Food and Bioproducts Processing 1 2 0 ( 2 0 2 0 ) 158–165

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Food and Bioproducts Processing

journal homepage: www.elsevier.com/locate/fbp

Microwave-assisted extraction of O. vulgare L. spp.

hirtum essential oil: Comparison with conventional
hydro-distillation

Zorica Drinić ∗ , Dejan Pljevljakušić, Jelena Živković, Dubravka Bigović,

Katarina Šavikin
Institute for Medicinal Plant Research “Dr. Josif Pančić”, Tadeuša Košćuška 1, 11000 Belgrade, Serbia

a r t i c l e i n f o a b s t r a c t

Article history: Microwave-assisted hydro-distillation (MAHD) at different power levels has been applied for
Received 8 May 2019 the extraction of essential oil from O. vulgare L. ssp. hirtum. Hydrolats and residual extracts,
Received in revised form 2 which present by-products, were also collected. A comparison was done with conventional
December 2019 hydro-distillation (HD) in terms of extraction time, extraction yield, the chemical composi-
Accepted 23 January 2020 tion of essential oils and environmental impact. MAHD proved to be a technique with many
Available online 30 January 2020 advantages compared to conventional HD. MAHD had shorter extraction time (24–45 min)
compared to HD (136 min), better yields of essential oil (2.55–7.10 % for MAHD compared to
Keywords: 5.81 % for HD), higher content of oxygenated compounds (78.89–85.15 % for MAHD compared
Microwave-assisted to 76.82 for HD), and a more environmentally friendly approach (electrical consumption was
hydro-distillation from 0.135–0.240 kW h for MAHD compared to 1.360 kW h for HD). The main component in
Oregano all essential oils, as well as in hydrolats, was carvacrol. MAHD at the power level of 600 W
Essential oil may be considered as the most promising among the tested power levels with respect to
Hydrolat all investigated parameters. Residual extracts after HD and MAHD showed to be valuable
by-products rich in phenolic compounds, with rosmarinic acid as dominant.
© 2020 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.

1. Introduction Plants belonging to the genus Origanum are widely distributed in

Eurasia and North Africa and have been used for centuries as a spice,
Medicinal plants are a valuable source of bioactive compounds. World as well as in ethnomedicine (Ietswaart,1980; Fleisher and Fleisher,
Health Organization reported that medicinal plants have been used 1988). Various biological activities have been reported for Origanum
in every country in the world. Interest for use plant-based natural species such as antifungal, antimicrobial, antioxidant, antispasmodic,
products increases over years (Al-Kalaldeh et al., 2010; Chishti et al., antitumoral, expectorant, antiparasitic, antihelminthic (Kokkini, 1997;
2013; Ahmad Khan and Ahmad, 2019). Medicinal plants are used in the Hummer et al., 1999; Elgayyar et al., 2001; Baser, 2002; Puertas et al.,
pharmaceutical, food and cosmetic industry. The major compounds 2002; Sokovic et al., 2002; Baydar et al., 2004; Kulisic et al., 2004; Sari
of plants that humans have used due to their beneficial role are ter- et al., 2006; Bakkali et al., 2008; Dundar et al., 2008). Still, oregano is
penoids, alkaloids, and phenolic compounds (Harborne, 1999). The mainly used as a spice for a variety of dishes in Mediterranean cui-
many aromatic and medicinal plants have become attractive as natu- sine, while for medicinal purposes, its essential oil is used due to its
ral sources of bioactive compounds. Aromatic plants are naturally rich antimicrobial properties.
sources of essential oils and their components have a potential multi- Monoterpenoid phenolic isomers carvacrol and thymol are the
purpose functional use in different fields such as medicine, pharmacy, main components of the essential oil of all O. vulgare subspecies
cosmetic and food (Mohammadhosseini et al., 2017). (D’Antuono et al., 2000; Kulisic et al., 2004; Menaker et al., 2004) and
those two compounds are primarily responsible for diverse biological
activities (Aeschbach et al., 1994; Yanishlieva et al., 1999; Mastelic et al.,
2008). Besides its essential oil, oregano is also rich in various biologi-
cally active phenolic compounds such as flavonoids and phenolic acids.
∗
Corresponding author. Moreover, Koukoulitsa et al. (2006) reported that the inhibitory activity
E-mail address: zdrinic@mocbilja.rs (Z. Drinić).
https://doi.org/10.1016/j.fbp.2020.01.011
0960-3085/© 2020 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
 Food and Bioproducts Processing 1 2 0 ( 2 0 2 0 ) 158–165 159

