Evaluation of environmental factors on essential oil and forage value of Cymbopogon olivieri by Journal of Research in Ecology

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Original Research

Evaluation of environmental factors on essential oil and forage value of

Cymbopogon olivieri Authors: Azadeh Afrigan1, Hossein Azarnivand2, Fatemeh Sefidkon3, Mohammad Jafari2, Mohammad Ali Zare Chahouki2 Institution: 1. Department of Reng Management Science and Research Branch, Islamic Azad University, Tehran, Iran 2. Natural Resources Faculty, Tehran University, P.O. Box, Tehran 315854314, Iran 3. Research Institute of Forests and Rangelands, Tehran, Iran

ABSTRACT: This study was performed to evaluate the environmental factors on essential oil and forage values of Cymbopogon olivieri. In this study, the aerial parts of Cymbopogon olivieri were collected after a time of flowering from 10 natural areas located in the Khuzestan province at two altitudes, and in three replications in the year 2016. Areas included were Chal Gandali, Talkhab e Kalat, Bardmar, Morad Abad, Tembi, Dezful, Indika, Lali, Shoushtar and Izeh. The essential oil compositions were analyzed using gas chromatography-mass spectrometry (GC-MS) analysis. Also, due to phytochemical studies, the plant composition showed Crude Protein (CP), Water Soluble Carbohydrates (WSC), Crude Fiber (CF), Acid detergent fiber (ADF), Neutral Detergent Fiber (NDF) and Total Ash (TA). According to the results, Lali area showed highest mean of essential oil. Analyses of the essential oils showed 21 main identified constituents, including Verbenen, 8/1-Dihydrocodeine , and -2-carene, p-cymene, limonene, p-cymene, Cis-p-mentha-2,8-dien-1-ol, Trans-p-menth, p-mentha-1,5diene, Methylacetophenone, p-Cymen-8-ol, terpineol, piperitone, germacrene, bselinene, valencene, f-eti-x-selinene, elemol, Intermedol and y-eudesmot etc. However, Piperitone was present in each area samples more than 50 percentage . The results indicated that essential oils and their chemical compositions of Cymbopogon olivieri are strongly affected by the environmental conditions. Also, according to the results, it was found that essential oil extraction had significant effects on WSC, NDF, CF and total ash. Also there were significant differences between areas. Keywords: Cymbopogon olivieri, essential oil, forage value

Corresponding author: Hossein Azarnivand

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Article Citation: Azadeh Afrigan, Hossein Azarnivand, Fatemeh Sefidkon, Mohammad Jafari and Mohammad Ali Zare Chahouki Evaluation of environmental factors on essential oil and forage value of Cymbopogon olivieri Journal of Research in Ecology (2017) 5(2): 1095-1112 Dates: Received: 20 Aug 2017 Accepted: 25 Aug 2017

Web Address: http://ecologyresearch.info/ documents/EC0481.pdf Journal of Research in Ecology An International Scientific Research Journal

Published: 15 Sep 2017

This article is governed by the Creative Commons Attribution License (http://creativecommons.org/ licenses/by/4.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.

1095-1112 | JRE | 2017 | Vol 5 | No 2

www.ecologyresearch.info

Afrigan et al., 2017 INTRODICTION

feed quality is critical for the predictum of animal per-

The genus Cymbopogon Spreng as aromatic

formance (Seven and ร erรง, 2006). Herbage assessment

grasses is belonging to the Poaceae family (Nair, 1982)

infers the depiction of feedstuffs as for their ability to

and the genus Cymbopogon comprises two perennial

ensure diverse kinds and levels of production (Juรกrez et

species in the Flora of Iran, C. Olivieri (Boiss.) Bor and

al., 2004). From a ruminant nutrition point of view, en-

C. parkeri Stapf., which are distributed in the tropical

ergy, dry matter digestibility and unrefined protein con-

regions of Iran including southern parts of Fars, Ker-

tents are three of the most imperative segments of herb-

man, Hormozgan, Khuzestan, Bushehr and Baluchestan

age quality (Malau-Aduli, 2007). It is necessary to ob-

provinces (Sonboli et al., 2010). C. Olivieri is known as

tain information about its chemical composition to im-

KahMakki, Potar and Nagerd in different areas which

prove the quality of the herbage consumed by grazing

this plant is gathered. In traditional medicine, leaves and

animals, which thereafter could be related to its capacity

roots are widely used as antiseptic and for the treatment

to satisfy the requirements of the grazing animals.

