2-amino-3-methylpentanoic acid (L-isoleu-
cine), (S)-2-methyl-1-butanol in fusel oil, (S)-6-methyloctano-
ic acid from “polymyxin”, and (S)-10-methyldodecanoic acid
in wool wax. The present results indicated that the presence of
methyl branching at the 6 position in known pheromones of
sawflies was not an essential requirement for the sawfly phero-
mones. The variation of the alkyl end structure has been
proved to be more versatile than that expected based on the
early studies. The presence of pheromone mimics indicates
Fig. 2. MS of EAD active fraction of female sawfly extract
and synthetic pheromone candidates.

A. Tai et al.
Bull. Chem. Soc. Jpn., 75, No. 1 (2002) 115
Table 2. Field Response of D. nipponica ( ) to Traps Baited with Synthetic Pheromone Candidates
No
Date of field bioassay
Aug 12–21, 1997
Aug 13–Sep 3, 1998
Jul 28–Aug 4, 1999
Jul 24–Aug 10, 2000
Mean (catch/trap) SE Mean (catch/trap) SE Mean (catch/trap) SE* Mean (catch/trap) SE*
Amount baited
Compound
on a trap/µg
n = 6
12.1 4.6
n = 5
3.8 0.9
n = 6
—
—
—
n = 5
—
—
—
1
2
3
4
5
6
7
8
9
(1S,2R,6RS)-4P
(1S,2S,6RS)-4P
(1R,2S,6RS)-4P
(1R,2R,6RS)-4P
(1S,2R,6S)-4P
(1S,2R,6R)-4P
(1S,2R,5RS)-5P
(1S,2R,7S)-6P
(1S,2R,7R)-6P
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
0
0
0
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
9.6 3.4
0
7.2 4.0 b
7.2 2.3 b
0 a
—
—
—
—
—
—
—
—
—
—
0 a
0.5 0.1 a
4.0 1.6 a,b
0 a
—
10 (1S,2R,8S)-7P
11 (1S,2R,8R)-7P
12 (1S,2S,8S)-7P
13 (1S,2R)-8P
14 (1S,2S)-8P
16.5 4.8 c
51.4 19.7 c
2.0 1.2 a,b
14.0 5.3 b
—
0 a
0 a
—
0 a
—
—
0 a
15 (1S,2R)-9P
—
16 (1S,2R,6S)-4P
17 (1S,2R,7S)-6P
18 (1S,2R,8S)-7P
19 (1S,2R,8R)-7P
2
2
2
2
—
—
—
—
0
0.6 0.4
—
1.6 0.6
—
—
—
—
0
2.1 1.4
0
*Means followed by the same letter (a,b, or c) are not significantly different according to Fisher’s protected least significant
difference (p < 0.05).
hydropyranyloxypropyl Tosylate (11). To a solution of (S)-
methyl 3-hydroxy-2-methylpropioate (10) (40 g) and dehydropyr-
ane (35 g) in 70 mL of ether was added p-toluenesulfonic acid (0.2
g) with stirring. After an exothermic reaction completed, the mix-
ture was kept for 5 h at room temperature, washed with aq
NaHCO3, and dried over K2CO3 overnight. The resulting solution
was added to a stirred and ice-cooled suspension of LiAlH4 (18 g)
in 1000 mL of ether. The mixture was refluxed for 3 h, cooled
down with an ice bath, and hydrolyzed with 33 mL of water. A
white precipitate was filtered on a Celite pad and washed with two
200 mL portions of ether. The combined filtrate was concentrated
in vacuo. Distillation of the residue gave 53.6 g of a diasteromeric
mixture of (R)-2-methyl-3-tetrahydropyranyloxypropan-1-ol
(90.9%, bp 86–90 °C/6 mm Hg (1 mmHg = 133.322 Pa)). This
material (50 g) was added to an ice-cooled solution of p-toluene-
sulfonyl chloride (64 g) in dry pyridine (220 mL) under stirring,
and then kept for 4 h to complete the reaction. The mixture was
poured into ice-water (500 g) and extracted with three 300 mL
portions of ether. The combined extract was washed with aq
CuSO4, aq NaHCO3, and brine, successively, and then dried over
Na2SO4. Evaporation of the solvent from the solution in vacuo
gave (S)-11 (93.9 g, 93% crude yield). This material was used for
the next step without further purification.
that the olfactory receptor of D. nipponica is not as rigorous as
to specify a single compound, but is sufficiently rigorous to
avoid interbreeding and to maintain reproductive isolation
from an ecological point of view.
Experimental
Instrumental. 1H NMR spectra were recorded at 400 MHz
on a JEOL EXcaliber-400 spectrometer with CDCl3 as a solvent.
The chemical shifts and coupling constants for new compounds
are shown in ppm and Hz, respectively. IR spectra were recorded
on a JASCO IR-800 spectrometer. Optical rotation was measured
on a Perkin-Elmer 241B polarimeter. Analytical GC for the syn-
thetic study was carried out with a Shimadzu GC-6A (TCD)
equipped (pass 1) with a HP-INNOWAX bonded wide-bore silica
capillary column (30 m × 0.53 mm) and (pass 2) with a stainless-
steel column (2 m × 3 mm) packed with 3% SE-30 on Chro-
mosorb-W. For chiral analysis, a Shimadzu GC-14A equipped
with a BETA DEX 225-coated silica capillary column (30 m ×
0.25 mm) was employed. Preparative GC was carried out with a
Shimadzu GC-6A (TCD) equipped (pass 1) with a stainless-steel
column (5 m × 5 mm) packed with 5% NPGS on Chromosorb-W,
and (pass 2) with the same stainless steel column (5 m × 5 mm)
packed with 5% OV-101 on Chromosorb-W. GC-EAD was car-
ried out with a Hewlett-Packerd 6890 (TCD) equipped with a HP-
INNOWAX coated wide bore capillary column (30 m × 0.53
mm). The oven temperatures were fixed at 60 °C for the first 2
min, then elevated at a constant rate (1 °C/min) up to 230 °C.
