NVEO 2018, Volume 5, Issue 2nveo.org/wp-content/uploads/2019/03/NVEO-2018-Volume-5... · 2019-06-17 · Nat. Volatiles & Essent. Oils, 2018; 5(2): 1-9 Shaimerdenova et al. 2 A. rupestris

NVEO 2018, Volume 5, Issue 2

CONTENTS

1. Chemical composition of essential oils from Artemisia glabella Kar. et Kir. and Artemisia

rupestris L. obtained by different methods / Pages: 1-9

Zhanar R. Shaimerdenova, Aigerim I. Makubayeva, Temel Özek, Gülmira Özek, Süleyman Yur,

Gayane A. Atazhanova, Sergazi M. Adekenov

2. Valorization of Citrus Peel Waste / Pages: 10-18

Merve Deniz Köse, Oğuz Bayraktar

3. Essential oil composition of Zosima absinthifolia (Vent.) Link from Northern Cyprus / Pages:

19-23

Kemal Hüsnü Can Başer, Omar Aburwais, Azmi Hanoğlu, Betul Demirci

Nat. Volatiles & Essent. Oils, 2018; 5(2): 1-9 Shaimerdenova et al.

1

RESEARCH ARTICLE

Chemical composition of essential oils from Artemisia glabella

Kar. et Kir. and Artemisia rupestris L. obtained by different

extraction methods

Zhanar R. Shaimerdenova1,*, Aigerim I. Makubayeva1, Temel Özek2,3, Gülmira Özek2, Süleyman

Yur2, Gayane A. Atazhanova1, Sergazi M. Adekenov1

1 JSC International Research and Production Holding “Phytochemistry”, Karaganda, Republic of KAZAKHSTAN 2 Department of Pharmacognosy, Faculty of Pharmacy, Anadolu University, 26470, Eskişehir, TURKEY 3 Medicinal Plants, Drugs and Scientific Research Center, Anadolu University, 26470, Eskişehir, TURKEY

*Corresponding author. Email: [email protected]

Abstract

The aim of the research was to investigate the chemical composition of essential oils and volatiles from two species of Artemisia

glabella and Artemisia rupestris growing in Kazakhstan. Two different techniques, the conventional hydrodistillation (HD) and modern

fast micro-steam distillation-solid-phase microextraction (MSD-SPME) have been used to obtain the volatiles. Chemical profiles of

the volatiles were comparatively analyzed with GC-FID/MS techniques. The yields of essential oils from A. glabella and A. rupestris

obtained by a hydrodistillation were 0.2% and 0.1%, respectively. The major components of A. rupestris essential oil were myrcene

(9.5%), β-elemene (5.4%) and capric acid (5.1%). The oil of A. glabella was found to be rich with 1,8-cineole (12.2%), cumin aldehyde

(9.4%), α-terpineol (5.7%) and borneol (5.2%).

Keywords: Artemisia glabella, Artemisia rupestris, essential oil, GC/FID, GC/MS, MSD-SPME.

Introduction

The genus Artemisia L. (Compositae) is one of the most widespread in the flora of Kazakhstan. Eighty-one

species of Artemisia L. grow on the territory of Kazakhstan, of which 16 are endemic species (Pavlov, 1966).

Wormwoods are of great interest as the object of research due to their wide distribution throughout the

territory of Kazakhstan. A high content of essential oils with a valuable chemical composition in Artemisia

makes it possible to produce medicinal substances with original pharmacological properties based on them.

Artemisia glabella Kar. et Kir. is found on stony mountain and hill slopes, on dry pebbled river beds and among

rocks (Pavlov, 1966). It is known that A. glabella contains a number of biologically active compounds including

sesquiterpene lactones, arglabin, argolide, dihydroargolide, matricarin, 1β,10α-dihydroxyarglabin, and

flavonoids, cirsilineol, pectolinarigenin, casticin, bonanzin (Adekenov et al., 1982, 1983, 1993,

Kulmagambetova et al., 2000). According to Atazhanova et al., (1999) the main constituents of essential oil

were 1,8-cineole (12.0%), linalool (8.0%), 4-terpineol (6.5%), α-terpineol (5.0%) and sabinol derivatives

(5.0%). Due to the fact that essential oil from A. glabella has antibacterial, antifungal and antiviral activities

(Seidakhmetova et al., 2002) the Epherol spray was developed on its basis, which has antimicrobial and anti-

inflammatory effects and improves the mucus drainage. The drug is recommended at treatment of upper

airway diseases, chronic obstructive bronchitis and pneumonia in the complex therapy that includes

traditional and officinal drugs. The main biologically active ingredient of the Epherol spray is 1,8-cineole

(Atazhanova, 2008).


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A. rupestris grows in a steppe zone on saline and solonetzic meadows, in stony and sandy dry river beds,

sometimes on non-saline steppe meadows, on stony and crushed rock slopes of the subalpine and alpine

mountain belts (Pavlov, 1966). A. rupestris is used in Traditional Chinese herbal medicine as an antibacterial,

antiviral, anti-tumor agent (Xiao, et al., 2008). It is also a source of rupestric acid which exhibits an antiviral

activity. The essential oil from A. rupestris collected in China has previously been reported to contain 1-

hexadecanol (18.1%), hexadecanoic acid (11.2%) (Bicchi, et al., 1985), α-terpinyl acetate (37.2%), spatulenol

(10.7%), α-terpineol (10.1%), linalool (7.6%), 4-terpineol (3.9%) as major constituents (Liu, et al., 2013).

The main goal of the present work was to make comparative study on the volatiles chemical profiles of A.

glabella and A. rupestris collected in Kazakhstan. The essential oil of A. rupestris. growing in Kazakhstan was

never investigated until now. The conventional hydrodistillation (HD) and rapid modern Microsteam

Distillation - Solid Phase Microextraction (MSD-SPME) procedures were applied for obtaining of the volatiles

from the plant material. MSD-SPME is a modern and rapid volatile sampling and concentration technique

introduced for the extraction of the volatiles from small amount of aromatic plant materials in a short time.

This technique involved concurrent solid-phase microextraction combined with continuous hydrodistillation

of the volatiles. This method offered important advantages in time (even less than a minute) and energy

saving for the isolation of the volatiles. MSD-SPME combined with GC/FID and GC/MS has been proven to be

simple, sensitive, rapid, solventless and non-toxic “green” technique for volatile constituents analysis at the

microscale level.

Materials and Methods

Plant material

The aerial parts of A. glabella (leaves, calathids, flower buds) were collected during the budding phase in

June, 2016 at a pilot pharm of Karaganda Pharmaceutical Plant near the Bereznyaki village of Bukhar-Zhyrau

district, Karaganda region (Kazakhstan). The aerial parts of A. rupestris (leaves, flower buds) were harvested

during the budding phase in May, 2016 in the vicinity of Karkaralinsk, along the Zhyrym River floodplain

(Kazakhstan). The herbs were dried under the shade. Botanical identifications of the both species were

performed by Dr.A.N. Kupriyanov. The voucher specimens (AG and AR) were deposited in the Herbarium of

Laboratory of Terpenoids Chemistry in Karaganda International Research Production Holding (IRPH).

Hydrodistillation

Air-dried aerial parts of A. glabella and A. rupestris (50.0 g) were ground and hydrodistilled in a Clevenger-

type apparatus (3 hours) (European Pharmacopoeia-2017). The oil yields were calculated on a dry weight

basis. The oils were dried over anhydrous sodium sulfate and stored in sealed vials in refrigerator (4°C), until

GC-FID/MS analyses. The oils were dissolved in n-hexane (10 %, v/v) to conduct chromatographic

determination of the compositions.

Microsteam distillation - solid phase microextraction

The ground plant material (1.0 g) was put into the flask (25 mL) together with water (3.0 mL) and heated. The

flask was fitted with a Claisen distillation head with plug and a condenser set up for refluxing rather than

distillation. Heating was achieved using electric heater, and threaded plug was used for SPME fiber assembly.