of stomachache (Singh et al., 2011). Essential oils of the

Mountousis et al. (2011) contemplated altitudinal and

Cymbopogon spp. are principally made out of cyclic and

seasonal differences in herbage composition and energy

non-cyclic monoterpenes like citral (3,7-dimethyl-2,6-

and protein content of grasslands on Mount Varnoudas,

octadienal; a blend of two isomer geranial and neral),

North-West Greece. They revealed that Crude Protein

geraniol, citronellol, citronellal, linalool, elemol, 1,8-

(CP) diminished from 106.65 to 72.03 g/kg dry matter

cineole, limonene, b-carophyllene, methylheptenone,

(DM) in the lowlands, from 133.95 to 80.38 g/kg DM in

geranyl acetate derivation and geranyl formate (Ito and

the center zone and from 127.13 to 74.47 g/kg DM in

Honda, 2007). It is well known that yield and yield

sub-alpine grasslands. Metabolizable Energy (ME) con-

components of plants are determined by a series of fac-

tent of the herbage diminished as the growing season

tors including plant genetic (Pirbalouti et al., 2013) cli-

advanced around 19%, 32% and 23% in the lower, mid-

mate, edaphic, elevation, and topography (Loziene and

dle and upper altitudinal zone, separately (Mountousis

Venskutonis, 2005) and also an interaction of various

et al., 2011). According to report of Mountousis et al.,

factors (Basu et al., 2009). In an investigation of the

(2011) ME content of the herbage decreased about 19%,

effect of environmental factors on Cymbopogon olivieri

32% and 23% in the lower, middle and upper altitudinal

(Boiss.) Bor (Poaceae) in four regions, Mirjalili and

zone, respectively (Mountousis et al., 2011). So, the aim

Omidbeigi (2005) concluded that the nearby altitudes of

of the present study is to the determine the environmen-

300-600 m in the regions of Masjid Soleiman and Jiruft

tal effects on the essential oil and forage value of

had a greater effect on the function of the essential oil in

Cymbopogon olivieri.

the lemongrass (Mirjalili and Omidbeigi, 2005) . Yazdani et al. (2002) reported that the percentage essential oil

MATERIALS AND METHODS

of Mentha piperita Stokes (Lamiaceae) varied from

Plant collection and essential oil extraction

1.45% to 3.2%, and was influenced by different envi-

In this study, the aerial parts of Cymbopogon

ronmental factors, such as altitude and the daylight peri-

olivieri were collected after a time of flowering from 10

od (Yazdani et al., 2002). Soil type, climatic regime,

natural areas located in Khuzestan Province at two alti-

botanical composition and soil improvement practices

tudes, and in three replications at 2016. Territories in-

as biotic environmental factors have effects on nutri-

cluded were ChalGandali, Talkhab e Kalat, Bardmar,

tional quality (Corona et al., 1998). The assessment of

Morad Abad, Tembi, Dezful, Indika, Lali, Shoushtar

1096

Journal of Research in Ecology 2017) 5(2):1095-1112

Essential oil (g)

Afrigan et al., 2017

Figure 1. Comparison of the means of essential oil between the regions of the study and Izeh. Basic characterization of region of the present

using the formula described by Rao et al. (2005) where

study shown in the Table 1.

the amount of essential oil recovered (g) was deter-

The freshly extracted herb was dried in the la-

mined by weighing the oil after moisture was removed

boratory setting. A sample (90 g) of the aerial parts of

(Rao et al., 2005).

the plant was extracted using a Clevenger apparatus

Gas Chromatography–Mass Spectrometry (GC–MS)

through water distillation for three hours. Samples were

analysis

dried using anhydrous sodium sulphate (Merck Co. Ger-

The essential oil analysis was donned using an

many) and then kept in amber vials at 4±1ºC prior to

analytical gas chromatograph. The GC apparatus was

use. The percentage yield of essential oil is determined

HP-5MS (5% phenyl methyl silicone and 95% dime-

Figure 2. Comparison of means for piperitone percentage between the studied regions Journal of Research in Ecology 2017) 5(2):1095-1112

1097

Afrigan et al., 2017 Table 1. Basic characterization of region of the present study (Meteorological Organization of Khuzestan Province Data).

