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Evaluation of several Rosa damascena varieties andRosa bourboniana accession for essential oil contentand composition in western HimalayasRakesh Kumara, Saurabh Sharmaa, Swati Sooda, Vijai K. Agnihotria, Virendra Singha & BikramSingha

a CSIR-Institute of Himalayan Bioresource Technology (Council of Scientific and IndustrialResearch), Palampur, 176 061, India.Published online: 12 Aug 2013.

To cite this article: Rakesh Kumar, Saurabh Sharma, Swati Sood, Vijai K. Agnihotri, Virendra Singh & Bikram Singh (2014)Evaluation of several Rosa damascena varieties and Rosa bourboniana accession for essential oil content and composition inwestern Himalayas, Journal of Essential Oil Research, 26:3, 147-152, DOI: 10.1080/10412905.2013.829004

To link to this article: http://dx.doi.org/10.1080/10412905.2013.829004

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Evaluation of several Rosa damascena varieties and Rosa bourboniana accession for essentialoil content and composition in western Himalayas

Rakesh Kumar*, Saurabh Sharma, Swati Sood, Vijai K. Agnihotri, Virendra Singh and Bikram Singh

CSIR-Institute of Himalayan Bioresource Technology (Council of Scientific and Industrial Research), Palampur 176 061, India

(Received 29 September 2012; accepted 6 July 2013)

Field experiments were conducted at CSIR-Institute of Himalayan Bioresource Technology, Palampur, India, to studythe performance of different varieties of rose oil species in western Himalayas. Five varieties of Rosa damascena,namely Indica, Jwala, Super jwala, Himroz, Hot himroz and one accession of Rosa bourboniana were investigatedfor essential oil content and chemical composition. The rose oil was obtained by hydrodistillation in Clevenger-typeapparatus and the components in the oil were analyzed by gas chromatography–flame ionization detector (GC–FID)and GC–mass spectrometry (GC–MS) analysis of volatile compounds. The essential oil content of the varieties ofR. damascena varied from 0.037% to 0.051% and that of R. bourboniana was 0.017%. Super jwala recorded thehighest oil content (0.051%). A total of 32 components were identified in the different varieties of rose oil species.These components constituted 78.1–93.5% of the total rose oil. The main components of rose oil were citronellol +nerol (16.3–30.1%), geraniol (15.8–29.3%), linalool (0.7–1.9%), rose oxide (0.9–2.6%), phenyl ethyl alcohol (0.1–0.4%), eugenol (0.3–2.2%), nonadecane (7.3–14.7%). The content of citronellol + nerol (30.1%) and geraniol(29.3%) was the highest in Himroz compared with other varieties.

Keywords: Rosa damascena; Rosa bourboniana; varieties; essential oil; composition

Introduction

The genus Rosa, which belongs to the Rosaceae family,includes around 200 species and more than 18,000cultivars (1). Roses have been also used in food,perfumery and cosmetic industries for many years.Different species of rose, namely Rosa damascena(Damask rose) and Rosa bourboniana (Edward rose)are commonly used for extraction of essential oil androse water production. The flowering in R. bourbonianais sporadic and it flowers three times a year, i.e.April–May, July–August and mid-October; however,April–May is the main flowering period (2). It is exclu-sively grown for rose water extraction, as the oil contentin its flowers is very low. The essential oil of this plantsoothes the mind and helps to relief depression, grief,nervous tension and stress, and because of this it is alsoused in the aromatherapy all over the world. Apart fromits perfumery uses, R. damascena essential oil has alsoreported antioxidant, antibacterial and antimicrobial(3–7), anti-HIV properties (8). The oil is also used inpoor circulation, asthma and coughs, irregular menstrua-tion, leucorrhoea, and uterine disorders. On the skin, itcan be used for dry skin, eczema and sensitive skin (9).Vapor therapy of rose oil is helpful for some allergies,headaches, migraine, etc. (10, 11).

