2016

NPC Natural Product Communications Vol. 11

No. 4
Comparative Chemical Analysis of Mentha piperita and M. spicata 551 - 555
and a Fast Assessment of Commercial Peppermint Teas
Mihaela Buleandraa, Eliza Opreab, Dana Elena Popaa,*, Iulia Gabriela Davida, Zenovia Moldovana,
Iuliana Mihaia and Irinel Adriana Badeaa
a
Department of Analytical Chemistry, Faculty of Chemistry, University of Bucharest, 2-14 Blvd. Regina Elisabeta,
030018, Bucharest, Romania
b
Department of Organic Chemistry, Biochemistry and Catalysis, Faculty of Chemistry, University of Bucharest,
2-14 Blvd. Regina Elisabeta, 030018, Bucharest, Romania

dana_lena1978@yahoo.com

Received: January 9th, 2016; Accepted: February 9th, 2016

The hydrodistilled essential oils and volatile compounds (by static headspaces technique) of Mentha piperita L. and M. spicata L. were characterized by
GC-MS. Headspace analysis of Mentha piperita revealed the existence of menthone (25.4%), 1,8-cineole (17.7%) and menthol (12.1%) as the main
components, while the essential oil contained high amounts of menthol (46.8%) and menthone (25.6%). By contrast, headspace analysis of M. spicata showed
a high content of limonene (37.0%) together with carvone (13.0%), -pinene (10.4%) and -pinene (9.8%), while the essential oil was reach in carvone
(51.7%), dihydrocarveol (11.5%) and cis-dihydrocarvone (9.1%). Eleven samples of peppermint tea available on the Romanian market were analysed by
headspace GC-MS. The volatile profile of the tea samples was compared with that of Mentha piperita L. and certain differences were emphasized and
discussed.

Keywords: Mentha spicata, Mentha piperita, Headspace, Hydrodistillation, GC-MS, Peppermint tea.

Mentha spicata L. (spearmint) and M. piperita L. (peppermint),
which is a natural hybrid of M. aquatica (watermint) and M. spicata
are cultivated throughout all regions of the world [1], including
Romania.
Mint tea, one of the most consumed herbal teas around the world,
and its essential oil, are well known for their therapeutic properties.
They are widely used in treating gastrointestinal disorders and
exhibit significant antioxidant, antimicrobial, antiallergenic,
antiviral and antitumor properties [2-6]. Major uses are in candies
and chewing gum, toothpaste and mouthwash, food flavouring,
aromatherapy, cosmetic and pharmaceutical products. Peppermint
has a stronger flavor, but spearmint, being more subtle and sweet,
is, due to its pleasant aroma, used to prepare mint tea, which is often
consumed just for pleasure. Fresh mint is a versatile kitchen herb
used in the preparation of different dishes (e.g. salads, sauces and
dressings), but also to infuse flavor in different beverages (e.g.
cocktails, coffee, juices, lemonades and other fresh drinks).
pastries; it is also an active ingredient in pesticide products and is
added as solvent in cleaning products for its lemon flavour [9].
Carvone is applied to flavor a number of foods, beverages, and oral
hygiene products [10]. Pulegone has a pleasant odour, midway
between that of peppermint and camphor [11].
The chemical composition of the essential oils of mint species may
depend on environmental and agronomical factors, e.g. soil,
climate, geographical variations, harvesting season, different organs
of the plant, fertilization, genetic factors and evolution [12], but the
extraction method must not be neglected. Different methods have
been used for the isolation of the volatile components from mint.
The most used technique for producing essential oils is
hydrodistillation (HD), but steam distillation [13], supercritical CO2
extraction [14,15], liquid-liquid extraction [16], Soxhlet extraction
[17,18], ultrasound extraction [19], microwave-assisted
hydrodistillation [20], and solid phase micro-extraction [21] are
some alternatives.