A manual SPME holder (57330-U, SUPELCO, Bellefonte, PA) and the PDMS-DVB (polydimethylsiloxane-

divinylbenzene) 65 μm fiber “bluetype” were used for SPME procedure of volatiles. The fiber was conditioned

at 250°C for 15 min before the experiment. After the SPME needle pierced the plug, the fiber was expressed

through the needle and exposed to the headspace above a plant sample. MSD-SPME procedure was carried


3

out at the boiling temperature of water used as solvent. The time of equilibrium was a period between

loading of SPME fiber into flask and starting of the extraction. Extraction time for 3.0 min was used as suitable

time after equilibrium. After the extraction (trapping) of the volatiles, the loaded SPME fiber was withdrawn

into the needle, and then the needle was removed from the plug and subsequently used for thermal

desorption in the injection port of GC-FID and GC/MS systems.

GC-FID and GC/MS analyses

The essential oils were dissolved in n-hexane (10%, v/v) before the chromatographic determination of their

compositions. In MSD-SPME technique, the termal desorption of the volatiles from the fiber coating was

performed by heating the fiber in the injection port at 250°C for 10 min. The SPME fiber was reconditioned

at 250°C for 15 min before the each extraction experiments. The fiber was subjected to a blank injection to

ensure fiber integrity and the absence of any analytes after each reconditioning period.

GC/MS analysis was carried out with an Agilent 5975 GC-MSD system (Agilent, USA; SEM Ltd., Istanbul,

Turkey). HP-Innowax FSC column (60 m × 0.25 mm, 0.25 μm film thickness, Agilent, USA) was used with a He

carrier gas at 0.8 mL/min. GC oven temperature was kept at 60°C for 10 min and programmed to 220°C at a

rate of 4°C/min, kept constant for 10 min at 220°C, and then programmed to increase at a rate of 1°C/min to

240°C. The oil was analyzed with a split ratio of 40:1 while the SPME experiments were in splitless mode. The

injector temperature was 250°C. Mass spectra were taken at 70 eV and the mass range was from m/z 35 to

450. The GC-FID analysis was carried out with capillary GC using an Agilent 6890N GC system (SEM Ltd.,

Istanbul, Turkey). Flame ionization detector (FID) temperature was set at 300°C in order to obtain the same

elution order with GC/MS. Simultaneous injection was performed using the same column and appropriate

operational conditions.

Identification and quantification of compounds

Compounds were identified by comparison of the chromatographic peaks retention times with those of

authentic compounds analyzed under the same conditions, and by comparison of the retention indices with

literature data. Comparisons of MS fragmentation patterns with those of standards and mass spectrum

database search were performed using the Wiley GC-MS Library (Wiley, New York, NY, USA), MassFinder

software 4.0 (Dr. Hochmuth Scientific Consulting, Hamburg), Adams Library, and NIST Library. Confirmation

was also achieved by using the in-house “Başer Library of Essential Oil Constituents” database, obtained from

chromatographic runs of pure compounds performed with the same equipment and conditions. A C8–C40 n-

alkane standard solution (Fluka, Buchs, Switzerland) was used to spike the samples for the determination of

relative retention indices (RRI). Percent composition was obtained for each constituent on the basis of GC-

FID analysis of the volatiles.

Results and Discussion

This is the first report on the composition of the volatiles obtained by HD and MSD-SPME techniques from A.

glabella and A. rupestris growing in Kazakhstan. GC/FID and GC/MS analysis performed simultaneously on

the isolated volatiles using HD and MSD-SPME from each Artemisia species showed that they have similar

compositions with varying percentages of some components depending on the technique applied. This

technique provided rapid recovery (3 min) of volatiles with the same composition as that obtained by

hydrodistillation (in Clevenger apparatus) from 1.0 g of plant material. MSD-SPME was therefore well suitable

for the extraction of aroma compounds from minute amounts (1.0 g) of aromatic plants. Furthermore, the

isolated product can be directly used for GC/FID and GC/MS analysis without further preparation. MSD-SPME


4

is useful for the analytical determination of volatiles and not for the preparation of essential oils.The aim of

the given research is to update data on the chemical composition of essential oils and volatiles from two

species of Artemisia L. found in Kazakhstan (A. glabella and A. rupestris) produced by means of the

conventional (hydrodistillation) and modern fast MSD-SPME extraction techniques. The yields of essential

oils from A. glabella and A. rupestris obtained by a hydrodistillation were 0.2% and 0.1%, respectively. The

comparative chemical composition analysis of A. glabella and A. rupestris is presented in Table 1 with their

RRI values and a relative percentage.

The essential oil from A. rupestris obtained by means of a hydrodistillation represented a yellow color liquid

with a pungent smell. Based on the gas chromatographic analysis, 69 components have been identified which

makes 78.10% of the total oil amount (Table 1). The main part of essential oil contains sesquiterpenes (49.5%)

and monoterpenes (13.9%), hydrocarbons (8.3%) and acids (6.4%). The major components of A. rupestris

essential oil were myrcene (9.5%), β-elemene (5.4%), capric acid (5.1%), γ-costol (3.9%), selin-11-en-4α-ol

(3.1%), spathulenol (3.8%), cedrol (3.3%), β-selinene (3.3%), valencene (3.7%).

Sixty-two volatile constituents received from A. rupestris by MSD-SPME have been determined. The main

compounds were found to be as valencene (9.2%), α-selinene (5.7%), myrcene (5.5%), β-selinene (5.4%), (Z)-

β-farnesene (4.9%) and β-elemene (3.6%).

The essential oil from A. glabella produced by means of a hydrodistillation represents the green mobile liquid

with a pleasant and persistent scent. Gas chromatographic analysis revealed 75 components which represent

81.7% of the total oil amount. The main components were the following: 1,8-cineole (12.2%), cumin aldehyde

(9.4%), α-terpineol (5.7%), borneol (5.2%), camphor (3.6%) and cumin alcohol (3.7%). Sixty-four volatile

components of A. glabella have been identified by means of GC-FID/MS analysis from the volatiles obtained

by MSD-SPME technique. The major components were cumin aldehyde (16.0%), 1,8-cineole (12.8%), cumin

alcohol (6.9%), borneol (5.4%), camphor (5.2%) and α-terpineol (4.8%).

The research results showed that A. glabella essential oil mainly contains monoterpenes (65.8%) with a

prevalence of the oxygenated ones (59.7%), and a low content of sesquiterpenes (12.1%), 6.5% of which are

oxygenated. The volatile components with monoterpenes content of 75.3%, the majority of which are the

oxygenated monoterpenes (71.6%), have been revealed during the MSD-SPME experiments.

The results from both species gave similar compound distributions except for their quantities.

Table 1. The chemical composition of essential oils and volatiles from A. glabella Kar. et Kir. and A. rupestris L. obtained

