Chemical characterization and synthesis of the major component of the sex pheromone of the sugarcane borer Diatraea saccharalis

Article abstract of DOI:10.1135/cccc20011682

(9Z,11E)-Hexadeca-9,11-dienal (1) was identified as a major component of the sex pheromone of the sugarcane borer Diatraea saccharalis. The identification is based on mass spectral examination of the extracts prepared from calling females, derivatization experiments with 4-methyl-3,5-dihydro-1,2,4-triazole-3,5-dione (2), stereospecific synthesis of dienal 1, and gas chromatography (GC) with electroantennographic detection (GC-EAD). In GC analysis, the antennaly active component and synthetic dienal 1 show identical retention times on several GC phases. However, the behavioral activity of synthetic 1 was much weaker than that of natural extracts. This suggests that an additional component(s) might be involved in sexual communication of this species.

Full text of DOI:10.1135/cccc20011682

1682
Svatoš et al.:
CHEMICAL CHARACTERIZATION AND SYNTHESIS OF THE MAJOR
COMPONENT OF THE SEX PHEROMONE OF THE SUGARCANE BORER
Dia tra ea sa ccha ra lis
a4
Aleš SVATOŠa1,*, Blanka KALINOVÁa2, Jiří KINDLa3, Jelena KULDOVÁ
,
Oldřich HOVORKAa5, Ruth Rufino DO NASCIMENTO and Neil J. OLDHAM
b
c
a
Department of Natural Products, Institute of Organic Chemistry and Biochemistry,
Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Prague 6, Czech Republic;
e-mail: 1 svatos@uochb.cas.cz, 2 kalinova@uochb.cas.cz, 3 kindl@uochb.cas.cz,
4 kuldova@uochb.cas.cz, 5 hovorka@uochb.cas.cz
b
c
Departamento de Química, Centro de Ciencias Exatas e Naturais,
Universidade Federal de Alagoas, BR 104 Norte Km 14 – Campus A. C. Simoes Tabuleiro
do Martins, Maceió, AL 57072970, Brazil; e-mail: rrn@ccen.ufal.br
The Dyson Perrins Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QY, U.K.;
e-mail: neil.oldham@chemistry.oxford.ac.uk
Received Septem ber 26, 2001
Accepted Novem ber 8, 2001
(9Z,11E)-Hexadeca-9,11-dien al (1) was iden tified as a m ajor com pon en t of th e sex ph ero-
m on e of th e sugarcan e borer Diatraea saccharalis. Th e iden tification is based on m ass spec-
tral exam in ation of th e extracts prepared from callin g fem ales, derivatization experim en ts
with 4-m eth yl-3,5-dih ydro-1,2,4-triazole-3,5-dion e (2), stereospecific syn th esis of dien al 1, an d
gas ch rom atograph y (GC) with electroan ten n ograph ic detection (GC-EAD). In GC an alysis,
th e an ten n aly active com pon en t an d syn th etic dien al 1 sh ow iden tical reten tion tim es on
several GC ph ases. However, th e beh avioral activity of syn th etic 1 was m uch weaker th an
th at of n atural extracts. Th is suggests th at an addition al com pon en t(s) m igh t be in volved in
sexual com m un ication of th is species.
Keyw ord s: Sex ph erom on es; Diatraea saccharalis; Isolation an d iden tification ; Con jugated
dien als; Aldeh ydes; Dien es; Stereoselective syn th esis.
Man y species of Lepidoptera h ave developed a rem arkably sen sitive com -
m un ication system between th e sexes wh ere m ales can respon d to m in ute
am oun ts of a ph erom on e produced by virgin fem ales1. To date, ca 500
Lepidopteran sex ph erom on es h ave been ch aracterized ch em ically an d syn -
th esized2. In th e sugarcan e borer Diatraea saccharalis Fabricius (Lepidoptera,
Pyralidae, Cram bin ae) th e sex ph erom on e was assign ed as (9Z,11E)-h exa-
deca-9,11-dien al (1) by Ham m on d in a sh ort con feren ce abstract3 an d a pat-
en t application 4 in 1982. However, th e described com poun d h as n ot been
Collect. Czech. Chem. Commun. (Vol. 66) (2001)
doi:10.1135/cccc20011682

Sex Pheromeone of the Sugarcane Borer
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foun d to be h igh ly attractive for con specific m ales in field experim en ts5.
