Nutmeg Seed
Nutmeg Seed
Nutmeg Seed
One of the centers of nutmeg seed commerce in Indonesia is 2.5 Physical analysis of essential oil
West Sumatera. In this area nutmeg seed are sold based on its The physical analysis that observed were specific gravity,
harvesting age, whether it is young seed, medium seed, or old refractive index, and optical rotation. These three parameters
seed. Each type has different price and in descending manner, were included in EP standard. The amount of samples
the most valuable would be mace, young seed, medium seed, required for this analysis as 20 ml.
and old seed. In the process of making nutmeg oil, entire
ingredients would be mixed via distillation. The chemical 2.6 Essential oil composition analysis by GC-MS
properties of nutmeg oil may be related to the harvesting age Nutmeg mace and seed oil compound profile analysis was
of nutmeg mace and seed. Several nutmeg related researches done by GC method. The equipment used was GC to
have been conducted, however research on relationship determine contents in area percentage and GC-MS to
between mace, different aged nutmeg seed, and determine essential oil compound composition. Essential oil
physicochemical characteristics from raw materials originated quality profile was determined by matching of sample
from West Sumatra had never been conducted before. This component fragmentation quality with library data on mass
prompts for research in studying physicochemical spectrum with 80% minimum. GC condition was set on:
characteristic of mace and different harvesting age of nutmeg injector temperature at 275 oC with split mode (split ratio
seed via hydrodistillation technique. This research is hope to 100:1) and nitrogen carrier gas with 0.5 ml/minute flow rate.
give useful information for farmers, distillers, and flavor and Oven condition was set as: 100 oC (10 minutes hold time)
fragrance industry. then 250-300 oC in 5 oC rate (15 minutes hold time). Detector
condition was set as: 275 oC, 30 ml/minute H2 flow, 400
2. Material and Methods ml/min air flow, and 25 ml/min make up flow. GC-MS
2.1 Plant Material condition for all essential oil sample was set as: 250 oC ion
Samples consist of mace and young, medium, and old nutmeg source temperature, 200 oC quadrupole temperature, 10-250
seed obtained directly from nutmeg sellers in Batang Kapas amu of scan mass, and around 35 μA of emission. Energy
Districts, South Pesisir Regency, West Sumatra Province, used was 70 eV with WMV < 2000 V. Carrier gas used
Indonesia. Each sample that had been sorted by nutmeg seller helium with 0.5 ml/min flow. Oven and injector conditioning
was weighted 420 grams. This research use laboratory was the same as GC. Compound identification was done by
analysis method with two repetition technique. comparing the Wiley 2008-NIST library and P. Adam data of
the peaks with those reported in literature, mass spectra of the
2.2 Physical Characterization of Nutmeg Seed peaks with literature data.
The characteristics of seed diameter and weight were
measured. Seed diameter was measured longitudinally and 2.6. Data analysis
transversally. The number of samples were 20 seeds (n=20). Data was analyzed by using analysis of variance (ANOVA).
Average value and deviation value were calculated. If the result were significantly different, analysis would
continue with Least Significant Difference (LSD) test on α =
2.3 Sample Preparation 0,05.
The Nutmeg and mace samples were crushed by Xi An
milling machine made in China, which followed by sieving 3. Result and Discussion
using 30 mesh sieve. Nutmeg mace and seed moisture was 3.1 Physical characteristics of nutmeg seed
measured by Sartorius Moisture Analyzer for both samples In observation, nutmeg mace is brownish red in color, has
that passed and did not the sieve. irregular shape, thin, and crumpled upon touch. Nutmeg seed
is mostly dark brown, differing in diameter, weight, and
2.4 Extraction of Essential Oil shape. The result of nutmeg seed diameter measurement and
This stage covers distillation, cooling, and separation of mace weight is presented in Table 1. Young nutmeg seed is marked
and seed oil from water. (1) Distillation was done in order to by the presence of wrinkles surrounding the seed and tender
separate mace and seed oil content using hydrodistillation when pressed by the tip of our nail. Graphic of the
technique, 200 grams samples were put inside round bottom relationship between nutmeg seed’s average weight, yield,
flask equipped with clevenger apparatus. Afterwards water moisture, and sieved is shown in Figure 1.
was added until the ratio of water to sample 6:1. The flask
was then placed on top of heating mantle or hot plate.
