Journal o f Chemical Ecology, Vol. 9, No.

3, 1983

DETERMINATION

OF THE VAPOR PRESSURES

OF MOTH SEX PHEROMONE

COMPONENTS

A GAS CHROMATOGRAPHIC

BY

METHOD

ANNE-MARIE OLSSON, 1 JAN AKE JONSSON, 1

BERNT THELIN, 2 and TOMMY LILJEFORS 2
~Department o f Analytical Chemistry, Chemical Center
ZDepartment o f Organic Chemistry 3, Chemical Center
University o f Lurid, P.O. Box 740, S-220 07 Lurid, Sweden
(Received April 27, 1982; revised August 10, 1982)

Abstract--The vapor pressures of decyl acetate, five decenyl acetate isomers,

(Z)-7-dodecenyl acetate, and (Z)-9-tetradecenyl acetate have been determined at three to six temperatures in the interval 25-45~ by a gas
chromatographic method suitable for accurate measurements of the low
vapor pressures of moth sex pheromone components at biologically
relevant temperatures. The vapor pressure values at 30.5 ~ C are 3.80 Pa for
decyl acetate, 4.08-5.40 Pa for the decenyl acetate isomers, 0.562 Pa for
(Z)-7-dodecenyl acetate, and 0.094 Pa for (Z)-9-tetradecenyl acetate. The
vapor pressures of the decenyl acetates show a small but significant
dependence on the double bond position. Four of the c o m p o u n d s in this
study, 10:Ac, Z 5 - 1 0 : A c , Z 7 - 1 2 : A c , and Z 9 - 1 4 : A c have recently been
identified as sex pheromone components of the turnip moth, Agrotis
segetum. Large differences between the mole percentages of the c o m p o n e n t
as found in liquid extracts of female abdominal tips and the corresponding
mole percentages in the vapor phase are predicted.
Key W o r d s - - V a p o r pressure, gas chromatography, sex pheromone, olefinic
acetates, turnip moth, Agrotis segeturn, liquid-vapor equilibrium.

INTRODUCTION

The volatility, measured as the saturated vapor pressure, of pheromone

components is an important factor in studies on the physicochemical aspects
of insect pheromones. Hirooka and Suwanai (1976) have, for instance,
derived an equation which relates the rate of pheromone release by a female
moth to the vapor pressure, the diffusion coefficient, and the size and form of
375
0098-0331/83/0300-0375503.00/09 1983PlenumPublishingCorporation

376

OLSSON ET AL.

the pheromone gland. For moth species the pheromone generally is a mixture
of several compounds, mainly olefinic acetates with varying chain lengths. In
many cases geometrical a n d / o r positional isomers are also present in the
pheromone blend (Ritter, 1979). The female moths produce a well-defined
ratio of the different compounds and the component ratio has been found to
play an important role for optimal attraction of males in the field (Roelofs,
1978). However, when the vapor pressures of the pheromone components
differ significantly, the ratio of the components in the vapor phase is different
from that on the female gland. The former ratio depends on the relative
saturated vapor pressures of the components and their relative mole fractions
in the liquid phase (Hirooka and Suwanai, 1978).
In connection with field trapping studies more basic work concerning the
release rates of pheromones from controlled release systems are needed
(Weatherston, 1981). In such investigations physical data on the volatility
relationships should be of great value.
In electrophysiological studies on dose-response relationships, the
volatilities of the test compound should be taken into account to make it
possible to determine the amount of substance to be used in the experiment.
Although the vapor pressures of the compounds used by insects as sex
pheromone components thus enter into many different types of pheromone
studies, very few experimental values are available. Hirooka and Suwanai
(1978) used a gas saturation method to determine the vapor pressure of a few
compounds used as pheromone components by moth species.
In this paper we report on vapor pressures for some pheromone
components and related compounds determined by a gas chromatographic
method, developed by two of us (Olsson et al., 1976). It has previously been
used for several applications (J6nsson et al., 1980; J6nsson and Pscheidl,
1981).
Four of the compounds chosen for this work, decyl, (Z)-5-decenyl,
(Z)-7-dodecenyl and (Z)-9-tetradecenyl acetate, have been identified as sex
pheromone components of the turnip moth, Agrotis segetum, (Tbth et al.,
1980; Arn et al.~ 1980; LOfstedt et al., 1981). To investigate the influence of the
position of the double bond on the vapor pressure, (Z)-3-, (Z)-4-, and
(Z)-6-decenyl acetate were included in the study. Finally, the vapor pressure
of a geometrical isomer, (E)-5-decenyl acetate, was determined.
METHODS AND MATERIALS