Region Chalgandali Talkhab-e Kalat Bardmar Morad Abad Tembi Dezful Indika Lali Shushtar Izeh

Chalgandali Talkhab-eKalat Bardmar Morad Abad Tembi Dezful Indika Lali Shushtar Izeh

Point 1 Latitude Longitude

Point 2 Latitude Longitude

Point 3 Latitude Longitude

Altitudes from sea level (m)

Slope %

Annual rainfall (mm)

Annual evaporation

Average Annual Temperature (C˚) 24

321487 3555621 323835 3553616 314284 3557151 346970 3526940 337497 3532764 293710 3596903 354573 3562422 326321 3578886 319954 3541019 377273 3537145

321495 3555627 323842 3553623 314293 3557156 346980 3526940 337505 3532759 293712 3596913 354582 3562427 326329 3578880 319961 3541012 377278 3537136

321485 3555611 323830 3553607 314274 3557151 346960 3526940 337490 3532771 293707 3596894 354565 3562417 326312 3578891 319946 3541025 377268 3537153

328

350

301

340

394

350

359

490

208

440

461

600

724

600

323

570

148

290

768

630

Point 1 Latitude Longitude

Point 2 Latitude Longitude

Point 3 Latitude Longitude

Altitudes from sea level (m)

Slope %

Annual rainfall (mm)

Annual evaporation

313459 3564137 315594 3560331 312163 3557295 354139 3562725 337815 3522292 275688 3614786 354084 3562773 326148 3578897 325263 3534175 376265 3528611

466

Average Annual Temperature (C˚) 23

313467 3564142 315596 3560341 312168 3557286 354143 3562734 337813 3522302 275695 3614779 354092 3562779 326154 3578889 325270 3534168 376270 3528603

313475 3564148 315601 3560349 312172 3557277 354149 3562742 337818 3522311 275703 3614773 354100 3562784 326161 3578882 325277 3534161 376276 3528595

390

345

370

425

350

778

610

238

440

628

670

742

600

393

580

170

340

852

620

thylpolysiloxane), that fitted with DB5 capillary column

increased to 120˚C at a 2˚C/min rate, held for three min,

(30 m×0.25 mm ID, 0.25 μm film thickness). The col-

and finally increased to 280˚C and held for 4˚C

umn temperature was initially 60˚C, and then gradually

(Shahbazi et al., 2015). The carrier gas was helium

1098

Journal of Research in Ecology 2017) 5(2):1095-1112

Afrigan et al., 2017 with 99.99% purity, consistent flow rate of 1ml for each

essential oil and significant differences were obtained

min, and a split proportion equivalent to 1:20 was uti-

between essential oil of Cymbopogon olivieri in Lali

lized. Additionally, the chemical investigation of the

with other area, Also, Tembi didn’t show significant

essential oil was done by analytical gas chromatograph

differences with Shushtar but showed against the three

combined with mass spectrometer detector. The capil-

area viz., Talkhab e Kalat, Morad Abad and Indika

lary column and temperature state of mass spectrometer

(Figure 1). Higher elevation and colder temperature

detector was comparable with that of gas chromato-

provided a better growing condition which led to the

graph that was portrayed previously. The procedure was

higher accumulation of oil in the leaves. The conse-

worked at 70 eV. The GC-MS investigation was done in

quences of different researches showed that altitude is

triplicate. Identification of the significant constituents of

the most imperative ecological factor affecting oil con-

the essential oil was done with in view of the correlation

tent (Kizil, 2010). Our result was obtained at the past

between their Retention Indices (RIs), and that of the

flowering stage but Tajidan et al. (2012) reported essen-

published data. Standard Mass Spectral fragmentation

tial oil content decreased with the increase in maturity

pattern (Wiley/NBS) and the NIST (National Institute of

stages at harvest, also Ganjewala and Luthra (2007)

Standards and Technology). The percentage of each

studied essential oil biosynthesis of Cymbopogon flexu-

essential oil compositions was calculated from the GC

osus (Nees ex Steud) incorporated with (2-14C) acetate.

peak areas.

Their discoveries demonstrated that the yield of essen-

Chemical Analysis

tial oil during the initial leaf growth of 10 to 15 days

Nitrogen (N) was determined on the basis of Kjeldahl

technique

(Bremner

and

was higher (110 to 135 pmol/10 leaves) contrasted with

Mulvaney,

the yield that was acquired toward the end of leaf

1982) ,Crude Protein (CP) was calculated as CP = N%

growth cycle of 40 to 50 days (40 to 50 pmol/10 leaves)

