Synthesis and characterization of hexadecadienyl compounds with a conjugated diene system, sex pheromone of the persimmon fruit moth and related compounds

Article abstract of DOI:10.1271/bbb.67.822

Hexadecadien-1-ol and the derivatives (acetate and aldehyde) with a conjugated diene system have recently been identified from a pheromone gland extract of the persimmon fruit moth (Stathmopoda masinissa), a pest insect of persimmon fruits distributed in

Full text of DOI:10.1271/bbb.67.822

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Synthesis and Characterization of Hexadecadienyl
Compounds with a Conjugated Diene System, Sex
Pheromone of the Persimmon Fruit Moth and Related
Compounds
Takanobu NISHIDAa, Le Van VANGa, Hiroyuki YAMAZAWAa, Ryuji YOSHIDAa, Hideshi NAKAb,
Koji TSUCHIDAb & Tetsu ANDOa
a Graduate School of Bio-Applications and Systems Engineering (BASE), Tokyo University
of Agriculture and TechnologyKoganei, Tokyo 184-8588, Japan
b Laboratory of Applied Entomology, Faculty of Agriculture, Gifu UniversityGifu
501-1193, Japan
Published online: 22 May 2014.
To cite this article: Takanobu NISHIDA, Le Van VANG, Hiroyuki YAMAZAWA, Ryuji YOSHIDA, Hideshi NAKA, Koji TSUCHIDA
& Tetsu ANDO (2003) Synthesis and Characterization of Hexadecadienyl Compounds with a Conjugated Diene System,
Sex Pheromone of the Persimmon Fruit Moth and Related Compounds, Bioscience, Biotechnology, and Biochemistry, 67:4,
822-829, DOI: 10.1271/bbb.67.822
To link to this article: http://dx.doi.org/10.1271/bbb.67.822
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Biosci. Biotechnol. Biochem., 67 (4), 822–829, 2003
Synthesis and Characterization of Hexadecadienyl Compounds
with a Conjugated Diene System, Sex Pheromone of the Persimmon
Fruit Moth and Related Compounds
1
Takanobu NISHIDA,1 Le Van VANG,1 Hiroyuki YAMAZAWA,1 Ryuji YOSHIDA
,
1,
†
Hideshi NAKA,2 Koji TSUCHIDA,2 and Tetsu ANDO
1Graduate School of Bio-Applications and Systems Engineering (BASE), Tokyo University of Agriculture
and Technology, Koganei, Tokyo 184-8588, Japan
2Laboratory of Applied Entomology, Faculty of Agriculture, Gifu University, Gifu 501-1193, Japan
Received November 7, 2002; Accepted December 4, 2002
Hexadecadien-1-ol and the derivatives (acetate and al-
dehyde) with a conjugated diene system have recently
been identiˆed from a pheromone gland extract of the
persimmon fruit moth (Stathmopoda masinissa), a pest
insect of persimmon fruits distributed in East Asia. The
This species has two generations a year and causes
serious damage from infestation by the larvae caus-
ing the fall of immature fruit. It is not easy to moni-
tor the population to predict the best time for pesti-
cide spraying because the adults are not su‹ciently
attracted by a light trap. If the synthetic pheromone
of S. masinissa is available, it will serve as a monitor-
ing tool and be further utilized to decrease the popu-
lation density directly by disrupting sexual communi-
cation instead of by killing with insecticides. New
programs of integrated pest management for S.
masinissa led us to search for the chemical structure
of the natural pheromone. We have recently
identiˆed hexadecadien-1-ol and its two derivatives,
acetate and aldehyde, from a pheromone gland ex-
tract of virgin females by using gas chromatography
combined with mass spectrometry (GC-MS). Com-
paring with some authentic dienyl compounds, the
chromatographic behavior of the three hex-
adecadienyl components indicated the conjugation of
two double bonds.1) Their exact positions, however,
have not been experimentally conˆrmed because the
content of the natural components was too low to be
applied to chemical reactions such as ozonolysis.
Lepidopteran sex pheromones have been identiˆed
in more than 500 species.2) Among them, compounds
with a conjugated diene system in a C10-C18 straight
chain are one of the most important groups. A series
of these conjugated dienes has been analyzed by GC-
MS, and some characteristic fragment ions, which
are informative for estimating the double-bond posi-
tions, have been observed in their spectra as meas-
ured by electron impact ionization (EI).3) However,
none of the spectra reported for the hexadecadienyl
compounds, including the diene structure between
the 9-position and the terminal methyl group, coin-
cided with those of the pheromone components of S.
alcohol and acetate showed their base peaks at m z 79 in
W
a GC-MS analysis by electron impact ionization, but the
aldehyde produced a unique base peak at m z 84, sug-
W
gesting a 4,6-diene structure. To conˆrm this inference,
four geometrical isomers of each 4,6-hexadecadienyl
compound were synthesized by two diŠerent routes in
which one of two double bonds was furnished in a high-
ly stereoselective manner. Separation of the two isomers
synthesized together by each route was facilely accom-
plished by preparative HPLC. Their mass spectra coin-
cided well with those of natural components, indicating
that they were available for use as authentic standards
for determining the conˆguration of the natural phero-
mone. Furthermore, other hexadecadienyl compounds,
including the conjugated diene system between the 3-
and 10-positions, were synthesized to accumulate the
spectral data of pheromone candidates. 5,7-Hexa-
decadienal interestingly showed the base peak at m z
W
80; meanwhile, the base peaks of its alcohol and acetate
were detected at m z 79 like the corresponding 4,6-
W
dienes. The base peaks of all 6,8-, 7,9-, and 8,10-dienes
universally appeared at m z 67 like 9,11-, 10,12-, and
W
13,15-dienes, the spectra of which have already been
published. Although 3,5-hexadecadienal was not pre-
pared, base peaks at m z 67 and 79 were recorded for
W
the alcohol and acetate, respectively.