by different methods

No RRI Components A. rupestris A. glabella

ID HD %

MSD-SPME %

HD %

MSD-SPME %

1 1032 -Pinene 0.3 - 2.0 1.0 a,b,c

2 1035 -Thujene - - 0.1 - a,b,c

3 1076 Camphene - - 0.4 0.2 a,b,c

4 1118 β-Pinene - - 0.3 0.1 a,b,c

5 1132 Sabinene - - 0.1 0.1 a,b,c

6 1138 Thuja-2,4(10)-diene - - 0.1 0.1 a,b,c

7 1174 Myrcene 9.5 5.5 - - a,b,c

8 1176 α-Phellandrene - - 0.3 0.2 a,b,c


5


ID HD %

MSD-SPME %

HD %

MSD-SPME %

9 1188 α-Terpinene - - 0.3 0.1 a,b,c

10 1195 Dehydro-1,8-cineole - - 0.1 - a,b,c

11 1213 1,8-Cineole 0.3 1.1 12.2 12.8 a,b,c

12 1218 β-Phellandrene - - - 0.1 a,b,c

13 1255 γ-Terpinene - - 0.6 0.2 a,b,c

14 1280 p-Cymene - - 1.5 1.1 a,b,c

15 1290 Terpinolene - - 0.1 0.1 a,b,c

16 1299 2-Methylbutyl isovalerate 0.2 0.2 - - a,b,c

17 1303 Pentyl 3-methylbutanoate

(= Amyl isovalerate) - 0.1 - - a,b,c

18 1420 Presilphiperfol-7-ene 0.2 0.6 - - b,c

19 1348 6-Methyl-5-hepten-2-one - 0.1 - - a,b,c

20 1350 1-Tridecene - 0.1 - - a,b,c

21 1400 Nonanal - 0.1 - - a,b,c

22 1437 α-Thujone 0.1 - - - a,b,c

23 1444 7--(H)-Silphiperfol-5-ene - 0.2 - - b,c

24 1506 Silphiperfol-6-ene 0.3 0.1 - - b,c

25 1450 trans-Linalool oxide (Furanoid) - - 0.1 0.1 a,b,c

26 1474 trans-Sabinene hydrate - - 0.3 0.3 a,b,c

27 1475 Acetic acid - 0.8 - - a,b,c

28 1478 cis-Linalool oxide (Furanoid) - - 0.1 0.1 a,b,c

29 1490 Siphin-1-ene - 0.3 - - b,c

30 1497 α-Copaene - - 0.3 0.1 a,b,c

31 1529 Dill ether - - 2.7 0.1 a,b,c

32 1532 Camphor 0.2 0.5 3.6 5.2 a,b,c

33 1553 Linalool 0.2 0.9 0.4 4.3 a,b,c

34 1556 cis-Sabinene hydrate - - 0.2 0.4 a,b,c

35 1565 Linalyl acetate - 1.1 - - a,b,c

36 1568 1-Methyl-4-acetyl-cyclohex-1-ene - 0.2 - - a,b,c

37 1571 trans-p-Menth-2-en-1-ol - - 0.1 0.2 a,b,c

38 1577 α-Cedrene 0.5 1.0 - - a,b,c

39 1582 cis-Chrysanthenyl acetate - 2,3 0.1 0.1 a,b,c

40 1586 Pinocarvone - - 0.1 - a,b,c

41 1590 Bornyl acetate 0.1 - 0.8 0.8 a,b,c

42 1600 β-Elemene 5.4 3.6 0.1 - a,b,c

43 1611 Terpinen-4-ol - - 1.9 1.5 a,b,c

44 1612 β-Caryophyllene 0.3 0.5 0.5 0.3 a,b,c

45 1613 β-Cedrene 0.4 0.9 - - a,b,c

46 1617 Lavandulyl acetate - - 0.4 - a,b,c

47 1620 3,9-Epoxy-p-menth-1-ene - - - 0.2 a,b,c


6


ID HD %

MSD-SPME %

HD %

MSD-SPME %

48 1628 Aromadendrene 0.2 0.7 - - a,b,c

49 1638 cis-p-Menth-2-en-1-ol - - 0.2 - a,b,c

50 1645 6-Acetoxy-3,7-dimethyleneoctene* - 0.9 - - c

51 1651 Bornyl isobutyrate - - 0.2 0.2 a,b,c

52 1655 Isobornyl propionate - - 0.3 0.3 a,b,c

53 1663 cis-Verbenol - - 0.2 - a,b,c

54 1664 trans-Pinocarveol - - 0.3 - a,b,c

55 1668 (Z)-β-Farnesene 2.3 4.9 - - a,b,c

56 1682 δ-Terpineol - - - 0.3 a,b,c

57 1683 trans-Verbenol - - 0.9 0.7 a,b,c

58 1686 Lavandulol - - 1.8 1.4 a,b,c

59 1688 Selina-4,11-diene

(=4,11-Eudesmadiene) 0.6 0.7 0.3 - a,b,c

60 1694 Drima-7,9(11)-diene - - 0.7 0.4 b,c

61 1704 γ-Muurolene 0.4 - - 0.4 a,b,c

62 1706 α-Terpineol 0.4 1.3 5.7 4.8 a,b,c

63 1709 α-Terpinyl acetate 0.2 - - - a,b,c

64 1719 Borneol 0.3 0.6 5.2 5.4 a,b,c

65 1725 Verbenone - - - 0.3 a,b,c

66 1726 Germacrene D - - 1.4 - a,b,c

67 1740 Valencene 3.7 9.2 - - a,b,c

68 1741 β-Bisabolene - - 1.2 0.5 a,b,c

69 1742 β-Selinene 3.3 5.4 - - a,b,c

70 1744 α-Selinene 2.9 5.7 - - a,b,c

71 1744 Phellandral - - 0.5 - a,b,c

72 1755 Bicyclogermacrene 0,3 - - - a,b,c

73 1764 cis-Chrysanthenol - 0.3 1.9 1.1 a,b,c

74 1765 Geranyl acetone - 0.6 - - a,b,c

75 1766 Decanol - - - t a,b,c

76 1773 δ-Cadinene 0.3 - 0.3 - a,b,c

77 1776 γ-Cadinene 0.2 - 0.1 - a,b,c

78 1783 -Sesquiphellandrene - - 0.1 - a,b,c

79 1785 7-epi-α-Selinene 0.2 - - - a,b,c

80 1786 Neryl propionate - - - 0.3 a,b,c

81 1788 ar-Curcumene - - 0.1 - a,b,c

82 1802 Cuminaldehyde 0.4 0.5 9.4 16.0 a,b,c

83 1804 Myrtenol - - 0.3 0.6 a,b,c

84 1806 Methyl salicylate - 0.4 - - a,b,c

85 1823 p-Mentha-1(7),5-dien-2-ol - - 0.1 0.2 a,b,c

86 1828 9-Decen-1-ol - - - 0.2 a,b,c


7


ID HD %

MSD-SPME %

HD %

MSD-SPME %

87 1830 Nootkatene - 0.9 - - a,b,c

88 1831 Citronellyl butyrate - 0.5 - - a,b,c

89 1834 Citronellyl isovalerate 1.2 - - - a,b,c

90 1840 trans-p-Menth-2-en-7-ol - - - 0.9 a,b,c

91 1857 Geraniol - - 0.3 0.3 a,b,c

92 1868 (E)-Geranyl acetone 0.1 - - - a,b,c

93 1870 Hexanoic acid - - - 0.2 a,b,c

94 1871 Neryl isovalerate - - - 0.3 a,b,c

95 1900 Nonadecane 0.3 0.4 - - a,b,c

96 1912 cis-Dihydrocarveol - - 0.9 0.2 a,b,c

97 1921 -Phellandrene epoxide - - 0.4 - a,b,c

98 1933 Neryl valerate - - - 0.1 a,b,c

99 1940 4-Isopropyl salicylaldehyde - - 0.7 0.2 a,b,c

100 1958 (E)--Ionone - 0.3 - - a,b,c

101 1990 Cameroonan-7-α-ol 0.4 - - - b,c

102 1973 1-Dodecanol - - 0.4 - a,b,c

103 1992 Neophytadiene - 0.2 - - a,b,c

104 2000 Eicosane t - - - a,b,c

105 2008 Caryophyllene oxide 1.2 - 1.1 1.7 a,b,c

106 2030 Methyl eugenol - 2,0 - - a,b,c

107 2037 Salvial-4(14)-en-1-one 1.2 - - 0.2 a,b,c

108 2050 (E)-Nerolidol 0.4 0.4 2.0 1.3 a,b,c

109 2055 (8R,8S)-8,8-Dimethyl-3,4,6,7,8,8-hexahydronaphthalen]-2(1H)-one*

- 1.7 - - c

110 2084 Octanoic acid 0.5 0.6 - - a,b,c

111 2100 Heneicosane 0.9 0.3 - - a,b,c

112 2110 Salviadienol - - - 0.4 a,b,c

113 2113 Cumin alcohol - 2.9 3.7 6.9 a,b,c

114 2131 Hexahydrofarnesyl acetone 0.7 - 0.5 - a,b,c

115 2135 Hexadecanal t - - - a,b,c

116 2143 Cedrol 3.3 - - - a,b,c

117 2144 Rosifoliol - - t t a,b,c

118 2146 Spathulenol 3.8 3.3 2.4 3.0 a,b,c

119 2149 -Cedrol - 2.0 - - a,b,c

120 2153 Neointermedeol 1.0 - - - a,b,c

121 2179 1-Tetradecanol - - 0.3 - a,b,c

