* Tokyo University of Agricultureand Technology, JapanJapan

JICA, Can Tho University, VietnamE-mail: antetsu@cc.tuat.ac.jp

Yuta Muraki, Hideshi Naka, Atsushi Honma, Johanna Mappes, Kaisa Suisto, and Tetsu Ando*

Yuta Muraki, Hideshi Naka, Atsushi Honma, Johanna Mappes, Kaisa Suisto, and Tetsu Ando*

2017 ISCE/APACE2017 ISCE/APACEKyoto, Japan (August 24, 2017)Kyoto, Japan (August 24, 2017)

GC / FT-IR analysis of a novel 2,4,6,9-tetraene occurring in a female pheromone gland of

Parasemia plantaginis (Lepidoptera: Arctiidae)

GC / FT-IR analysis of a novel 2,4,6,9-tetraene occurring in a female pheromone gland of

Parasemia plantaginis (Lepidoptera: Arctiidae)

3000 2000 1000 (cm-1)

OHH

O

HOOAc

OAc

silkworm moth smaller tea tortrix rice stem borer

Milionia basalis giant geometrid moth

bombykol

Type I

Type IIO O

epo3,Z6,Z9-19:H Z3,epo6,Z9-19:H

Sex pheromones have been identified from more than 670 species.Male attractants have been reported for other 1300 species.

Unsaturated fatty alcohols, acetates and aldehydes with a C10 – C18 chain

Found most commonly (75%)

Polyunsaturated hydrocarbons and their epoxides with a C17 – C23 chain

Identified from evolved-insect groups (15%)

Representative lepidopteran sex pheromones

Z3,Z6,Z9-21:H

plum cankerworm moth

Photos from http://www.jpmoth.org/

plum cankerworm mothZ3,Z6,Z9-21:HZ3,Z6,Z9-23:H

satin moth Z3,epo6,epo9-21:H

Pyraloidea

TortricoideaCossoidea

GelechioideaZygaenoidea

Tineoidea

Ditrysia

Monotrysia

Dacnonypha

Paramecoptera

Nepticuloidea

Papillionoidea

Pterophoroidea

Hepialoidea

EriocranioideaZeugloptera

Geometroidea

NoctuoideaBombycoidea

Incurvarioidea

Yponomeutoidea Castnioidea

HesperioideaSesioidea

Type I pheromoneType II pheromone

Phylogenetic tree of Lepidoptera

yellow peach mothE10-16:AldZ3,Z6,Z9-23:H

Linoleic acidZ6,Z9-dienes

E4,Z6,Z9-trienesZ6,Z9,E11-trienesZ6,Z9,Z12-trienesdesaturation

mono-epoxides

OH

O

OH

OLinolenic acidZ3,Z6,Z9-trienes

1,Z3,Z6,Z9-tetraenesZ3,Z6,Z9,E11-tetraenesZ3,Z6,Z9,Z11-tetraenesdesaturation

mono- and di-epoxides

CnH2n+1

CnH2n+1Δ1

secretion

secretion

Δ11

Δ4 Δ11

Δ12

The known chemical diversity is still limited. Novel compounds must be found.

Chemical structures of Type II pheromones

emerald moth

Four subspecies are recorded in Japan.P. p. macromera P. p. melanissimaP. p. jezoensis P. p. melanomera

Noctuoidea species and their sex pheromones

Parasemia plantaqinis, wood tiger moth(Arctiidae; Arctiinae)

Noctuoidea

LithosiinaeSyntominaeArctiinae

Super-family Family Sub-family Taxonomy

*Erebidae

<Trifinae> <Quadritinae>*

NotodontidaeNolidaeNoctuidae

Lymantriidae*Arctiidae* II, others

IIII, others

IIIIII, othersII, others

Type ofpheromone

79 [4]4 [1]

51 [3]

124 [0]107 [0]787 [34]489 [4]59 [16]

Japanesespecies number

[ ] indicate number of species whose sex pheromone has been identified.