Key words: sex pheromone; persimmon fruit moth;
4,6-hexadecadienyl acetate; 4,6-hexadeca-
dienal; conjugated diene
The persimmon fruit moth, Stathmopoda masinis-
sa Meyrick (Lepidoptera; Oecophoridae), is a pest in-
sect of persimmon fruits distributed in East Asia.
masinissa. In particular, the m z values of their base
W
peaks were diŠerent. As shown in Table 1, the base
†
To whom correspondence should be addressed. Tel Fax:
81-42-388-7278; E-mail: antetsu cc.tuat.ac.jp
＋ ＠
W

Sex Pheromone with a Conjugated Diene System
823
Table 1. Base Peaks in the Mass Spectra of Some Synthetic Hexadecadienyl Compounds with a Conjugated Diene System and of Natural
Pheromone Components of S. masinissa
Base peak (m z
)
W
Functional
group
Synthetic compounda
Natural pheromone
(4,6-diene)
9,11-diene
10,12-diene
11,13-diene
12,14-diene
13,15-diene
Alcohol
Acetate
Aldehyde
67
67
67
67
67
67
95
95
95
81
81
81
67
67
67
79
79
84
a
Spectral data of the compounds including (
E
)-double bonds previously measured with an ESCO EMD-05B spectrometer (70 eV).3)
the unknown conˆguration of the S. masinissa
pheromone. We are also interested in the mass spec-
tra of other positional isomers, not only to examine
the validity of the foregoing fragmentation pathway,
but also to support future research concerning a
dienyl pheromone. This paper deals with the synthe-
sis and characterization of a series of hexadecadienyl
compounds with a conjugated diene system on the
side of the terminal functional group.
Materials and Methods
Fig. 1. Fragmentation Pathways Proposed for the Characteristic
Fragment Ions Which Were Detected in the EI-MS Analysis of
Instruments. 1H- and 13C-NMR spectra were
recorded by a Jeol Alpha 500 Fourier transform spec-
trometer at 500.2 and 125.7 MHz, respectively, for
CDCl3 solutions containing TMS as an internal stan-
n
-Aldehydes with a Conjugate Diene System at the 4,6-Position
(A) or 5,7-Position (B).
1
peaks of the synthetic dienes were detected at m z 67,
dard. H–1H-COSY and HSQC spectra were meas-
W
81, or 95, and the three functional derivatives with
the diene structure at the same position showed the
ured with the same spectrometer, using the usual
pulse sequences and parameters. GC-MS was con-
ducted in the EI mode with an HP 5973 mass spec-
trometer equipped with a DB-23 capillary column
base peaks at the same m z value. The base peaks of
W
the natural components are diŠerent from those of
the synthetic dienes, indicating that the conjugated
diene system of the S. masinissa pheromone is locat-
ed at the 8,10-position or at a position lower than
that.1) Furthermore, the base peak of the natural hexa-
×
(0.25 mm ID 30 m, J & W Scientiˆc). The column
temperature program was 50 C for 2 min and
10 C min to 220 C. The ionization voltage was
9
9
9
W
70 eV. High-resolution (HR) mass spectra were meas-
ured by a Jeol JMS-700 mass spectrometer, using the
same GC column and conditions as those for the
measurements by the foregoing quadrupole machine.
HPLC involved a Jasco PU-980 liquid chromato-
graph equipped with an integrator (System Instru-
ment Chromatocorder 21J), a UV spectrometric
detector (Jasco UV-970) operated at 240 nm, and an
decadienal (m z 84), which diŠers from those of the
W
other two components (m z 79), suggests the pos-
W
sibility of a 4,6-diene structure for the aldehyde
component. 4,6-Hexadecadienal can be expected to
produce a stable fragment ion with a 2,3-di-
hydropyranyl structure (C5H8O＋, m z 84) by cycliza-
W
tion after cleavage of the C-C bond between the 5-
and 6-positions (Fig. 1A).
ODS column (Grand Pack ODS, 2.0 cm ID
×
25 cm;
Sex pheromones with the 4,6-diene structure have
not been identiˆed from lepidopteran insects and
their spectral data have not been reported to the best
of our knowledge.2) Another aldehyde with a 5,7-
diene structure can be expected to predominantly
produce a homologous fragment ion (C6H10O＋) at
Senshukagaku, Tokyo, Japan). As the solvent,
10
z H2O in methanol was used at a ‰ow rate of
4.5 ml min.
W
Abbreviations used for the chemical structures.
The chemical structures of the synthetic pheromone
＝
m z 98 (Fig. 1B). While we have not found spectral
candidates are abbreviated as follows: Z
(
Z
)-dou-
W
data for 5,7-hexadecadienal, the EI-mass spectrum
ble bond, E
＝
(
E
)-double bond, number before
of 5,7-dodecadienal has been published.4) An abun-
hyphen position of double bond, number after
＝
＝ ＝
hyphen carbon number of straight chain, OH
dant ion was actually detected at m z 98, supporting
W
＝ ＝
alcohol, OAc acetate, and Ald aldehyde.
the fragmentation speculated in Fig. 1. We therefore
decided to synthesize every geometrical isomer of
4,6-hexadecadienyl compounds in order to resolve

824
T. NISHIDA et al
.