Distillation was done for 12 hours or until the oil content
inside distilled water had run out, or considered done marked
by clear distillate water. (2). Cooling was conducted to
condense and cool the mixture of hot water steam and oil, so
that oil could be separated from water. In this stage, the
mixture of steam and oil was passed through spiral glass tube
which was cooled by running water. Oil and steam would be
cooled and condensed along the cooling tubes, in which
distillate water would be contained in a separating funnel with
oil. (3) Separation was conducted in the purpose of separating
oil from water. Oil which has lighter specific gravity than
water would be on top, while oil with heavier specific gravity Fig 1: Graphic of the relationship between nutmeg seed’s average
than water would be below water. Afterwards mace and seed weight, yield, moisture, and sieved
oil was weighted to calculate yield then analyzed, after then
stored inside tightly closed glass bottle in room temperature According Siregar et al. young nutmeg upon harvesting
for physical and compounds analysis. would then has its fruit flesh cut open to reveal greenish white
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Journal of Pharmacognosy and Phytochemistry
mace and seed, with is shell still tender on pressure [11]. with oval seed [12]. The average index of the seed’s diameter is
Young nutmeg seed has high moisture, and after drying it will 1.17. The average index was obtained from the ratio of
shrink and wrinkles. Observation on medium nutmeg seed longitudinal and transversal diameter. The measurement of
revealed that half of the seed has hardened while half of it old West Sumatera nutmeg seed showed 1.16 average index
wrinkles. This is different with old nutmeg seed where it has value. This gives hint that nutmeg seed originating from West
strong and rough shell in oval shape. According to Soenarsih Sumatera has genus likeness with the seed coming from
et al., the shape of Banda Myristica fragrans nutmeg is round Banda.
Nutmeg seed
Analysis description Mace EP Standard**
Young Medium Old
Mace and nutmeg seed
b b a
Longitudinal diameter (cm) - 1.45 ± 0.03 1.56 ± 0.08 2.34 ± 0.01 -
c b a
Transversal diameter (cm) - 1.02 ± 0.07 1.77 ± 0.05 2.02 ± 0.04 -
c b a
Weight (gr) - 0.77 ± 0.11 1.67 ± 0.01 2.71 ± 0.07 -
b a a a
Moisture (%) 10.36 ± 0.23 14.48 ± 0.66 13.75 ± 0.42 14.71 ± 0.08 -
d a b c
Sieved particles by mesh 30 (%)* 3.03 ± 0.03 10.23 ± 0.07 7.74 ± 0.32 6.83 ± 0.07 -
Nutmeg oil
a b c d
Yield (w/w %) 19.51 ± 0.50 8.91 ± 0.58 6.35 ± 0.17 5.04 ± 0.03 -
0 b c b a
Specivic gravity (25/25 C) 0.919 ± 0.002 0.902 ± 0.005 0.923 ± 0.001 0.930 ± 0.001 0.89 - 0.905
a b a
Refractory index (200C) 1.487 ± 0.001 1.481 ± 0.0017 1.486 ± 0.000 1.487 ± 0.000 a
1,475 - 1,485
a a a a
Optical rotation (+)6.07 ± (+)0.27 (+)7.51 ± (+)0.79 (+)6.61 ± (+)0.29 (+)6.58 ± (+)0.48 (+)8 - (+)18
Annotation: *finely grounded nutmeg seed **European pharmacopoeia. Superscripted letters on numbers indicated
significant difference (LSD α = 0.05).