The essential oil composition of different mint species is well
known in the literature. The main difference between the essential
oil compositions of different mint species consists of the number of
compounds with commercial importance (for example, menthol,
menthone, carvone, limonene, menthyl acetate, and pulegone).
Menthol, the main constituent of M. piperita and M. arvensis, is
used in medicines, cosmetics, aromatherapy, as a flavoring agent in
cigarettes, in confectionery and in different food products (candies,
chewing gum, cough drops, liquors) [7]. It is known as a local
anesthetic, antiseptic, and gastric sedative, as well as an insect
repellent [8]. Menthone is used as an additive in food [7], but also
in perfumery and cosmetics for its characteristic smell. Menthyl
acetate is employed in perfumery in toilet waters. Limonene is a
natural constituent of some foods and it is commonly employed as a
fragrance in perfumery, in cosmetic products and as a flavouring
agent in different beverages, chewing gum, ice, pudding and
A critical review of the literature regarding Romanian M. piperita
and M. spicata reveals that they have been the subject of several
studies. The essential oil composition was studied by Gherman et
al. [16], the ways to obtain Z-sabinene hydrate-rich essential oils by
Pop et al. [14], while Grumezescu et al. [22] have shown that the
essential oils of M. piperita could also enhance the antimicrobial
activity of antibiotics. Găinar et al. [23] studied the efficiency of
both supercritical fluid extraction and hydrodistillation for essential
oil isolation from peppermint leaves. Also, the feasibility of
controlled release systems based on cyclodextrins and cross-linked
cyclodextrin polymers for delivery of peppermint essential oil was
investigated [18].
However, the literature indicates that no comparison has been made
between M. piperita and M. spicata extracted by HD and HS.
Therefore, the aim of this study was to compare the aroma profile of
552 Natural Product Communications Vol. 11 (4) 2016 Buleandra et al.