of polar extracts of O. vulgare L. ssp. hirtum has a promising potential in apparatus according to the procedure I of the Yugoslavian
the prevention of diabetes complications in long-lasting treatments. Pharmacopoeia IV (1984). MAHD was performed using the spe-
Essential oils can be isolated using several classical and advanced cially created system, consisting of the microwave oven (HMT
approaches. Some of the conventional and most commonly used 72M450, Bosch, Gerlingen, Germany) connected to the Cle-
methods are hydro-distillation, steam distillation, and organic solvent
venger type apparatus. The water/plant ratio was the same
extraction. The essential oil yield, losses of some volatile compounds
for HD and MAHD, 20:1 (w/w). MAHD was performed at three
and the content of bioactive components depend on the method used
different power levels (180, 360, and 600 W) until no more
(Ferhat et al., 2007). Nevertheless, all of these conventional techniques
have some shortcomings. Some components of essential oils are sen- essential oil was obtained (ca. 20 min). The time required to
sitive to steam distillation conditions (Lo Presti et al., 2005; Karakaya reach the final amount of essential oil will be referred to’ total
et al., 2011), while, on the other hand, organic solvents such as n- extraction time’ in further discussion. The essential oils were
hexane and methylene chloride provide good recovery of oil and lipid collected, and dried over anhydrous sodium sulphate. The
compounds, but they are toxic (Mamidipally and Liu, 2004). Moreover, essential oil yield, expressed as a percentage, was calculated
conventional techniques are usually time- and cost-consuming (Pavlić on a moisture-free basis. Oil samples (20 ␮L) were dissolved in
et al., 2015). EtOH 96 % (2 mL).
Nowadays, alternative techniques for the isolation of essential
The hydrolats are defined as the distilled water from the
oils such as subcritical water extraction, supercritical carbon dioxide
production of essential oils obtained by steam water distilla-
extraction and microwave-assisted hydro-distillation have been devel-
tion from aromatic plants (flowers, leaves, stems, roots), so
oped (Karakaya et al., 2011; Pavlić et al., 2015). For these new extraction
techniques, it is expected to be economically and environmentally
the main components are water-soluble volatile compounds
friendly, sustainable, highly efficient and to provide good product (Rajeswara Rao, 2013). Hydrolats, which presented water from
quality. Microwave energy with a frequency of 2.45 GHz has attracted the production of essential oils obtained by steam distilla-
attention in analytical chemistry due to reduced analysis time, simpli- tion or hydro-distillation from aromatic plants, were collected
fied manipulation and work-up, and higher purity of the final product from the burette after HD and MAHD, extracted with n-hexane
(Kingston and Jassie, 1988; Zlotorzynski, 1995; Kingston and Haswell, (5 mL of hydrolats with 3 × 1 mL n-hexane), hexane fractions
1999). Many authors reported the main advantages of microwave- were collected and combined. Residual extracts after HD and
assisted hydro-distillation of essential oils regarding shorter extraction MAHD were also collected. Prepared samples of essential oil,
time, better yields, a higher amount of oxygenated compounds, envi-
hydrolats and residual extracts were stored in the freezer until
ronmentally friendly character and lower cost (Ferhat et al., 2006;
further analysis.
Golmakani and Rezaei, 2008; Abdellatif and Hassani, 2015; Hashemi-
Moghaddam et al., 2018).
In the present study, conventional hydro-distillation (HD) and alter-
native microwave-assisted hydro-distillation (MAHD) techniques were 2.4. Chemical analyses of essential oil and hydrolats
applied for the isolation of essential oil from O. vulgare L. subspecies hir-
tum and to obtain hydrolats. The qualitative and quantitative profiles
The chemical composition of the essential oil was analyzed
of essential oils and hydrolats were analyzed using GC/MS. Moreover,
using GC/MS technique. GC/MS analyses were performed on a
water extracts remaining after HD and MAHD were collected and ana-
Shimadzu GCMS-QP2010 ultra mass spectrometer fitted with
lyzed for total phenolics content as well as for individual phenolic
compounds. a flame ionic detector and coupled with a GC2010 gas chro-
matograph. The InertCap5 capillary column (60.0 m × 0.25
mm × 0.25 ␮m) was used for separation. Helium (He), at a split
2. Material and methods
ratio of 1:5 and a linear velocity of 35.2 cm/s was used as car-
rier gas. Initially, the oven temperature was 60 ◦ C, which was
2.1. Standards and reagents
held for 4 min, then increased to 280 ◦ C at a rate of 4 ◦ C/min,
and held for 10 min. The injector and detector temperatures
Ethanol, distilled water, and n-hexane were purchased from
were adjusted at 250 ◦ C and 300 ◦ C, respectively. The ion source
Zorka Pharma, Šabac (Serbia). Gallic acid, sodium carbonate
temperature was 200 ◦ C. The identification of the constituents
anhydrous (Na2 CO3 ), sodium sulfate anhydrous (Na2 SO4 ) and
was performed by comparing their mass spectra and reten-
Folin–Ciocalteu phenol reagent were purchased from Sigma
tion indices (RIs) with those obtained from authentic samples
Chemicals Co. (USA). The phenolic compounds standards
and/or listed in the NIST/Wiley mass-spectra libraries, using
were from Merck (Germany). All chemicals used in the experi-
different types of search (PBM/NIST/AMDIS) and available lit-
mental procedure were of analytical grade purity. For the HPLC
erature data (Hochmuth, 2006; Adams, 2007).
analysis solvents and formic acid were of HPLC grade.