Principles. The stibstance under study is used as the stationary liquid

phase (SLP) in a ga~,-liquid chromatographic column. A suitable sample
compound, the "probe," is repeatedly injected into the gas chromatograph
and the net retention volume VNiS carefully measured. According to basic gas
chromatographic theory, the following equation applies:

377

V A P O R PRESSURES OF P H E R O M O N E S

VN = VR -- VM = K" VL

(1)

Here VR is the total retention volume, usually calculated as the p r o d u c t of the

retention time and the carrier gas flow rate. VM is the volume o f the e m p t y
space in the column, VL is the volume of the SLP, and K i s a constant, the
partition coefficient. V ~ c a n be measured as the retention volume of m e t h a n e
as K for methane can be neglected under normal conditions.
D u e to the evaporation of the SLP, VL decreases linearly with the total
a m o u n t of gas, Vacc, which has passed the column since the start of the
experiment. As is seen f r o m equation (1), this means that VN also decreases in
the same way. A n experimental example of such a line is s h o w n in Figure I.
The slope, -dVN/dVaocis p r o p o r t i o n a l to the e v a p o r a t i o n rate and thus to the
v a p o r pressure of the S L P if the carrier gas is saturated with the evaporated

90

85

80

7,5

80

100

Vac c (11
FIG. 1. The retention volume, VN, of isobutyl acetate on a (Z)-7-dodecenyl acetate
column vs. the accumulated carrier gas volume, Vacc, at 35. I~

378

OLSSON ET AL.

substance. This is normally the case, as was shown by Olsson and coworkers
(1976) and Olsson and JOnsson (1981).
From a knowledge of the initial weight of the SLP WLs and the initial
retention volume VNs, the vapor pressurep of the SLP can be calculated from
the equation (Olsson et al., 1976):
dVN
P=dVacc

WLB R T
Vws M

(2)

Here, T is the absolute temperature, M is the molecular weight of the SLP,

and R is the gas constant. Equations (1) and (2) are valid if contributions from
adsorption effects to the retention volume of the probe can be neglected. This
can be achieved by selection of a probe which is chemically similar to the SLP.
Chemicals9 The following acetates were used in the study: decyl (10: Ac),
(Z)-3-decenyl (Z3-10: Ac), (Z)-4-decenyl (Z4-10:Ac), (Z)-5-decenyl (Z510: Ac), (E)-5-decenyl (E5-10:Ac), (Z)-6-decenyl (Z6-10: Ac), (Z)-7-dodecenyl (Z7-12: Ac), and (Z)-9-tetradecenyl (Z9-14: Ac).
The monoolefinic compounds were all synthesized from alkynes and
o~-bromoalkyl tetrahydropyranyl ethers9 The resulting alkynyl tetrahydropyranyl ethers were hydrogenated over Lindlar catalyst to give the corresponding (Z)-alkenyl compounds or were treated with sodium in liquid
ammonia to give the (E)-alkenyl compound. Finally they were converted to
acetates by acetyl chloride in acetic acid.
After purification by argentation chromatography using a cationexchange resin impregnated with silver nitrate (Houx et al., 1974), the
compounds were >99.5% pure except for Z9-14:Ac which was about 97%
pure.
Experimental Procedure. For each substance, one gas chromatographic
column was prepared, according to the method described by Conder and
Young (1979, p. 595). The solid support was Chromosorb W-AW-DMCS
(Johns-Manville, Denver, Colorado). About 600 mg of packing, containing
about 60 mg of the substance was filled into each column. The glass columns
were V-shaped and had the dimensions 800 mm 2 mm ID. The initial
amount of liquid was determined by careful weighings during the column
preparation procedure.
All retention volume measurements were performed using a high
precision gas chromatograph, coupled online with a mini-computer. This
system, described in detail elsewhere (J6nsson et al., 1975), permits the direct
measurement of retention volumes, without involving the concept of retention
time.
The carrier gas was hydrogen in order to minimize the pressure gradient
in the column. The flow rate was ca. 30 ml/min. The column volume VM,was
measured by injection of methane (99.95%). As "probe" we used isobutyl