As shown in the Table 1, the difference in percentage between are 1.487, 1.481, 1,486, and 1.487. The value of specific
particles that passed the sieve might be related to harvesting gravity and refractive index showed that the older the
age factor. Young nutmeg seed tissue is still tender and thus harvesting age, the higher the value of specific gravity and
after crushing it has more small particles compared to refractive index of nutmeg seed. Optical rotation is molecular
medium and old nutmeg seed. Medium and old nutmeg seed structure response against single light wave path. Table 1
has hard tissues, which caused their passable sized particles to showed that the older the seed, the smaller the value of optical
be fewer than young seed. Particles that were unable to pass rotation. If compared to EP standard, the value of nutmeg’s
the sieve were mostly shell shards specific gravity and refractive index meet the standard,
however its optical rotation value does not. Research
3.2 Extraction of Essential oil conducted by Ma’mun towards nutmeg oil sample from Papua
Figure 1 shows highest amount of nutmeg seed particles that obtained 0.909 specific index value, 1.487 refractive index
passed sieving has highest yield, while seeds which lower value, and (+)15.3 optical rotation value [7].
particles passing the sieve has lower amount of yield. This is
inversely proportional with weight, where old nutmeg seed 3.4 Chemical composition of Essential Oil
with highest weight produced the lower percentage of yield. A total of 40 compounds were identified for nutmeg mace and
Particles of old nutmeg seed that did not pass the sieving were seed oil. The result of the analysis is shown in Figure 2, while
mostly shards of old nutmeg seed’s shell, which influenced the type of volatile compound and percentage of total area are
the percentage of nutmeg oil yield. This is in line with of a shown in Table 2 and Table 3 of quantitative analysis result
research done by Suryatmi et al, which stated that distillation from young, medium, and old nutmeg seed.
with nutmeg shell as main ingredients does not produce
nutmeg essential oil [13]. Yield percentage obtained from that
research was almost the same with the one done by Pal et al,
where old nutmeg yield percentage from Andaman Nicobar
Island was 8.25 [14]. However, this research did not explain
harvesting age of nutmeg seed.
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Journal of Pharmacognosy and Phytochemistry
registered in EP standard were alpha pinene, sabinene, beta supporting standard in acceptance of nutmeg oil within flavor
pinene, 3-carene, limonene, gamma terpinene, 4-terpineol, and fragrance industry. They were eugenol, methyl eugenol,
safrole, and myristicine. The 4 other constituents were used as isoeugenol, and elimicine.
Table 2: Chemical compound profile in nutmeg mace and young, medium, and old seed oil
Peak Nutmeg mace oil Young nutmeg Medium nutmeg Old nutmeg seed
Chemical compound m/z
number (%) seed oil (%) seed oil (%) oil (%)
Alpha thunjene c a ab b
1 136 1.78 ± 0.03 2.27 ± 0.03 2.22 ± 0.04 2.11 ± 0.00
Alpha pinene* a a b c
2 136 19.77 ± 0.06 19.28 ± 0.12 17.75 ± 0.26 16.16 ± 0.25
Camphene a a b b
3 136 0.36 ± 0.01 0.37 ± 0.01 0.33 ± 0.01 0.31 ± 0.01