2.2. Plant material

2.5. Determination of total phenolic contents of
The aerial parts of O. vulgare L. subspecies hirtum were residual extracts
collected in July 2018 from the collection of the Institute
for Medicinal Plant Research “Dr. Josif Pančić “(20◦ 42 4.55 E, The total phenolic content of residual extracts was deter-
44◦ 52 20.54 N, altitude 74 m, Belgrade. Voucher samples are mined by Folin-Ciocalteu spectrophotometric procedure with
stored in the Herbarium of the Institute of Botany and Botan- slight modification (Harris, 2000). Two hundred microliters of
ical Garden “Jevremovac”, Faculty of Biology, University of extracts were added to 1 mL of 1:10 diluted Folin-Ciocalteu
Belgrade; voucher No. 181014. reagent. After 4 min, 800 ␮L of sodium carbonate (75 g/L) was
added. After 2 h of incubation at room temperature, the
2.3. Essential oils, hydrolats and residual extracts absorbance was measured at 765 nm. Gallic acid (0–100 mg/L)
was used for the calibration of a standard curve. The results
Isolation of essential oils was performed using HD and were expressed as milligrams of gallic acid equivalents per g
MAHD approaches. HD was performed using a Clevenger type of plant dry weight (mg GAE/g dw).
160 Food and Bioproducts Processing 1 2 0 ( 2 0 2 0 ) 158–165

2.6. HPLC analysis of residual extracts

The HPLC-DAD analysis of phenolic compounds was per-

formed using the Agilent 1100 Series (Agilent Technologies,
Palo Alto, CA, USA) comparing the retention times and
absorption spectra (200−400 nm) of unknown peaks with the
reference standards. The column was Lichrospher® 100 RP 18e
(5 ␮m, 250 × 4 mm). Mobile phase A was formic acid in water
(0.17 %), while mobile phase B was acetonitrile. The injection
volume was 10 ␮L, and the flow rate 0.8 mL/min with gradient
program (0–53 min 0–100 % B). The stop time of the analy-
sis was 55 min. The investigated samples were analyzed in
triplicate.

2.7. Environmental impact of applied techniques

Regarding environmental impact, electrical consumption and

CO2 emission were calculated. The electrical consumption (A)
of different methods was calculated as the electrical power for
a time, using the following equation:
Fig. 1 – Total extraction time for HD and MAHD at different
A = P×t, power levels.