EAD was assembled according to the design reported by Struble
et al.12 GC-MS was carried out with a Hewlett-Packerd 5971A in
the electron impact mode (EI, 70 ev). The separation column was
a HP-INNOWAX capillary column (30 m × 0.25 mm) and the
oven temperatures were controlled by the same program as that
applied to GC-EAD.
(R)-2-Methyl-3-tetrahydropyranyloxypropyl Tosylate (11).
By the same method as described above, (R)-10 (25 g) was con-
verted to (R)-11 (61.1 g).
(R)-2-Methy-1-hexanol (12). Grignard reagent prepared
from 1-bromopropane (15 g) and Mg (3.4 g) in THF (120 mL)
was cooled to −50 °C and mixed with CuI (0.5 g). To the solu-
tion, (S)-11 (30 g) dissolved in 30 mL of THF was added at −50
°C with stirring. The mixture was allowed to stand over night at
room temperature, and was then heated at 40–50 °C for 1 h. After
cooling, the reaction mixture was poured in a saturated and ice-
cooled aq NH4Cl solution (100 mL) and allowed to stand for 1 h
Chemicals. Synthesis of C8 Block. (S)-2-Methyl-3-tetra-

116 Bull. Chem. Soc. Jpn., 75, No. 1 (2002)
Sex Pheromone of Pine Sawfly; D. nipponica
under stirring. The insoluble materials in the mixture were filtered
on a Celite pad and washed with ether (100 mL). The ethereal
layer of the combined filtrate was separated, washed with 100 mL
each of aq NH4Cl, aq NaHCO3, and water, and then dried over
K2CO3 and concentrated in vacuo. The residue was mixed with
methanol (250 mL) containing a small amount of p-toluensulfonic
acid (ca. 100 mg) and allowed to stand for 5 h at room tempera-
ture. After neutralization with sodium methoxide (1 mL of 0.5 M
methanol solution), the solution was concentrated in vacuo. The
concentrate consisting of 12 and 2-methoxytetrahydropyrane was
subjected to fractional distillation under reduced pressure with a
Vigreux column to afford crude (R)-12 containing 2-methoxytet-
rahydropyrane in ca. 5% (7.3 g, 65% yield by calculation) as a
fraction of bp 75–82 °C at 45 mmHg. The product was used for
the next step without further purification. The preparative GC
(NPGS, 70 °C) of a small portion of this material afforded pure
(R)-12: [α]D20 = 12.2 (c 5.4, MeOH (Ref. 13, [α]D25 = 2.45 in etha-
nol)), GC (BETA DEX 225, 70 °C) 24.9 min, single peak (No
peak of the antipode (rt = 26.2 min) was observed.), MS m/z (M+
− H), Found: 115.1122, Calcd for C7H15O: 115.1123, IR 3325,
2925, 1460, 1380, 1040 cm−1, 1H NMR (400 MHz, CDCl3) δ 0.88
(t, J = 6.8 Hz, 3H), 0.90 (d, J = 6.3 Hz, 3H), 1.05–1.63 (m, 7H),
3.38–3.51 (m, 2H).
over K2CO3 and concentrated in vacuo. Distillation of the residue
under reduced pressure gave (R)-18 (3.3 g, 70% yield from 15): bp
85–86 °C/13 mmHg, [α]D20 = +3.09 (c 2.14, MeOH), GC (BETA
DEX 225, 70 °C) 29.3 min, single peak, (No peak of the antipode
(rt = 29.8 min) was observed.), MS m/z (M+ − H), Found:
129.1273, Calcd for C8H17O: 129.1279, IR 3350, 2925, 1460,
1370, 1050 cm−1, 1H NMR (400 MHz, CDCl3) δ 0.872 (t, J = 6.8
Hz, 3H), 0.874 (d, J = 6.3 Hz, 3H), 1.11–1.61 (m, 10H), 3.64 –
3.67 (m, 2H).
(S)-3-Methyl-1-heptanol (18). By the same method as de-
scribed above, (S)-15 (6.5 g) was converted to (S)-18 (4.1 g, 86%
yield): [α]D20 = −3.76 (c 2.34, MeOH (Ref. 13, [α]D25 = −2.76)),
GC (BETA DEX 225, 70 °C) 29.8 min, single peak, (No peak of
the antipode (rt = 29.3) was observed.). The other analytical data
were identical with those of (R)-18.
(R)-1-Bromo-3-methylheptane (23). (R)-18 (2.1 g) was add-
ed to an ice-cooled solution of p-toluenesulfonyl chloride (3.9 g)
in dry pyridine (8 mL) under stirring and then kept for 4 h to com-
plete the reaction. The mixture was poured into ice-suspended 2
M aq HCl (50 mL) with 50 mL of ether. After being well shaken,
the ethereal layer was separated and washed with 50 mL each of 2
M aq HCl, brine, aq NaHCO3, and brine, successively, and then
dried over MgSO4 and concentrated in vacuo. The residue was
added to a solution of anhydrous LiBr (6.6 g) dissolved in DMF
(35 mL) at 40 °C, and the mixture was stirred overnight at room
temperature. The mixture was poured into water (35 mL), and ex-
tracted three times with a 30 mL portion of pentane. The com-
bined extract was washed with water, dried over MgSO4 and con-
centrated. The concentrate was passed through a short silica gel
column with pentane as an eluent. The elute was concentrated in
vacuo and the residue distilled under reduced pressure to give (R)-
23 (2.9 g, 50% yield): bp 76–77 °C/22 mmHg, GC (SE-30, 90 °C)
4.1 min (single peak), 1H NMR (400 MHz, CDCl3) δ 0.87 (d, J =
6.4 Hz, 3H), 0.88 (t, J = 6.3 Hz, 3H), 1.10–1.30 (m, 6H), 1.55–
1.69 (m, 2H), 1.85 (m, 1H), 3.37–3.48 (m, 2H).
(S)-1-Bromo-3-methylheptane (23). By the same method as
described above, (S)-18 (3.7 g) was converted to (S)-23 (3.53 g,
63% yield). The analytical data were identical to those of (R)-23.