Synthesis of (4E,6Z)- and (4E,6E )-4,6-hexa-
decadienyl compounds (Scheme 1). After one
hydroxyl group of 1,4-butanediol 1 had been protect-
ed as a tetrahydropyranyl (THP) ether, another
hydroxyl group was oxidized with pyridinium chlo-
rochromate (PCC) in CH2Cl2 to produce aldehyde
2, which was converted into (E )-2-alkenal 3 in
Scheme 1. Synthetic Route to the (4E,6Z)- and (4E,6E )-Isomers
of 4,6-Hexadecadien-1-ol and Its Derivatives, Acetate and Alde-
hyde.
three steps: a coupling reaction with methoxycar-
bonylmethylenetriphenylphosphorane, reduction
with LiAl(OC2H5)2H2, and oxidation with PCC. In
dry THF, 3 was coupled with a phosphorane derived
a, 2,3-dihydropyran
p
-TsOH; b, PCC CH2Cl2; c, Ph3P
＝
＝
W
W
CHCO2CH3 benzene; d, LiAl(OC2H5)2H2 ether; e, Ph3P
W
W
CH(CH2)8CH3 THF; f,
pyridine.
p
-TsOH EtOH; g, HPLC; h, Ac2O
W
W
W
from
n-decyltriphenylphosphonium bromide, using
n
-butyl lithium as a base, to obtain a mixture of THP
ethers of E4,Z6-16:OH and E4,E6-16:OH in a ratio
of ca. 3:2. After removing the THP group by heating
the lithium acetylide of 5 gave 2-alkynal 6, which was
coupled with -decylidenetriphenylphosphorane in
with catalytic
p
-toluenesulfonic acid in ethanol, two
n
geometrical isomers were separated by preparative
HPLC ( Rs: E4,Z6-16:OH, 33.9 min; E4,E6-16:OH,
45.2 min). An aliquot of each isolated isomer was
separately acetylated with acetic anhydride in pyri-
dine to yield E4,Z6-16:OAc and E4,E6-16:OAc, and
then oxidized with PCC to yield E4,Z6-16:Ald and
THF to synthesize enyne compound 7 with a C16
straight chain. Highly selective and speciˆc conver-
sion of the triple bond was accomplished by
hydroboration with dicyclohexylborane and succes-
sive protonolysis, and a mixture of THP ethers of
Z4,Z6-16:OH and Z4,E6-16:OH in a ratio of ca. 9:1
was obtained. After removing the THP group, two
geometrical isomers were separated by preparative
t
E4,E6-16:Ald. 1H-NMR (
d
, ppm): E4,Z6-16:OH,
1.27 (14H, broad s), 1.68
7, 7 Hz), 2.15 (2H, dt, 7, 7 Hz), 2.20
7 Hz), 5.33 (1H,
＝
6.5 Hz), ¿
0.88 (3H, t,
J
(2H, tt,
(2H, dt,
J
＝
J
＝
HPLC (tRs: Z4,Z6-16:OH, 41.0 min; Z4,E6-16:OH,
＝
＝
J
J
7, 7 Hz), 3.67 (2H, t,
J
34.2 min). An aliquot of each puriˆed isomer was
separately acetylated with acetic anhydride in pyri-
dine to yield Z4,Z6-16:OAc and Z4,E6-16:OAc, and
then oxidized with PCC to yield Z4,Z6-16:Ald and
＝
＝
dt,
(1H, dd,
E4,E6-16:OH, 0.88 (3H, t,
J
11, 7 Hz), 5.66 (1H, dt,
15, 7 Hz), 5.94
J
＝
11, 11 Hz), 6.35 (1H, dd,
J
＝
¿
15, 11 Hz);
1.26 (14H,
＝
J
6.5 Hz),
broad s), 1.67 (2H, tt,
J
＝
7, 7 Hz), 2.05 (2H, dt,
＝
Z4,E6-16:Ald. 1H-NMR (
d
, ppm): Z4,Z6-16:OH,
＝
＝
＝
0.88 (3H, t, J
J
J
7, 7 Hz), 2.16 (2H, dt,
6.5 Hz), 5.58 (2H, m), 6.02 (2H, m); E4,Z6-
6.5 Hz), 1.27 (14H, broad
7, 7 Hz), 2.05 (3H, s), 2.16 (4H,
J
7, 7 Hz), 3.66 (2H, t,
6.5 Hz),
7, 7 Hz), 2.16 (2H, dt, 7, 7 Hz), 2.27
＝
7, 7 Hz), 3.65 (2H, t, J 6.5 Hz), 5.46
¿1.26 (14H, broad s), 1.66
＝
＝
(2H, tt,
(2H, dt,
J
J
16:OAc, 0.88 (3H, t,
J
＝
¿
J
＝
＝
s), 1.74 (2H, tt,
J
(2H, m), 6.27 (2H, m); Z4,E6-16:OH, 0.88 (3H, t,
tt,
J
＝
7, 7 Hz), 4.08 (2H, t,
J
＝
J
6.5 Hz), 5.34 (1H, dt,
15, 7 Hz), 5.93 (1H,
J
＝
6.5 Hz),
¿
1.27 (14H, broad s), 1.68 (2H, tt,
J
J
＝
＝
11,
＝
＝
＝
＝
7, 7 Hz), 2.20 (2H, dt,
J
11, 7 Hz), 5.63 (1H, dt,
7, 7 Hz), 2.15 (2H, dt,
J
＝
＝
＝
＝
dd,
J
11, 11 Hz), 6.33 (1H, dd,
J