× 6.25(Jones, 1981). Neutral Detergent Fiber (NDF) and

(Ganjewala and Luthra, 2007). Research reports demon-

Acid Detergent Fiber (ADF) were analyzed by the

strated that overall essential oil synthesis is related with

method of AOAC (2000). Dry Matter Digestibility

the early growth stage in plants, for example, Cymbopo-

(DMD) was estimated using the formula of DMD% =

gon flexuosus (Singh et al., 1989), Cymbopogon martini

83.58 – 0.824 ADF% + 2.626N%, suggested by Arzani

(Sangwan et al., 1982) and Mentha (McCaskill and Cro-

et al. (2006). Metabolizable Energy (ME) was calculat-

teau, 1995). In general, the yield of essential oil is high-

ed with the equation of ME=0.17 DMD% - 2 reported

ly correlated with the yield of biomass.

by AOAC (2000). Statistical analysis

As indicated by Lewinsohn et al. (1998), the oil cells of lemongrass were situated inside the paren-

The data was statistically analyzed by SPSS

chymatous cells. Different examinations on aromatic

(19.0) software (SPSS, 2010), Means of the traits were

plants showed that essential oils deposited inside the

compared by Duncan’s multiple range test at p < 0.05

glandular trichomes (Serrato-Valenti et al., 1997), how-

level.

ever it had been watched that the surface of lemongrass does not contain glandular trichomes (Lewinsohn et al.,

RESUTS AND DISCUSSION

1998).

The means of the essential oil levels in the

Analyses of the essential oils showed 21 main

Cymbopogon olivieri samples are shown in Figure 1;

identified constituents, we can clearly see from table 2

according to results, Lali area showed highest mean of

that Verbenen, 8/1-Dihydrocodeine , and -2-carene, p-

Journal of Research in Ecology 2017) 5(2):1095-1112

1099

1100

99.01

p-mentha-5/1-diem Methylacetophenone Cymen-8-01 terpineol piperitone yermacrene b-selinene valencene f-eti-x-selinene elemol y-eudesmot Intermedol

0.84

Sum

Trams-p-menth

0.59

58.88 0.15 0.41 0.18 1.00 0.64 7.51

Cis-p-memth-2/8

Cis-p-menth-2-en

0.25

0.60

p-cymenene

8/1-dihydrocinede &-2-carene p-cyemene limonene

0.27 23.30 0.31 2.43

0.48

Verbenen

1.18

piperitone b-selinene valencene f-eti-x-selinene elemol y-eudesmol Intermedol

Trams-p-menth p-mentha-5/1diem Methylacetophenone

p-cymenene Cis-p-memth2/8

Verbenen 8/1dihydrocodeine, -2-carene p-cymene limonene

0.90 0.19 0.24 57.59 0.18 0.34 0.47 0.20 1.66 1.19 9.36 98.84

0.46

0.45

0.29

0.12

0.45

0.12 21.37 0.24 2.25

0.78

Trams-p-menth p-mentha-5/1diem Methylacetophens Cymen-8-01 terpineol piperitone valencene elemol y-eudesmot Intermedol

p-cymenene Cis-p-menth-2en Cis-p-memth2/8

Verbenen 8/1dihydrocodeine &-2-carene p-cymene limonene

99.45

0.82 0.25 0.26 59.25 0.26 1.38 0.89 8.99

0.75

0.59

0.29

1.33

0.49

0.21 20.27 0.34 2.06

1.02

Cymen-8-01 terpineol piperitone b-selinene valencene f-eti-x-selinene elemol y-eudesmot

Trams-p-menth p-mentha-5/1diem Methylacetophens

Cis-p-memth-2/8

Cis-p-menth-2-en

&-2-carene p-cymene limonene p-cymenene

Verbenen

92.01

0.22 0.26 58.20 0.14 0.40 0.19 0.99 0.56

0.85

0.51

0.63

0.28

0.18

24.31 0.25 2.42 0.49

1.13

Continue ......

Trams-p-menth p-mentha-5/1diem Methylacetophens Cymen-8-01 terpineol piperitone b-selinene valencene f-eti-x-selinene elemol y-eudesmot Intermedol

Cis-p-memth-2/8

Cis-p-menth-2-en

p-cymenene

8/1-dihydrocinede &-2-carene p-cymene limonene

Verbenen

Table 2. Identification of essential oil compositions of Cymbopogon olivieriat the studied regions (Altitudes 1) Chal-GTalkhab Morad Andali e Kalat Bardmar Abad