Table 1: Volatile compounds identified by GC-MS in Mentha piperita and M. spicata The principal components of M. spicata identified using HS
using HS and HD extraction techniques
techniques were limonene (37.0%), carvone (13.0%), -pinene
Mentha Mentha Mentha Mentha
No RI Compound piperita piperita spicata spicata
(10.4%) and -pinene (9.8%) from a total of forty-one compounds
plant (%) oil (%) plant (%) oil (%) (99.7%). Using the HD extraction technique twenty-nine
1 935 α-Thujene 0.2 - 0.3 - compounds (99.7%) were identified, the main ones being carvone
2 943 α-Pinene 5.6 0.2 9.8 0.3 (51.7%), dihydrocarveol (11.5%) and cis-dihydrocarvone (9.1%).
3 959 Camphene 0.2 - 2.5 -
4 982 Sabinene 2.9 tr. 3.1 -
5 987 β-Pinene 5.5 0.3 10.4 0.4 The difference between the two volatile profiles using the
6 996 β-Myrcene 1.9 tr. 2.1 - mentioned extraction techniques is attributed to the fact that part of
7 1002 3-Octanol 0.2 - tr. -
8 1012 α-Phellandrene 0.1 - 0.3 - the most volatile components could be lost during the essential oil
9 1019 3-Carene tr. - tr. - distillation. In the headspace method the highly volatile compounds
10 1024 α-Terpinene tr. tr. tr. -
11 1033 p-Cymene 0.1 - - -
(monoterpenes and oxygenated hydrocarbons) are mainly released
12 1040 Limonene 9.1 0.6 37.0 4.5 from the plant material. On the other hand, sesquiterpene
13 1043 1,8-Cineole 17.7 2.4 2.9 0.3 hydrocarbons and oxygenated sesquiterpenes, which are less
14 1054 trans-β-Ocimene 0.1 - 0.8 tr.
15 1067 γ-Terpinene 0.1 0.1 tr. - volatile, are either retained or only slowly released due to their low
16 1078 trans-Sabinene hydrate 1.0 0.2 tr. - partition in the gas headspace volume [24].
17 1097 α-Terpinolene 0.1 - tr. -
18 1106 Linalool tr. tr. tr. -
19 1110 cis-p-Menth-2-en-1-ol 0.1 tr. tr. tr. In this way, headspace analysis minimizes the obtaining of artifacts
20 1117 α-Thujone tr. - - - due to the hydrodistillation and establishes the real aroma profile of
21 1158 Isopulegol tr. 0.1 - -
22 1169 Menthone 25.4 25.6 - - the plant, taking into account that some of the volatile aromatic
23 1176 Menthofuran 5.8 3.9 - - compounds are biosynthesized during the normal metabolic process
24 1178 Isomenthone 4.7 6.0 - - in plants or may be modified by processing [25]. Menthol is
25 1180 Borneol - - tr. 0.2
26 1185 Menthol 12.1 46.8 - - produced through chemical derivatization from menthone, limonene
27 1191 Terpinen-4-ol 0.1 - tr. tr. and pinene [26,27].This may explain the decreasing contents of -
28 1196 Isomenthol 0.2 0.5 - -
29 1203 α-Terpineol - - 0.2 0.4 pinene (from 5.6 to 0.2%), -pinene (from 5.5 to 0.3%), limonene
30 1208 Dihydrocarveol - - 4.2 11.5 (from 9.1 to 0.6%) and 1,8-cineole (from 17.7 to 2.4%) obtained by
31 1210 cis-Dihydrocarvone - - 5.2 9.1
32 1246 cis-Carveol - - tr. 0.3
headspace analysis of M. piperita and the increasing content of
33 1255 Pulegone 1.8 3.0 - - menthol (from 12.1 to 46.8%) in hydrodistilled essential oil. It
34 1261 Carvone - - 13.0 51.7 should be mentioned that the menthone content of the two different
35 1271 Piperitone 0.3 1.1 tr. tr.
36 1299 Bornyl acetate - - 0.6 1.3 extract types of M. piperita is not significantly different. The
37 1304 Menthyl acetate 2.4 6.2 - - difference in the percentages of 1,8-cineole obtained using the two
38 1338 trans-Dihydrocarvyl acetate - - 2.3 6.1 extraction methods could be due to the loss of this component in the
39 1347 trans-Carvyl acetate - - 0.4 1.4