(R)-2-Methyl-1-pentanol (13). By the same procedure as
that applied to the preparation of 12, Grignard reagent prepared
from bromoethane (12 g) and Mg (2.6 g) in THF (100 mL) was
subjected to a reaction with (S)-11 (30 g). Removal of the protect-
ing group from the product gave (R)-13 containing 2-methoxytet-
rahydropyrane in ca. 5% (7.2 g, 78% yield by calculation) as a
fraction at bp 70–76 °C at 45 mmHg. This material was used in
the next step without further purification. The preparative GC
(NPGS) of a small portion of this material afforded pure (R)-13:
[α]D20 = 12.3 (c 5.0, MeOH), GC (BETA DEX 225, 65 °C) 17.3
min, single peak, (No peak of the antipode (rt = 19.0 min) was
observed.), MS m/z, Found: 101.0958, Calcd for C6H13O:
101.0966, IR 3375, 2925, 1450, 1380, 1050 cm−1, 1H NMR (400
MHz, CDCl3) δ 0.886 (t, J = 6.8 Hz, 3H), 0.895 (d, J = 6.8 Hz,
3H), 1.04–1.64 (m, 5H), 3.37–3.52 (m, 2H).
(S)-2-Methyl-1-hexanol (12). By the same method as de-
scribed above, (R)-11 (30 g) was converted to (S)-12 containing 2-
methoxytetrahydropyrane in ca. 5% (7.7 g, 68% yield by calcula-
tion): [α]D20 = −12.7 (c 5.2, MeOH), GC (BETA DEX 229, 70 °C)
26.2 min, single peak (No peak of the antipode (rt = 24.9 min)
was observed.). The other analytical data were identical with
those of (R)-12.
(R)-1-Bromo-2-methylhexane (15). Crude (R)-12 (7.1 g)
was added to an ice-cooled solution of p-toluenesulfonyl chloride
(14 g) in dry pyridine (25 mL) under stirring, and was then kept
for 4 h to complete the reaction. The mixture was poured into ice-
water (50 mL) and extracted with three 50 mL portions of ether.
The combined extract was washed with aq CuSO4 solution, aq
NaHCO3, and brine, successively, and dried over Na2SO4. The
solvent was evaporated from the solution in vacuo, and the residue
was kept overnight under a high vacuum to give 2-methylhexyl
tosylate (16.8 g). This material (16.5 g) was added to a solution of
anhydrous LiBr (22 g) dissolved in DMF (150 mL) at 40 °C, and
the mixture was stirred overnight at room temperature. The mix-
ture was poured into water (150 mL) and extracted three times
with a 150 mL portion of pentane. The combined extract was
washed with water, dried over MgSO4, and concentrated. The res-
idue was passed through a short silica gel column with pentane as
an eluent. The elute was evaporated in vacuo and distilled under
reduced pressure to give (R)-15 (7.3 g, 67% yield): bp 73–78 °C/
40 mmHg. The analytical data were compatible with those report-
ed in the literature.14
(S)-1-Bromo-2-methylhexane (15). By the same method as
described above, (R)-12 (7.1 g) was converted to (S)-15 (7.2 g,
66% yield). The analytical data were compatible with those re-
ported in the literature.14
(R)-3-Methyl-1-heptanol (18). To an ice-cooled solution of
Grignard reagent prepared from (R)-15 (6.5 g) and Mg (1 g) in 50
mL of ether, gaseous CH2O generated from well-dried paraform-
aldehyde (1.3 g) was introduced by the aid of N2 stream. After
praformaldehyde was consumed, the mixture was heated under re-
flux for 30 min, then cooled down and poured into ice-suspended
aq 2 M HCl (50 mL). The resulting ethereal layer was washed
with brine, aq NaHCO3, and brine, successively, and then dried
(S)-2-Methyl-1-pentanol (13). By the same method as de-
scribed above, (S)-11 (30 g) was converted to (S)-3 containing 2-
methoxytetrahydropyrane in ca. 5% (6.8 g, 74% yield): [α]D20
=
−13.1 (c 5.0, MeOH), GC (BETA DEX 225, 65 °C) 19.0 min, sin-
gle peak, (No peak of the antipode (rt = 17.3 min) was observed).
The other analytical data were identical to those of (R)-13.
(R)-1-Bromo-2-methylpentane (16). By the same proce-
dure as that applied to the preparation of 15, (R)-13 (6.6 g) was
converted to (R)-16 (5.6, 55% yield): bp 77–80 °C/89 mmHg, GC

A. Tai et al.
Bull. Chem. Soc. Jpn., 75, No. 1 (2002) 117
(SE30, 70 °C) 4.1 min, 1H NMR (400 MHz, CDCl3) δ 0.89 (t, J =
7.3 Hz, 3H), 0.99 (d, J = 6.8 Hz, 3H) 1.18–1.18 (m, 4H), 1.78 (m,
1H), 3.30 (dd, J = 9.8, 6.3 Hz, 1H), 3.37 (dd, J = 9.8, 4.9 Hz,
1H).
ture of (R)-14 and 2-methoxytetrahydropyrane in a ratio of 57:43
was obtained (11 g, 76% yield from 11 by calculation) as a frac-
tion of bp 125–130 °C. This material was used in the next step
without further purification. The preparative GC (NPGS, 60 °C)
(S)-1-Bromo-2-methylpentane (16). By the same method as
described above, (S)-13 (6.5 g) was converted to (S)-16 (6.3 g,
60% yield). The analytical data were identical to those of (R)-16.