15, 11 Hz);
6.5 Hz), 1.26
7, 7 Hz), 2.04 (3H,
7, 7 Hz), 4.06 (2H,
7, 7 Hz), 3.67 (2H, t,
7 Hz), 5.66 (1H, dt,
J
7 Hz), 5.30 (1H, dt,
J
E4,E6-16:OAc, 0.88 (3H, t,
J
＝
¿
J
＝
15, 7 Hz), 5.98 (1H, dd,
J
＝
J
＝
15, 11 Hz); Z4,Z6-
(14H, broad s), 1.71 (2H, tt,
J
11, 11 Hz), 6.29 (1H, dd,
16:OAc, 0.88 (3H, t, 6.5 Hz), ¿1.27 (14H, broad
J
s), 2.05 (2H, m), 2.13 (2H, dt,
＝
J
＝
＝
＝
7, 7 Hz), 2.05 (3H, s), 2.16 (2H,
t,
16:Ald, 0.88 (3H, t,
s), 2.15 (2H, dt,
7, 7 Hz), 2.56 (2H, td,
J
6.5 Hz), 5.56 (2H, m), 6.00 (2H, m); E4,Z6-
s), 1.72 (2H, tt,
J
＝
＝
＝
7, 7 Hz), 4.07 (2H,
J
6.5 Hz),
¿
1.27 (14H, broad
dt,
t,
J
7, 7 Hz), 2.25 (2H, dt,
J
J
＝
7, 7 Hz), 2.44 (2H, dt,
J
J
＝
＝
J
＝
6.5 Hz), 5.44 (2H, m), 6.25 (2H, m); Z4,E6-
＝
＝
6.5 Hz),
J
7, 2 Hz), 5.36 (1H, dt,
16:OAc. 0.88 (3H, t,
J
¿1.27 (14H, broad
11, 7 Hz), 5.63 (1H, dt,
J
＝
15, 7 Hz), 5.92 (1H, dd,
s), 1.74 (2H, tt, 7, 7 Hz), 2.05 (3H, s), 2.16 (4H,
J
＝
＝
＝
＝
＝
J
J
11, 11 Hz), 6.35 (1H, dd,
J
15, 11 Hz), 9.78
tt,
J
7, 7 Hz), 4.08 (2H, t, 6.5 Hz), 5.28 (1H, dt,
＝
11, 7 Hz), 5.66 (1H, dt, J 15, 7 Hz), 5.96 (1H,
＝
＝
＝
(1H, t,
J
2 Hz); E4,E6-16:Ald, 0.88 (3H, t,
J
6.5
J
Hz),
¿
1.26 (14H, broad s), 2.18 (2H, dt,
J
＝
7, 7
dd,
J
＝
11, 11 Hz), 6.29 (1H, dd,
J
＝
15, 11 Hz);
＝
＝
＝
Z4,Z6-16:Ald, 0.88 (3H, t, J
Hz), 2.46 (2H, dt,
Hz), 4.06 (2H, t,
J
J
7, 7 Hz), 2.62 (2H, td,
J
7, 2
6.5 Hz),
＝
7, 7 Hz), 2.44 (2H, dt, J
¿1.27 (14H,
＝
＝
7 Hz), 5.56 (2H, m), 6.00 (2H,
broad s), 2.13 (2H, dt,
7, 7 Hz), 2.58 (2H, td,
6.25 (2H, m), 9.78 (1H, t,
0.88 (3H, t, 6.5 Hz),
7, 7 Hz), 2.44 (2H, dt,
J
＝
＝
7, 2 Hz), 5.44 (2H, m),
m), 9.78 (1H, t,
J
2 Hz).
J
＝
J
2 Hz); Z4,E6-16:Ald,
Synthesis of (4Z,6Z)- and (4Z,6E )-4,6-hexa-
decadienyl compounds (Scheme 2). After THP pro-
tection of the hydroxyl group, 3-chloro-1-propanol 4
J
＝
¿1.27 (14H, broad s), 2.15
＝
＝
＝
(2H, dt,
(2H, td,
(1H, dt,
J
J
7, 7 Hz), 2.56
＝
11, 7 Hz), 5.66
J
7, 2 Hz), 5.28 (1H, dt, J
＝
＝
11, 11 Hz),
was converted into acetylene compound 5 via
coupling reaction with lithium acetylide-
ethylenediamine complex in DMSO. Formylation of
a
J
15, 7 Hz), 5.96 (1H, dd,
J
＝
＝
2 Hz).
a
6.29 (1H, dd,
J
15, 11 Hz), 9.78 (1H, t,
J

Sex Pheromone with a Conjugated Diene System
825
Synthesis of 3,5-, 5,7-, 6,8-, 7,9-, and 8,10-hexa-
decadienyl compounds. According to a procedure
similar to that shown in Scheme 1, each positional
\
isomer of the 4,6-hexadecadienyl compounds was
synthesized by changing the carbon chain length of
the starting diol and that of the alkyl halides for
phosphoranes used in the Wittig reaction as follows:
1,3-propanediol and 1-bromoundecane for the 3,5-
dienes, 1,5-pentanediol and 1-bromononane for the
5,7-dienes, 1,6-hexanediol and 1-bromooctane for
the 6,8-dienes, 1,7-heptanediol and 1-bromoheptane
for the 7,9-dienes, and 1,8-octanediol and 1-bromo-
Scheme 2. Synthetic Route to the (4
Z
,6
Z
)- and (4
Z
,6
E
)-4,6-
Isomers of 4,6-Hexadecadien-1-ol and Its Derivatives, Acetate
and Aldehyde.
a, 2,3-dihydropyran; b, LiC
-BuLi THF; d, Ph3P CH(CH2)8CH3 THF; e, BH(C6H11)
2
øCH EDA DMSO; c, HCO2Et
･
W
n
＝
W
W
W
hexane for the 8,10-dienes. Separation of the (
and ( )-isomers of each hexadecadien-1-ol was
also achieved by preparative HPLC. The hex-
adecadien-1-ols with conˆguration were convert-
E,Z)-
THF; f,
p
-TsOH EtOH; g, HPLC; h, Ac2O pyridine; i, PCC
W
W
W
W
E,E
CH2Cl2.