98.96

0.89 0.21 0.28 59.73 0.14 0.37 0.19 0.99 0.59 7.11

0.57

0.48

0.39

0.18

0.53

0.18 22.36 0.28 2.47

1.03

Tembi

Afrigan et al., 2017

Journal of Research in Ecology 2017) 5(2):1095-1112

sum

b-selinene valencene f-eti-x-selinene elemol y-eudesmot Intermedol

99.01

0.15 0.27 0.20 0.81 0.54 7.20

0.26 0.56 0.62 0.88 0.22 58.22

2.83 0.49

limonene p-cymenene

Cis-p-memth-2/8 Trams-p-menth p-mentha-5/1-diem Methyl-acetophens Cymen-8-01 piperitone

23.87 0.37

1.27 0.26

Dezful

&-2-carene p-cymene

Verbenen 8/1-dihydrocodeine

Continue ...

terpineol piperitone valencene elemol Intermedol

Cis-p-menth-2-en Cis-p-memth-2/8 Trams-p-menth p-mentha-5/1-diem Methyl-acetophens Cymen-8-01

limonene p-cymenene

&-2-carene p-cymene

Verbenen 8/1-dihydrocodeine

98.70

0.29 62.64 0.88 0.70 7.49

0.13 0.32 0.59 0.56 0.88 0.20

2.04 0.55

20.04 0.25

1.01 0.12

Indika

terpineol piperitone y-eudesmot Intermedol

Cis-p-menth-2-en Cis-p-memth-2/8 Trams-p-menth p-mentha-5/1-diem Methyl-acetophens Cymen-8-01

limonene p-cymenene

&-2-carene p-cymene

Verbenen 8/1-dihydrocodeine

97.86

0.26 66.61 0.37 3.75

0.21 0.34 0.43 0.64 1.01 0.24

2.32 0.57

19.92 0.30

0.69 0.20

Lali

terpineol piperitone b-selinene valencene f-eti-x-selinene elemol y-eudesmot Intermedol

Cis-p-menth-2-en Cis-p-memth-2/8 Trams-p-menth p-mentha-5/1-diem Methyl-acetophens Cymen-8-01

limonene p-cymenene

&-2-carene p-cymene

Verbenen 8/1-dihydrocodeine

0.27 64.62 0.12 0.33 0.17 0.73 0.55 7.19 98.83

0.24 0.34 0.57 0.63 0.95 0.29

1.97 0.48

18.01 0.28

0.92 0.17

Shushtar

terpineol piperitone valencene elemol y-eudesmot Intermedol

Cis-p-menth-2-en Cis-p-memth-2/8 Trams-p-menth p-mentha-5/1-diem Methyl-acetophens Cymen-8-01

limonene p-cymenene

&-2-carene p-cymene

Verbenen 8/1-dihydrocodeine

99.44

0.82 66.53 0.22 0.40 0.32 5.60

0.22 0.36 0.41 0.53 0.97 0.28

2.21 0.55

18.52 0.31

1.04 0.14

Izeh

Afrigan et al., 2017

1101

1102

ChalGandali 1.00 0.19 16.30 0.29 1.82 0.51 0.22 0.34 0.66

0.77 0.87 0.30 0.29 64.95 0.14 0.42 0.20 0.70 0.53 7.80 98.29

Verbenen 8/1-dihydrocodeine &-2-carene p-cymene limonene p-cymenene Cis-p-menth-2-en Cis-p-memth-2/8 Trams-p-menth

p-mentha-5/1-diem Methyl-acetophens Cymen-8-01 terpineol piperitone b-selinene valencene f-eti-x-selinene elemol y-eudesmot Intermedol Sum

p-mentha-5/1-diem Methyl-acetophens Cymen-8-01 terpineol piperitone b-selinene valencene f-eti-x-selinene elemol y-eudesmot Intermedol

Verbenen 8/1-dihydrocodeine &-2-carene p-cymene limonene p-cymenene Cis-p-menth-2-en Cis-p-memth-2/8 Trams-p-menth 0.49 1.00 0.26 0.29 63.32 0.11 0.31 0.16 0.89 0.59 7.39 99.00

Talkh-ab- e Kalat 0.76 0.20 19.59 0.15 1.79 0.59 0.15 0.37 0.60 p-mentha-5/1-diem Methyl-acetophens Cymen-8-01 terpineol piperitone valencene elemol y-eudesmot Intermedol

Bardmar Verbenen 8/1-dihydrocodeine &-2-carene p-cymene limonene p-cymenene Cis-p-menth-2-en Cis-p-memth-2/8 Trams-p-menth

99.27

0.68 0.96 0.27 0.70 60.28 0.30 1.29 0.76 11.17

Morad Abad 1.01 0.18 17.84 0.21 1.90 0.53 0.15 0.33 0.71

Continue...