(R)-4-Methyl-1-heptanol (19). To a Grignard reagent pre-
pared from (R)-16 (5.1 g) and Mg (0.9 g) in 30 mL of ether was
added a 20% solution of ethylene oxide in ether (10 mL) with stir-
ring at lower than 5 °C. The reaction mixture was allowed to
stand overnight and added into ice-suspended 2 M aq HCl (50
mL). The resulting ethereal layer was washed with brine, aq
NaHCO3, and brine successively, dried over K2CO3, and then con-
centrated in vacuo. Distillation of the residue under reduced pres-
sure gave a crude fraction of (R)-19 containing 4,7-dimethlyde-
cane in ca 3%, (2.1 g, 48.4% yield from 16 by calculation): bp 77–
80 °C/98 mmHg. This material was used in the next step without
further purification. The preparative GC (NPGS) of a small por-
tion of this material afforded pure (R)-19: GC (BETA DEX 225,
75 °C) 26.0 min, single peak, (No peak of the antipode (rt = 26.3
min) was observed). The [α]D20 value was too small to determine,
MS m/z, Found: 129.1270, Calcd for C8H17O: 129.1279, IR 3325,
2950, 1460, 1380, 1060 cm−1, 1H NMR (400 MHz, CDCl3) δ 0.83
(t, J = 6.3 Hz, 3H), 0.85 (d, J = 6.8 Hz, 3H), 1.06–1.61 (m, 10H),
3.61 (t, J = 6.3 Hz, 2H).
of a small portion of this material afforded pure (R)-14: [α]D20
=
6.45 (c 0.95, MeOH), GC (BETA DEX 225, 60 °C) 16.7 min, sin-
gle peak (No peak of the antipode (rt = 17.6 min) was observed).
1H NMR and IR spectral data were compatible with those of (S)-
14 obtained from a commercial source.
(R)-1-Bromo-2-methylbutane (17). The mixture of (R)-14
and 2-methoxytetrahydropyrane (10.0 g containing ca. 5.7 g of
14) was added to an ice-cooled solution of p-toluenesulfonyl chlo-
ride (15 g) in dry pyridine (30 mL) under stirring and the mixture
was kept for 4 h. The mixture was poured into ice-water (50 mL)
and extracted with three 50 mL portions of ether. The combined
extract was washed with aq CuSO4 solution, aq NaHCO3, brine,
and dried over NaSO4. Evaporation of the solvent and 2-methox-
ytetrahydropyrane from the solution under high vacuum at room
temperature gave (R)-2-methyllbutyl tosylate (10.2 g, 65% yield).
This product was subjected to a reaction with anhydrous LiBr (20
g) in DMF (100 mL) by the same method as that applied to the
preparation of 15 to yield (R)-17 (4.3 g, 68% yield): bp 91–93 °C,
1H NMR and IR spectral data were compatible with those of (S)-
17 obtained from commercial source.
(R)-5-Methyl-1-heptanol (20). Grignard reagent prepared
from (R)-17 (4.0 g) and Mg (0.78 g) in 20 mL of ether was cooled
to −50 °C and mixed with CuI (0.1 g). To the solution was added
3-bromo-1-propene (3.2 mL) dissolved in 10 mL of ether with
stirring. The mixture was kept for 1 h at −50 °C, allowed to stand
over night at room temperature, and then mixed with an ice-cooled
saturated aq. solution of NH4Cl (70 mL). The separated ethereal
layer was washed with 70 mL portions of saturated aq. solution of
NH4Cl, aq NaHCO3, brine, successively, and dried over MgSO4.
From the resulting solution of 5-methyl-1-heptene, ether was care-
fully distilled off through a Vigreux column under atmospheric
pressure. To a solution of the residue in 25 mL of THF was added
a 1 M solution of BH3 in THF (27 mL), and the mixture was al-
lowed to stand at room temperature for 5 h, and was then cooled
with an ice-bath. To this mixture, 30 mL of a 1 M aq solution of
NaOH was added, followed by 3.5 mL of a 35% solution of H2O2.
After stirring for 2 h at room temperature, the mixture was extract-
ed twice with 50 mL portions of ether. The combined extracts
were washed with brine, dried over K2CO3, and concentrated in
vacuo. Distillation of the residue under reduced pressure gave
(S)-4-Methyl-1-heptanol (19). By the same method as de-
scribed above, (S)-16 (4.5 g) was converted to (S)-19 containing 2-
methoxytetrahydropyrane in ca. 5%, (1.7 g, 45% yield): GC (BE-
TA DEX 225, 75 °C) 26.3 min, single peak, (No peak of the anti-
pode (rt = 26.0 min) was observed). The [α]D20 value was too
small to determine. Other analytical data were identical with
those of (R)-19.
(R)-1-Bromo-4-methylheptane (24). (R)-19 containing 5%
of 4,7-dimethyldecane (1.6 g) was converted to tosylate by the
same procedure as that applied to (R)-18. The obtained crude to-
sylate was chromatographed over silica gel. 4,7-Dimethyldecane
eluted with hexane was discarded and tosylate eluted with 25%
ethyl acetate in hexane was collected. The concentrate of the elu-
ent (2.6 g) was subjected to a reaction with LiBr in DMF by the
same method as applied to (R)-18 to give (R)-24 (1.23 g, 53%
yield from 19): bp 77–78 °C/20 mmHg, GC (SE-30, 80 °C) 8.5
1
min (single peak), H NMR (400 MHz, CDCl3) δ 0.84, (d, J =
7.3, 3H), 0.86 (t, J = 6.8 Hz, 3H), 1.05–1.45 (m, 7H), 1.83 (m,
2H), 3.61 (t, J = 6.2 Hz, 2H).
(R)-20 (2.39 g, 69% yield): bp 90–92 °C/26 mmHg, [α]D20
=
(S)-1-Bromo-4-methylheptane (24). By the same method as
described above, (S)-19 (1.4 g) was converted to (S)-24 (0.75 g,
38% yield). The analytical data were identical to those of (R)-24.