where A is electrical consumption (kWh), P is electrical power

it is directly connected with the dielectric constant of the used
(kW) and t is time (h).
medium (Chemat et al., 2009). Water is a solvent with a high
According to Ferhat et al. (2006) to obtain 1 kW h from coal
dielectric constant which leads to rapid heating in the MAHD
or fuel, 800 g of CO2 will be rejected in the atmosphere during
process. Differences in the MAHD extraction time at different
the combustion of fossil fuel. CO2 emission can be described
power levels are correlated to the density of the waves.
by the equation:
The reduction in the extraction time of essential oil by
ECO2 = (A × 800)/1000 microwaves was significantly higher. Even at the level of the
lowest microwave power (180 W), the total extraction time was
where ECO2 is CO2 emission (kg) and A is electric consumption significantly shorter than in hydro-distillation (Fig. 1). Similar
(kWh). to our results, many authors reported that MAHD significantly
reduces extraction time in comparison to conventional HD
2.8. Statistical analysis (Ferhat et al., 2007; Golmakani and Rezaei, 2008; Rezvanpanah
et al., 2008).
All experiments were made in triplicate and the obtained
results are expressed as mean values. Regarding phenolic 3.2. Essential oils yield
compounds, a null hypothesis was tested by one-way ANOVA
and differences among mean values subsequently estimated Essential oil yield was 7.10, 5.67, and 2.55 % for MAHD at
by post hoc Duncan’s MR test. Statistical analysis was per- 600, 360 and 180 W, respectively, while for HD it was 5.81 %
formed using the MS Office Excel v. 2010, while charts were (Fig. 2). The highest yield of the essential oil was obtained
produced in R Base Plot environment (CRAN). after MAHD at 600 W, while the yield using MAHD at 360 W
was similar to HD. The lowest oil yield was obtained by MAHD
3. Resultion and discussion at 180 W. Higher yield of extracted essential oil at higher
microwave power has been previously explained by Chemat
3.1. Extraction time et al. (2009) as synergistic effect of two transport phenom-
ena, such as temperature gradient and mass gradient, acting
From the results obtained after comparison of HD and MAHD in the same direction. In contrast, at lower levels of microwave
at different power levels, it can be concluded that the total power, the density of microwaves is also lower, leading to
extraction time differed among applied techniques and con- lower heating ability and consequently to lower extraction
ditions: total extraction time was 24, 29, and 45 min for MAHD yield. In a nutshell, the extraction of active substances from
at 600, 360, and 180 W, respectively, while for HD it was 136 min plant material using microwave extraction occurs as a result
(Fig. 1). The time required to start the extraction was 4, 9, and of a change in cell structure caused by electromagnetic waves
25 min for MAHD at 600, 360, and 180 W, respectively, while for (Veggi et al., 2013). At higher power levels, water can absorb
HD it was 16 min. These differences can be explained by a dif- more microwave beams in that way disrupting cell structure
ferent mechanism of extraction with microwaves compared which leads to increased release of the essential oil (Chen and
to hydro-distillation or in other words by the mechanism of Spiro, 1995). Higher power levels can lead to an increase in
heat penetration into the plant material. Veggi et al. (2013) system temperature and extraction yield are directly depen-
highlighted that heating during HD is achieved by gradual dent on the extraction temperature (Veggi et al., 2013). The
heat transfer from the heating medium to the interior of the extraction efficiency increases with increasing temperature to
samples, while microwave heating is volumetrically dispersed the optimum temperature, which depends on the stability of
within the radiation. Furthermore, microwave heating is based the target compounds. Therefore, it is important to select the
on two phenomena: ionic conduction and dipole rotation, and appropriate microwave power because the high microwave
 Food and Bioproducts Processing 1 2 0 ( 2 0 2 0 ) 158–165 161

as in the essential oils, the main component in all hydrolats

was carvacrol and its content represented 99.00, 99.58, 99.40,
and 94.71 % of all identified components by HD, MAHD at 600,
360, and 180 W, respectively. Some authors reported that the
main component in essential oil was also the main compo-
nent of corresponding hydrolats (Paolini et al., 2008; Shahani
et al., 2011). Since, carvacrol was reported as a key compo-
nent for the biological activity of Origanum species essential
oil, the high content of carvacrol in obtained hydrolats indi-
cates the possibility for their use in the pharmaceutical and
cosmetic industry. The number of identified compounds in
hydrolat obtained by MAHD at 180 W power level was signifi-
cantly higher than in other hydrolats. This discrepancy is most
probably the result of prolonged time of water extraction of the
plant material before medium boiling and evaporation of the
essential oil.