40 1373 cis-Carvyl acetate - - 0.1 0.3 water (HD extraction technique) because of its solubility (3500
41 1393 α-Copaene - - tr. tr. mg/1000 mL).
42 1403 β-Bourbonene - - 1.3 1.8
43 1406 β-Elemene - - tr. 0.2
44 1429 α-Gurjunene - - 0.1 0.2 The same thing happened when M. spicata was analysed. The
45 1441 β-Caryophyllene 1.6 1.7 2.4 6.4 higher content of oxygenated monoterpenes could be explained by
46 1450 β-Gurjunene - - 0.1 0.3
47 1476 Humulene - - tr. 0.3 the oxidation of limonene, which is an important substrate for the
48 1483 γ-Muurolene - - tr. tr. production of carvone, cis/trans-carveol and cis/trans-1,2-limonene
49 1489 Germacrene D - - tr. 0.1
50 1503 α-Muurolene 0.4 0.5 0.1 0.4
oxide [28].
51 1614 Caryophyllene oxide - - tr. 1.5
tr. < 0.1 % Given the higher proportion of terpenes in headspace volatiles it
may be assumed that the chemical and, therefore, biological
the hydrodistilled essential oils and headspace volatiles of these properties of mint oil and tea are not the same.
species of mint highlighting the advantages of headspace extraction.
Moreover, in order to establish the tea composition, a comparative The volatile aroma components of tea represent one of the most
analysis of volatile components of different commercial peppermint important factors affecting its quality [29]. To comply with the
teas was realized, taking into account that tea infusion is the most European Pharmacopoeia [30], the monoterpene content of
consumed beverage due to both its beneficial properties and its peppermint oil determined by gas chromatography should be: 1,8-
pleasant taste. cineole (6–14%), limonene (1–5%), -pinene (1.0–1.5%), -pinene
(1–2%), menthone (14–32%), menthofuran (1–9%), isomenthone
The oil contents of M. piperita (sample M7) and M. spicata were (2–10%), menthyl acetate (3–5%), menthol (30–55%), pulegone
found to be 0.3% and 0.4%, respectively. Results obtained by GS- (not more than 4%) and carvone (not more than 1.0%). The ratio of
MS for the two species of mint using HS and HD extraction 1,8-cineole to limonene should be greater than 2 [31]. However,
techniques are presented in Table 1. considering the previous discussion on the comparative analysis of
essential oils and headspace volatiles some observations regarding
The obtained results showed that the number of components was the analysis of commercial peppermint teas can be made.
different using the two extraction techniques, the headspace leading
to a greater number of compounds. In M. piperita, using headspace The analysis of headspace volatiles of the eleven commercial tea
extraction, thirty-two compounds were identified accounting for samples has highlighted 54 compounds, the peppermint aroma
99.9% of the total identified components, with menthone (25.4%), extracts showing different quantitative compositions (Table 2). It
eucalyptol (17.7%) and menthol (12.1%) as the main components. should be mentioned that we have no information about the M.
The essential oil of M. piperita contained only twenty-two piperita cultivars used in our study. This feature could be important
components (99.6%). Menthol (46.8%) and menthone (25.6%) were because Grulova et al. [32] have recently reported a new cultivar
the main components of this oil (Table 1). of M. piperita entitled ‘Kristínka’ with high content of menthol
Two Mentha species comparative evaluation Natural Product Communications Vol. 11 (4) 2016 553