379

VAPOR PRESSURES OF PHEROMONES

acetate (Eastman, Rochester, New York) which is chemically similar to the

substances studied, thus minimizing adsorption.
Vapor samples of the probe were repeatedly injected at specified time
intervals under computer control. For each injection the retention volume was
automatically calculated by the computer and stored for further computer
processing. The evaporation of a substance was traced at each temperature,
until the 95% confidence interval for the slope dVN/dVacc was less than 1%
relative. This is of the same magnitude as other sources of error and,
consequently, the precision cannot be improved by further measurements.
Depending on the vapor pressure, the time needed to reach enough high
precision was 5-75 hr. The same column was used for several temperatures as
in an earlier work (JOnsson et al., 1980) and the temperatures were run in
random order.

RESULTS AND DISCUSSION

In Table 1 and Figures 2 and 3 the results of the vapor pressure

measurements are summarized. In the figures, the values of lnp are plotted vs.
the reciprocal temperatures. In a limited temperature interval, a straight line
for each substance should be obtained according to Clausius-Claperon's
equation:
d In p = _ A Hvap

d(1/T)

(3)

In a few cases the measurements were repeated with a new column. Such data
are indicated in Table 1 and are not plotted. It can be seen that repeated
measurements give values which closely agree with each other. This reflects
the good reproducibility in the entire measurement procedure, including the
preparation of the column.
In an earlier study (J6nsson et al., 1980) vapor pressures for 1chloroalkanes were found to agree well with literature data, generally within
5%, and we conclude that the method gives reliable v a p o r pressure values. It is
also a suitable method for the study of moth sex pheromone components as it
is possible to accurately measure the low vapor pressures of such compounds
at biologically relevant temperatures. Extrapolations of vapor pressure data
over large temperature intervals may give grossly erroneous values. The
method is best suited for the determination of vapor pressures in the range
0.01-100 Pa.
In Figures 2 and 3 the slopes of the lines for all decyl and decenyl acetates
are approximately the same, and the slope increases with increasing chain
length. F r o m this slope, the enthalpy of vaporization, A Hvap, can be calculated
according to equation (3).

380

OLSSON ET AL.
TABLE 1. VAPOR PRESSURES ( p ) OF SUBSTANCES STUDIED a

Substance
10:Ac

Z3-10:Ac

Z4-10:Ac

Z5-10:Ac

E5-10:Ac

Z6-10:Ac

Z7-12:Ac

Z9-14:Ac

Temperature (~

p (Pa) b

25.9
26.1
30.4
34.8
39.4
39.5
26.0
30.4
34.8
39.4
34.9
34.9

2.48
2.52
3.80
5.66
8.86
8.70
3.02
4.65
6.76
10.40
8.26
7.95

+ 0.01
+ 0.04 c
-+ 0.05
+ 0.03
+ 0.03
+- 0.11 c
-+ 0.04
- 0.04
_+ 0.07
+ 0.11
+ 0.06
-+ 0.06 c

39.4
26.2
30.6
35.1
39.6
26.0
30.4
34.9
39.3
25.9
30.4
34.8
39.4
30.5
34.8
35.1

12.12
2.93
4.65
6.91
10.13
2.84
4.08
6.44
9.58
2.91
4.38
6.83
10.08
0.562
0.83
0.89

_+ 0.09
+ 0.04
+ 0.04
- 0.06
+ 0.11
_+ 0.02
+ 0.04
+ 0.08
+ 0.10
+ 0.03
- 0.03
+ 0.06
- 0.06
+ 0.005
+ 0.01
- 0.01 c

39.2
44.0
44.1
30.5
35.0
39.5
44.0

1.33
2.01
2.07
0.094
0.163
0.267
0.428

-+
+
+
+
+
+
+

0.01
0.02
0.01 c
0.001
0.001
0.002
0.004

~The u n c e r t a i n t y is g i v e n as a 9 5 % c o n f i d e n c e interval. T h e a b b r e v i a t i o n s o f the n a m e s o f the

s u b s t a n c e s are e x p l a i n e d in the text u n d e r C h e m i c a l s .
~1 P a = 0.987 10-5 a t m = 7.50 10 -3 m m H g .
~New, i n d e p e n d e n t m e a s u r e m e n t . See text.