3.4. Total phenolic content and composition of residual

extracts

Fig. 2 – Total essential oil yield for HD and MAHD at

Residual extracts after isolation of essential oils are usually
different power levels.
recognized as waste. On the other hand, such extracts have
a remarkable potential as valuable by-products in the dis-
power can cause degradation of temperature sensitive com-
tillation process. Total phenols content in obtained extracts
pounds. Raner et al. (1993) reported that variation of power
ranged from 9.34 to 17.74 ␮g GAE/g dw. The most abundant
level from 500 to 1000 W had no significant effect on the yield
phenolic compound in all obtained extracts was rosmarinic
of flavonoids, while Mohamadia et al. (2013) reported that
acid (Table 3). Some authors reported that rosmarinic acid is
powers greater than 600 W had no significant effect on the
the dominant phenolic acid of family Lamiaceae in general
extraction efficiency of essential oil.
(Wang et al., 2004; Lee, 2010; Embuscado, 2015). The highest
rosmarinic acid content was found in the extract obtained by
3.3. Composition of essential oils and hydrolats
MAHD at 600 W. Other phenolic components determined using
HPLC were chlorogenic acid (0.34–1.50 ␮g/g dw), caffeic acid
The chemical constituents of obtained essential oils by both
(1.61–5.37 ␮g/g dw) and luteolin-7-O-glucoside (0.57–1.51 ␮g/g
applied methods are presented in Table 1. A total of 42
dw). The highest phenolic content was noticed in the extract
compounds have been identified, representing more than
obtained by MAHD at 600 W which is in accordance with
99.97 % of the oil. The dominant compound in all samples
previously mentioned fact that extraction with microwaves
was carvacrol followed by ␥-terpinene, p-cymene, trans-
achieves a higher yield than the classic HD method. Among
caryophyllene, ␣-terpinene, and myrcene. In respect to this
all tested distillation models, MAHD at 600 W yielded signifi-
order, the chemical composition of the essential oils obtained
cantly higher amounts of individual phenolic compounds. To
by HD and MAHD at 600 W were similar, while essential oils
the best of our knowledge, there are no previously published
obtained by MAHD at 360 and 180 W showed slight differences.
results on the phenolic compounds of the O. vulgare L. ssp.
The content of carvacrol in essential oils was higher when
hirtum residual extract after distillation.
MAHD was applied (76.02, 86.13, and 82.65 % for power lev-
els 600, 360, and 180 W, respectively), while in HD it was 71.71
%. Carvacrol was also dominant compounds in O. vulgare L. 3.5. Environmental impact
ssp. hirtum essential oil analyzed by other authors (Kokkini
et al., 1991; Adam et al., 1998). Essential oils obtained by MAHD Electrical consumption and CO2 emission were smaller for
had a higher content of oxygenated monoterpenes and lower MAHD compared to HD (Fig. 3). HD electrical consumption was
content of monoterpenes hydrocarbons. The amounts of oxy- 1.360 kW h, while for MAHD at levels 600 W, 360 W, and 180 W
genated monoterpenes in essential oils obtained by MAHD at it was 0.240, 0.174, and 0.135 kW h, respectively. CO2 emission
600, 360, and 180 W was 78.89, 87.83, and 85.15 %, respectively, for HD was 1.09 kg, while for MAHD at 600 W, 360 W, and 180 W
while the content in HD obtained essential oil was 76.02 %. it was 0.19, 0.14, and 0.11 kg, respectively. Since the yield of
Since hydrocarbon monoterpenes are the product of thermal essential oils obtained by HD and MAHD at different power
and hydrolytic decomposition of oxygenated monoterpenes, levels was not the same, in order to compare environmen-
higher content of oxygenated monoterpenes in essential oils tal impact, the total electrical consumption and CO2 emission
obtained by MAHD is probably due to shorter time need for were converted to electrical consumption and CO2 emission
MAHD compared to HD. This is in agreement with previously for the yield of 1 % of essential oil. Electrical consumption
reported studies (Filly et al., 2014; Ferhat et al., 2006). Also, the for yield of 1 % was 0.2341, 0.0338, 0.0307, 0.0529 kW h, while
different chemical composition in obtained essential oil, in CO2 emission was 0.1873, 0.0270, 0.0246, and 0.0424 kg for
addition to hydrolytic processes, can be related to the possible HD, MAHD at 600 W, 360 W and 180 W, respectively. Using the
degradation of products by oxidation and trans-esterification MAHD method energy saving was on average about 75 % com-
due to different extraction time (Benkaci–Ali et al., 2007). pared to the hydro-distillation, and the emission CO2 was
Chemical constituents in the hydrolats obtained by HD and reduced in the same percentage. Electrical consumption and
MAHD at different power levels are given in Table 2. Same CO2 emission are related to the extraction time. Since MAHD
162 Food and Bioproducts Processing 1 2 0 ( 2 0 2 0 ) 158–165