Th ese n egative results raise doubts about th e correct ch aracterization of th e
sugarcan e borer sex ph erom on e.
O
1
Here we presen t, for th e first tim e, firm structural eviden ce, supported by
biological experim en ts, wh ich con firm th e origin al fin din gs3,4 an d we dis-
cuss th e possible reason s for th e low attractiven ess of dien al 1 for con -
specific m ales in field experim en ts.
EXPERIMENTAL
Sam ple Preparation an d Extraction
Laboratory reared D. saccharalis callin g virgin fem ales (1 to 2 days old) were cooled to –20 °C
for 5 m in . Fem ale 7th an d 8th abdom in al segm en ts were excised an d wash ed in h exan e for
30 s (ca 10 µl per ovipositor, vial with solven t was kept on dry ice) or extracted with di-
ch lorom eth an e at room tem perature (10 µl per ovipositor). Th us 10 µl of extracts represen t
on e fem ale equivalen t (FE). Extracts were stored at –20 °C.
Preparation of Diels–Alder Adducts6 (3)
A h exan e extract of callin g fem ales prepared above (100 µl, 10 FE) was treated with a di-
ch lorom eth an e solution of dion e 2 (5% w/v, 50 µl) at room tem perature for 0.5 h . Th e re-
sultin g violet solution con tain in g 3 was con cen trated to a m in im um volum e in a stream of
argon , recon stituted in h exan e (5 µl) an d 1–2 µl of th e solution was in jected on GC-MS in -
strum en t.
Th e dich lorom eth an e extract (100 µl, 10 FE) was gradually treated with acetic an h ydride
(10 µl) an d pyridin e (10 µl) an d after 1 h at room tem perature, th e obtain ed solution was
con cen trated to m in im um volum e in a stream of argon an d treated with a dich lorom eth an e
solution of dion e 2 (5%, 50 µl) at room tem perature for 0.5 h . Th e obtain ed solution con -
tain in g 3 was con cen trated to m in im um volum e in a stream of argon , recon stituted in h ex-
an e (5 µl) an d 1–2 µl of th e solution was in jected on GC-MS in strum en t.
GC-EAD (ref.7)
Gas ch rom atograph ic an alysis coupled with electroan ten n ograph ic detection (GC-EAD) was
perform ed on a Hewlett–Packard HP 5890 gas ch rom atograph equipped with BPX5 colum n
(30 m × 0.33 m m , i.d.). For EAD detection , th e isolated an ten n al preparation was used. Th e
excised m ale an ten n a was placed between two glass Ag/AgCl electrodes filled with in sect
h aem olym ph salin e. Th e colum n was split at th e en d by a four-arm splitter (Graph pack
3D/2, U.S.A.). Th e secon d arm of th e splitter was con n ected to flam e ion isation detector
(FID), by m ean s of deactivated silica colum n (0.5 m × 0.33 m m , i.d.). Th e th ird arm (0.5 m )
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Svatoš et al.:
was led th rough h eated tran sfer lin e (Effluen t Con dition in g Assem bly, Syn tech , Hilversum ,
Th e Neth erlan ds) outside th e GC an d en ded in a m ixin g ch am ber, from wh ich a con tin ual
air stream (1 l m in –1) blew on to th e an ten n al preparation via PTFE-coated stain less steel flow
tube. Th rough th e fourth arm , a m ake-up gas (N2, 20 m l m in –1) was in troduced in to th e
splitter. Th e split ratio FID/EAD was set to 1 : 1. Th e capillary colum n tran sfer tube was
h eated up to 180 °C. Approxim ately 2 µl of con cen trated extract (5 to 10 FE) were in jected
splittless an d an alysed un der a tem perature program : Th e tem perature program started at
50 °C h eld for 2 m in , th en th e tem perature of th e oven was in creased to 140 °C (rate 50 °C
m in–1), to 240 °C (rate 5 °C m in–1) and then to 280 °C (rate 10 °C m in–1; 5 m in delay at 280 °C).