(R)-2-Methyl-1-butanol (14). Grignard reagent prepared
from bromomethane (2 M solution in ether, 125 mL) and Mg (6.4
g) was diluted with THF (100 mL) and cooled to −50 °C. To the
solution were added CuI (0.5 g) and then a solution of (S)-11 (30
g) in 30 mL of THF with stirring. The mixture was kept for 1 h at
−50 °C, allowed to stand overnight at room temperature, and then
heated at 40–50 °C for 1 h. By the same work up as that applied to
the preparation of 12, 2-methyl-1-tetrahydropyanylbutane (17.6 g)
was obtained from the reaction. This material was mixed with
methanol (250 mL) containing a small amount of p-toluensulfonic
acid (ca. 100 mg), and allowed to stand overnight at room temper-
ature. After neutralization with sodium methoxide (1 mL of 0.5 M
methanol solution), the mixture was subjected to fractional distil-
lation under atmospheric pressure with a Vigreux column. A mix-
−7.16 (neat), GC (SE-30, 80 °C) 7.9 min. MS m/z (M+ − H),
Found: 129.1277, Calcd for C8H17O: 129.1279, IR 3350, 2920,
1460, 1370, 1060 cm−1, 1H NMR (400 MHz, CDCl3) δ 0.83 (d, J
= 7.3 Hz, 3H), 0.84 (t, J = 7.3 Hz, 3H), 1.09–1.57 (m, 10H), 3.63
(t, J = 6.3 Hz, 2H).
(S)-5-Methyl-1-heptanol (20). By the same method as de-
scribed above, commercial (S)-1-bromo-2-methylbutane (17) (5.0
g) was converted to (S)-20 (2.7 g, 62% yield), [α]D20 = 6.94 (neat
(Ref. 13, [α]D24 = 2.99)). The analytical data were identical to
those of (R)-20. This compound has recently become commer-
cially available from TCI Co.
(R)-1-Bromo-5-methylheptane (25). This compound (2.1 g,
74% yield) was obtained from (R)-20 (1.9 g) by the same proce-
dure as that employed for the preparation of 23: bp 78 °C/20 mm-
1
Hg, GC (SE-30, 90 °C) 4.2 min (single peak), H NMR (400
MHz, CDCl3) δ 0.87 (d, J = 6.4 Hz, 3H), 0.88 (t, J = 6.3 Hz, 3H),
1.10–1.30 (m, 6H), 1.15–1.69 (m, 2H), 1.85 (m, 1H), 3.37–3.48

118 Bull. Chem. Soc. Jpn., 75, No. 1 (2002)
Sex Pheromone of Pine Sawfly; D. nipponica
(m, 2H).
pling of C5 and C8 blocks. The common procedure was as fol-
lows: Grignard reagent was prepared from C8 block (1 g, 5.1
mmol) and Mg (0.15 g, 1.2 equiv) in 5 ml of THF. The Grignard
reagent was cooled to −70 °C and mixed with a catalytic amount
of CuI (ca. 5 mg) under stirring. To the solution, a 1 M solution of
the C5 block in THF (5 mL, 5 mmol) was added. The mixture was
kept at −70 °C for 1 h, followed by stirring overnight at room
temperature. The mixture was poured into an ice-cooled saturate
aq solution of NH4Cl (20 mL) with 10 ml of ether. The separated
ethereal layer was washed with 20 mL portions of aq NaHCO3,
brine successively, dried over MgSO4, and concentrated under re-
duced pressure. The residue was mixed with methanol (10 mL)
containing p-toluenesulfonic acid (ca. 1 mg) and allowed to stand
over night. After removing methanol from the mixture, the resi-
due was dissoluble in ether (15 mL) and washed with aq NaHCO3,
brine, dried over K2CO3, and concentrated in vacuo. The residue
was dissolved in hexane (5 mL) and chromatographed over a silica
gel column. Evaporation of the solvent from a fraction eluted with
15% of ethyl acetate in hexane in vacuo gave a C13 alcohol. Syn-
thetic data of each C13 alcohol are listed in Table 3.
(S)-1-Bromo-5-methylheptane (25). By the same method as
described above, (S)-20 (1.4 g) was converted to (S)-25 (0.75 g,
38% yield). The analytical data were identical with those of (R)-
25.
6-Methyl-1-heptanol (21). Commercial 1-bromo-4-methyl-
pentane (5.5 g) was converted to Grignard reagent with Mg (1 g)
in ether (50 mL). To the ice-cooled Grignard reagent was added a
20% solution of ethylene oxide in ether (22 mL) with stirring.
The mixture was kept stirring overnight at room temperature, and
then worked up in the same manner as that applied to the prepara-
tion of 16 to yield crude 21 containing ca. 5% of 2,9-dimethylde-
cane (1.58 g, 34% yield by calculation): bp 84–89 °C/15 mmHg
(Ref. 13, 87–89 °C/14 mmHg). This material was used for the
next step without further purification.
1-Bromo-6-methylheptane (26). This compound was ob-
tained from 21 (1.5 g) by the same procedure as that employed for
the preparation of 24. 26 (1.35 g, 61% yield by calculation): bp
77–78 °C/20 mmHg, GC (3% SE-30, at 90 °C) 4.3 min (single
1
peak), H NMR (400 MHz, CDCl3): δ 0.85 (d, J = 6.8 Hz, 6H),
1.15–1.30 (m, 2H), 1.32–1.42 (m, 4H), 1.51 (m, 2H), 1.84 (septet,
J = 6.3 Hz, 1H) 3.39 (t, J = 6.8 Hz, 2H).
Preparation of C13 Propionates (4P–9P). All C13 alco-
hols listed in Table 3 were converted to propionate by the common
procedure and employed for a field bioassay. A C13 alcohol (0.1
mL, 86.5 mg) and pyridine (0.1 mL) were mixed with ether (2
mL) and cooled with an ice-bath. To this solution was added pro-
pionyl chloride (0.06 mL) under stirring. The mixture was al-
lowed to stand overnight at room temperature, and was then
poured into ice-suspended water (2 mL). The ethereal layer was
washed with solutions of 2 M aq HCl, aq NaHCO3, and brine, suc-
cessively, and dried over MgSO4. The solvent was removed, and
the residue was purified by preparative GC (OV-101, 150 °C).