122 2183 (E)-Sesquilavandulol - - 1.0 0.5 a,b,c

123 2185 1,3,5-Trimethoxybenzene - 0.3 - - a,b,c

124 2186 Eugenol - 0.3 0.6 - a,b,c

125 2188 13-nor-Valenc-1(10)-en-11-one - 1.3 - - c


8


ID HD %

MSD-SPME %

HD %

MSD-SPME %

126 2192 Nonanoic acid - 0.8 0.4 0.5 a,b,c

127 2225 Geranyl--terpinene* - 0.6 0.5 - a,b,c

128 2239 Carvacrol - - - 0.8 a,b,c

129 2241 p-Isopropyl phenol - 1.8 - - a,b,c

130 2245 4-Isopropyl phenol - - 3.0 4.1 a,b,c

131 2247 trans-α-Bergamotol 0.3 - 0.1 - a,b,c

132 2255 α-Cadinol 0.2 - - - a,b,c

133 2257 -Eudesmol - - 0.2 - a,b,c

134 2304 Torilenol (= 1-Hydroxy-6,8-cyclo-4(14)-eudesmene)

0.4 0.3 0.2 - a,b,c

135 2264 Intermedeol (=11-Eudesmol-4) 0.8 - - - a,b,c

136 2273 Selin-11-en-4α-ol 3.1 1.8 - - a,b,c

137 2289 Alismol

(=Guaia-6,10(14)-diene-4-β-ol) 2.3 - - - a,b,c

138 2298 Decanoic acid 5.1 3.1 - - a,b,c

139 2300 Tricosane 1.1 - - - a,b,c

140 2306 9-Geranyl-p-cymene 0.8 0.5 0.3 - a,b,c

141 2316 (Z)-9-Tricosene 2.6 - - - a,b,c

142 2324 Caryophylla-2(12),6(13)-dien-5α-ol (=Caryophylladienol II)

- - - 0.2 a,b,c

143 2325 13-nor-7,8-Epoxy-eremophil-1(10)-en-11-one

- - 0.2 - a,b,c

144 2355 (Z)-Nuciferyl acetate - 0.8 - - a,b,c

145 2368 Eudesma-4(15),7-diene-1-β-ol 0.5 0.3 0.4 0.4 a,b,c

146 2389 Caryophyllenol I

(=Caryophylla-2(12),6-dien-5α-ol) - - - 0.2 a,b,c

147 2396 Caryophyllenol-II

(=Caryophylla-2(12),6-dien-5--ol) - 0.4 - - a,b,c

148 2466 Costol isomer 2.5 - - - c

149 2500 Pentacosane 1.1 - - - a,b,c

150 2510 9-Pentacosene 0.3 - - - a,b,c

151 2515 Coumarin - 1.0 - - b,c

152 2533 γ-Costol 3.9 - - - a,b,c

153 2604 -Costol - - 0.2 - a,b,c

154 2606 β-Costol 1.2 - - - a,b,c

155 2607 1-Octadecanol t - - - a,b,c

156 2607 14-Hydroxy-δ-cadinene t - - - a,b,c

157 2610 Benzophenone t - - - a,b,c

158 2617 Tridecanoic acid t - - - a,b,c

159 2622 Phytol 0.9 - - - a,b,c

160 2655 Benzyl benzoate 0.4 - - - a,b,c

161 2670 Tetradecanoic acid 0.8 - - - a,b,c


9


ID HD %

MSD-SPME %

HD %

MSD-SPME %

(= Myristic acid)

162 2700 Heptacosane 0.5 - - - a,b,c

Total 78.1 80.2 81.7 85.3

RRI: Relative Retention Indices calculated against n-alkanes (C8-C40) on HP-Innowax column; % calculated from FID data; a: Identification based on retention index of genuine compounds on the HP-Innowax column; b: Identification on the basis of computer matching of the mass spectra from Başer Library; c: Tentative identified on the basis of computer matching of the mass spectra from Adams, Mass Finder, Wiley and NIST libraries; t: Trace (< 0.1 %).

Comparison of the obtained results with the literature data (Xiao, et al., 2008; Bicchi, et al., 1985) showed

the considerable differences in the chemical composition of A. rupestris essential oil extracted by various

hydrodistillation methods. This difference in chemical composition of A. rupestris essential oil could be due

to the growth locations of the wormwood. However, there was no significant difference observed in the

composition of essential oil from A. glabella extracted by various methods.

Thus, the plant volatile compounds from A. glabella and A. rupestris have been extracted for the first time

using the MSD-SPME technique. This method is the fast, cost-effective and economical in terms of raw

materials; it also reduces the extraction time, while, allowing us to extract the maximum amount of volatiles.

REFERENCES

Adekenov, S.M., Mukhametzhanov, M.N., Kagarlitskii, A.D. & Kupriyanov, A.N. (1982). Arglabin — A new sesquiterpene lactone from Artemisia glabella. Chemistry of Natural Compounds, 18(5), 623-624.

Adekenov, S.M., Mukhametzhanov, M.N., Kagarlitsky, A.D. & Agashkin, O.V. (1983). Sesquiterpene lactones of Artemisia glabella. Bulletin of AS of KazSSR (chemical series), 2, 54-60. (in Russian)

Adekenov, S.M., Turdybekov, K.M., Aituganoav, K.A., Lindeman, S.V., Struchkov Yu.T. & Shaltakov, S.M., (1993). 1β,10α-Dihydroxyarglabin-a new sesquiterpene lactone from Artemisia glabella. Chemistry of Natural Compounds, 29(6), 735-736.

Atazhanova, G.A., Dembitskii, A.D., Zhizhin, N.I. & Adekenov, S.M. (1999). Chemical composition of the essential oil of Artemisia glabella. Chemistry of Natural Compounds, 35(2), 172-175.

Atazhanova, G.A. (2008). Terpenoids of plant essential oils. Moscow, 286. (in Russian)

Bicchi, C., Fratmni, C.& Sacco, T. (1985). Essential oils of three Asiatic Artemisia Species. Phytochemistry, 24(10), 2440-2442.

European Pharmacopoeia (2017). Determination of Essential Oils in Herbal Drugs, 2.8.12. 9th ed. European Pharmacopoeia. European Directorate for the Quality of Medicines and Healthcare. Strasbourg: 285-286.

Kulmagambetova, E.A., Pribytkova, L.N. & Adekenov, S.M. (2000). Flavonoids of Artemisia glabella. Chemistry of Natural Compounds, 36(1), 95-96.

Liu, X.C., Li, Y.P., Li, H.Q., Deng, Z.W., Zhou, L., Liu, Z.L. & Du, S.S. (2013). Identification of repellent and insecticidal constituents of the essential oil of Artemisia rupestris L. aerial parts against Liposcelis bostrychophila badonnel. Molecules, 18, 10733-10746.

Pavlov N.V. (1966). Flora of Kazakhstan, Vol. 9. – Almaty. 127. (in Russian)

Seidakhmetova, R.B., Beisenbaeva, A.A., Atazhanova, G.A., Suleimenov, E.M., Pak, R.N., Kulyyasov, A.T. & Adekenov, S.M (2002). Chemical composition and biological activity of the essential oil from Artemisia glabella. Pharmaceutical Chemistry Journal, 36(3), 135-138.