Cymen-8-01 terpineol piperitone valencene f-eti-x-selinene elemol y-eudesmot Intermedol

Verbenen 8/1-dihydrocodeine &-2-carene limonene p-cymenene Cis-p-memth-2/8 Trams-p-menth p-mentha-5/1-diem Methyl-acetophens

Table 3. Identification of essential oil compositions of Cymbopogon olivieri at studied regions (Altitudes 2)

99.28

0.20 0.27 56.25 0.23 0.15 0.67 0.39 5.04

Tembi 1.11 0.27 29.26 2.48 0.58 0.35 0.63 0.47 0.92

Afrigan et al., 2017 cymene, limonene, p-cymene, Cis-p-mentha-2,8-dien-1b-selinene, valencene, f-eti-x-selinene, elemol, Inter-

ol, Trans-p-menth, p-mentha-1,5-diene, Methylacetomedol and y-eudesmot were obtained. A study on the

phens, p-cymen-8-ol, terpineol, piperitone, germacrene, quality and quantity of essential oil showed

Journal of Research in Ecology 2017) 5(2):1095-1112

Dezful 1.03

0.21 19.97 0.35 2.29

0.51

0.22

0.32 0.43

0.74

0.92 0.27 0.27 62.66 0.25 0.14

0.80 0.62 6.91

98.91

Verbenen

8/1-dihydrocodeine &-2-carene p-cymene limonene

p-cymenene

Cis-p-menth-2-en

Cis-p-memth-2/8 Trams-p-menth

p-mentha-5/1-diem

Methyl-acetophens Cymen-8-01 terpineol piperitone valencene f-eti-x-selinene

elemol y-eudesmot Intermedol

Sum

Continue â&#x20AC;Ś

Journal of Research in Ecology 2017) 5(2):1095-1112 valencene f-eti-x-selinene elemol y-eudesmot Intermedol

p-cymenene Cis-p-menth-2en Cis-p-memth2/8 Trams-p-menth p-mentha-5/1diem Methylacetophens Cymen-8-01 terpineol piperitone yermacrene b-selinene

Verbenen 8/1dihydrocodeine &-2-carene p-cymene limonene

0.48 0.21 1.35 0.94 8.38 99.02

1.00 0.25 0.30 65.92 0.37 0.36

0.54

0.34 0.53

0.15

0.54

0.13 14.59 0.20 1.63

Indika 0.81

Methyl-acetophens piperitone b-selinene valencene y-eudesmot Intermedol

p-mentha-5/1-diem

Cis-p-memth-2/8 Trams-p-menth

Cis-p-menth-2-en

p-cymenene

8/1-dihydrocodeine &-2-carene p-cymene limonene

Verbenen

97.02

6.13 61.70 0.29 0.24 0.48 4.81

0.60

0.31 1.17

0.19

0.16

18.17 0.32 1.97 0.28

Lali 0.19

f-eti-x-selinene elemol y-eudesmot Intermedol

Methyl-acetophens Cymen-8-01 terpineol piperitone b-selinene valencene

p-mentha-5/1-diem

Cis-p-memth-2/8 Trams-p-menth

Cis-p-menth-2-en

p-cymenene

8/1-dihydrocodeine &-2-carene p-cymene limonene

Verbenen

98.83

0.17 0.73 0.55 7.19

0.95 0.29 0.27 64.62 0.12 0.33

0.63

0.34 0.57

0.24

0.48

0.17 18.01 0.28 1.97

Shushtar 0.92

y-eudesmot Intermedol

Methyl-acetophens Cymen-8-01 terpineol piperitone valencene elemol

p-mentha-5/1-diem

Cis-p-memth-2/8 Trams-p-menth

Cis-p-menth-2-en

p-cymenene

8/1-dihydrocodeine &-2-carene p-cymene limonene

Verbenen

99.06

0.46 5.67

1.02 0.29 0.28 65.66 0.20 0.58

0.55

0.38 0.63

0.16

0.57

0.10 18.97 0.32 2.17

Izeh 1.03

Afrigan et al., 2017

1103

Afrigan et al., 2017 that similar observation was reported in the production

concentration of some constituents (Raina et al., 2003).

of essential oil of lemongrass in relation to the leaf age

Gas Chromatograph (Thermo-UFM) for the regions

(Singh et al., 1989).

showen at figure 3 to 12.