The GC-purified material was made up to a 1 mg/mL hexane solu-
tion, and stored in sealed glass ampoules (1 mL each) as a stock
solution for bioassays. The analytical data of each purified mate-
rial are listed in Table 4.
rac-2-Methyl-1-heptanol (22). A solution of BH3 (1 M in
THF, 32 mL) was added to commercial 2-methyl-1-heptene (3.2
g) diluted with 25 mL of THF with stirring at 5 °C. The mixture
was allowed to stand at room temperature for 5 h, and was then
cooled with an ice bath. To this mixture, 30 mL of 3 M aq NaOH
was added, followed by 20 mL of 30% H2O2. After stirring at
room temperature for 2 h, the mixture was extracted twice with 50
mL portions of ether. The combined extracts were washed with
brine, dried over K2CO3, and concentrated in vacuo. Distillation
of the residue under reduced pressure gave 22 (4.0 g, yield 95%):
bp 83–90 °C/15 mmHg. The analytical data were compatible with
those reported in literatures.15
rac-1-Bromo-2-methylheptane (28). This compound was
obtained from 22 (3.5 g) by the same procedure as that employed
for the preparation of 24. 28 (3.9 g, 74% yield): bp 77–78 °C/ 20
mmHg. The analytical data were compatible with those reported
in the literature.6
Biomaterials and Analysis. Breeding of Insects.
In the
late summer of 1965, larvae were collected at Apoi Mountain,
eastern Hidaka Hokkaido. The larvae were fed on fresh pine
blanches (Pinus strobus) until forming cocoons. The cocoons
were held during the winter at outdoor temperature. In late spring,
large female cocoons and small male cocoons were separated and
the individuals were kept in plastic boxes. The cocoons in plastic
boxes were stored indoor under regulated temperature to let the
adults emerge around mid summer. The females used for the ex-
traction were stored in a freezer within a few hours after emer-
gence. The males used for EAD were collected shortly after
emergence and stored at 10 °C for 3 to 5 days before testing.
Preparation and Analysis of Female Extract. The whole
bodies of 30 females were immersed in 20 mL of hexane and kept
2 days in a refrigerator. After removing the insects, the extract
was filtered and concentrated to 100 µL. The concentrated extract
was chromatographed over Florisil with an ether/hexane eluent
following a procedure reported by Carroll.16 A 0.3 female equiva-
lent aliquot of each fraction was subjected to GC–EAD analysis
using an antenna of one of the males. The EAD-active material
was found in the fraction of 5% ether in hexane (corresponding to
ester fraction). After selecting isopropyl dodecanoate (32) as a
suitable internal standard for monitoring the rt of the EAD-active
material in the GC system, all other of active fractions (25 female
equivalent) were fractionated by the same GC system. The frac-
tion corresponding to the EAD-active material accompanying 32
as the internal standard was subjected to a GC-MS analysis.
Synthesis of C5 Block. Methyl (2S,3S)-3-Hydroxy-2-meth-
ylbutanoate and Methyl (2R,3S)-3-Hydroxy-2-methylbu-
tanoate (29). These compounds were prepared by the same pro-
cedure reported before from optically pure (S)-butyl 3-hydroxybu-
tanoate (100 g, [α]D20 = 14.6 (neat)).9 (2S,3S)-29 (14.8 g): [α]D20
=
36.8 (c 5.0, MeOH), (2R,3S)-29 (6.2 g): [α]D20 = −14.3 (c 5.0,
MeOH).
(2R,3S)-2-Methyl-3-tetrahydropyranyloxybutyl
Tosylate
(31) and (2S,3S)-2-Methyl-3-tetrahydropyranyloxybutyl Tosyl-
ate (31). These compounds were obtained from 29 by the same
procedure as that applied to the preparation of 11. (2R,3S)-31
(21.7 g, 67%) was obtained from (2S,3S)-29 (12.5 g) via (2R,3S)-
2-methyl-3-tetrahydropyranyloxybutanol (30) (bp 86–90 °C/6
mmHg, Found: C, 63.91, H, 10.91%; Calcd for C10H20O3: C,
63.79; H, 10.71%). This compound (20.5 g, 60 mmol) was made
up to a 1 M solution in THF and used for the coupling reaction.
(2S,3S)-31 (7.7 g, 71% yield) was obtained from (2R,3S)-29 (4.2
g) via (2S,3S)-30 (bp 86–90 °C/6 mmHg, Found: C, 63.78 H,
10.89%; Calcd for C10H20O3: C, 63.79; H, 10.71%). This com-
pound was made up to a 1 M solution in THF and used for the
coupling reaction.
Coupling of C5 and C8 Blocks. Preparation of C13 Alco-
hols (4H–9H). All C13 alcohols were obtained by cross cou-