Xiao, W., Sirafil, A., Li, Z. & Lishizhen, J. (2008). Rapid screening of anti-HIV ingredients in Artemisia rupestris L. extracts interacting with V3 loop region of HIV1 gp120 and reverse transcriptase by affinity capillary electrophoresis and capillary zone electrophoresis. Med. Mater. Med. Res. 19, 2836-2838.

https://link.springer.com/article/10.1007/BF02234927



Nat. Volatiles & Essent. Oils, 2018; 5(2): 10-18 Köse and Bayraktar

10

RESEARCH ARTICLE

Valorization of Citrus Peel Waste

Merve Deniz Köse1 and Oguz Bayraktar1*

1 Chemical Engineering Department, Ege University, 35100, İzmir, TURKEY


Abstract

Citrus sinensis commercially known as an orange tree has a high pectin ratio and valuable essential oils. The huge amount of orange

peel is generated in industries and they can be used as a raw material for essential oils and pectin. To obtain essential oils from

orange peels distillation method is used and collected oil is analyzed to determine its d- limonene amount, also the yield of the

essential oil production is calculated. The essential oil yield was found as 0.19%. GC results revealed that the sample had 94% d-

limonene, and the data from the literature indicated that d-limonene amount of orange essential oil may vary between 32% and 98%

depending on the variety of orange. In addition to essential oil, pectin was also extracted from the orange peels using two types of

acids. First one was hydrochloric acid (HCl) as an inorganic acid, and the other was tartaric acid (TA) as an organic acid. The purpose

of selecting the several types of acid was to observe the effects of acid type on pectin yield. The pectin amount obtained with HCl

usage (4.72 % g pectin/orange fresh peel) was higher than that with TA usage (4.037 % g pectin/orange fresh peel). The experimental

results for pectin contents of samples were also confirmed with the colorimetric test results. According to these results,

concentrations of galacturonic acid in the samples obtained with HCl (0.509 mg galacturonic acid/mg) was found to be higher than

the concentrations of galacturonic acid in the samples obtained with TA (0.103 mg galacturonic acid/mg).

Keywords: Essential oils, valorization, extraction of pectin, orange peel

Introduction

Citrus is one of the most important fruit crops in the world. Production of citrus fruits has increased

enormously in the last few decades, going from an average of 62 million tons a year in the period 1987–1989

to about 100 million tons in the year 2010. Citrus is grown in more than 100 countries all over the world,

mainly in tropical and subtropical areas, where favorable soil and climatic conditions prevail for citrus

cultivation. Citrus fruits are marketed mainly as fresh fruit or as processed juice. During processing of citrus

fruits, a huge amount of peels is generated as a by-product, which does not add value to the product as these

are discarded or dumped. The potential use of citrus peels as value-added products has been widely studied

because it contains numerous biologically active compounds including natural antioxidants such as phenolic

compounds (Hayat et al, 2010).

Citrus waste includes more than half of the whole fruit when processed for juice extraction and mainly

consists of:

• waste generated by the juice manufacturing industry, consisting of peel and pressed pulp

• fruit discarded for commercial reasons (damaged fruit, as an example)

• fruit discarded due to regulations that limit production

All these materials considered as waste because they are not part of food chain. In juice extraction process

produces 500 tons of waste per 1000 tons of fruit processed. The percentage of fruit discarded due to

commercial or regulatory issues are more difficult to calculate, but it ranges from 2% to 10% depending on

the type of citrus considered and environmental aspects, such as weather conditions. Citrus waste generally


11

has a low pH (3-4), high water content (80-90%) and high organic matter content (95% of total solids) (Ruiz

and Flotats, 2014).

Orange waste is produced in high quantities all over the world. In orange juice production, only half of the

fresh orange weight is transformed into juice. Generating excessive amounts of residue (peel, pulp, seeds,

orange leaves and whole orange fruits that do not reach the quality requirements), which accounts for the

other 50% of the weight of the fruit and has a moisture content of approximately 82 g /100 g.

These wastes cause contamination in areas adjacent to the production locations, for its final use as a raw

material in animal feed, or else it is burned. Management of this waste is important. One alternative to

improve the management of these residues is the implementation of new processes for their recovery, for

instance, through the production of organic fertilizers, pectin, bio-oil, essential oils, and antioxidant

compounds, or as a substrate to produce several compounds with high added value, such as microbial

proteins, organic acids, ethanol, enzymes and biologically active secondary metabolites and adsorbent

materials. These are excellent alternatives to avoid environmental pollution and to add value to these

substances (Rezzadoria et al., 2012).

Citrus Essential Oils (CEO) are liquids that contain, among other components, the volatile aroma compounds

of citrus plants. The essential oils present in small vesicles which located in the flavedo (the upper shell of

fruit) or exocarp of citrus fruit. Antimicrobial properties of CEOs are discovered in old times and citrus

essential oils are used as natural healing (Ruiz and Flotats, 2014). Citrus fruits have oval, balloon-shaped oil

sacs, glands or vesicles, the diameter of which varies from 0.4 to 0.6 mm. The essential oil presents in that.

Ductless, and without communication with surrounding cells or the exterior, they have no proper walls but

are simply bounded by the debris of degraded tissue (Board, 2011). Citrus essential oils have many

components (more than 200) including terpenes, sesquiterpenes, aldehydes, alcohols and esters, and can be

described as a mixture of terpene hydrocarbons, oxygenated compounds, and non-volatile residues.

Terpenes are unsaturated compounds that readily decay by light, heat, and oxygen. Removing of terpenes

avoids unpleasant flavours; they make up about 80-98% in most citrus peel oils (Diaz et al., 2005). Limonene

is the main volatile component of CEOs because of this the chemical, physical and biological properties of

this compound influence the properties of the essential oil. Its concentration in the essential oil may vary

between 32% and 98%, depend on the variety: 32-45% in bergamot, 45-76% in lemon and 68-98% in sweet

orange (Ruiz and Flotats, 2014).

In addition to the citrus essential oils, citrus sinensis has high pectin content. Pectin, which is a valuable

product, can be used for different areas for different purposes. It has long been used for its gel-forming,

thickening and stabilizing properties in a wide range of applications from food to the pharmaceutical and

cosmetic industries.

Pectin is naturally found in the structure of cell walls of all higher plants. The outer surface of plants is

especially rich in pectin. Fruit peels are well known and used as pectin sources for industrial applications.

Fruit peels are a rich source of rough dietary fibers. Pectin, hemicellulose, tannins, gum can be given as

examples of these fibers. The fiber compounds give bulkiness into the food and help preventing constipation

by reducing gastrointestinal transit time. Besides, they link to toxin chemicals in the food, in this way they

protect them contacting with gut mucosa and thereby help cut-down colon cancer risks. Moreover, they link

tightly to bile salts, which are produced from cholesterol, and eliminate the salts from the gut, thus,

accordingly help lower serum LDL cholesterol levels (Joye and Luzio, 2000).


12

Pectin is a complex mixture of acidic structural polysaccharides in cell walls of all land plants, which is mainly

formed of d-galacturonic acid and neutral sugars, such as l-rhamnose, l-arabinose, and d-galactose. D-

galacturonic acid units that are partially esterified with methanol or acetic acid at the carboxylic acid

(BeMiller, 1986). In this study, the valorization of orange peel wastes for the recovery of essential oil and

pectin were aimed. Extraction of pectin in orange peel was studied using two different acids including

hydrochloric acid and tartaric acid. Pectin yields were compared. The obtained pectin samples were further

characterized.


Materials

The fresh oranges (Citrus sinensis) were bought from local grocery stores in Izmir/Turkey, in January to March

2016. All the reagents used in the experiment were in analytical grade and used without further purification.

Ethanol %96 (Analiz Kimya, Turkey), HCl %37 (Sigma-Aldrich), tartaric acid (Sigma-Aldrich), H2S04 %98 (Merck,

Germany), D-(+)-Galacturonic acid (Sigma – Aldrich), 3-Phenylphenol, 85% (Aldrich), NaOH (Aldrich), sodium

tetraborate (Borax) (Merck, Germany).

Methods

Preparation of orange peel for hydrodistillation

Experimental set up was insulated before to reduce heat losses. Orange peel waste was subjected to

hydrodistillation. Distillation was performed twice to enhance the recovery of volatile fraction at the end of

the process. In the first run, distillate was collected then collected distillate was distilled again.