In jector an d detector tem perature was set at 200 an d 290 °C, respectively.
Win d Tun n el Bioassay8
Th e biological activity of extracts an d syn th etic ch em icals was evaluated in win d tun n el.
Th e experim en ts were perform ed durin g 6–7 h of th e scotoph ase. Males of D. saccharalis
(1–2 days old) were used. Males (pre-con dition ed in th e laboratory for 2 h ) were released in -
dividually in to an odor plum e created by pin n in g th e filter paper disc with test odor on to a
h older placed cen trally at th e upwin d en d of th e win d tun n el (air velocity was 0.2 m s–1).
Males were observed durin g a 5-m in period. Male beh avior was categorized: 0, n o respon se;
1, activation on startin g platform ; 2, take-off from th e platform ; 3, orien ted fligh t towards
th e odor source; 4, source location ; 5, lan din g on th e source an d attem pted copulation . Th e
percen tage of m ales exh ibitin g a particular beh avior was evaluated. In each experim en t,
m ales were allowed to respon d to h exan e extracts (3 FE) an d to a test stim ulus. Altogeth er
25 m ales were flown in 5 replicates for each treatm en t.
Ch em ical Syn th esis
Spectral methods. NMR spectra were determ in ed in CDCl3 solution s on
a Varian
UNITY-500 spectrom eter operatin g at 499.5 MHz for 1H an d at 125.7 MHz for 13C NMR, re-
spectively. Ch em ical sh ifts are expressed in ppm (δ-scale) relative to tetram eth ylsilan e for 1H
an d relative to CDCl3 sign al (77.00 ppm ) for 13C NMR, respectively. Couplin g con stan ts (J)
are reported in Hz.
Chromatography. GC an alyses were perform ed on
a Hewlett–Packard HP 6890 gas
ch rom atograph equipped with a FID detector (290 °C), split/splitless in jection port (200 °C),
electron ic pressure con trol (EPC) an d HP 6890 autom atic in jector. A Hewlett–Packard HP-5
capillary colum n (30 m × 0.32 m m ; film th ickn ess 0.25 µm ) an d h elium gas (con stan t pres-
sure 66 kPa) were used for th e separation s. Th e tem perature program started at 50 °C h eld
for 2 m in , th en th e tem perature of th e oven was in creased to 140 °C (rate 40 °C m in –1) an d
th en to 280 °C (rate 10 °C m in –1; 5 m in delay at 280 °C). Preparative m edium -pressure liquid
ch rom atograph y (PMPLC) separation s were m ade on Merck 60 silica gel (0.040–0.063 m m ) us-
in g a Büch i B-680 Prep LC System with eth yl acetate or ben zen e in h exan e. Mobile ph ase for
ch rom atograph y of th e sen sitive con jugated dien ic aldeh yde 1 was m odified with trieth yl-
am in e (0.3% v/v).
Chemicals. All ch em ical reaction s were run in oven -dried glassware un der an argon atm o-
sph ere. Tetrah ydrofuran (THF), h exan e an d ben zen e were distilled from sodium ben zoph e-
n on e ketyl in argon atm osph ere. Dich lorom eth an e was distilled from calcium h ydride an d
stored over m olecular sieves. All oth er ch em icals were used as purch ased. NMR spectra an d
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Sex Pheromeone of the Sugarcane Borer
1685
elem en tal an alyses of all syn th esized com poun ds were fully con sisten t with th e proposed
structures. 10-Tetrah ydropyran yloxydec-1-yn e (5) was previously prepared in our laboratory9.
(E)-1-Iodoh ex-1-en e10 (4)
A solution of h ex-1-yn e 3.28 g (40 m m ol, 4.58 m l) in dry h exan e (20 m l) was treated with
diisobutylalum in um h ydride solution (1.5 m ol l–1 in toluen e, 30 m l) on ice bath an d th en
th e m ixture was h eated to 55 °C for 5 h , cooled to –55 °C an d a solution of iodin e (11.16 g,
44 m m ol) in dry THF (40 m l) was added durin g 5 m in . After stirrin g overn igh t at room tem -
perature th e m ixture was poured in to a separatory fun n el con tain in g ice-cold m ixture of sat-
urated aqueous solution s of am m on ium ch loride an d sodium th iosulfate (3 : 1, 80 m l). Th e
form ed salts were rem oved on a Celite pad (15 g), wh ich was wash ed with pen tan e (3 × 40 m l).