At level 1, The percentages of identified compo-

In this study, piperitone as main composition

nents in the essential of the plant at the ChalGandali,

varied between 57.99 and 66.10%, the mean comparison

Talkhab e Kalat, Bardmar, Morad Abad, Tembi, Dezful,

conducted through the Duncan test and results showed

Indika, Lali, Shoushtar and Izeh were 99.01, 98.84,

significant differences between regions for piperitone

99.45, 92.01, 98.96, 99.01, 98.7, 97.86, 98.83 and

percentage Figure 2. Highest mean was observed by

99.44%, respectively Table 2.

Izeh samples and lowest means were obtained by Tembi

Also at level 2, identified components showed

region. In this order, Tajidan et al. (2012) reported that

98.29, 99.0, 99.27, 99.28, 98.91, 99.02, 97.02, 98.83

among 13 compounds, only 7 compounds (β-myrcene, 3

and 99.06 for ChalGandali, Talkhab e Kalat, Bardmar,

-undecyne, neral, geranial, nerol, geranyl acetate and

Morad Abad, Tembi, Dezful, Indika, Lali, Shoushtar

juniper camphor) had concentration greater than 1%. As

and Izeh, respectively Table 3. According to Raina et al.

indicated by Schaneberg and Khan (2002), the essential

(2003) on the essential oil composition of Cymbopogon

oil of lemongrass contains for the most part geranial and

martinii from the different places of India, three sam-

neral. Other compounds isolated, for example, β-

ples of geraniol (67.6–83.6%) was the major constituent

myrcene, ocimene, β-ocimene, linalool, citronellal, cit-

and, although the composition of the three oils were

ronellol, caryophyllene and β-pinene, were available as

similar, quantitative differences were observed in the

minor segments (Torres and Ragadio, 1996).

Areas ChalGandali TalkhabK alat Bardmar Morad Abad Tembi Dezful Indika Lali Shushtar Izeh

1104

Table 4. Means comparison between areas and essential oil extraction stages Essential oil extraction CP WSC DMD ADF NDF CF

ASH

Before

5.34

10.47

46.13

45.11

65.01

60.89

3.93

cde

After Before After

4.78 3.98 5.14

c e b

4.08 8.10 11.37

g e cd

46.69 50.32 48.22

hi a e

45.27 41.40 40.80

cd gh h

75.27 70.00 60.12

e g i

70.76 61.10 56.25

ab f h

3.25 4.08 5.36

efg b-e a

Before

4.90

11.36

47.71

42.61

64.40

60.28

3.88

After Before

4.37 3.31

d g

6.59 10.57

f d

49.32 47.13

cd gh

43.15 42.80

ef fg

77.05 57.87

d j

68.83 56.57

c h

3.56 5.26

efg a

After

3.31

4.80

46.71

46.15

81.00

67.64

4.62

a-d

Before

2.75

10.72

48.96

40.41

57.38

54.03

4.93

abc

After

1.43

6.40

43.63

50.00

81.70

70.60

2.88

Before

3.53

12.49

49.91

40.22

55.69

55.37

4.48

a-e

After

3.60

4.70

46.46

46.21

80.29

71.27

2.95

efg

Before

1.32

15.01

48.77

37.66

55.40

53.80

3.60

After

1.21

8.05

46.83

42.98

71.58

66.06

2.68

Before

4.21

4.49

44.52

47.68

78.04

71.12

3.19

efg

After

5.18

10.80

47.27

43.35

60.79

57.16

4.84

abc

Before

2.27

13.53

47.66

40.52

51.20

54.18

4.13

b-e

After

4.82

5.33

48.92

44.68

78.09

69.15

3.97

cde

Before

3.79

11.99

bcd

49.44

42.13

59.83

57.93

4.54

a-d

After

3.00

6.26

46.21

46.71

75.25

68.56

3.30

efg

Journal of Research in Ecology 2017) 5(2):1095-1112

Afrigan et al., 2017

Figure 3. Gas Chromatograph (Thermo-UFM) for the Indika sample

Figure 4. Gas Chromatograph (Thermo-UFM) for the Dezful sample Crude Protein (CP)

reported that crude protein declines with the stage of

Results showed that that there was no signifi-

maturation. Consequently, the reduction of crude pro-

cant difference between before and after essential oil

tein content, which was observed in the lower zone,

extraction, but studied areas showed significant differ-

could be attributed to the different phonological stages

ences at 5% statistical level and the highest CP content

of vegetation. It has been recommended (Bxuton, 1996)