In the product emulsion of water and essential oil was formed and after a short period of time essential oil

and water were separated into two distinct phases. The amount of essential oil was measured with the help

of Clevenger apparatus’s measurement section. D-limonene amount in the essential oil was determined by

using Gas Chromatography (GC) equipment (Agilent 7890A) with a FID. The samples (20µl) were injected into

the injection port. A capillary column HP-5 (30m x 320μm x 0.25 μm film thickness) (Agilent) was used for

chromatographic separation. The used temperature program was 5 min at 50 °C isothermal and an increase

of 5 °C/min to 200 °C. Helium was used at 2ml/min as the carrier gas. The temperature of injector and

detector was 250 and 270 °C, respectively.

Preparation of orange peel for extraction

Fresh oranges were washed with deionized water and then dried. Cleaned and dried oranges were peeled

off. After that, orange peels were diced into small and fine pieces. The diced pieces were treated with 96%

ethanol, which was preheated to 65 °C, in the ratio of 1:2.5 (w/v). In this treatment procedure, firstly, the

diced orange peels and 96% ethanol are mixed in a beaker for one hour at 65 °C. Then, the diced peel –

ethanol mixture was kept at room temperature overnight. After that, the mixture was filtered by hand

through muslin cloth, after which the insoluble materials were washed twice with warm 96% ethanol. The

remaining solids or alcohol insoluble solids (AIS) given in Figure 1 were dried at 60 °C in an oven and stored

until use.


13

Figure 1. The picture of prepared alcohol insoluble solids (AIS)

Extraction and purification of pectin

The extraction of orange peel and its purification methods was adapted from the method for isolation,

characterization and modification of citrus pectin, as described in the literature (Georgiev et al.,2012).

The extraction steps were done in two parts. The first part was done with water, and then the second part

was done with selected acid, which was either hydrochloric acid or tartaric acid. Firstly, AIS was treated with

hot deionized water (1:25), (w/v) at 82 °C for 1 hour with continuous stirring and then filtered. Then, the

retentate was treated with hot deionized water (1:10), (w/v) at 82 °C for 10 minutes with continuous stirring

and then filtered.

To obtain water extracted pectin (WEP), the solutes from water extracted operation was purified. Since

pectin dissolves in hot water, the solutes were coagulated with cold acidic 96% ethanol (0.5% HCl), in the

ratio of 1:2 (v/v). The precipitated crude pectin was separated by filtration, washed once with 100 mL of 70%

acidic ethanol, then with 70% ethanol to a neutral pH and finally with 100 mL of 96% ethanol. Pectin samples

were dried at room temperature in fume hood. Thus, water extracted pectin (WEP) was obtained.

Also, for the recovery of acid extracted pectin (AEP), similar steps for the recovery of WEP were followed.

Acid-extracted pectin continued with the solutes obtained from water extraction. The residue was treated

with 0.5% acid (HCl and TA), with 1:20 ratio (w/v) at 82 °C for 50 min and continuously stirred at pH 1.7. Then,

the acidic mixture was filtered and the solid retentate was treated with 0.5% HCl (1:8), (w/v) at 82 °C for 10

min and continuously stirred at pH 1.7. The only difference in that the ethanol in purification method was

not acidic. Since the solutes came from the acidic extraction step, there was no need to acidify the ethanol

in Figure 2 end products of extracted and purified pectin was given.

Figure 2. The picture of extracted and purified pectin samples


14

Determination of pectin substances

To determine the pectin content of extracted orange peels, the colorimetric method was used. The principal

of this method is based on the reaction of the galacturonic acid with a reagent material, m-hydroxydiphenyl.

If there is galacturonic acid in the sample, the reagent gives a pinkish color to the sample. Due to this reaction,

pectin content of the samples can be determined by measuring the absorbance values using a

spectrophotometer.

In the experiments, anhydrogalacturonic acid (AGA) content was determined by the m-hydroxydiphenyl

method, using D-GalA as a standard.

Firstly 0.2 ml of sample, which contained 0.5 to 20 μg uronic acids, was prepared. Then, 1.2 ml of sulfuric

acid/tetraborate was added. The tubes were refrigerated in crushed ice. The mixture was shaken regularly,

and the tubes were heated in a water bath at 100°C for 5 min. Then, the tubes were cooled in a water-ice

bath. After the tubes are cooled, 20 μl of the m-hydroxydiphenyl reagent was added into each of them. The

tubes were shaken properly, and within 5 min, absorbance measurements made at 525 nm in a UV

spectrophotometer. As carbohydrates produce a pinkish chromogen with sulfuric acid/ tetraborate at 100

°C, a blank sample was run without addition of the reagent, which was replaced by 20 μl of 0.5% NaOH. So,

the absorbance of the blank sample was subtracted from the total absorbance (Blumenkrantz and Asboe-

hansen, 1973).

Figure 3. Different samples of colorimetric methods.

FT-IR analysis

The pectin from orange peel was further investigated by using FT-IR analysis and the resulting spectrum was

studied in order to understand the functional groups present. The dried pectin samples were ground with

KBr at a 1/100 ratio (w/w). The powders were pelletized, and then the infrared spectra were obtained. The

spectra of the samples were recorded in the 4000 - 650 cm-1 region at room temperature.


Determination of yield of essential oil

One of the aims of valorization of orange peel waste was obtaining essential oil from orange peel with a high

yield as much as possible. From 73 g fresh orange peel and 200 ml water, 0.14 ml orange oil was obtained.


15

The yield was calculated as 0.19%. Calculated yield value was in the range of values reported by other

researchers in the literature.

Yield depends on the season of harvesting, plant variety, the plant parts sampled, and the conditions under

which the plant is grown. So, obtained yield can be affected by all these parameters. Also, composition of

essential oil can vary with these parameters.

Figure 4. Gas Chromatography-Mass Spectrometry of Orange Peel Essential Oil

Figure 5. Gas chromatogram of Orange Peel Essential Oil

Results show that sample has 94% d-limonene, and from the literature survey d-limonene amount of orange

oil may vary between 32% and 98% depending on variety of the orange.

The size of the waste peel was changing a parameter in the experiment, in the first run peels were cut into

small pieces but results were ineffective. Because oil sacs place on the peel and when the peels were cut,


16

they were destroyed, and oil would escape. With this reason, the remaining runs of the experiment, peels

were separated from white part and the orange outer surface was used as possible. Results were better than

the first run.

For a collection of essential oil, the distillate from first distillation run was fed back into the system and at

the end of second distillation, oil droplets were observed clearly. After waiting for a brief period, oil droplets

have created a layer on the water surface. Separation was achieved easily due to the density differences

between oil and water.

Characterization of extracted pectin samples

In order to characterize the pectin samples, FT-IR spectra were obtained for acid and water extracted pectin

material and results are given in Figure 6.

Figure 6. FT-IR Spectra of Extracted Pectin Samples: (ahcl: Acid Extracted Pectin with HCl; ata: Acid Extracted Pectin

with TA; wta: Water Extracted Pectin with TA; whcl: Water Extracted Pectin with HCl)

The broad, strong area of absorption between 3600 and 2500 cm-1 refers to O-H stretching absorption due

to intermolecular and intramolecular hydrogen bonds. The O-H stretching vibrations occur within a broad

range of frequencies and indicate several features of a compound, including free hydroxyl groups stretching

bands which occur in samples in the vapour phase and bonded O-H bands of carboxylic acid (Silverstein et

al., 1991). In the case of pectin samples, absorption in the O-H region is due to the intermolecular and

intramolecular hydrogen bonding of the galacturonic acid polymer. Finer bands appearing at the longer end

of the O-H region indicate overtones and combination of tones. Bands around 2950 cm-1 (3000-2800 cm-1)

refer to C-H absorption. These include CH, CH2, and CH3 stretching and bending vibrations. Typically, two

moderately intense bands are observed in the C-H region of aliphatic compounds (Gnanasambandam and

Proctor, 2000).