The genetic and morphological diversity of rosecultivars have been studied earlier in Bulgaria (12, 13),

India (14, 15), Iran (16, 17) and Pakistan (18). Thechemical evaluation of damask rose oil has been doneby many researchers (19–21). In India, rose oil ismostly cultivated in northern states including HimachalPradesh, Jammu and Kashmir, Rajasthan, Haryana,Uttar Pradesh and some areas of Punjab in 2500–3000hectares of land and then, 200 kg of rose oil isproduced annually (22). Few attempts have been doneto compare the performance of varieties of rose oil forchemical composition (23–25). The objectives of thisstudy were to evaluate the variation among five ofR. damascena and one R. bourboniana varieties foressential oil content and composition in the mid-hills ofthe western Himalayas.

Experimental

Experimental site

The present study was conducted at the experimentalfarm of the CSIR-Institute of Himalayan BioresourceTechnology, Palampur (1325 m amsl, 32°06´05”N,76°34´10”E), India, in 2011. Weather data during thecrop development period of damask rose is shown inTable 1. Minimum temperature ranges from 3.5° to 19.8°C, maximum temperature ranges from 15.2° to 31.4°C,relative humidity varies between 62.2% and 94.1% inthe morning and 45.0% and 87.2% in the evening, andbright sunshine hour ranges from 2.9 to 8.9 hours.

*Corresponding author. Email: rakeshkumar@ihbt.res.in

Journal of Essential Oil Research, 2014Vol. 26, No. 3, 147–152, http://dx.doi.org/10.1080/10412905.2013.829004

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Plant material

A population of approximately 50,000 plants raisedfrom mixed stem cuttings collected from perennial roseplantations at the University of Agriculture, Udaipur,Rajasthan, India, and maintained in the field of theCSIR-Institute of Himalayan Bioresource Technology,Palampur, Himachal Pradesh, India, were utilized as anoriginal gene pool of R. damascena. Two varieties,Jwala and Himroz were diversified through selectionsof desirable traits (morphological/oil content) across25,000 plants. The five elites, three of R. damascenavar. Jwala, (Indica, Super jwala and Jwala) and two ofR. damascena var. Himroz (Hot himroz and Himroz)were developed through field selections and maintainedat the Natural Plant Products Division ExperimentalFarm of the Institute. Rosa bourboniana plants werecollected from the Fragrance and Flavour DevelopmentCentre, Kannauj, UP, India, during 1992 and main-tained at the Natural Plant Products Division Experi-mental Farm of the Institute. The plant material wasauthenticated and deposited in the herbarium (PLP) ofthe institute. A representative sample of authentic mate-rial was deposited, with voucher numbers PLP16505(Indica), PLP16511 (Jwala), PLP16509 (Super Jwala),PLP16507 (Himroz), PLP16510 (Hot himroz) andPLP16508 (R. bourboniana). From each variety, threesamples each weighing 1.0 kg representing one replica-tion were used for the study; thus three replicationswere maintained.

Essential oil extraction

Essential oil content was determined by distilling arepresentative flower sample per replication inClevenger’s apparatus. 1.0 kg of rose flowers wereplaced in a distillation apparatus with 3 L of water andhydrodistilled for 5 hours. The oil content was measuredand reported as v/w percentage. All the essential oilsamples were dried over anhydrous sodium sulfate, and

stored at 4°C in dark glass bottles until analyzed by gaschromatography–flame ionization detector (GC–FID)and GC–mass spectrometry (GC–MS).

Gas chromatography analysis

GC analysis was carried out on a Shimadzu GC–2010gas chromatograph fitted with an FID detector and aDB-5 capillary column (SGE International, Ringwood,Australia) of 30 m length, 0.25 mm i.d. and 0.25 μmfilm thickness, 5% diphenyl, 95% methylpolsiloxanestationary phase. The operating conditions were asfollows: carrier gas was helium (99.9% pure) with aflow rate of 1.05 mL/minute, the oven temperature wasprogrammed as follows: 70°C (4 minutes) and then220°C at the rate of 4°C/minute, injector and detectortemperatures were set at 240° and 260°C, respectively.