381

VAPOR PRESSURES OF PHEROMONES

2.00

decyl and
decenyl acetate=
( see Fig. 3 )

1.00

O.
,,,.,.

0.00

Z7 - 1 2 : A c

-1.00

-2.00

Z9 -14:Ac

W~

3.15

3.25

3.35

1 0 3 / T (K)
FIG. 2. The logarithm of the vapor pressure for pheromone substances vs. the
reciprocal temperature. The abbreviations for the names of the substances are
explained in the text under Chemicals.

382

2.50

OLSSON ET AL.

I
"x

2.00

O.
t,,,m

1.50

1.00

/ L ~

- ] U : AC

- IO:Ac

~--~-E5 _ IO:Ac
~IO:Ac

....1
3.20

3.25

3.30

103IT

I
3.35

(K)

FIG. 3. Expansion of the upper part of Figure 2.

T h e results calculated from our data are: 72 k J / m o l for decyl and decenyl
acetates, 77.5 k J / m o l for Z7-12:Ac, and 90 k J / m o l for Z9-14:Ac. These
data agree well with vaporization enthalpy data obtained for 1-chloroalkanes
of similar molecular sizes (JOnsson et al., 1980).
Hirooka and Suwanai (1978) measured vapor pressures of compounds
similar to those in this study. Only one of them is the same (Z9-14:Ac). The
agreement with our values for this compound is within 10% at 40~ and 50 ~C,
but at 30~ their value is ca. 50% higher than ours. This compound is one of
the least volatile in their study. The vaporization enthalpy for Z 9 - 1 4 : A c
calculated from the data of Hirooka and Suwanai is 72 k J / m o l , which is
unrealistically low for a molecule of this size. Their method seems to suffer
f r o m systematic errors, especially at low v a p o r pressures. No other sources of
literature data of vapor pressures at biologically relevant temperatures for the

VAPOR PRESSURES OF PHEROMONES

383

substances under study have been found, not even in a very comprehensive
compilation (Dykyj and Rep~tg, 1979).
Effects of Position of Double Bond. From measurements on computergenerated models of the different (Z)-decenyl acetates, the midpoint of the
molecule is found to be near the 4 position. Thus Z 4 - 1 0 : A c is the most
symmetrical compound with respect to the double bond position. As (Z)olefins are folded at the double bond, a more centrally located double bond
gives a smaller molecular size. The vapor pressure mainly depends on
intramolecular forces in the liquid phase. Therefore, a molecule with a smaller
effective size should have a higher vapor pressure than one which is effectively
larger. Thus Z4-10: Ac should have the highest vapor pressure, followed by
Z3-10: Ac and Z5-10: Ac which is in agreement with the data obtained.
This effect can also be seen in vapor pressure data for small olefins as
hexenes (Camin and Rossini, 1956) and heptenes (Eisen and Orav, 1970).
(Z)-3~Hexene and (Z)-3-heptene both have higher vapor pressures than (Z)2-hexene and (Z)-2-heptene, respectively. Also, Butler and McDonough
(1979) found higher evaporation rates for more symmetric Z-olefin acetates
from rubber septa. This was explained by less retardation by the rubber
matrix. Our results indicate that at least part of this effect may be explained by
differences in vapor pressure. The logarithms of the half-lives (t89 as
determined by Butler and McDonough and the corresponding vapor
pressures reported in the present work show an excellent linear relationship.
Effects of Chain Length. The vapor pressure ratio at 35~ for the
homologs studied in this work are 7.8 for Z5-10:Ac/Z7-12:Ac and 5.4 for
Z7-12:Ac/Z9-14:Ac. Thus there is no strict additivity in In p for the
homologs in this series as is found for more simple compounds, e.g., normal
alkanes. As was shown above, the vapor pressures of olefinic compounds
depend on the position of the double bond. Strict additivity should thus only
be expected for a series of homologs in which the compounds have nearly the
same relative length of the two hydrocarbon chains separated by the double
bond.
Effects of Double Bond Configuration. In the only pair which we have
studied, Z5-10:Ac has a higher vapor pressure than E5-10:Ac. The
difference is less than 10%.
Effects of Double Bond. As expected, all decenyl acetates have higher
vapor pressures than the saturated decyl acetate. Discussions of the relative
amounts of compounds in a pheromone blend in terms of major and minor
components are generally based on the relative amounts found in extracts of
female glands. When the vapor pressures of the pheromone components are
significantly different, the relative amounts in the vapor phase differ from
those in the liquid phase. As an example, four of the compounds in this work,
10:Ac, Z5-10:Ac, Z7-12:Ac, and Z9-14:Ac, identified as sex pheromone