In pectin samples, the C-H stretching and bending vibrations were seen, usually, as a band superimposed

upon the broader O-H band that ranges from 2500 to 3600 cm-1. This was observed with all pectin samples

studied. In pectin samples, the weaker symmetric COO- stretching is followed by moderately intense

absorption patterns between 1300 and 800 cm-1 collectively referred to as the “fingerprint” region that is


17

unique to a compound. These bands are usually difficult to interpret. As seen from the Figure 6 all the

extracted pectin samples are compatible with each other and with the literature survey.

Determination of pectin content

For 500 grams fresh orange peel, 77 grams alcohol insoluble material (AIS) was obtained. For 100 g fresh

orange peel, 15.4 g AIS was obtained. These values are in accordance with the results reported in the

literature (Georgiev et al., 2012, Kaya et al., 2014).

At the end of the extraction and purification steps of each set, purified pectin samples were dried in a fume

hood at room temperature. After drying process, pectin samples were used for the preparation of pellets.

These pellets form of pectin samples were weighed, and with this weight measurement values, g pectin / g

fresh orange peel ratio was calculated for each set. The data are tabulated in Table 1.

Table 1 Pelletized pectin samples prepared by using hydrochloric acid and tartaric acid.

HCl TA

WEP, g 1.773 1.360

AEP, g 4.124 3.685

Total Pectin, g 5.896 5.046

%(g pectin)/(orange fresh peel) 4.7170 4.0367

According to data given in Table 1, it was shown that amount of pectin extracted with HCl (4.72 % g

pectin/orange fresh peel) was higher than amount of pectin extracted with TA (4.037 % g pectin/orange fresh

peel). The colorimetric method was used to determine the amount of pectin substances in WEP and AEP

samples in terms of galacturonic acid equivalent concentrations.

In the colorimetric method, the coloring reagent, which was m-hydroxydiphenyl, gives a reaction in the

presence of galacturonic acid resulting in a pinkish color in the sample. The concentration of pectin

substances in the samples was calculated by using a UV spectrophotometer. One g AEP sample contains

0.509 g galacturonic acid equivalent and 0.103 g galacturonic acid equivalent for HCl and TA sets,

respectively. Also, one g of WEP sample contains approximately 0.383 g galacturonic acid equivalent (gae).

All these values determined as galacturonic acid equivalent are given in Table 2.

Table 2. Pectin substances content of pectin samples

g gae/ g sample

WEP from HCl set 0.424

0.342

0.509

0.103

WEP from TA set

AEP from HCl set

AEP from TA set

The yields of extraction with inorganic acids were higher than those with organic acids. However, considering

the undesired properties of inorganic acids, the results obtained with tartaric acid were comparable to the

ones obtained with hydrochloric acid. In addition to the less harmful and toxic properties, tartaric acid which

occurs naturally in many plants and can be recovered from various natural resources, mainly from winery

byproducts. Other sources of tartaric acid are biotechnological processes or synthesis via the peroxidation of

maleic anhydride (Kontogiannopoulos et al., 2016), another waste product of many industries. It is possible


18

to utilize the waste of food industry such as tartaric acid for the feasible production of value-added products

such as pectin from waste peel of oranges in juice industry.

Conclusion

Produced waste amounts increase with urbanization, correspondingly consumption growth. Waste recovery

is an important and urgent issue. Moreover, since fruit industry grows day by day, generated fruit wastes

increase. Large amounts of fruit wastes are probably caused a pollution problem in the case of improper

management of these wastes. If the wastes are effectively managed to produce value-added products

economic benefits along with reduced environmental problems can be achieved. Orange juice is one of the

big juice sectors. From these sectors, large amount of wastes especially orange peels are discarded. However,

orange peel wastes can effectively be utilized for the economic production of value-added products such as

pectin. Our results revealed that it was possible to produce value-added products from the orange peel

wastes such as essential oil and pectin.

REFERENCES

BeMiller, J. N. (1986). An Introduction to Pectins: Structure and Properties. Chemistry and Function of Pectins Fishman

and Jen; Chapter 1, 2–12.

Blumenkrantz, N., Asboe-Hansen, G. (1973). New method for quantitative determination of uronic acids. Analytical

Biochemistry 54, 484-489.

Board, N., 2011. The Complete Technology Book of Essential Oils (aromatic Chemicals). India: National Institute of

Industrial Research.

Diaz, S., Espinosa, S., Brignole, E., (2005). Citrus peel oil deterpenation with supercritical fluids optimal process and

solvent cycle design. Supercritical Fluids, 35, 49-61.

Hayat, K., Zhang, X., Chen, H., Xia, S., Jia, C., Zhong, F., (2010). Liberation and separation of phenolic compounds from

citrus mandarin peels by microwave heating and its effect on antioxidant activity. Separation and Purification

Technology 73, (3), 371–376.

Georgiev, Y., Ognyanov, M., Yanakieva, I., Kussovski, V., Kratchanova, M. (2012). Isolation, characterization, and

modification of citrus pectins. Journal of BioScience and Biotechnology, 1 (3), 223-233.

Gnanasambandam, R., Proctor, A., (2000). Determination of pectin degree of esterification by diffuse reflectance Fourier

transform infrared spectroscopy. Food Chemistry 68 (3), 327–332.

Joye, D.D., Luzio, G.A., (2000). Process for Selective Extraction of Pectins from Plant Material by Different pH.

Carbohydrate Polymers 43, 337-342.

Kaya, M., Sousa, A. G., Crépeau, M.-J., Sørensen, S. O., & Ralet, M.-C. (2014). Characterization of citrus pectin samples

extracted under different conditions: influence of acid type and pH of extraction. Annals of Botany, 114 (6), 1319–1326.

Kontogiannopoulos, K.N., Patsios, S. I., Karabelas, A. J. (2016). Tartaric Acid Recovery from Winery Lees Using Cation

Exchange Resin: Optimization by Response Surface Methodology. Separation and Purification Technology 165, 32–41.

Rezzadoria, K., Benedettia, S., Amanteb., E.R. (2012). Proposals for the residues recovery: Orange waste as raw material

for new products. Food and Bioproducts Processing, 90, 606-614.

Ruiz, B., Flotats, X., (2014). Citrus Essential Oils and Their Influence on the Anaerobic Digestion Process: An Overview.

Waste Management, 34, 2063-2079.

Silverstein, R. M., Bassler, G. C., Morril, T. C. (1991). Infrared spectroscopy. In Spectrometric identification of organic

compounds (5th ed., pp. 91-164). New York: JohnWiley & Sons, Inc. (Chapter3).

Nat. Volatiles & Essent. Oils, 2018; 5(2): 19-23 Aburwais et al.

19

RESEARCH ARTICLE

Essential oil composition of Zosima absinthifolia (Vent.) Link from Northern Cyprus

Omar Aburwais1, Azmi Hanoglu1, Betül Demirci2 and K. Hüsnü Can Başer1,*

1 Department of Pharmacognosy, Near East University, Faculty of Pharmacy, Lefkoşa, N. Cyprus, Mersin 10, TURKEY 2 Department of Pharmacognosy, Anadolu University, Faculty of Pharmacy, Eskişehir, TURKEY


Abstract

Hydrodistilled essential oils from dried fruits of Zosima absithifolia (Vent.) Link (Apiaceae) collected from Northern Cyprus was

analyzed by Gas Chromatography-Flame Ionization Detector (GC-FID) and Gas Chromatography/Mass Spectrometry (GC/MS). Octyl

acetate (63.2-59.5%), octyl hexanoate (19.8-18.6%), octyl octanoate (9.9-9.2%) and octanol (7.1-2.2%) were characterized as main

constituents.

Keywords: Zosima, Apiaceae, Essential Oil

Introduction

Zosima absinthifolia (Vent.) Link is a perennial herb found in the family Apiaceae. This plant is a widely

distributed from Iran to Turkey, Central Asia, Afghanistan and Pakistan. It normally grows in fields, steppe,

and lime stone slopes at an altitude of the range 400-2000 m (Davis, 1972).