E,Z
ed into their acetate and aldehyde derivatives in the
same manner as that used for the 4,6-diene, except
＝
(1H, t,
J
2 Hz); E6,Z8-16:Ald, E7,Z9-16:Ald and
＝
6.5 Hz), ¿1.28
for E3,Z5-16:Ald. This aldehyde was not yielded by
E8,Z10-16:Ald, 0.88 (3H, t,
J
1
the PCC oxidation of E3,Z5-16:OH. H-NMR (
d
,
(10H, broad s), 1.42–1.44 (2H, m), 1.63 (2H, tt,
＝
＝
＝
7, 7 Hz), 2.13–2.14 (4H, m), 2.44 (2H, td, J
ppm): E3,Z5-16:OH, 0.88 (3H, t,
J
6.5 Hz),
¿
1.26
J
＝
＝
＝
＝
＝
7, 2 Hz), 5.31–5.32 (1H, dt, J 11, 7Hz), 5.62–5.63
(16H), 2.13 (4H, tt,
J
7, 7 Hz), 2.38 (2H, dt,
J
7, 7
＝
＝
＝
15, 7 Hz), 5.93–5.94 (1H, dd, J
Hz), 3.68 (2H, t,
Hz), 5.62 (1H, dt,
J
6.5 Hz), 5.37 (1H, dt,
J
11, 7
(1H, dt,
Hz), 6.30–6.31 (1H, dd,
(1H, t, 2 Hz).
J
11, 11
＝
J 15, 11 Hz), 9.76–9.95
＝
J
15, 7 Hz), 5.96 (1H, dd,
J
11,
11 Hz), 6.43 (1H, dd,
J
＝
15, 11 Hz); E5,Z7-16:OH,
¿1.27 (12H), 1.47 (2H, tt,
J
＝
＝
0.88 (3H, t,
J
6.5 Hz),
7, 7 Hz), 1.59 (2H, tt,
3.64 (2H, t, 6.5 Hz), 5.31 (1H, dt,
5.64 (1H, dt,
Hz), 6.31 (1H, dd,
E7,Z9-16:OH and E8,Z10-16:OH, 0.88 (3H, t,
＝
＝
J
J
7, 7 Hz), 2.15 (4H, m),
11, 7 Hz),
Results
J
＝
J
＝
＝
＝
11, 11
J
15, 7 Hz), 5.93 (1H, dd,
J
Synthesis
J
＝
15, 11 Hz); E6,Z8-16:OH,
Four geometrical isomers of 4,6-hexadecadien-1-ol
and their functional derivatives, acetate and alde-
hyde, were synthesized by two routes, modifying the
previous methods for dodecadienyl compounds with
a conjugated diene system,5) in which one of two dou-
ble bonds was furnished in a highly stereoselective
manner and the other by a Wittig reaction rather
nonspeciˆcally. Namely, in the route for Scheme 1
＝
J
6.5 Hz),
tt,
6.5 Hz), 5.30–5.31 (1H, dt,
(1H, dt,
6.29–6.30 (1H, dd,
¿1.3 (12H), 1.41 (2H, m), 1.57–1.58 (2H,
7, 7 Hz), 2.13 (4H, m), 3.64 (2H, t,
J
＝
J
＝
＝
J
11, 7 Hz), 5.63–5.64
11, 11 Hz),
15, 11 Hz); E3,Z5-16:OAc,
J
15, 7 Hz), 5.94 (1H, dd, J
＝ ＝
＝
J
＝
0.88 (3H, t,
2.15 (4H, tt,
4.11 (2H, t,
J
6.5 Hz),
7, 7 Hz), 2.43 (2H, dt,
¿1.26 (16H), 2.05 (3H, s),
J
＝
＝
J
＝
7, 7 Hz),
for the (4
ration at the 4-position was derived from stable (
2-alkenal 3. In the route for Scheme 2 for the
(4 ,6 )- and (4 ,6 )-isomers, the conˆguration at
E
,6
Z
)- and (4
E
,6
E
)-isomers, the E conˆgu-
＝
11, 7 Hz),
J
6.5 Hz), 5.37 (1H, dt,
15, 7 Hz), 5.95 (1H, dd, J 11, 11
＝
J
E
)-
5.60 (1H, dt,
J
＝
＝
Hz), 6.39 (1H, dd,
J
15, 11 Hz); E5,Z7-16:OAc,
1.27 (12H), 1.49 (2H, tt,
7, 7 Hz), 2.05 (3H, s),
Z
Z
Z
E
Z
＝
0.88 (3H, t,
J
6.5 Hz),
¿
the 4-position was strictly established by hydrobora-
tion of the triple bond in enyne compound 7. By a
Wittig reaction to form the double bond at the 6-
positon, the conˆguration was more facilely
formed than the E conˆguration. Separation of the
two geometrical isomers was accomplished by
preparative HPLC with an ODS column as shown in
Fig. 2. The tRs of the (4E,6Z)- and (4Z,6E )-isomers
J
＝
7, 7 Hz), 1.65 (2H, tt, J
＝
＝
J
＝
2.15 (4H, dt,
J
7, 7 Hz), 4.07 (2H, t,
J
6.5 Hz),
15,
11, 11 Hz), 6.31 (1H, dd,
15, 11 Hz); E6,Z8-16:OAc, E7,Z9-16:OAc and
E8,Z10-16:OAc, 0.88–0.89 (3H, t, 6.5 Hz), 1.3
(12H), 1.39 (2H, m), 1.61–1.63 (2H, tt, 7, 7 Hz),
2.04 (3H, s), 2.12–2.13 (4H, m), 4.05 (2H, t,
5.32 (1H, dt,
＝
11, 7 Hz), 5.63 (1H, dt,
J
＝
Z
＝
7 Hz), 5.94 (1H, dd,
J
＝
J
J
＝
¿
＝
J
＝
J
were almost the same, but the chromatographic be-
havior of the two isomers prepared by each route was
quite diŠerent. By the route similar to that in Scheme
1, the 3,5-, 5,7-, 6,8-, 7,9-, and 8,10-hexadecadienyl
compounds were synthesized, and two geometrical
isomers were also separated by the ODS column. The
chemical structure of each isomer was conˆrmed by
＝
11, 7 Hz), 5.63–5.64
6.5 Hz), 5.30–5.31 (1H, dt,
J
＝
＝
11, 11 Hz),
(1H, dt,
J
15, 7 Hz), 5.94 (1H, dd,
15, 11 Hz); E5,Z7-16:Ald,
1.27 (12H), 1.54 (2H, tt,
J
6.29–6.30 (1H, dd,
J
＝
＝
6.5 Hz), ¿
0.88 (3H, t,
J
＝
＝
＝
＝
J
J
J
J
J
7, 7 Hz), 1.65 (2H, tt,
7, 7 Hz), 2.44 (2H, td,
7, 7 Hz), 2.16 (4H, dt,
7, 2 Hz), 5.33 (1H, dt,
＝
11, 7 Hz), 5.62 (1H, dt,
J
15, 7 Hz), 5.93 (1H,
15, 11 Hz), 9.76
1H-NMR data, these being analyzed by the H–1H-
1
＝
dd,
J
＝
11, 11 Hz), 6.32 (1H, dd,
J
＝
COSY spectrum.