384

OLSSON ET AL.

c o m p o n e n t s of Agrothr s e g e t u m , m a y be considered. The a m o u n t s of

substances f o u n d in female a b d o m i n a l tip extracts ( L 6 f s t e d t et al., 1981)
c o r r e s p o n d to the m o l e p e r c e n t a g e s 5.7, 4.3, 62.7, a n d 27.3 f o r 10: Ac, Z S 10 : Ac, Z 7 - 1 2 : Ac, a n d Z 9 - 1 4 : Ac, respectively, if only these f o u r c o m p o n e n t s
are considered. Using R a o u l t ' s law a n d the v a p o r pressures of the c o m p o u n d s
at 20 ~ o b t a i n e d by e x t r a p o l a t i o n o f the d a t a in T a b l e 1, the c o r r e s p o n d i n g
m o l e percentages in the v a p o r p h a s e b e c o m e 28.6, 25.5, 43.2, a n d 2.7,
respectively. Thus the m i n o r c o m p o n e n t s in the extract, 1 0 : A c a n d Z 5 10: Ac, d r a s t i c a l l y increase their relative a m o u n t s in the v a p o r phase, due to
higher v a p o r pressures. I n s t e a d Z 9 - 1 4 : Ac (27.3 mole % in the liquid phase)
with a lower v a p o r p r e s s u r e b e c o m e s a m i n o r c o m p o n e n t s (2.8 m o l e %) in the
v a p o r phase.
If the t e c h n i c a l p r o b l e m s involved in q u a n t i f y i n g sex p h e r o m o n e s
v o l a t i l i z e d f r o m female glands can be solved, a n analysis of the female
effluvium directly gives the v a p o r phase c o m p o s i t i o n of the p h e r o m o n e blend.
This m e t h o d s h o u l d give a m o r e b i o l o g i c a l l y relevant picture of p h e r o m o n e
c o m p o s i t i o n s t h a n the generally used analysis of female g l a n d extracts. The
v a p o r p r e s s u r e d a t a of p h e r o m o n e c o m p o n e n t s p r e s e n t e d in this w o r k s h o u l d
then be useful in c o n n e c t i o n with research on dispensing p h e r o m o n e
c o m p o n e n t s as an o p t i m a l blend.
Acknowledgments--We thank Dr. J. L6fqvist, Dr. J. Byers, C. L/Sfstedt, and F. Schlyter,
Department of Animal Ecology, University of Lund, for valuable criticism of the manuscript.
This work was partly funded by the Swedish Natural Science Research Council which is gratefully
acknowledged.