Th e com bin ed extracts were wash ed with 10% sulfuric acid (30 m l), water (2 × 30 m l) an d
aqueous sodium ch loride solution (30 m l). Solven ts were evaporated an d th e residue was
distilled, iodide 4, b.p.75–78 °C/150 Pa, yield 4.0 g (45%) was obtain ed in 95% isom eric pu-
rity (GC).
(E)-16-Tetrah ydropyran yloxy-h exadec-9-en -11-yn e (6)
A suspen sion of [Pd(PPh 3)4] (0.17 g, 5 m ole %) in dry an d deoxygen ated ben zen e (10 m l) in
3-n ecked roun d bottom flask purged with argon was treated with (E)-1-iodoh ex-1-en e (4; 1.0 g,
4.46 m m ol), an d after 1 h of stirrin g at room tem perature with CuI (0.1 g, 0.54 m m ol) an d
with a solution of 10-(tetrah ydropyran yloxy)dec-1-yn e (1 g, 4.42 m m ol) in butylam in e,
freeze-deoxygen ated at a reduced pressure (3 m l, 30 m m ol). Th e dark green solution was
stirred at room tem perature for 16 h (GC con trol, sam ple quen ch ed with 2 M HCl–h exan e–
MgSO4) an d th e reaction m ixture was diluted with eth er (10 m l) an d quen ch ed with a m ix-
ture of am m on ia an d aqueous am m on ium ch loride saturated solution s (1 : 2, 10 m l). Th e
organ ic ph ase was wash ed with th e am m on ia–am m on im ch loride m ixture (10 m l) an d brin e
(10 m l) an d th e obtain ed crude oil was dissolved in h exan e (10 m l) an d filtered th rough
silica gel/Celite/active ch arcoal pad. Th e pad was wash ed with h exan e–eth er m ixture (5%,
50 m l). Th e obtain ed oil (0.70 g) was purified on
a silica gel colum n (100 g) an d
AgNO3–SiO2 yieldin g 0.25 g of th e title com poun d 6 (26%). 1H NMR (CDCl3): 0.98 t, 3 H,
J = 7.0 (CH3CH2-); 1.24–1.80 m , 18 H (9 × CH2); 1.50 m , 2 H (CH3CH2-); 1.52 m , 2 H
(-CH2CH2OR); 2.07 dq, 2 H, J = 1.5, 3 × 7.1 (-CH=CHCH2-); 2.26 dt, 2 H, J = 1.2, 2 × 1.7
(-CH2C≡C-); 3.40 t, 2 H, J = 6.7 (-CH2OR); 3.80 m , 2 H (CH2[6′]); 4.58 bs, 1 H (CH2[2′]); 5.45 m ,
1 H, J = 4 × 1.7, 15.8 (-CH=CH-); 6.03 m , 1 H, J = 4 × 1.7, 15.8 (-CH=CH-). For C21H36O2 (320.5)
calculated: 78.70% C, 11.32% H; foun d: 79.00% C, 11.08% H.
(9Z,11E)-Hexadeca-9,11-dien -1-ol (7)
A solution of cycloh exen e (0.6 m l, 6 m m ol) in dry THF (3 m l) was treated with a solution of
boran e–dim eth yl sulfide com plex in THF (1 m ol l–1, 3 m l, 3 m m ol) on ice bath for 0.3 h .
Th e m ixture was stirred on ice bath for 0.3 h an d at room tem perature for 2 h . Th e wh ite
slurry was treated with THF solution of 6 (80 m g, 0.25 m m ol, 1 m l) on ice bath . Th e m ix-
ture was warm ed to room tem perature an d stirred for 4 h . Th e form ed vin ylboran e was h y-
drolyzed with glacial acetic acid (0.2 m l) at 20 °C for 12 h . Th e reaction m ixture was
n eutralized with NaOH (20%, 0.2 m l) an d carefully treated with aqueous H2O2 (30%, 0.2 m l).
