130 M. Alibabaie and M. H. Safaralizadeh
Test on Toxicity
All tests were done in a climate chamber at
27 ± 1° C with a 12-h photoperiod at ambient RH
Untreated beans were used as control for every
experiment. Different amounts of essential oils,
at 0, 1.5, 2.21, 3.31, 4.95, and 7.80 μL/L in air
were placed on to Whatman No. 1 filter-paper
disks of 3 cm diameter. Each filter-paper disk
was then air dried for 2 min and placed on the un-
derside of the screw cap of a glass vial (300 mL).
Caps were screwed tightly on the vials, each of
which contained 20 unsexed adults (1–2 days
old). Cap was screwed tightly and the lid was
sealed with parafilm. Each concentration and
control was replicated four times. Insect mortal-
ity was checked after 24 h. Test insects were con-
sidered dead if appendages did not move when
prodded with a fine brush. Each treatment was
replicated three times.Statistical Analyses
Using SPSS (SPSS Inc. 1989–2002), data
were analyzed by one-way analysis of variance
(ANOVA) and means were compared using
Tukey test at P < 0.05. (If significant differences
between the treatments were found, they were
tested by Tukey’s tes.) Mortality of adults was
arcsine square root transformed prior to analysis.
The number of dead and live insects was counted
and probit analysis was used to estimate LC 50
and LC 95 values with their Wducial limits by
SAS 8.0 (SAS Institute 2000 ). One-way ANOVA
using Statistica (Statsoft 1998 ) was performed on
the data. A Duncan test was applied to the means
to detect significant differences of repellency
among concentrations and oils at the 0.05 % level.
Data are presented in tables as means with stan-
dard errors. A median lethal concentration (LC 50 )
was considered significantly different when the
respective 95 % fiducial limits did not overlap.Results and Discussion
The essential oil of nutmeg seeds was isolated
with a yield of 6.85 % w/w. Thirty-two compo-
nents were identified. The major compounds
in the oil were sabinene (21.38 %), 4-terpineol
(13.92 %), and myristicin (13.57 %). On the other
hand, allylbenzene and propylbenzene deriva-
tives (myristicin, safrole, eugenol, and deriva-
tives thereof) were the predominant compounds
in nutmeg seeds. The data describing select com-
pounds extracted from nutmeg seeds are present-
ed in Table 1.Table 1 Chemical composition of essential oil of nutmeg
seeds
No. Retention
timeLRI Compounds Percentage1 5.967 920 α-thujene 0.78
2 6.246 931 α-prinene 10.23
3 6.583 943 Camphene 0.16
4 7.508 978 Sabinene 21.38
5 7.792 989 α-myrcena 2.38
6 8.493 1017 α-terpinene 2.72
7 8.843 1032 Limonene 5.57
8 9.142 1045 β-ocimene 0.03
9 9.525 1061 y-terpinene 3.98
10 9.728 1070 trans-sabinene
hydrate0.0311 10.097 1085 Terpinolene 1.62
12 10.393 1098 Linalool 0.75
13 10.821 1119 Fenchyl alcohol 0.05
14 11.603 1158 cis-sabinene
hydrate0.0615 12.306 1193 4-terpineol 13.92
16 12.492 1203 α-terpineol 3.11
17 12.67 1212 Citronellol 0.77
18 13.297 1247 Linalyl acetate 0.06
19 13.949 1282 Bornyl acetate 0.24
20 14.158 1293 Safrole 4.28
21 15.76 1392 Methyl eugeunol 0.77
22 16.568 1447 Isoeugeunol 1.74
23 17.773 1530 Myristicin 13.57
24 18 1551 Elimicin 1.42
25 18.573 1586 Metoxyeugeunol 0.1
26 18.742 1595 β-asaron 0.03
27 20.625 1767 Myristic acid 0.11
28 21.01 1789 Ethyl miristate 0.04
29 21.352 1716 Palmitic acid 0.03
30 22.946 1954 Ethyl palmitate 0.07
31 25.103 2181 Stearic acid 0.01
32 25.183 2193 Ethyl oleate 0.01
LRI linear retention indices