A. Tai et al.
Bull. Chem. Soc. Jpn., 75, No. 1 (2002) 119
Table 3. Syntheses and Properties of the Alcoholic Parts (4H–9H) of the Pheromone Candidates
No Materials and amount of use
C8 block C5 block
Product of
[α]D20
Elemental analysis
1H NMR (400 MHz, CDCl3)
C13 alcohol
(c in MeOH) Calcd for C13H28O;
C,77.92; H,14.08
(g, mmol) (1 M solution/mL)
(g, yield from C5 block)
(2S,3R,6RS)-3,6-dimethyl-2-
undecanol (5H) (0.10, 23%)
Found:
1
2
rac-28
(2R,3S)-31
—
C,77.68; H,14.40 δ 0.82–0.88 (d, J = 6.8 Hz, 6H),
0.99–1.48 (m, 17H),
(0.56, 2.9)
(2.2)
1.11 (d, J = 6.8 Hz, 3H),
3.64–3.66 (m, 1H)
C,77.58; H,14.30 δ 0.83 (d, J = 6.8 Hz, 3H),
0.86 (d, J = 6.8 Hz, 3H),
(R)-23
(1.00, 5.1)
(2R,3S)-31
(2S,3R,7R)-3,7-dimethyl-2-
undecanol (4H) (0.16, 16%)
20.3
(2.8)
(5.0)
0.88 (t, J = 7.0 Hz, 3H)
1.10 (d, J = 6.8 Hz, 3H),
1.01–1.52 (m, 14H),
3.64 (m, 1H)
3
(S)-23
(1.00, 5.1)
(2R,3S)-31
(2S,3R,7S)-3,7-dimethyl-2-
undecanol (4H) (0.28, 28%)
22.7
(2.6)
C,77.83; H,14.56 δ 0.83 (d, J = 6.8 Hz, 3H),
0.85 (d, J = 6.8 Hz, 3H),
(5.0)
0.87 (t, J = 7.0 Hz, 3H)
1.10 (d, J = 6.8 Hz, 3H),
1.02–1.51 (m, 14H), 3.64 (m, 1H)
C,77.84; H,14.74 δ 0.83 (d, J = 6.8 Hz, 3H),
0.85–0.87 (m, 6H),
4
5
(R)-24
(1.00, 5.1)
(2R,3S)-31
(2S,3R,8R)-3,8-dimethyl-2-
undecanol (6H) (0.33, 33%)
18.6
(3.8)
(5.0)
1.11 (d, J = 6.7 Hz, 3H)
1.03–1.48 (m, 14H), 3.63 (m, 1H)
C,77.58; H,14.30 δ 0.82 (d, J = 6.8 Hz, 3H),
0.85 (d, J = 6.8 Hz, 3H),
(S)-24
(0.7, 3.6)
(2R,3S)-31
(2S,3R,8S)-3,8-dimethyl-2-
undecanol (6H) (0.17, 21%)
23.2
(3.3)
(4.0)
0.87 (t, J = 7.0 Hz, 3H)
1.10 (d, J = 6.8 Hz, 3H),
1.03–1.45 (m, 14H), 3.65 (m, 1H)
C,77.94; H,14.29 δ 0.83 (d, J = 6.8 Hz, 3H),
0.85 (d, J = 6.8 Hz, 3H),
0.87 (t, J = 7.0 Hz, 3H)
1.10 (d, J = 6.8 Hz, 3H),
1.00–1.50 (m, 14H), 3.64 (m, 1H)
C,77.53; H,14.62 δ 0.83 (d, J = 6.8 Hz, 3H),
0.86 (d, J = 6.8 Hz, 3H),
6
7
(R)-25
(1.00, 5.1)
(2R,3S)-31
(2S,3R,9R)-3,9-dimethyl-2-
undecanol (7H) (0.25, 25%)
11.9
(4.2)
(5.0)
(S)-25
(1.00, 5.1)
(2R,3S)-31
(2S,3R,9S)-3,9-dimethyl-2-
undecanol (7H) (0.59, 59%)
29.1
(3.4)
(5.0)
0.88 (t, J = 7.0 Hz, 3H)
1.10 (d, J = 6.8 Hz, 3H),
1.01–1.52 (m, 14H), 3.64 (m, 1H)
δ 0.81–0.85 (m, 6H),
a)
a)
8
9
(S)-25
(0.19, 1.0)
(2S,3S)-31
(2S,3S,9S)-3,9-dimethyl-2-
undecanol (7H) (0.03, 15%)
—
—
(1.0)
0.86 (d, J = 6.8 Hz, 3H),
1.13 (d, J = 6.4 Hz, 3H),
1.08–1.52 (m, 14H), 3.69 (m, 1H)
26
(2R,3S)-31
(2S,3R)-3,10-dimethyl-2-
undecanol (8H) (0.31, 44%)
20.4
(4.9)
C,77.83; H,14.50 δ 0.84 (d, J = 6.8 Hz, 3H),
0.86 (d, J = 6.8 Hz, 3H),
(0.65, 3.3)
(3.5)
1.10 (d, J = 6.4 Hz, 6H)
1.24–1.51 (m, 14H), 3.64 (m, 1H)
C,77.63; H,14.20 δ 0.84 (d, J = 6.8 Hz, 3H),
0.86 (d, J = 6.8 Hz, 3H),
10
11
26
(2S,3S)-31
(2S,3S)-3,10-dimethyl-2-
undecanol (8H) (0.22, 31%)
−17.9
(2.0)
(0.65, 3.3)
(3.5)
1.13 (d, J = 6.4 Hz, 6H)
1.24–1.51 (m, 14H), 3.64 (m, 1H)
C,77.53; H,14.62 δ 0.84 (d, J = 6.8 Hz, 3H),
0.86 (t, J = 6.8 Hz, 3H),
27
(2R,3S)-31
(2S,3R)-3-methyl-2-
dodecanol (9H) (0.57, 57%)
20.4
(2.5)
(1.00, 5.1)
(5.0)
1.10 (d, J = 6.4 Hz, 3H)
1.04–1.49 (m, 17H), 3.64 (m, 1H)
a) An amount of the sample was insufficient for analyses
Field Tests. Stock solutions of pheromone candidates (1 mg/
mL in hexane) were each diluted 10 times with hexane. The de-
sired amount of the resulting solution (100 µg/mL in hexane) of
each compound was loaded onto a cotton-roll dispenser (1 × 4
cm, Nitiei Co.). By 100 µL or 20 µL loading, a dispenser baited
with 10 µg or 2 µg of pheromone candidate was prepared.