826
T. NISHIDA et al
.
Table 2. 13C-NMR Assignments for Hexadecadienyl Compounds with a Conjugated Diene System
a
b
c
d
e
f
a
＝
＝
[–C H2–C H C H–C H C H–C H2–]
Chemical shift (
d
ppm)
Cc
Compound
C1
C2
C3
Ca
—
Cb
＝
—
Cd
＝
Ce
—
Cf
E4,Z6-16:OH
Z4,Z6-16:OH
Dd [(E4,Z6)
a
a
＝
＝
(Z4,Z6)]
＝
＝
(Z4,E6)]
3
3
62.5
62.4
32.3
32.5
—
—
—
—
—
—
—
—
25.7
25.5
25.8
25.9
—
29.2
23.8
5.4
28.9
24.1
4.8
32.2
28.3
32.4
32.7
32.8
32.8
25.4
133.4
130.8
2.6
131.1
128.7
2.4
131.6
132.6
133.6
134.0
134.3
134.5
131.5
126.3
124.4
1.9
131.1
129.4
1.7
128.1
126.5
126.2
126.0
125.8
125.8
126.9
128.3
123.3
5.0
130.0
125.3
4.7
128.2
128.2
128.4
128.5
128.5
128.6
127.9
130.7
132.8
27.7
27.5
0.2
32.6
32.9
－
－
1.9
E4,E6-16:OH
Z4,E6-16:OH
a
a
3
3
62.5
62.4
32.3
32.6
133.1
135.4
Dd [(E4,E6)
－
－
2.3
－
0.3
27.7
27.7
27.7
27.7
27.7
27.7
27.7
E3,Z5-16:OAc
E4,Z6-16:OAc
E5,Z7-16:OAc
E6,Z8-16:OAc
E7,Z9-16:OAc
E8,Z10-16:OAc
E4,Z6-16:Ald
a
a
a
a
a
a
a
＝
＝
＝
＝
＝
＝
＝
2
3
4
5
6
7
3
63.8
64.0
64.4
64.5
64.6
64.6
201.9
—
128.6
130.9
130.6
130.4
130.3
130.2
131.3
29.2
28.2
28.5
28.6
28.6
43.4
a
In each compound, the chemical shifts of C14, C15, and C16 are 31.5–31.9, 22.6–22.7, and 14.1 ppm, respectively.
This table also lists the 13C assignments proposed for
six positional isomers of the hexadecadienyl acetates
with an
E,Z conˆguration. The spectra of the 3,5-,
4,6-, and 5,7-dienes are distinguishable by consider-
ing an aŠect of the diŠerent distances between the
acetoxyl group and the diene system, while the spec-
tra of the 6,8-, 7,9-, and 8,10-dienes resemble each
other. The acetoxyl group of the latter three dienes is
located far from the conjugated system.
Fig. 2. Separation of the Geometrical Isomers of 4,6-Hexa-
Mass spectra of hexadecadienyl compounds
Four geometrical isomers of the 4,6-hexa-
decadienyl compounds show almost the same mass
decadien-1-ol in an ODS Column; (4
E,6Z)- and (4E,6E )-
Isomers (A) and (4 ,6 )- and (4 ,6 )-Isomers (B).
Z
E
Z Z
spectra, these being similar to those of the natural
1)
pheromone components of S. masinissa
.
The base
13C-NMR analysis of hexadecadienyl compounds
The signals of four oleˆnic protons in the con-
jugate diene system were separately recorded with the
NMR measurements of E4,Z6-16:OH and Z4,E6-
16:OH. Therefore, their corresponding oleˆnic car-
bon signals were unambiguously assigned by the
HSQC spectra. For E4,E6-16:OH and Z4,Z6-16:OH,
however, the 13C assignments could not be estab-
lished even by 2D NMR experiments because the sig-
nals of H4 and H7 overlapped each other and those of
H5 and H6 also overlapped. Therefore, the oleˆnic
carbon signals were assigned by utilizing an empirical
rule proposed for unbranched conjugated dienes;5,6)
peak of the synthetic aldehyde was detected at m z
W
84, as expected by the fragmentation indicated in
Fig. 1A, and the acetate and alcohol produced their
base peaks at m z 79, the same value as that for the
W
natural components. Table 3 shows GC-MS data for
the (E,Z)-isomers of the hexadecadienyl compounds
synthesized in this experiment, spectra for the ˆve
hexadecadienals being shown in Fig. 3. Although 3,5-
hexadecadienal was not synthesized, the base peaks
of the alcohol and acetate derivatives were observed
at m z 67 and 79, respectively. 5,7-Hexadecadienal
W
shows an interesting spectrum with the base peak at
m z 80, while the alcohol and acetate produced the
W
b
c
＝
i.e., when an
E
＝
conˆguration of the C C bond in a
base peak at m z 79. The base peaks of the 6,8-, 7,9-,
W
Ca–Cb Cc–Cd
＝
Ce diene system is changed to a
Z
and 8,10-dienes were universally detected at m z 67,
W
d
e
＝
conˆguration without the conversion of the C
C
as was true for the 9,11-, 10,12-, and 13,15-dienes
bond, the signals of Ca, Cb, Cc, and Cd shift upˆeld
by ca. 5, 2.5, 2, and 5 ppm, respectively, and the sig-
nal of Ce shifts downˆeld by ca. 2 ppm. Table 2
shows, in addition to the signal assignments for the
(Table 1).3)
As a homologue of the ion at m z 84 of 4,6-hexa-
W
decadienal, 5,7-hexadecadienal produced an abun-
dant ion at m z 98 (Fig. 1B). Another fragment ion
W
four isomers, diŠerences in the chemical shifts (Dd
)
at m z 112, further enlarged by 14 mass units, was
W
between the two geometrical isomers, these being ob-
tained by subtracting the values of Z4,Z6-16:OH
from those of E4,Z6-16:OH and by subtracting the
values of Z4,E6-16:OH from those of E4,E6-16:OH.