Product was extracted with eth er (4 × 5 m l). Th e crude m aterial was dissolved in m eth an ol
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Svatoš et al.:
(1 m l) an d treated with ion exch an ge resin (Dowex 50WX, 10 m g) for 5 h . Th e crude alco-
h ol 7 was purified by ch rom atograph y (PMPLC) in h exan e yieldin g dien ol 7 (42 m g, 70%)
in 95% isom eric purity (GC). 1H NMR (CDCl3): 0.90 t, 3 H, J = 7.3 (CH3CH2-); 1.24–1.50 m ,
12 H (6 × CH2); 1.41 m , 2 H (CH3CH2-); 1.58 m , 2 H (-CH2CH2OH); 2.05 dq, 2 H, J = 1.4, 3 ×
7.1 (-CH=CHCH2-); 2.10 dq, 2 H, J = 1.4, 3 × 7.5 (-CH2CH=CH-); 3.63 t, 2 H, J = 6.5
(-CH2OH); 5.30 bdt, 1 H, J = 3 × 7.5, 10.8 (-CH=CH-CH=); 5.65 dq, 1 H, J = 2 × 7.1, 15.1
(=CH-CH=CH); 5.94 dtt, 1 H, J = 3 × 1.4, 11.0 (-CH=CH-CH=); 6.29 ddq, 1 H, J = 2 × 1.5,
11.0, 15.1 (=CH-CH=CH). HR EI: for C16H30O calculated 238.2297; foun d 238.2330.
(9Z,11E)-Hexadeca-9,11-dien al (1)
Dien ol 7 (20 m g, 0,084 m m ol) was in jected in to a stirred suspen sion of pyridin ium ch loro-
ch rom ate (PCC; 25.8 m g, 0.12 m m ol) an d an h ydrous sodium acetate (2 m g) in dich loro-
m eth an e (1 m l). Th e m ixture was stirred at room tem perature for 90 m in , th en poured in to
50 m l of eth er an d filtered th rough com bin ed layer of n eutral alum in a/ch arcoal/Celite.
Subsequent evaporation of the solvents and PMPLC (0.3% triethylamine in benzene–hexane 1 : 1)
afforded 14 m g (70%) of th e pure (96%, GC) title com poun d 1. 1H NMR (CDCl3): 0.92 t, 3 H,
J = 7.1 (CH3CH2-); 1.24–1.50 m , 12 H (6 × CH2); 1.41 m , 2 H (CH3CH2-); 2.12 dq, 4 H, J = 1.4,
3 × 7.2 (2 × -CH=CHCH2-); 2.41 m , 2 H (-CH2CHO); 5.35–5.45 m , 2 H (-CH=CH-CH=CH-);
6.20–6.29 m , 2 H (-CH=CH-CH=); 9.75 t, 1 H, J = 1.8 (-CH=O). EI-MS (m/z, rel.%): 236 (12),
135 (8), 109 (15), 96 (35), 81 (76), 79 (39), 67 (100), 55 (29), 54 (28), 41 (43). HR EI: for
C
16H28O calculated 236.2140, foun d 236.2117.
RESULTS AND DISCUSSION
Th e prelim in ary exam in ation s of h exan e extracts of th e abdom en s of call-
in g sugarcan e fem ales usin g gas ch rom atograph y with m ass spectroscopic
detection (GC-MS) sh ow, beside h ydrocarbon s an d fatty acids m eth yl es-
ters, on e peak possessin g a m ass spectrum with rath er stron g m olecular ion
at m/z 236 Da. Th e observed fragm en tation pattern resem bled th at of a
h exadecadien al11. In dich lorom eth an e extracts of callin g fem ales in addi-
tion to aldeh yde 1, th e correspon din g h exadecadien -1-ol was detected.
In th e m ass spectrum of th e dien al, a stron g m olecular ion was observed
an d it was reason able to expect12 th at th e double bon ds are con jugated. For
th e assign m en t of th e position s of con jugated double bon ds, a Diels–Alder
reaction of th e h exadecadien al with 4-m eth yl-3,5-dih ydro-1,2,4-triazole-
3,5-dion e (2) was perform ed6 (Sch em e 1).