Sabinene* a a a a
4 136 12.45 ± 0.14 12.23 ± 0.65 11.09 ± 0.07 11.07 ± 0.35
Beta pinene * ab a b c
5 136 14.77 ± 0.13 14.78 ± 0.37 13.37 ± 0.02 12.26 ± 0.08
Beta myrcene a a b b
6 136 2.30 ± 0.02 2.30 ± 0.06 2.06 ± 0.01 1.94 ± 0.02
Alpha phelandrene a b c c
7 136 0.93 ± 0.03 0.54 ± 0.01 0.49 ± 0.01 0.49 ± 0.02
3-carene* a b b b
8 136 1.53 ± 0.03 0.42 ± 0.01 0.40 ± 0.01 0.40 ± 0.02
Alpha terpinene b a b b
9 136 3.21 ± 0.05 3.56 ± 0.09 3.25 ± 0.05 3.18 ± 0.01
Beta ocimene c a b b
10 136 0.41 ± 0.01 1.73 ± 0.05 1.35 ± 0.06 1.24 ± 0.04
Limonene* a a b b
11 136 5.63 ± 0.10 5.84 ± 0.18 5.12 ± 0.04 4.93 ± 0.03
Gamma terpinene* b a ab b
12 136 4.98 ± 0.08 5.54 ± 0.16 5.19 ± 0.06 5.12 ± 0.00
Cis sabinene hydrat c a a b
13 136 0.16 ± 0.00 1.08 ± 0.04 1.05 ± 0.02 0.91 ± 0.03
Cymenene b a a a
14 132 0.05 ± 0.00 0.07 ± 0.00 0.07 ± 0.01 0.07 ± 0.00
Alpha terpinolen a b b b
15 136 2.24 ± 0.04 1.49 ± 0.05 1.41 ± 0.01 1.40 ± 0.01
Trans sabinene hydrat c a a b
16 154 0.13 ± 0.00 0.99 ± 0.05 1.00 ± 0.02 0.85 ± 0.03
17 4-Isopropyl-1-methyl-2-cyclohexen-1-ol c a ab b
154 0.35 ± 0.00 0.65 ± 0.02 0.63 ± 0.00 0.60 ± 0.01
18 1-Methyl-4-isopropyl-3-cyclohexen-1-ol c a ab b
154 0.25 ± 0.00 0.44 ± 0.01 0.42 ± 0.00 0.40 ± 0.01
4-terpineol* c a b b
19 154 7.87 ± 0.20 11.47 ± 0.24 10.35 ± 0.08 10.39 ± 0.04
Beta fhencol c a b b
20 154 0.94 ± 0.02 1.23 ± 0.04 1.12 ± 0.01 1.14 ± 0.01
Trans piperitol b a a a
21 154 0.11 ± 0.00 0.20 ± 0.01 0.21 ± 0.00 0.21 ± 0.00
Safrole* a c c b
22 162 2.30 ± 0.00 1.23 ± 0.06 1.24 ± 0.01 1.40 ± 0.00
Alpha bornyl acetate b a ab ab
23 196 0.13 ± 0.01 0.20 ± 0.01 0.19 ± 0.02 0.15 ± 0.00
p-Pentylanisole b b a a
24 178 0.13 ± 0.01 0.14 ± 0.01 0.27 ± 0.02 0.30 ± 0.01
Eugenol** a b b a
25 164 0.39 ± 0.01 0.26 ± 0.01 0.29 ± 0.02 0.35 ± 0.00
Citronelyl acetate b b a a
26 198 0.19 ± 0.01 0.19 ± 0.01 0.22 ± 0.01 0.23 ± 0.00
Alpha terpenyl acetate c b a b
27 196 0.02 ± 0.00 0.07 ± 0.00 0.09 ± 0.01 0.07 ± 0.00
Alpha cubebene c b a ab
28 204 0.06 ± 0.01 0.12 ± 0.00 0.16 ± 0.01 0.13 ± 0.00
Methyl Eugenol** a b a a
29 178 0.35 ± 0.00 0.24 ± 0.03 0.34 ± 0.01 0.38 ± 0.01
Neryl acetate b a a a
30 196 0.02 ± 0.01 0.07 ± 0.01 0.09 ± 0.01 0.07 ± 0.02
Beta caryophyllene c b ab a
31 204 0.01 ± 0.00 0.03 ± 0.00 0.04 ± 0.00 0.05 ± 0.01
Isoeugenol** a b a a
32 164 1.38 ± 0.12 0.44 ± 0.18 0.98 ± 0.05 1.21 ± 0.00
Alpha bergamotene c b a ab
33 204 0.02 ± 0.00 0.11 ± 0.01 0.14 ± 0.01 0.13 ± 0.01
Beta farnesene ni ni ni ni
34 204 0.00 ± 0.00 0.03 ± 0.00 0.05 ± 0.00 0.04 ± 0.00
Methyl isoeugenol b b a a
35 178 0.04 ± 0.01 0.04 ± 0.01 0.07 ± 0.00 0.07 ± 0.00
Germacrene c b a ab
36 204 0.01 ± 0.01 0.05 ± 0.00 0.07 ± 0.01 0.06 ± 0.00
Myristicine* a b a a
37 192 13.83 ± 0.71 8.67 ± 1.90 14.02 ± 0.17 15.61 ± 0.33
Beta bisabolene d c a b
38 204 0.02 ± 0.00 0.09 ± 0.00 0.13 ± 0.01 0.11 ± 0.00
Elemicine** b b a a
39 208 0.25 ± 0.03 0.24 ± 0.09 0.57 ± 0.02 0.66 ± 0.01
Methoxy eugenol c c b a
40 194 0.14 ± 0.06 0.02 ± 0.01 0.55 ± 0.15 2.12 ± 0.06
Total 99.41 ± 0.05 98.63 ± 0.11 98.26 ± 0.03 98.24 ± 0.04
Annotation: *European Pharmacopoeia standard, *flavor and fragrance industry standard. m/z = molecular mass compared to protonated
molecular charge. Superscripted letters on numbers indicated significant difference (LSD α = 0.05).