Gas chromatography–Mass spectrometry

GC–MS analysis was carried out by GC–MS (QP2010Shimadzu, Tokyo, Japan) equipped with AOC-5000Auto injector and DB-5 capillary column (SGE Interna-tional, Ringwood, Australia) of 30 m length, 0.25 mmi.d. and 0.25 μm film thickness, 5% diphenyl, 95%methylpolsiloxane stationary phase. Temperature wasprogrammed from 70°C for 4 minutes and then at therate of 4°C/minute to 220°C and held for 5 minutes.Mass spectrometer injector temperature, 240°C andinterface temperature, 250°C. Sample injection volume2 μL (dilution: 10 μL oil in 2 mL dichloromethane, GCgrade); split ratio 1:50 and mass scan 50–800 amu;ionization energy 70 eV. Helium (99.9% pure) wasused as a carrier gas with flow rate of 1.1 mL/minute.

Identification of volatile components

The retention index (RI) was calculated for all volatileconstituents using homologous series of n-alkanes(C8–C24). The components of oil were identified bymatching their RI and mass spectra with those of

Table 1. Meteorological conditions during growth and development of oil-bearing rose species.

Months Maximum temperature (°C) Minimum temperature (°C) Total rainfall (mm)

Relative humidity(%) Sunshine hours

Morning Evening (h)

Jun-10 30.8 19.1 217.0 62.2 46.8 8.1Jul-10 28.9 19.6 995.1 75.3 65.4 5.5Aug-10 26.0 19.8 1047.9 94.1 87.2 2.9Sep-10 25.4 17.5 322.0 91.4 83.7 4.4Oct-10 25.6 13.3 32.9 79.6 59.5 7.7Nov-10 22.9 9.4 5.2 74.9 48.2 8.7Dec-10 18.3 4.8 91.2 71.0 47.0 7.7Jan-11 15.2 3.5 65.0 71.0 51.5 6.6Feb-11 16.9 6.6 139.4 74.6 58.9 4.1Mar-11 22.6 10.2 45.3 67.1 45.0 7.1Apr-11 25.2 13.1 90.7 69.3 46.9 7.6May-11 31.4 19.3 120.4 78.2 51.8 8.9

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reported in literature (26) and stored in the computerlibrary namely Wiley, New York mass spectral (MS)library and National Institute of Standards andTechnology, NIST (27).

Statistical analysis

The experiment was set up according to randomizedblock design with three replications and each packageweighing 1.0 kg rose flowers was regarded asreplication. Statistical analysis of data was done as perthe standard analysis of variance technique (ANOVA)for the design used, and treatment means were comparedat p<0.05 level of significance. The data was analyzedby software SYSTAT-12 (SYSTAT Software Inc.,Chicago, IL).

Results and discussion

Oil content

The essential oil content and oil composition of fivevarieties of R. damascena and one accession ofR. bourboniana are shown in Table 2. Varieties had astatistically significant effect on the oil content. Thehighest oil content was found in Jwala and Super jwala(0.051%) compared with other varieties. Rosabourboniana had the lowest oil content (0.017%).Flower oil content from different rose species rangedfrom 0.036% to 0.147% in Pakistan (18). Genetic vari-ation may be one of the reasons for the variation in theessential oil content in the examined species. In Iranianrose oil, the content from different rose oil varietiesgrown under irrigated conditions ranged from 0.015%to 0.035% (28). Studying different varieties is indis-pensable for the selection of superior accession withhigher yield and quality for a particular region.

Oil composition

A perusal of data presented in Tables 2 and 3 revealedthat a total of thirty-two components were identified indifferent varieties of rose oil. These componentsconstituted 78.1–93.5% of the oil. The main componentsof rose oil were citronellol + nerol (16.3–30.1%),geraniol (15.8–29.3%), linalool (0.7–1.9%), rose oxide(0.9–2.6%), phenyl ethyl alcohol (0.1–0.4%), eugenol(0.3–2.2%) and nonadecane (7.3–14.7%). Manyresearchers have reported high percentage of the mono-terpene alcohols including citronellol, nerol, geraniol,linalool and phenyl ethyl alcohol in oil rose species (29,30). Linalool concentration was higher in R. damascenavar. Indica (1.9%), followed by R. damascena var. Superjwala (1.6%), R. damascena var. Jwala and R. damasce-na var. Himroz (1.3%). Citronellol + nerol (30.1%) andgeraniol (29.3%) content was found to be higher inR. damascena var. Himroz compared with others. Phenylethyl alcohol was more in R. bourboniana (0.4%)

compared with other varieties of R. damascena. cis-Roseoxide was more in Indica (2.6%) followed by Himroz(2.0%), Jwala (1.4%), Hot himroz (1.1%), Super jwala(0.9%) of R. damascena.