Table 1 – Chemical constituents in the O. vulgare L. ssp. hirtum essential oil obtained with hydro-distillation and
microwave-assisted hydro-distillation at different power levels.
Peak Compound RIa Hydro-distillation Microwave assisted hydro-distillation

600 W 360 W 180 W

1. ␣-thujene 923.6 0.91 0.60 0.21 0.26

2. ␣-pinene 931.6 0.39 0.32 0.10 0.13
3. camphene 948.5 0.11 0.09 – 0.02
4. 1-octen-3-ol 973.8 – 0.41 0.18 0.32
5. sabinene 973.9 0.40 – – –
6. ␤-pinene 978.6 0.11 0.09 0.02 0.03
7. 3-octanone 981.3 0.11 0.12 0.06 0.09
8. myrcene 987.5 1.38 1.31 0.59 0.63
9. ␣-phellandrene 1002.7 0.19 0.16 0.05 0.06
10. ␦-3-carene 1008.4 0.06 0.04 – –
11. ␣-terpinene 1013.6 1.67 1.46 0.69 0.78
12. p-cymene 1019.6 4.36 4.08 2.43 3.10
13. limonene 1024.0 0.19 0.19 0.06 0.07
14. ␤-phellandrene 1026.3 0.14 0.12 0.04 0.05
15. cis-␤-ocimene 1028.4 0.19 0.20 0.07 0.08
16. trans-␤-ocimene 1039.1 0.10 0.08 0.02 0.03
17. ␥-terpinene 1053.3 10.14 9.51 6.39 7.16
18. cis-sabinene hydrate 1061.8 0.37 0.48 0.26 0.24
19. terpinolene 1083.3 0.07 0.06 – 0.04
20. linalool 1090.7 0.10 0.07 0.02 0.05
21. trans-sabinene hydrate 1093.8 0.24 0.29 0.10 0.11
22. borneol 1164.5 0.57 0.45 0.15 0.29
23. 4-terpinenol 1175.3 0.50 0.41 0.22 0.57
24. ␣-terpineol 1188.7 0.18 0.05 – 0.06
25. cis-dihydrocarvone 1195.7 0.02 0.05 – –
26. trans-dihydrocarvone 1198.6 0.11 0.01 – 0.03
27. carvacrol methyl ether 1241.9 0.48 0.46 0.25 0.40
28. linalool acetate 1254.6 0.04 – – –
29. carvone 1252.0 – 0.03 – –
30. thymol 1289.6 1.60 – 0.70 0.75
31. carvacrol 1297.4 71.71 76.14 86.13 82.65
32. dihydro-carveol acetate 1307.7 0.04 – – –
33. n.i.b 1319.9 0.03 – – –
34. linalool propanoate 1338.9 – 0.17 – –
35. carvacrol acetate 1358.8 0.05 0.28 – –
36. trans-caryophyllene 1416.4 2.26 1.47 0.94 1.46
37. ␣-humulene 1455.7 0.40 0.23 0.12 0.20
38. ␤-acoradiene 1467.1 – 0.03 – –
39. ␣-curcumene 1470.0 – 0.09 – –
40. germacrene D 1487.0 0.05 0.03 – –
41. bicyclogermacrene 1496.7 – 0.03 – –
42. ␣-farnesene 1497.1 0.06 – – –
43. ␤-bisabolene 1504.6 0.66 0.38 0.20 0.34
Monoterpene hydrocarbons 20.41 18.31 10.67 12.44
Oxygenated monoterpenes 76.02 78.89 87.83 85.15
Sesquiterpene hydrocarbons 3.43 2.26 1.26 2.00
Other 0.11 0.53 0.24 0.41
n.i. 0.03 – – –

a
RI, retention indices as determined on HP-5 column using homologous series of C8 -C30 alkanes.
b
n.i. stands for Not identified.