Table 2: GC MS results for analysis of commercial peppermint teas

No RI Compound M1 % M2 % M3 % M4 % M5 % M6 % M7 % M8 % M9 % M10 % M11%
1 935 α-Thujene 0.2 tr. 0.2 0.2 4.3 0.2 0.2 tr. 0.1 tr.
2 943 α-Pinene 5.5 1.4 4.7 3.5 5.6 6.6 5.6 4.6 6.7 0.4 0.7
3 959 Camphene 0.3 tr. 0.2 0.1 0.2 0.6 0.2 0.2 0.1 - -
4 982 Sabinene 2.2 0.6 1.8 1.3 4.6 3.0 2.9 1.7 2.4 0.1 0.3
5 987 β-Pinene 5.3 1.4 4.5 3.6 5.6 5.7 5.5 4.1 6.4 0.4 7.2
6 996 β-Myrcene 0.7 0.4 0.4 0.5 3.1 1.9 1.9 0.7 0.8 0.2 0.2
7 1002 3-Octanol tr. 0.1 0.1 0.1 - 0.3 0.2 0.1 0.1 0.1 0.1
8 1012 α-Phellandrene tr. - tr. tr. - 0.2 tr. tr. tr. tr. -
9 1019 3-Carene - - - tr. - - tr. tr. tr. - -
10 1024 α-Terpinene 0.2 - 0.2 0.5 2.4 tr. tr. tr. tr. 0.2 tr.
11 1033 p-Cymene 0.5 tr. 0.2 1.3 5.4 - 0.1 0.1 0.2 0.2 tr.
12 1040 Limonene 10.5 1.2 3.3 4.0 4.3 19.2 9.1 4.4 1.7 0.7 0.6
13 1043 1,8-Cineole 17.5 6.9 17.8 17.2 26.3 17.0 17.7 15.0 20.6 5.0 4.7
14 1054 trans-β-Ocimene - tr. - tr. 0.1 0.1 0.1 tr. tr. tr. tr.
15 1067 γ-Terpinene 0.3 - 0.3 0.5 6.9 0.1 0.1 tr. tr. 0.3 tr.
16 1078 trans-Sabinene hydrate 1.2 0.1 1.7 0.8 3.3 0.5 1.0 0.3 tr. 1.0 0.2
17 1097 α-Terpinolene tr. tr. tr. tr. 0.1 tr. 0.1 tr. tr. tr. tr.
18 1106 Linalool tr. 0.2 0.2 0.1 2.5 tr. tr. tr. tr. 0.2 0.1
19 1110 cis-p-Menth-2-en-1-ol 0.1 - 0.1 tr. 0.2 0.1 0.1 tr. tr. 0.3 -
20 1117 α-Thujone - - - - - - tr. - - - -
21 1158 Isopulegol - - - 0.1 - - tr. 0.1 tr. 0.2 0.1
22 1169 Menthone 13.9 48.6 24.3 31.3 7.0 8.0 25.4 37.8 32.0 44.5 49.3
23 1176 Menthofuran 7.9 0.3 4.2 3.8 - - 5.8 0.6 1.2 0.3 -
24 1178 Isomenthone 2.4 11.3 4.2 5.3 4.9 3.6 4.7 7.8 8.1 10.6 11.8
25 1185 Menthol 13.0 15.5 19.7 15.9 0.1 1.1 12.1 11.8 13.9 28.5 14.3
26 1191 Terpinen-4-ol 0.4 - 0.7 0.7 0.2 0.6 0.1 0.5 0.1 - -
27 1196 Isomenthol 0.3 0.3 0.3 0.3 - - 0.2 0.2 0.5 0.4 0.2
28 1203 α-Terpineol - - - tr. 0.2 0.1 - tr. tr. - -
29 1208 Dihydrocarveol - - - - tr. 0.3 - - - - -
30 1210 cis-Dihydrocarvone - - - - 2.7 0.3 - - - - -
31 1219 trans-Dihydrocarvone - - - - 1.8 - - - - - -
32 1246 cis-Carveol - - - - 0.5 0.2 - - - - -
33 1255 Pulegone 1.1 0.2 0.5 0.5 0.8 4.2 1.8 1.1 0.2 2.1 0.3
34 1261 Carvone 4.5 - 1.9 0.9 - 22.5 - 2.5 tr. - -
35 1271 Piperitone 0.1 0.4 0.3 0.2 - 0.3 0.3 0.4 0.4 - 0.4
36 1272 Piperitone oxide - - - - 3.0 - - - - - -
37 1299 Bornyl acetate - - - - - tr. - - - - -
38 1304 Menthyl acetate 9.0 8.3 3.5 4.5 0.3 2.4 3.5 2.3 2.9 6.7
39 1311 Thymol - - - - 0.3 - - - - - -
40 1338 trans-Dihydrocarvyl acetate - - - - - 0.1 - - - - -
41 1347 trans-Carvyl acetate - - - - - tr. - - - - -
42 1373 cis-Carvyl acetate - - - - - 0.2 - - - - -
43 1393 α-Copaene tr. tr. tr. tr. tr. tr. - tr. tr. - 0.1
44 1403 β-Bourbonene 0.2 0.4 0.5 0.2 0.1 0.6 - 0.2 0.1 tr. 0.4
45 1406 β-Elemene tr. 0.1 0.1 tr. tr. 0.2 - tr. - tr. 0.2
46 1429 α-Gurjunene - - - - - - - - - - tr.
47 1441 β-Caryophyllene 0.9 1.1 2.3 1.4 2.0 0.9 1.6 1.1 0.5 0.5 1.2
48 1450 β-Gurjunene tr. tr. 0.1 tr. - tr. - tr. tr. tr. 0.1
49 1476 Humulene tr. - - tr. tr. - - tr. tr. - tr.
50 1483 γ-Muurolene 0.1 tr. 0.1 tr. 0.4 tr. 0.1
51 1503 α-Muurolene 0.2 0.4 0.7 0.4 0.7 0.3 0.4 0.4 0.6 0.2 0.4
52 1535 γ-Cadinene - - - - - - - tr. - - -
53 1544 Calamenene tr. - - - - 0.2 - tr. - - -
54 1614 Caryophyllene oxide - - - - - - - - tr. 0.1 -
Total identified components 98.9 99.5 99.2 99.4 99.4 99.9 99.9 99.8 99.5 99.6 99.5
tr. < 0.1%