120 Bull. Chem. Soc. Jpn., 75, No. 1 (2002)
Sex Pheromone of Pine Sawfly; D. nipponica
Table 4. 1H NMR and GC-MS Data of Pheromone Candidates
No
Compound
1H NMR (400 MHz, CDCl3)
GC-MS
Retention time and that Major fragments in
of internal standard
(in parentheses)
EI mode
1 (1S,2R,5RS)-1,2,5-trimethyl- δ 0.81–0.88 (m, 9H), 1.00–1.37 (m, 20H),
decyl propionate (5P) 2.29 (q, J = 7.2 Hz, 2H), 4.78–4.82 (m, 1H)
23.16 and 23.34 (24.16) m/z 57, 69, 86, 101,
112, 130, 153, 165,
182
2 (1S,2R,6R)-1,2,6-trimethyl- δ 0.82 (d, J = 6.8 Hz, 3H), 0.85 (t, J = 7.5 Hz, 3H),
28.57 (29.15)
28.60 (29.15)
28.90 (29.15)
28.92 (29.15)
m/z 57, 70, 86, 101,
111, 125, 140, 158,
165, 182
decyl propionate (4P)
0.87 (d, J = 6.8 Hz, 3H), 1.11 (d, J = 7.5 Hz, 3H),
1.13 (t, J = 7.5 Hz, 3H), 1.05–1.40 (m, 13H),
1.63 (m, 1H), 2.28 (q, J = 7.5 Hz, 2H), 4.80 (m, 1H)
3 (1S,2R,6S)-1,2,6-trimethyl- δ 0.82 (d, J = 6.8 Hz, 3H), 0.85 (t, J = 7.5 Hz, 3H),
See Fig. 2D
decyl propionate (4P)
0.85 (t, J = 7.5 Hz, 3H) 0.87 (d, J = 6.8 Hz, 3H),
1.12 (t, J = 7.5 Hz, 3H), 1.02–1.35 (m, 13H),
1.63 (m, 1H), 2.28 (q, J = 7.5 Hz, 2H), 4.78 (m, 1H)
4 (1S,2R,7R)-1,2,7-trimethyl- δ 0.82 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.8 Hz, 3H),
m/z 57, 70, 86, 101,
111, 130, 139, 158,
182
decyl propionate (6P)
0.85 (t, J = 7.5 Hz, 3H), 1.10 (d, J = 6.8 Hz, 3H),
1.11 (t, J = 7.5 Hz, 3H), 1.01–1.48 (m, 13H),
1.63 (m, 1H), 2.29 (q, J = 7.5 Hz, 2H), 4.79 (m, 1H)
5 (1S,2R,7S)-1,2,7-trimethyl- δ 0.82 (d, J = 6.8 Hz, 3H), 0.87 (d, J = 6.8 Hz, 3H),
See Fig. 2C
decyl propionate (6P)
0.87 (t, J = 7.5 Hz, 3H), 1.11 (d, J = 6.8 Hz, 3H),
1.12 (t, J = 7.5 Hz, 3H), 1.00–1.38 (m, 13H),
1.63 (m, 1H), 2.28 (q, J = 7.5 Hz, 2H), 4.79 (m, 1H)
6 (1S,2R,8R)-1,2,8-trimethyl- δ 0.81–0.86 (m, 9H), 1.11 (d, J = 6.3 Hz, 3H),
24.17 (24.14)
28.65 (28.60)
28.49 (28.59)
m/z 57, 70, 86, 101,
111, 130, 153, 182
decyl propionate (7P)
1.12 (t, J = 7.3 Hz, 3H), 1.07–1.34 (m, 14H),
2.28 (q, J = 7.5 Hz, 2H), 4.80 (m, 1H)
7 (1S,2R,8S)-1,2,8-trimethyl- δ 0.81–0.86 (m, 9H), 1.12 (d, J = 6.3 Hz, 3H),
See Fig. 2B
decyl propionate (7P)
1.07–1.33 (m, 17H), 2.28 (q, J = 7.4 Hz, 2H),
4.80 (m, 1H)
8 (1S,2S,8S)-1,2,8-trimethyl- δ 0.82 (d, J = 6.3 Hz, 3H), 0.87 (d, J = 6.8 Hz, 3H),
m/z 57, 70, 86, 101,
111, 130, 153, 182
decyl propionate (7P)
1.12 (t, J = 7.3 Hz, 3H), 1.14 (d, J = 6.3 Hz, 3H),
1.06–1.40 (m, 17H), 2.28 (q, J = 7.3 Hz, 2H),
4.82 (m, 1H)
9 (1S,2R)-1,2,9-trimethyl-
decyl propionate (8P)
δ 0.79 (d, J = 6.8 Hz, 6H), 0.80 (d, J = 6.8 Hz, 3H),
1.07 (t, J = 7.5 Hz, 3H), 1.00–1.28 (m, 17H),
2.28 (q, J = 7.5 Hz, 2H), 4.79 (m, 1H)
δ 0.79 (d, J = 6.8 Hz, 6H), 0.81 (d, J = 6.8 Hz, 3H),
1.07 (t, J = 7.5 Hz, 3H), 1.09 (d, J = 6.4 Hz, 3H),
0.98 –1.26 (m, 14H), 2.23 (q, J = 7.5 Hz, 2H),
4.77 (m, 1H)
23.89 (24.14)
23.74 (24.14)
m/z 57, 70, 86, 101,
121, 130, 153, 167,
182
m/z 57, 70, 86, 101,
121, 130, 153, 167,
182
10 (1S,2S)-1,2,9-trimethyl-
decyl propionate (8P)
11 (1S,2R)-1,2-trimethyl-
undecyl propionate (9P)
δ 0.85 (d, J = 6.8 Hz, 3H), 1.12 (t, J = 7.6 Hz, 3H),
1.11 (d, J = 6.8 Hz, 3H), 1.03–1.33 (m, 20H),
2.29 (q, J = 7.2 Hz, 2H), 4.77–4.83 (m, 1H)
25.07 (24.23)
m/z 57, 70, 86, 101,
111, 130, 153, 182
The field attraction of males to pheromone candidates was test-
ed on Apoi Mountain, Hidaka, Hokkaido, Japan. The test areas
were mixed stands of Pinus parviflora var, Picea glehenii, and
Rhododendron spp. The sawfly attacked those P. parviflora in the
previous summer. A cockroach trap supplied by Earth Pharma-
ceutical Co. was used throughout the field tests. A cotton-roll dis-
penser baited with a synthetic compound was placed at the center
of the sticky surface of the trap. The baited traps were randomly
used and attached to pine branches 1.5–2 m high and about 10 m
apart. The dates of test period and the results are listed in Table 2
in the text.
Osaka University for the high-resolution MS analysis and the
elementary analysis, and to Mr. N. Hayashi, Mrs. M.
Higashiura, Mr. K. Higashiura, and Mr. T. Higashiura of Hok-
kaido Forestry Research Institute for their help in field studies.
Our thanks are also due to Dr. J. Hasegawa of Kaneka Co. for a
gift of optically pure (R)- and (S)-methyl 3-hydroxy-2-methyl-
propioate, and to Mr. M. Daino of Earth Pharmaceutical Co.
for the supplying a large number of cockroach traps.
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