Taking into account the concentration found forthose components included in the ISO 9842-2003 stan-dards (31), the percentage of heptadecane (1.2–1.8%)was found to be at par the ISO 9842-2003 standards.The amount of citronellol + nerol in all genotypes ofR. damascena were ranged in between 25.1% and30.1%; however, it was low in R. bourboniana (16.3%).Geraniol is the major constituent of rose-like odor,which is important for perfumery and flavoring. It wasobserved to be higher than the desired limit in varietiesof R. damascena (23.2–29.3%) and R. bourboniana(15.8%) of the recommended values of ISO 9842-2003standards. Nonadecane was found to be within therecommended amount in Jwala (13.4%), Super jwala(14.7%), Himroz (11.3%), Hot himroz (13.8%) ofR. damascena and R. bourboniana (14.4%); however, itwas lower in Indica (7.3%) of R. damascena. Thepercentage of phenyl ethyl alcohol (ISO<3.5%) was wellwithin the desired limit in all varieties (0.1–0.4%).Lower concentration of Heneicosane is desirable (3.0–5.5%) in rose oil for perfumery. Heneicosane was foundin accordance with the ISO 9842-2003 standards inIndica (3.5%), Jwala (5.5%), Himroz (4.9%), slightlyhigher in Hot himroz (7.4%) and Super jwala (7.9%)genotypes of R. damascena and found much higher inR. bourboniana (13.8%). While comparing theR. damascena and R. canina seed oil hips, the contentof linoleic acid in R. damascena (54.18%) was found tobe higher than that of R. canina (48.84%) (32). Rosacentifolia was found higher for all the chemical constitu-ents studied except phenyl ethyl alcohol, which wasgreater in case of R. damascena than R. centifolia. Fortylandraces of damask rose were collected fromtwenty-eight provinces (thirteen origin sites) in Iran andevaluated to determine the diversity among them (16).The characteristic odor of the fresh flowers came fromphenyl ethyl alcohol (33) the principal component ofrose water. Rosa bourboniana contains a higher amountof phenyl ethyl alcohol (0.4%) than other genotypes ofR. damascena; thus this species is good for rose water.The oil content in this species was low (0.017%). Whilestudying the chemical composition of rose oil of Noorja-han, Ranisahiba and Kannouj varieties of R. damascenaunder the mountainous conditions of Uttarakhand, India,it was reported that the major components of the oilwere citronellol (15.9–35.3%), heneicosane (2.6–7.9%)and linalool (0.7–2.8%) (25). In Lebanon, seven culti-vars of R. damascena were studied, which are growingin different ecosystems for the first time (34). The studyshows remarkable differences in quantity and qualitybetween all samples and oils from other countries.

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Methyl eugenol is a high value aroma chemicalused in cosmetic products and flavoring agents, how-ever, it was not desired above a certain concentrationin the essential oils due to side effects on human health(35). It ranged from 0.4% to 1.1% among the differentvarieties shown in Table 2.

Conclusion

Among six varieties of two rose oil species, Superjwala of R. damascena yielded highest oil content(0.051%) and R. bourboniana yielded lowest oil con-tent (0.017%). The content of citronellol + nerol(30.1%) and geraniol (29.3%) was higher in Himroz

Table 2. Oil content (%) and composition of different varieties of oil-bearing rose by gas chromatography–flame ionizationdetector (GC–FID) and GC–mass spectrometry (GC–MS).