R1
R2
R1
R2
+
N
N
N
N
O
O
N
O
O
N
CH3
CH3
3
2
SCHEME 1
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Sex Pheromeone of the Sugarcane Borer
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Electron -im pact m ass spectra of th e form ed adducts 3 exh ibit very
characteristic fragmentation patterns, from which the position of carbon–carbon
double bon ds is directly deducible6. Wh en th e h exan e extract was treated
with triazole 2, th e m easured m ass spectrum sh ows m olecular ion at m/z
349 Da an d distin ct fragm en t ion s at m/z 222 an d 292. Th is spectrum was
assign ed to an adduct of h exadeca-9,11-dien al (Fig. 1A). Addition ally, th e
dich lorom eth an e extract of callin g fem ales, presum ably con tain in g beside
aldeh yde 1 its biosyn th etic precursor alcoh ol 7, was first treated with an
acetic an h ydride–pyridin e m ixture to con vert th e h exadecadien -1-ol to th e
acetate, an d th en th e reaction m ixture was treated with 2. Again , th e 9,11
position of th e con jugated carbon –carbon double bon ds was clearly as-
sign ed (Fig. 1B).
Th e deduced h exadeca-9,11-dien al can exist as four stereoisom ers. Direct
spectral ch aracterization of such dien al isom ers in a com plex m atrix is on ly
feasible with a gas-ph ase Fourier-tran sform in frared spectroscopy (FTIR) us-
in g GC-FTIR in strum en ts at detection lim its in th e n an ogram ran ge13. In
our case, we dealt with picogram s an d, th erefore, h ad to rely on less
straigh tforward com parison s of prepared syn th etic stan dards with th e n atu-
ral com poun d. We h ave design ed a sim ple syn th etic approach 14,15 to
prepare aldeh yde 1 based on Pd-catalyzed Son ogash ira couplin g an d h ydro-
boration an d h ydrogen olysis of th e form ed en yn e 6 (Sch em e 2).
(i)
CH3(CH2)3
CH3(CH2)3
+
(CH2)8-O-THP
I
(CH2)8-O-THP
4
5
6
(iv)
(ii), (iii)
CH3(CH2)3
(CH2)8-OH
1
7
SCHEME 2
Reten tion param eters of th e syn th etic dien al 1 were com pared with th ose
of th e n atural sam ple on several GC ph ases (DB1, BPX5, DB WAX) an d in
all co-in jection experim en ts com poun d 1 sh ows iden tical reten tion tim e to
th e n atural h exadeca-9,11-dien al. Th e MS of syn th etic 1 an d th e h exadeca-
9,11-dien al in th e h exan e extract of callin g fem ales were in distin guish able.
Am oun ts of th e dien al produced by a callin g fem ale were estim ated, usin g a
calibration with auth en tic stan dard, to 10–50 pg per callin g fem ale.
Th e structural iden tity of th e m ajor com pon en t was furth er supported
with electroan ten n ograph ic m easurem en ts perform ed on a GC–EAD system ,
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1688
Svatoš et al.:
wh ere both h exan e extract an d syn th etic stan dard 1 sh owed pron oun ced
an ten n al activity on th e EAD trace, an d th eir reten tion beh avior expressed
as Kovats’ in dices was iden tical (Fig. 2; KI = 1 842 ± 1 for th e h exan e extract
an d 1 843 ± 1 for 1).