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Journal of Pharmacognosy and Phytochemistry
Table 3 showed the amount of alpha pinene, sabinene, beta safrole, eugenol, methyl eugenol, and isoeugenol. Finally, old
pinene, 3-carene, limonene, and gamma terpinene gradually seed oil the value was 46% or 6 out of 13 compounds which
decreases as the harvesting age gets older while the value of are alpha pinene, limonene, gamma terpinene, safrole,
safrole, myristicine, eugenol, methyl eugenol, isoeugenol, and eugenol, and methyl eugenol. From here we observe that
elimicine increase. This shows that the older nutmeg seed young nutmeg seed oil has a quality near EP and flavor and
turned into, the lower the development of secondary fragrance industry standard, nearer compared to mace oil,
metabolites and what afterwards increase is primary medium seed oil, and old seed oil.
metabolites, which resulted to production of compound
essential for plant growth [16]. This analysis shows the value of 4. Conclusion
safrole, isoeugenol, myristicine, and elimicine, to be higher Weight and size of nutmeg seed is directly proportional to
than standard value of EP and flavor and fragrance industry. harvesting age; whereas nutmeg oil yield is inversely
According to Schenk and Lamparsky, safrole, isoeugenol, proportional. Nutmeg oil physical characteristics showed that
myristicine, and elimicine compound are among aromatic the older the seed, the higher the value of specific gravity and
compound group consisting oxygenated hydrocarbon content, refractive index, while optical rotation will lower. As many as
thus having high value of specific gravity and refractive index 40 chemical compounds have been identified as constituents
[17]
. The amount of monoterpenoid hydrocarbon compounds of nutmeg mace, young seed, medium seed, and old seed oil.
such as alpha pinene, beta pinene, sabinene, and 3-carene The main chemical compounds of nutmeg oil were alpha
were lower than the amount shown in EP standard. This pinene, sabinene, beta pinene, 3-carene, limonene, gamma
influenced optical rotation value to be small. terpinene, 4-terpineol, safrole, myiristicine, eugenol, methyl
According to Riyadi et al., the chemical compounds of eugenol, isoeugenol, and elimicine with different compound
nutmeg oil originated from Sulawesi and Java meets the composition for every nutmeg type. Terpenes group
quality standard by 85% and 100%, respectively [9]. Research compound decrease as nutmeg seed gets older however
conducted by Pal et al. using main ingredient originated from aromatic component will increase. Various nutmeg oil does
Andaman Nicobar meets the quality standard by 69% via not qualify EP and flavor and fragrance industry
hydrodistillation process [14]. Nutmeg oil from Java and physicochemical quality standard. The quality of young
Sulawesi have a better quality. This opens the possibility of nutmeg seed oil is the closest to EP and flavor and fragrance
nutmeg oil extraction by using steam distillation while industry standard, compared to mace oil, medium seed oil,
influenced by environmental factors and plant genetics. and old seed oil. Further study is needed in order to obtain
Chemical compounds of mace oil that meets EP standard and quality that meets the standard, whether of processing or of
flavor and fragrance industry standard were 62%, or 8 out of raw materials composition.
13 compounds; which are alpha pinene, beta pinene, 3-carene,
limonene, gamma terpinene, eugenol, methyl eugenol, and 5. Acknowledgment
elimicine. For young seed oil the value was 77%, or 10 out of The authors are grateful to PT Indesso Aroma Indonesia for
13 compounds; which are alpha pinene, beta pinene, providing the research facility.
limonene, gamma terpinene, safrole, myristicine, eugenol,
methyl eugenol, isoeugenol, and elimicine. For medium seed 6. References
oil the value was 62% or 8 out of 13 compounds, which are 1. Arrijani. Biology and conservation of Myristica family in
alpha pinene, beta pinene, limonene, gamma terpinene, Indonesia. Biodiversitas. 2005; 6(2):147-151.
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