characteristically detected in the spectrum of 6,8-hexa-
decadienal. Analysis by an HR-MS instrument con-
ˆrmed the formulae of these ions as follows: C5H8O,
obs. m z 84.0517 (calc. 84.0575); C6H10O, obs. m z
W
W

Sex Pheromone with a Conjugated Diene System
827
Table 3. GC-MS Data of Synthetic Hexadecadienyl Compounds with an
E
,
Z
Conˆguration in the Conjugated Diene Systema
Relative intensity (
z
) of ions at indicated m z
W
tR
(min)
67
79
80
81
84
95
98
109
112
M-60
M-18
M
Alcohol
3,5-diene
4,6-diene
5,7-diene
6,8-diene
7,9-diene
8,10-diene
20.64
20.93
21.03
21.01
21.00
21.07
100
45
86
100
100
100
68
100
100
81
72
58
41
32
67
44
38
31
64
35
59
72
71
69
25
11
24
9
8
6
46
28
46
44
48
48
36
18
29
26
21
14
22
9
13
21
19
19
4
8
5
5
3
3
—
—
—
—
—
—
1
2
14
2
2
1
58
23
10
28
26
25
Acetate
3,5-diene
4,6-diene
5,7-diene
6,8-diene
7,9-diene
8,10-diene
Aldehyde
4,6-diene
5,7-diene
6,8-diene
7,9-diene
8,10-diene
20.25
20.31
20.46
20.48
20.58
20.64
28
25
51
100
100
100
100
100
100
88
88
75
96
98
98
60
60
50
26
31
32
59
67
74
5
1
4
2
1
1
11
14
21
45
47
52
1
1
1
2
1
1
5
4
6
13
14
14
0
0
0
1
0
0
21
25
6
6
8
—
—
—
—
—
—
10
10
28
16
36
28
5
18.95
18.94
19.16
19.20
19.25
85
52
100
100
100
92
78
68
51
42
28
100
22
20
20
55
31
61
68
72
100
5
56
11
4
64
26
43
42
40
10
61
21
76
23
16
9
18
18
15
1
2
19
3
—
—
—
—
—
1
1
1
1
1
19
18
20
20
21
6
a
Capillary column (0.25 mm ID
×
30 m); temperature program: 50 C for 2 min, 10 C min to 220 C, and 220 C for 10 min.
9 9 9 9
W
98.0710 (calc. 98.0732); and C7H12 O, obs. m z
16:OAc, and E4,Z6-16:Ald. The results of elec-
trophysiological experiments support this conclu-
sion. Antennae of the male moths more strongly
responded to these compounds than to the other ge-
ometrical isomers. The synthetic compounds induced
mating behavior of the males in a laboratory bioas-
say, and furthermore, a preliminary ˆeld test interes-
tingly revealed that a lure baited only with E4,Z6-
16:OAc could attract the males. We will report these
results in detail in another paper.1) It should be em-
phasized here that successful determination of the S.
masinissa pheromone is attributable to the new syn-
thetic standards under an experimental situation con-
ducted with a limited amount of the female extract.
In order to determine the double-bond positions, a
GC-MS analysis used to be carried out on the com-
pound(s) derived by a chemical reaction of the phero-
mone component, such as aldehydes by ozonolysis
and an adduct of dimethyl disulˆde for a monoene
compound.7) For a conjugated diene, conversion
with 4-methyl-1,2,4-triazoline-3,5-dione has been
proposed.8) The Diels-Alder cyclo-addition product
between a diene and dienophile shows fragment ions
re‰ecting the double-bond positions of the parent
diene. However, when the pheromone content of the
females is very low or a rearing method for the insect
has not been established, it is di‹cult to supply a
su‹cient amount of the natural pheromone for the
reaction. In this case, it is preferable to presume the
structure by a GC-MS analysis without any chemical
reactions, and we have found that each compound,
including a conjugated diene system near the termi-
nal methyl group, produced diagnostic ions unrelated
W
112.0832 (calc. 112.0888). The 6,8-diene also pro-
duced the ion at m z 84 with moderate intensity. The
W
ion at m z 98 also appeared in the spectrum of 7,9-
W
hexadecadienal with a slightly stronger intensity than
that of the 5,7-diene. These ions, shown in Fig. 1
modiˆed by extra dehydrogenation, might have been
produced along a similar pathway.
5,7-Hexadecadienal produced another characteris-
tic fragment ion at m z 192 (M
－
44) as the corre-
W
sponding ion to 5,7-dodecadienal at m z 136,4)
W
－
indicating the diagnostic ion at m z
M
44 for 5,7-
W
dienals. The abundance of the ions at m z 193 of 6,8-
W
hexadecadienal and at m z 137 of 6,8-dodecadienal4)
W
indicates that the ion at m z
M 43 is diagnostic for
－
W
6,8-dienals. These fragment ions are helpful leads for
ˆnding the double-bond positions of alkdienals. On
the other hand, hexadecadien-1-ols and their acetates
lack such a diagnostic ion. Mass spectra of the 4,6-
dienes resemble those of the 5,7-dienes, and spectra
of the 6,8-dienes resemble those of the 7,9- and 8,10-
dienes.