(5.06%) was in Chalgandali samples, also Indika

that the aging of plants deduces in the decrease of crude

showed lowest content (1.26%). Arzani et al. (2001)

protein content of leaves and stems, however it addi-

revealed that locations had tremendous impart on forage

tionally prompts a more noteworthy extent of stems,

quality due to the distinctions in soil and climate attrib-

which contains less crude protein than leaves.

utes. The CP content declined through the growing sea-

Water Soluble Carbohydrates (WSC)

son, as a response to tissue ageing, Corona et al. (1998) Journal of Research in Ecology 2017) 5(2):1095-1112

WSC value differed among two stages of before 1105

Afrigan et al., 2017

Figure 5. Gas Chromatograph (Thermo-UFM) for the Izehsample

Figure 6. Gas Chromatograph (Thermo-UFM) for the Lali sample and after essential oil extraction (P<0.05). The amount

ence on forage quality, plant growth and its develop-

of WSC before extraction was 2% higher than after ex-

ment, and plant productivity (Beni et al., 2014).

traction. Between area, it was founded that highest and

Acid Detergent Fiber (ADF) and Neutral Detergent

lowest means were 49.27% and 45.89% for Talkhab-e-

Fiber (NDF)

Kalat and Lali, respectively (Table 4). Beni et al. (2014)

Essential oil extraction did not show significant

investigated the temperature effect on forage quality and

effect on ADF (P>0.05), but Significant differences

found that effects of plant maturity is most serious on

were obtained between ADF of areas, Lali and Chal-

forage quality which is related to plant environment

gandali with 45.5% and 45.19 showing highest value of

including temperature. They reported that in addition to

ADF and lowest mean was observed by Indika area

temperature, other environ mental factors including so-

(40.32%). There were significant differences in NDF

lar radiation, soil characteristics, water and insects influ-

content of between before and after essential oil extrac

1106

Journal of Research in Ecology 2017) 5(2):1095-1112

Afrigan et al., 2017

Figure 7. Gas Chromatograph (Thermo-UFM) for the Bardmarsample

Figure 8. Gas Chromatograph (Thermo-UFM) for the Chal-G-Andali sample tion (P<0.05), essential oil extraction led to 20% in-

increases result from an increase in the herbage cell wall

creasing in NDF, highest NDF (70.72%) was obtained

contents with maturity compared to cell content

at Bardmar samples, also it was founded that Indika had

(Skapetas et al., 2004).

lowest mean (63.49%) between areas (Table 4). Altitude

Crude Fiber (CF)

has effects on maturity time, also, the fiber concentra-

Essential oil extraction had significant effect on

tion increases as plants mature, which is the most im-

CF (P<0.05), this treatment led to 13% increase in CF

portant factor affecting dry matter digestibility. These

value, Between areas, Calgandali and Talkhab-e-Kalat

Journal of Research in Ecology 2017) 5(2):1095-1112

1107

Afrigan et al., 2017

Figure 9. Gas Chromatograph (Thermo-UFM) for the MoradAbad sample

Figure 10. Gas Chromatograph (Thermo-UFM) for the Talkhab -e-Kalat sample they showed highest (65.82%) and lowest (58.6) means,

(P<0.05), essential oil extraction led to 12% reduction

respectively. In this order, Marshal et al. (2005) showed

in ash, Also, Morad Abad and Indika showed highest

that CP was positively associated with forage growth (P

(4.94%) and lowest (3.14%) means, respectively. This is

< 0.001) and with rainfall (P ≤ 0.025) (Marshal et al.,

similar to the finding of other researchers such as Arza-

2005).

ni et al. (2006) and Mountousis et al. (2011)

Total ash There were significant differences in ash content of between before and after essential oil extraction

1108

CONCLUSION The essential oil content showed significant

Journal of Research in Ecology 2017) 5(2):1095-1112

Afrigan et al., 2017

Figure 11. Gas Chromatograph (Thermo-UFM) for the Tembi sample

Figure 12. Gas Chromatograph (Thermo-UFM) for the Shushtar sample differences when lemongrass was harvested from differ-

than 50 percentage. According to the results, it was

ent areas. The highest percentage of essential oil was

founded that essential oil had significant effects on

obtained when lemongrass was collected from Lali area.

WSC, NDF,CF and total ash. Also there were signifi-

There were 21 chemical compounds detected in the es-

cant differences between areas. Altitude has effects on

sential oil of lemongrass. However, piperitone of the

maturity time, so, chemical component change accord-

compounds was present in each area samples at more

ing to growth stages.

Journal of Research in Ecology 2017) 5(2):1095-1112

1109

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