(58.8 - 76.7%). In addition, Sústriková and Šalamon [33] have
shown that peppermint oil originating from different countries has
various percentages of menthol. In addition, as reported in the
literature, limonene and menthone are the major components in
young peppermint leaves. The proportion of limonene decreases
rapidly with plant development, while menthone increases, being
converted into menthol in mature peppermint leaves. Also, the
proportions of menthofuran and pulegone decrease as leaves age,
while those of 1,8-cineole increase substantially [34].
Therefore, in the present study, the difference in monoterpene
contents in the composition of the teas could be attributed to the
harvesting time, taking into consideration that the total content of
monoterpenes depends on the age of leaves. The variation of the
main components of peppermint tea, namely limonene, menthone,
menthofuran, isomenthone, menthyl acetate and menthol is
presented in Figure 1. Thus, the different monoterpene contents
obtained could be attributed to the degree of maturity of the plants
or the part of the plants (all plant or leaves) harvested for the tea
production.
However, the lower content of menthone and menthol, which are
characteristic aroma components of M. piperita in the samples M5
and M6 and the higher carvone content, especially in sample M6,
which could be attributed to another mint species, namely M.
spicata, suggest that these two samples also contain other species
than those declared by the producer.
Previous papers showed that the menthofuran level was at its
highest at full bloom because of the relatively higher flower portion
of the plant raw material and probably this is the case with M1 and
M7, which had higher percentages of menthofurane (7.9% and
9.1%, respectively) [21, 35]. Grulova et al. [32] have shown that
menthol levels increased from early to late bloom, whereas the
menthone levels decreased. Simultaneously, the level of limonene
has slightly decreased. The composition of the main compounds of
peppermint, menthol and menthone, had changed in the essential oil
due to internal and external factors, environmental conditions
appearing to be very important.
Böttcher et al. [36] have reported essential oil compositional
changes based on the physiological postharvest response during
80 hours of storage and that storing the harvested plant material at
554 Natural Product Communications Vol. 11 (4) 2016
60 Limonene
1,8-Cineole
50 Menthone
Menthofuran
Relative peak area, %
Isomenthone
40
Menthol
Menthyl acetate
30
20
10
0
M1 M2 M3 M4 M5 M6
Figure 1: Variation in the concentrations of the main components of peppermint aroma
isolated from different commercial mint samples.
10°C was the best way for maintaining external quality traits of
peppermint essential oil. For some components (menthol,
menthofuran, menthylacetate, isomenthone) they registered
increased percentages, which could be explained by a biosynthesis
of the components during the postharvest period. For menthone,
pulegone and carvone they found decreased percentages during
storage. Moreover, the adulteration of peppermint essential oil, as
well as tea, is not a new problem. The most common practice for
essential oils is dilution with cornmint oil obtained from M.
arvensis, which is less expensive than peppermint oil [37]. The
main components of peppermint and cornmint are menthol,
menthone, menthyl acetate and other esters. One way to
discriminate between the two mints is the presence of trans-
sabinene hydrate (at the 1% level), menthofuran and viridiflorol,
which are present only in M. piperita [38]. The other easier way to
detect the adulteration is to look at the menthone / isomenthone
ratio that has to be between 5.0 and 9.2 [39]. A great difference
from this ratio value is observed for samples M5 and M6, too.
Wildung and Croteau [40] have created transgenic peppermint
plants with improved oil composition and yield, using the genes
encoding enzymes of the eight-step pathway to (-)-menthol, but, for
the moment, genetically modified peppermint is unlikely to exist in
Romania.
The data presented in Table 2 were statistically analysed by means
of principal component analysis (PCA) in order to identify the
relationships among the studied tea samples. In the loading plot,
limonene and 1,8-cineole have similar important loadings for the
first component, and menthol, menthone, methyl acetate,
isomenthone and isomenthol for the second component. The score
plot presented in Figure 2 shows a compact group of eight samples,
while three of them, namely M5, M6 and M11, are not grouped at
all. The compact group contains samples which are differentiated by
their rich composition in menthol, menthone, methyl acetate,
isomenthone and isomenthol. Moreover, these compounds are also
highly correlated in this group. Thus, the data obtained by GC-MS
analysed by PCA may contribute to the identification and
classification of commercially available mint teas.
Experimental
Plant material: Mentha spicata L. (specimen voucher deposited in
the Herbarium of the University of Bucharest – BUC 402041) was
References
[1] WHO. (2002) WHO Monographs on Selected Medicinal Plants, Vol, 2, World Health Organization, Geneva, 188, 199.
[2] McKay DL, Blumberg JB. (2006) A review of the bioactivity and potential health benefits of peppermint tea (Mentha piperita L.). Phytotherapy
Research, 20, 619-633.
Buleandra et al.
M7 M8 M9 M10 M11
Figure 2: Score plot for the first two components.
collected in July 2014. The leaves were separated from branches,
dried at room temperature, manually ground and further stored in
paper, dark, hermetically tight bags to protect them from humidity
and light.
M. piperita L. peppermint teas from different commercial suppliers
were purchased from the supermarket. The samples were named
M1-M11. The source of the tea declared by the producer was
Mentha piperita for M1-M7 and M11, while M8-M10 were sold as
peppermint tea. Except for M3, M5, M7, and M9, which were bulk
teas packed in boxes, the others consisted of tea bags.
Essential oil extraction: Plant material (20 g) was hydrodistilled in
a Clevenger-type apparatus for 4 h. The essential oil was dried over
anhydrous Na2SO4, stored in a dark glass bottle, and kept at 4C
until analysis [41]. The oil sample was diluted in n-hexane (1:10)
for GC-MS analysis.
Headspace volatiles extraction: For headspace analysis, dry plant
material (1 g) was placed in a 20 mL headspace vial sealed with a
silicone rubber septum and aluminum cap. The headspace was
connected to a Triplus HS Autosampler (Thermo Electron). The
headspace vial containing the plant material was heated to 80C for
10 min and 500 L of headspace gas was injected into the column.
GC-MS analysis: A Thermo Electron system - Focus GC
chromatograph coupled with a Polaris Q ion trap mass detector with
a DB-5MS capillary column (25 m x 0.25 mm; 0.25 μm) was used.
The GC oven temperature program was: 60C (3 min) with an
increase of 10C/min up to 200C (2 min) and then 12C/min to
240C (2 min). Helium was used as carrier gas (1.0 mL/min); the
source and interface temperatures were 200C and 250C,
respectively. The detector worked in the electron impact mode (70
eV) and detection was performed in the range of m/z 35-300. All
peaks of the chromatograms were analysed using Xcalibur®
software and the NIST Mass Spectral Library was used in order to
identify the corresponding compounds. A standard alkanes solution
for GC (C8-C20 in n-hexane) was used for retention indexes (RI)
calculation. Relative percent of individual components was
calculated based on the GC peak areas without correction factors.
Statistical analysis and principal component analysis (PCA) were
performed using Minitab 17 software.
Two Mentha species comparative evaluation Natural Product Communications Vol. 11 (4) 2016 555

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