It is known as “Peynir otu” or “Ayı eli” in Turkey and used as digestive, antiinflammatory and carminative in

Turkey (Bahadir et al., 2010). It is a cheese ingredient and is eaten as food after cooked (Aksakal & Kaya,

2008; Ozcelik, 1994). In Pakistan, it is used for bowel disorders and in the treatment of cough (Goodman &

Ghafoor, 1992).

While an ethanolic extract showed antimicrobial activity (Al-Shamma & Mitscher 1979), a methanolic extract

possessed anti-oxidative, phytotoxic and anti-proliferative activities (Razavi et al., 2008). Essential oil was

also shown to have antibacterial activity (Najed-Ebrahim & Razavi, 2008).

Isolation of coumarins and alkaloids have been reported (Crowden et al., 1969; Razavi & Samad, 2009; Razavi

et al., 2013).

Previously, Başer et al. (2000) and Razavi et al. (2009) reported essential oil composition of fruits from Turkey

and Iran, respectively (Başer et al., 2000) (Razavi et al., 2009).

Here, we report on the chemical composition of fruit essential oils collected from Alevkayası near Girne

(Kyrenia) from Northern Cyprus. To the best of our knowledge, this is the first report of Zosima from Cyprus.


Fruits of Zosima absinthifolia were collected from Alevkayası on June 1, 2016 and from the garden of

Armenian Monastery at Alevkayasi on May 3, 2916. The plants were identified by one of us (KHCB). Voucher

specimens were deposited in the Herbarium of Near East University (NEUN 6887 and 6894). Dried fruits were

hydrodistilled using a Clevenger-type apparatus for 3 h. Oil yield was 0.05% on moisture-free basis.


20

Gas Chromatography Mass Spectroscopy (GC-MS)

The essential oils of Zosima absinthifolia were characterized using GC-MS system. The GC-MS analysis was

carried out with an Agilent 5975 GC-MSD system. Innowax FSC column (60 m x 0.25 mm, 0.25 mm film

thickness) was utilized with helium as carrier gas (0.8 ml/min). GC oven temperature was kept at 60°C for 10

min and programmed to 220°C at a rate of 4°C/min, and kept constant at 220°C for 10 min and then modified

to 240°C at a rate of 1°C/min. Split ratio was adjusted at 40:1. The injector temperature was set at 250°C.

Mass spectra were recorded at 70 eV. Mass range was from m/z 35 to 450.

Gas Chromatography Flame Ionization Detector (GC-FID)

The GC analysis was carried out using an Agilent 6890N GC system. FID detector temperature was 300°C. To

obtain the same elution order with GC-MS, simultaneous auto-injection was done on a duplicate of the same

column applying the same operational conditions. Relative percentage amounts of the separated compounds

were calculated from FID chromatograms Identification of the essential oil components were carried out by

comparison of their relative retention times with those of authentic samples or by comparison of their

relative retention index (RRI) to series of n-alkanes. Computer matching against commercial (Wiley GC/MS

Library, MassFinder 3 Library) (Mac Lafferty & Stauffer, 1989) and (Koenig et al., 2004) and in-house “Başer

Library of Essential Oil Constituents” built up by genuine compounds and components of known oils, as well

as MS literature data (Koenig et al., 1998; Boelens, 1999) were used for the identification.


The oils were analyzed by GC-FID and GC/MS. The Girne/ Alevkayası Armenian Monastery sample showed 32

compounds representing 99.5 percent of the oil. Octyl acetate (63.2%), octyl hexanoate (18.6%), octyl

octanoate (9.2%) and octanol (2.2%) were characterized as main constituents. Octyl acetate (63.2%), octyl

hexanoate (18.6%), octyl octanoate (9.2%) and octanol (2.2%) were characterized as main constituents.

Girne/ Alevkayası sample similarly showed 14 compounds representing 99.6 percent of the total oil. Octyl

acetate (59.5%), octyl hexanoate (19.8%), octyl octanoate (9.9%) and octanol (7.1%) were characterized as

major constituents (Table 1 and 2).

There are two previous reports on the essential oil composition of Z. absinthifolia fruits. Başer et al. (2000)

reported that octyl acetate (38.9%), octyl hexanoate (31.9%) and octanol (12.9%) were the main constituents

in the oil of Z. absinthifolia collected in Turkey. Razavi et al. (2009) reported the essential oil composition of

Z. absinthifolia fruits of Iran origin as octyl acetate (87.5), Octyl octanoate (5.0%), octanol (2.4%), hexyl

hexanoate (1.5%) and octanoic acid (1.1%) as main constituents. Our results are in accordance with the

previous reports.

Major constituents octyl acetate, octyl hexanoate, octyl octanoate and octanol have previously been

reported as main constituents from other Heracleum oils. Hajhashemi et al. (2009) reported octyl acetate

(16.5%) as main constituent of fruit oil of H. persicum Desf. ex Fisch. (Hajhashemi et al., 2009).

1-Octanol (13.6% and octyl hexanoate (8.1%) were reported as main constituents of the Heracleum sibiricum

L. oil. Its antibacterial activity was also investigated (Miladinovic et al., 2013).

Antibacterial activity and chemical composition showing octyl acetate (93.7%) as main constituent of the

essential oils of Heracleum sphondylium L. subsp. ternatum (Velen) Brummit were reported (Iscan et al.,

2004).


21

Table 1: Essential oil composition of Z. absinthifolia collected from Girne/ Alevkayası in Armenian Monastery location

LRI Compound %

1032 α-Pinene tr

1048 2-Methyl-3-buten-2-ol 0.1

1093 Hexanal 0.1

1194 Heptanal 0.1

1244 2-Pentyl furan tr

1255 -Terpinene 0.1

1280 p-Cymene tr

1296 Octanal 0.3

1327 3-Methyl-2-butenol tr

1400 Nonanal 0.1

1483 Octyl acetate 63.2

1516 (Z)-4-Octenyl acetate 0.5

1535 -Bourbonene tr

1562 Octanol 2.2

1612 -Caryophyllene 0.5

1623 Octyl butyrate 1.0

1634 Octyl 2-methyl butyrate 0.1

1726 Germacrene D 0.1

1755 Bicyclogermacrene 0.1

1829 Octyl hexanoate 18.6

1856 (Z)-4-Octenyl hexanoate 0.1

1893 Octyl heptanoate tr

2008 Caryophyllene oxide 0.2

2020 Octyl octanoate 9.2

2084 Octanoic acid 0.3

2144 Spathulenol 0.1

2298 Decanoic acid 0.1

2300 Tricosane 0.4

2500 Pentacosane 0.1

2503 Dodecanoic acid 0.7

2670 Tetradecanoic acid 0.8

2931 Hexadecanoic acid 0.4

Total 99.5

LRI: Linear retention indices calculated against n-alkanes. %: calculated from FID data. tr: Trace (<0.1%)


22

Table 2: Essential oil composition of Z. absinthifolia fruits collected from Girne/ Alevkayası location

LRI Compound name %

1303 Octanal 0.3

1487 Octyl acetate 59.5

1522 (Z)-4-octenyl acetate 0.5

1567 Octanol 7.1

1611 (Z)-3-octen-1-ol tr

1628 Octyl butyrate 1.7

1638 Octyl-2-methyl butyrate tr

1743 7-epi-1,2-dehydro-sesquicineole 0.1

1757 Octyl angelate 0.1

1824 Octyl hexanoate 19.8

1862 (Z)-4-octenyl hexanoate 0.1

2022 Octyl octanoate 9.9

2063 3-Octyl octenoate 0.3

2243 α-Bisabolol 0.1

Total 99.6

LRI: Linear retention indices calculated against n-alkanes. %: calculated from FID data. tr: Trace (<0.1%)

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NVEO 2018, Volume 5, Issue 2nveo.org/wp-content/uploads/2019/03/NVEO-2018-Volume-5... · 2019-06-17 · Nat. Volatiles & Essent. Oils, 2018; 5(2): 1-9 Shaimerdenova et al. 2 A. rupestris