Four subspecies are recorded in Japan.P. p. macromera P. p. melanissimaP. p. jezoensis P. p. melanomera

EAD

GC-EAD analysis of the pheromone extract

FID

0.1 mV

Comp. IComp. II

Mixing ratioI : II : III : IV ≑ 30:3:5:1

Comp. III

Comp. IV

GC column: DB-23 (0.25 mm X 30 m)

Temperature program:80ºC (1 min) →

210ºC (10ºC/min)

20.0 22.018.0 Rt (min)

Comp. IZ3,Z6,Z9-21:H

Crude pheromoneextract (1.0 FE)

Parasemia plantaqinis, wood tiger moth(Arctiidae; Arctiinae)

Comp. II100

0

121135

149107

93

79

GC / MS analysis of the pheromone extract

247194

GC column: DB-23(0.25 mm X 30 m)

Temperature program:80ºC (1 min) →

210ºC (8ºC/min)

I

IIIII

IV

TIC

E4,Z6,Z9-21:H

135121

79

247 194

290M+

Crude pheromoneextract (1.0 FE)

20.018.0 Rt (min)

25020015010050 300 m/z

234

25020015010050 300

79 91

106 119133147 288

m/z

M+

Comp. III100

0

1,Z3,Z6,Z9-21:H

234

133

79

10691

100

025020015010050 300

220

79

94

105119133

147 288

m/z

M+

91

Comp. IV 133105

94 220

912,4,6,9-tetraene ?

No information on the configurationsGC / FT-IR

OH

Functional group OH Carbon chain length 16

Double bond Number 2Position 10, 12Configuration E, Z

Determined by the IR spectrum,if a large amount of the pure pheromone is available.

IR analysis is not utilized, becausethe species-specific pheromones are composed of multiple components, which are produced at μg level, at most.

O-Htrans

cis

Merits of IR analysis

Lq. N2GC / FT-IR

<Spectra Analysis, Inc.>

Capillary GC High separation

FT-IR (solid phase) High sensitivity

Familiarized spectrum

® Discover IR

IR beamGCcapillary

Detector

ZnSe diskcooled at −30ºC

Comp. I3007

29652918

2851 14721396 935

728

30132958

2919

2852 1473

30102954

2919

2853899

IR spectra of four EAG-active components

Comp. II

Comp. III

Measured by GC / FT-IR Disk temperature: −30ºCDisk speed: 2 mm/minCrude extract (4 FE)

Comp. IV includes E,Z conjugated systems, and no Z,Z conjugation.

983951

1000

E4,Z6,Z9-21:H

30202957

2921

2853

3000 2000 1000 (cm-1)

Comp. IV 1470 765992

941

Z2,E4,Z6,Z9-21:H

3040

ConjugatedZ,Z-diene

970Z3,Z6,Z9-21:H

1,Z3,Z6,Z9-21:H

H

O

H

O

_Br Ph3

+P P+Ph3 Br_

••

Ph3P PPh3

H

O

Synthesis of 2,4,6,9-tetraene

Br Br

Poor yield (< 5%), but a very simple route

GC / FT-IR analyses of natural and synthetic 2,4,6,9-tetraene

20.0 22.0Rt (min)

20.0 22.0

[Peak chromatogram]

I

IIIII

IV

Synthetic

E2,E43020

2921

Comp. IV

3020

2921

993942766

993942766

Crude extract

Synthetic

3000 2000 1000 (cm-1)

3017

2921

752996

SyntheticE2,E4,Z6,Z9-21:H

good coincident

Z2,E4,Z6,Z9-21:H

Crude extract(4 FE)

Z2,E4

Column: HP-5(0.25 mm X 30 m)

Summary & discussion

1) Four EAG-active components were found in the pheromone grand of P. p. melanomera and a Finnish subspecies.

2) GC/MS and GC/FT-IR analyses showed the following structures.Comp. I Z3,Z6,Z9-21:H ( ← linolenic acid)Comp. II E4.Z6,Z9-21:H ( ← linoleic acid)Comp. III 1,Z3,Z6,Z9-21:H ( ← linolenic acid)Comp. IV Z2,E4,Z6,Z9-21:H ( ← linoleic acid)

Mixing ratio; I : II : III : IV ≑ 30 : 3 : 5 : 1

3) An IR spectrum of the minor Comp. IV measured by a high-sensitive GC/FT-IR instrument indicated its 2Z,4E configuration.

4) The structure of Comp. IV was confirmed by synthesis utilizing the double Wittig reaction.

5) Male attraction by synthetic lures in the field has not succeeded, indicating necessity of additional pheromone component(s).

Thank you for attention !! Thank you for attention !!

I retired Tokyo University of Agriculture and Technology (TUAT) in 2015.

I am grateful to all member of Chemical Ecology Laboratory of TUAT, who studied lepidopteran sex pheromones together.

I retired Tokyo University of Agriculture and Technology (TUAT) in 2015.

I am grateful to all member of Chemical Ecology Laboratory of TUAT for study of lepidopteran sex pheromones.

Present address: Can Tho University (CTU) in Can Tho City, Vietnam Chief advisor of JICA’s project for “Building Capacity for CTU”

https://lepipheromone.sakura.ne.jp/index_eng.html