Discussion
Mass spectra of the synthetic dienes conˆrmed that
the natural pheromone components produced by
S. masinissa were 4,6-hexadecadienyl compounds.
Among the four geometrical isomers, the (4E,6Z)-
isomers showed the same chromatographic behavior
as the natural components did in two kinds of capil-
lary GC columns, indicating that the female sex
pheromone consisted of E4,Z6-16:OH, E4,Z6-

828
T. NISHIDA et al.
Fig. 3. Mass Spectra of Synthetic 4,6-, 5,7-, 6,8-, 7,9- and 8,10-Hexadecadienals with an
E,Z Conˆguration (A-E).
to the functional group at the opposite site in a C12-
16 chain.3) Additionally, this study has revealed that
mulae of the ions at m z 84 (C5H8O) derived from
W
C
4,6-hexadecadienal and at m z 98 (C6H10 O) derived
W
the positional isomers of C16 aldehydes, including the
diene structure near the formyl group, show some
from 5,7-hexadecadienal. These ions include an
oxygen atom and might be made by the fragmenta-
tion occurring at the side of the functional group as
shown in Fig. 1. Therefore, it is expected that these
fragment ions are also abundantly produced from
4,6- and 5,7-dienes with another chain length. In
addition to the conjugated dienes, we measured the
mass spectra of some monounsaturated compounds
characteristic fragment ions such as at m z 84 and
W
98, although C16 alcohols and the acetate derivatives,
including the diene structure near the functional
group, did not present any diagnostic fragment ions.
Hexadecadienals are long-chain compounds, and
there are many positional isomers. It is not easy to
distinguish the mass spectra of the 8,10- and 9,11-
hexadecadienals, the diene structure of which is
located around the center of the C16 chain, but other
positional isomers can be identiˆed by their distinct
spectra.
and observed the base peaks at m z 84 and 98 for 4-
W
hexadecenal and 5-hexadecenal, respectively. Abun-
dant ions at m z 84 and 98 have been also recorded
W
for 4-dodecenal and 5-dodecenal, respectively,
among a series of positional isomers of C12 alde-
hydes.9) These ions are characteristic of the aldehydes
The HR-MS analysis conˆrmed the fragment for-

829
tra of lepidopterous sex pheromones with a conjugat-
ed diene system. Agric. Biol. Chem., 52, 1415–1423
(1988).
and not of the alcohol and acetate derivatives. In this
study, mass spectra were measured by EI at 70 eV,
and the positional isomers showed diŠerent spectra.
This result suggests that migration of the double
bond did not occur easily after this rather drastic
ionization. It is important in future work to deter-
mine how strong instrumental conditions in‰uence
the intensity of the diagnostic ions.
While 4,6,10- and 4,6,11-hexadecatrienyl com-
pounds have been identiˆed from female moths as
pheromone components,10,11) the 4,6-hexadecadienyl
compounds are new pheromone components. Many
conjugated dienes have been identiˆed as lepidopte-
ran sex pheromones, and their double bonds are usu-
ally located on one side of the terminal methyl
group.2) The chemical structure of the S. masinissa
pheromone, however, suggests the possibility of
more pheromone compounds with a conjugated
diene system near the functional group. The EI mass
spectra reported in this study are informative for
future pheromone research.
4) Ando, T., Katagiri, Y., and Uchiyama, M., Mass
spectra of dodecadienic compounds with a conjugat-
ed double bond, lepidopterous sex pheromones.
Agric. Biol. Chem., 49, 413–421 (1985).
5) Ando, T., Kurotsu, Y., Kaiya, M., and Uchiyama,
M., Systematic syntheses and characterization of
dodecadien-1-ols with conjugated double bond,
lepidopterous sex pheromones. Agric. Biol. Chem.
,
49, 141–148 (1985).
6) Ando, T., Kusa, K., Uchiyama, M., Yoshida, S., and
Takahashi, N., 13C NMR analyses on conjugated
dienic pheromones of Lepidoptera. Agric. Biol.
Chem., 47, 2849–2853 (1983).
7) Buser, H.-R., Arn, H., Guerin, P., and Rauscher, S.,
Determination of double bond position in mono-
unsaturated acetates by mass spectrometry of
dimethyl disulˆde adducts. Anal. Chem., 55, 818–822
(1983).
8) Young, D. C., Vouros, P., and Holick, M. F., Gas
chromatography-mass spectrometry of conjugated
dienes by derivatization with 4-methyl-1,2,4-tria-
zoline-3,5-dione. J. Chromato., 522, 295–302 (1990).
9) Horiike, M., and Hirano, C., Mass spectrometric
indication of double bond position in dodecenals
without any chemical derivatization. Biomed. Envi-
ron. Mass Spectrom., 17, 301–306 (1988).
10) Beevor, P. S., Cork, A., Hall, D. R., Nesbitt, B. F.,
Day, R. K., and Mumford, J. D., Components of
female sex pheromone of cocoa pod borer moth,
Conopomorpha cramerella. J. Chem. Ecol., 12, 1–23
(1986).
References
1) Naka, H., Vang, L. V., Inomata, S., Ando, T.,
Kimura, T., Honda, H., Tsuchida, K., and Sakurai,
H., Sex pheromone of the Japanese persimmon
moth,
Oecophoridae): identiˆcation and laboratory bioas-
say of (4 ,6 )-4,6-hexadecadienyl compounds. J.
Chem. Ecol., submitted.
2) Arn, H., Toth, M., and Priesner, E., List of sex
pheromones of Lepidoptera and related attractants.
OILB-SROP IOBC-WPRS, Secretariat General,
Stathmopoda
masinissa
(Lepidoptera:
E
Z
á
11) Bestmann, H. J., Attygalle, A. B., Schwarz, J.,
Garbe, W., Vostrowsky, O., and Tomida, I., Phero-
mones, 71. Identiˆcation and synthesis of female sex
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