REFERENCES
ARN, H., ST.g.DLER,E., RAUSCHER,S., BUSER,H.R., MUSTAPARTA,H., ESBJERG,P., PH1LIPSEN,
H., ZETHNER,O., STRUBLE,D.L., and BUES, R. 1980. Multicomponent sex pheromone in
Agrotis segetum: Preliminary analysis and field evaluation. Z. Naturforsch. 35c:986-989.
BUTLER, L.I., and McDoNOUGH, L.M. 1979. Insect sex pheromones. Evaporation rates of
acetates from natural rubber septa. J. Chem. Ecol. 5:825-837.
CAMIN,D.L., and ROSSINI,F.D. 1956. Physical properties of the 17 isomeric hexenes of the API
research series. J. Phys. Chem. 60:1446-1457.
CONDER,J.R., and YOUNG,C.L, 1979. Physicochemical Measurement by Gas Chromatography.
Wiley-lnterscience, New York.
DYKYJ,J., and REP~, M. 1979. The Vapour Pressure of Organic Compounds (in Slovak). Veda,
Bratislava.
EISEN, O., and ORAV,A. 1970. Determination of boiling points and vapour pressures of some
unsaturated hydrocarbons (in Russian). Eesti N S V Tead. Acad. Tiom., Keem., Geol.
19:202-205.
HIROOKA, Y., and SUWANALM. 1976. Role of insect sex pheromone in mating behavior. 1.
Theoretical consideration on release and diffusion of sex pheromone in the air. Appl.
Entomol. Zool. 1I:126-132.

VAPOR PRESSURESOF PHEROMONES

385

HIROOKA,Y., and SUWANAI,M. 1978. Role of insect sex pheromone in mating behaviour. IL An
aspect of sex pheromone as a volatile material. Appl. Entomol. Zool. 13:38-43.
Houx, N.W.H., VOERMAN,S., and JONGEN,W,M.F. 1974. Purification and analysis of synthetic
insect sex attractants by liquid chromatography on a silver-loaded resin. J. Chromatogr.
96:25-32.
J()NsSoN, J.A., and PSCHEIDL, H., 1982. Chromatographische Reaktionsf/ihrung: Stabilit~it der
gaschromatographischen station~iren LSsungsmittelphase. Z. Chem. (Leipzig) 22:105-106.
JONSSON, J.A., JONSSON, R., and MALM, K. 1975. Direct measurement of the gas chromatographic retention yolume using an on-line mini-computer, lI. Apparatus: J. Chromatogr.
115:57-64.
JONgSON,J.A., MATHIASSON,L., and OLSSON,A.-M. 1980. A GLC method for the determination
of low vapour pressures applied to l-Chloroalkanes. Acta Chem. Scand. A34:147-150.
LOFSTEDT, C., VANDER PERS,J.N.C., LOFQVIST,J., LANNE,B.S., APPELGREN,M., BEGSTROM,G.,
and THELIN, B. 1983. Sex pheromone components of the turnip moth, Agrotis segetum:
Chemical identification, electrophysiological evaluation and behavioural activity. J. Chem.
Ecol. 8:1305-1321.
OLSSON,A.-M., and JONSSON,3.fl~. 1982. Distribution of the stationary phase in the GLC column.
J. High Resolution Chromatogr. Chromatogr. Commun. 5:55-56.
OLSSON,A.-M., MATmASSON,L., JONSSON,J.~., and HARALDSON,L. 1976. Determination of the
loss of the stationary phase in gas-liquid chromatography from the change in retention
volume. J. Chromatogr. 128:35-43.
RITTER, F.J. 1979. Chemical ecology: Odour communication in animals, general introduction
and overview, in F.J. Ritter (ed.). Chemical Ecology: Odour communciation in Animals.
Elsevier/North-Holland Biomedical Press, Amsterdam.
ROELO~S, W.L. 1978. Threshold hypothesis for pheromone perception. J. Chem. Ecol. 4:685-689.
TbTH, M., JAKAB, J., and NOVs L. 1980. Identification of two components from the sex
pheromone system of the white-line dart moth, Scotia segetum (Schiff.) (Lep., Noctuidae).
Z. Angew. Entomol. 90:505-510.
WEATnERSTON, J., GOLUB, M.A., BROOKS, T.W., HUANG, Y.Y., and BENN, M.H. 1981.
Methodology for determining the release rates of pheromones from hollow fibers, in E.R.
Mitchell (ed.). Management of Insect Pests with Semiochemicals. Concepts and Practice.
Plenum Press, New York.