Rosa damascena

Rosa bourbonianaCompounds RI Indica Jwala Super jwala Himroz Hot himroz

Oil content (%) 0.037 0.034 0.051 0.043 0.045 0.017α-Pinene 932 1.3 0.4 0.8 0.3 nd ndβ-Pinene 974 0.4 0.1 0.3 0.1 nd ndβ-Myrcene 988 1.3 0.7 0.8 0.7 0.5 0.5Linalool 1095 1.9 1.3 1.6 1.3 0.7 1.1Phenyl ethyl alcohol 1110 0.2 0.1 0.1 0.1 0.1 0.4cis-Rose oxide 1110 2.6 1.4 0.9 2.0 1.1 0.9trans-Rose oxide 1125 tr 0.2 0.1 0.1 tr 0.2γ-Terpineol 1180 0.4 0.2 0.3 0.3 0.2 trα-Terpineol 1186 0.4 0.3 0.3 0.4 0.2 0.2β-Citronellol + nerol 1229 28.7 27.7 25.1 30.1 26.9 16.3Neral 1238 0.6 0.4 0.3 0.4 0.3 0.5Geraniol 1256 24.5 27.5 23.2 29.3 27.0 15.8Geranial 1268 0.4 0.1 0.2 0.2 0.2 0.6Citronellyl acetate 1352 0.3 0.3 0.4 0.3 0.3 0.5Eugenol 1356 1.8 1.3 0.7 2.2 1.2 0.3Neryl acetate 1359 0.3 0.3 0.3 tr 0.3 0.4Geranyl acetate 1380 1.1 2.1 2.5 1.0 1.7 3.7β-Bourbonene 1387 tr 0.1 0.2 0.1 0.1 0.5Methyl eugenol 1400 0.8 0.7 0.7 1.1 0.8 0.4Caryophyllene 1417 0.9 0.5 0.8 0.7 0.4 0.3α-Guaiene 1437 0.4 0.4 0.6 0.4 0.3 0.5α-Humulene 1452 0.5 0.5 0.6 0.5 0.3 0.4Germacrene D 1484 1.4 1.1 1.8 1.5 1.2 1.4Pentadecane 1500 0.4 0.4 0.5 0.4 0.4 0.6E,E-Farnesene 1506 0.7 0.5 0.7 0.5 0.5 0.9Hexadecane 1600 0.2 0.1 0.1 0.1 0.1 TrHeptadecane 1700 1.8 1.5 1.5 1.3 1.7 1.2E,E-Farnesol 1720 0.5 1.1 1.3 0.7 1.7 0.5Octadecane 1800 tr 0.2 0.2 0.2 0.2 0.2Nonadecane 1904 7.3 13.4 14.7 11.3 13.8 14.4Eicosane 2000 0.6 1.2 1.3 1 1.3 1.6Heneicosane 2112 3.5 5.5 7.8 4.9 7.4 13.8Total 85.2 91.6 90.7 93.5 90.9 78.1

Note: RI, retention index; tr, traces; nd, not detected.

Table 3. Percentage composition of major compound groups of rose oil.

Rosa damascena

Rosa bourbonianaSr. No. Compound group Indica Jwala Super jwala Himroz Hot himroz

1 Hydrocarbons 13.8 22.3 26.1 19.2 24.9 31.82 Monoterpene hydrocarbons 3.0 1.2 1.9 1.1 0.5 0.53 Oxygenated monoterpenes 64.0 63.9 56.7 68.8 61.0 41.34 Sesquiterpene hydrocarbons 3.9 3.1 4.7 3.7 2.8 4.05 Oxygenated sesquiterpenes 0.5 1.1 1.3 0.7 1.7 0.5

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compared with other genotypes. The composition ofrose oil produced by the varieties of R. damascena inthe western Himalayan hills was found to be wellmatched with international standards, so these varietiescan be used for the production of rose oil of interna-tional quality.

AcknowledgementsWe are grateful to the Director, IHBT, Palampur, forproviding the necessary facilities during the course of study.We are also thankful to Mr. Sushil Kumar and VijaylataPathania, technical assistants, for field management andchemical analysis. The authors are also grateful to Council ofScientific and Industrial Research, New Delhi. This is IHBTPublication No. 2336.

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