FIG. 1
Spectra of prepared Diels–Alder adducts (3) of 4-m eth yl-3,5-dih ydro-1,2,4-triazole-3,5-dion e
(2) with : h exadecadien al in th e h exan e extract (A) an d h exadecadien yl acetate prepared from
th e dich lorom eth an e extract (B). Both extracts were prepared from callin g D. saccharalis fe-
m ales
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Sex Pheromeone of the Sugarcane Borer
1689
TABLE I
Beh avioral reaction s of D. saccharalis m ales (n = 25) observed in win d tun n el to h exan e ex-
tracts of excised ph erom on e glan ds expressed as fem ale equivalen ts (FE) an d to differen t
doses of (9Z,11E)-h exadeca-9,11-dien al (1)
No. Beh avioral category
3 FE
1 pg
1 n g
1 µg
0
1
2
3
4
5
n o respon se
activation
0
25
25
15
15
15
0
25
21
11
9
0
25
25
9
3
22
20
0
takeoff
orien ted fligh t
source location
lan din g
1
0
7
0
0
EAD
A
C
16
C
C
17
18
C
19
GC
B
EAD
C
16
C
17
C
18
C
19
GC
1011
12
13
14
15
Time, min
FIG. 2
Sections of GC–EAD–FID traces: hexane extract of D. saccharalis fem ale pherom one glands (4 FE)
(A) an d syn th etic (9Z,11E)-h exadeca-9,11-dien al (1) (100 pg) on BPX5 ph ase (B), both
co-in jected with h ydrocarbon stan dards. Th e vertical bar in section a represen ts 0.2 m V for
EAD an d 10 m V for FID traces, respectively
Collect. Czech. Chem. Commun. (Vol. 66) (2001)

1690
Svatoš et al.:
In win d tun n el experim en ts, 1 pg of syn th etic 1 displayed h igh attrac-
tiven ess for D. saccharalis m ales; h owever, relatively few m ales foun d th e
source an d tried to copulate in com parison with m ales respon din g to 3 FE
of glan d extract (Table I). Th is observation m ay in dicate th at th e ph erom on e
produced by fem ales of D. saccharalis is very likely a m ulti-com pon en t blen d.
Our future experim en ts are aim ed towards determ in ation of th e m issin g
com pon en t(s).
W e are highly indebted to Ms J. Titzenthalerová and to Mr O. Blažek for skillful technical
assistance in insect handling and electrophysiological experiments. Financial support from the
Institute (Z4 055905) is acknowledged with a pleasure.
REFERENCES
1. Kaissling K. E.: R. H. Wright Lectures on Insect Olfaction. Simon Fraser University, Burnaby
(Canada) 1987.
2. Arn H., Tóth M. in: List of Sex Pheromones of Lepidoptera and Related Attractants (H. Arn,
M. Tóth and E. Priesner, Eds), The Pherolist. Internet database: www-pherolist.slu.se.
3. Hammond A. M., Fisher N. H: Sugar Azacar 1982, 77, 32.
4. Carney R. L., Lui A. S. T.: U.S. Pat. Appl. 1982, 4357474.
5. Mendonça A. F. (Ed.): Pragas da cana-de-açúcar. Insetos & Cia, Maceió 1996.
6. Young D. C., Vouros P., Decosta B., Holick M. F.: Anal. Chem. 1987, 59, 1954.
7. Struble D. L., Arn H. in: Combined Gas Chromatography and Electroantennogram Recording
of Insect Olfactory Response (H. Hummel and A. K. Minks, Eds), p. 161. Springer-Verlag,
New York 1984.
8. Hoskovec M., Hovorka O., Kalinová B., Koutek B., Streinz L., Svatoš A., Šebek P., Šaman D.,
Vrkoč J.: Bioorg. Med. Chem. 1996, 4, 479.
9. Svatoš A., Attygalle A. B., Meinwald J.: Tetrahedron Lett. 1994, 35, 9497.
10. Stille J. K., Simpson J. H.: J. Am. Chem. Soc. 1987, 109, 2138.
11. Ando T., Koike M., UchigamuM., Kuroko H.: Agric. Biol. Chem. 1987, 51, 2691.
12. Millar J. G., Haynes K. F. (Eds): Methods in Chemical Ecology, Vol. 1, p. 390. Kluwer Academic
Publishers, Norwell (MA) 1998.
13. Attygalle A. B., Svatoš A., Wilcox C., Voerman S.: Anal. Chem. 1995, 67, 1558.
14. Svatoš A., Šaman D.: Collect. Czech. Chem. Commun. 1997, 62, 1457.
15. Hoskovec M., Šaman D., Svatoš A.: Collect. Czech. Chem. Commun. 2000, 65, 511.
Collect. Czech. Chem. Commun. (Vol. 66) (2001)