was faster method for the isolation of the essential oils than hydro-distillation (HD). MAHD had shorter extraction time,
HD, it turns out as more “environmentally friendly” approach. 24 min compared to 136 min, better yields of essential oil,
7.10 % compared to 5.81 %, higher content of oxygenated
4. Conclusion compounds, 78.89 % compared to 76.82 %, and a more envi-
ronmentally friendly approach with electrical consumption
Microwave-assisted hydro-distillation (MAHD) at different 0.240 kW h compared to 1.360 kW h and CO2 emission 0.19 kg
power levels has been compared with conventional hydro- compared to 1.09 kg. The main component in all essential
distillation (HD) method for the extraction of essential oil oils, as well as in obtained hydrolats, was carvacrol. Residual
from O. vulgare L. ssp. hirtum. MAHD at the power level of extracts after HD and MAHD showed to be valuable by-
600 W may be considered as the most promising among the products, rich in phenolic compounds with rosmarinic acid as
tested power levels with respect to all investigated param- the most abundant compound. The higher amount of pheno-
eters. MAHD at the power level of 600 W proved to be a lic compounds was detected in residual extracts after MAHD.
technique with many advantages compared to conventional MAHD at the power level of 600 W may be considered as the
 Food and Bioproducts Processing 1 2 0 ( 2 0 2 0 ) 158–165 163

Table 2 – Chemical constituents in the O. vulgare L. ssp. hirtum hydrolat obtained using hydro-distillation and
microwave-assisted hydro-distillation at different power levels.
Peak Compound RIa Hydro-distillation Microwave assisted hydro-distillation

600 W 360 W 180 W

1 ␣-thujene 923.9 – – – 0.05

2 1-octen-3-ol 973.8 – – – 0.09
3 myrcene 987.7 – – – 0.15
4 ␣-terpinene 1013.8 – – – 0.18
5 p-cymene 1019.6 0.11 – – 0.83
6 ␥-terpinene 1052.9 0.06 – – 1.97
7 cis-sabinene hydrate 1061.9 – – – 0.03
8 camphor 1143.1 0.04 – – –
10 4-terpinenol 1175.6 – – – 0.31
11 carvacrol methyl ether 1242.2 – – – 0.10
12 p-8-cymenol 1179.8 0.07 – – –
13 thymol 1289.1 0.72 0.42 0.60 0.88
14 carvacrol 1296.7 99.00 99.58 99.40 98.22
15 trans-caryophyllene 1416.5 – – – 0.71
16 ␣-humulene 1455.9 – – – 0.06
17 ␤-bisabolene 1504.7 – – – 0.13

a
RI, retention indices as determined on HP-5 column using homologous series of C8 -C30 alkanes.

Table 3 – Chemical constituents in the O. vulgare L. ssp. hirtum extracts obtained after hydro-distillation and
microwave-assisted hydro-distillation at different power levels.
Total phenols contentb Chlorogenic acid Caffeic acid Rosmarinic acid Luteolin-7-O-glucoside
[␮g GAE/g dw] [␮g/g dw] [␮g/g dw] [␮g/g dw] [␮g/g dw]

Hydro-distillation 10.59 0.92 ± 0.03 b 3.30 ± 0.08 b 31.76 ± 1.10 b 1.10 ± 0.04 b
MAHD 600 Wa 17.74 1.50 ± 0.04 a 5.37 ± 0.12 a 56.86 ± 1.42 a 1.51 ± 0.04 a
MAHD 360 W 9.34 0.74 ± 0.02 c 2.43 ± 0.07 c 26.59 ± 1.00 c 0.77 ± 0.03 c
MAHD 180 W 9.58 0.34 ± 0.02 d 1.61 ± 0.05 d 19.14 ± 0.40 d 0.57 ± 0.02 d

a
MAHD stands for Microwave Assisted Hydro-Distillation.
b
Means followed by different letters are significantly different according to the post hoc Duncan’s test al level P < 0.05.

Fig. 3 – Environmental impact: A) Electrical consumption for yield of 1 % (kWh), and B) CO2 emission (kg).

most promising among the tested power levels with respect Conflict of interest
to all investigated parameters. Therefore, MAHD at the power
level of 600 W can be good alternative technique for the extrac- None.
tion of essential oil from O. vulgare ssp. hirtum.
164 Food and Bioproducts Processing 1 2 0 ( 2 0 2 0 ) 158–165

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